title: Foot-and-Mouth Disease Virus Evolution: Exploring Pathways Towards Virus Extinction

text: Foot-and-mouth disease virus (FMDV) is genetically and phenotypically variable. As a typical RNA virus, FMDV follows a quasispecies dynamics, with the many biological implications of such a dynamics. Mutant spectra provide a reservoir of FMDV variants, and minority subpopulations may become dominant in response to environmental demands or as a result of statistical fluctuations in population size. Accumulation of mutations in the FMDV genome occurs upon subjecting viral populations to repeated bottleneck events and upon viral replication in the presence of mutagenic base or nucleoside analogs. During serial bottleneck passages, FMDV survive during extended rounds of replication maintaining low average relative fitness, despite linear accumulation of mutations in the consensus genomic sequence. The critical event is the occurrence of a low frequency of compensatory mutations. In contrast, upon replication in the presence of mutagens, the complexity of mutant spectra increases, apparently no compensatory mutations can express their fitness-enhancing potential, and the virus can cross an error threshold for maintenance of genetic information, resulting in virus extinction. Low relative fitness and low viral load favor FMDV extinction in cell culture. The comparison of the molecular basis of resistance to extinction upon bottleneck passage and extinction by enhanced mutagenesis is providing new insights in the understanding of quasispecies dynamics. Such a comparison is contributing to the development of new antiviral strategies based on the transition of viral replication into error catastrophe.


title: Men, Primates, and Germs: An Ongoing Affair

text: Humans and nonhuman primates are phylogenetically (i.e., genetically) related and share pathogens that can jump from one species to another. The specific strategies of three groups of pathogens for crossing the species barrier among primates will be discussed. In Africa, gorillas and chimpanzees have succumbed for years to simultaneous epizootics (i.e.. “multi-emergence”) of Ebola virus in places where they are in contact with Chiropters, which could be animal reservoirs of these viruses. Human epidemics often follow these major outbreaks. Simian immunodeficiency viruses (SIVs) have an ancient history of coevolution and many interspecific exchanges with their natural hosts. Chimpanzee and gorilla SIVs have crossed the species barrier at different times and places, leading to the emergence of HIV-1 and HIV-2. Other retroviruses, such as the Simian T-Lymphotropic Viruses and Foamiviruses, have also a unique ancient or recent history of crossing the species barrier. The identification of gorilla Plasmodium parasites that are genetically close to P. falciparum suggests that gorillas were the source of the deadly human P. falciparum. Nonhuman plasmodium species that can infect humans represent an underestimated risk.


title: Common Pancreatic Disease

text: Pancreatitis is an inflammatory disease of the pancreas characterized by acute and chronic condition as well as varying duration and severity. Acute pancreatitis (AP) is a severe abdominal inflammation, characterized by parenchymal edema, necrosis with occasional presence of pseudocysts, abscess, hemorrhage, and inflammatory cell infiltration. Its disease spectrum varies from mild edematous to severe disease with fatal complications. Pancreatic acinar cells are very susceptible to inflammation during an episode of AP attack. It is believed that some initiating factors can lead to the premature transformation of inactive pro-proteases into active proteases within the pancreas. The active proteases degrade a number of cellular proteins such as the structural protein F-actin, thus leading to the collapse and malfunction of acinar and pancreatic damage, a process known as autodigestion of the pancreas.


title: The Role of Protein Structural Analysis in the Next Generation Sequencing Era

text: Proteins are macromolecules that serve a cell’s myriad processes and functions in all living organisms via dynamic interactions with other proteins, small molecules and cellular components. Genetic variations in the protein-encoding regions of the human genome account for >85% of all known Mendelian diseases, and play an influential role in shaping complex polygenic diseases. Proteins also serve as the predominant target class for the design of small molecule drugs to modulate their activity. Knowledge of the shape and form of proteins, by means of their three-dimensional structures, is therefore instrumental to understanding their roles in disease and their potentials for drug development. In this chapter we outline, with the wide readership of non-structural biologists in mind, the various experimental and computational methods available for protein structure determination. We summarize how the wealth of structure information, contributed to a large extent by the technological advances in structure determination to date, serves as a useful tool to decipher the molecular basis of genetic variations for disease characterization and diagnosis, particularly in the emerging era of genomic medicine, and becomes an integral component in the modern day approach towards rational drug development.


title: Therapeutics Against Influenza

text: Despite 75 years of research into prevention and treatment of influenza, the viruses that cause this disease continue to rank as some of the most important pathogens afflicting humans today. Progress in development of therapeutics for influenza has been slow for much of that time, but has accelerated in pace over the last two decades. Two classes of antiviral medications are used in humans at present, but each has limitations in scope and effectiveness of use. New strategies involving these licensed agents, including alternate forms of delivery and combination therapy with other drugs, are currently being explored. In addition, several novel antiviral compounds are in various clinical phases of development. Together with strategies designed to target the virus itself, new approaches to interrupt host–pathogen interactions or modulate detrimental aspects of the immune response have been proposed. Therapy for influenza will likely undergo substantial changes in the decades to come, evolving with our knowledge of pathogenesis as new approaches become viable and are validated clinically.


title: The Human Environment Interface: Applying Ecosystem Concepts to Health

text: One Health approaches have tended to focus on closer collaboration among veterinarians and medical professionals, but remain unclear about how ecological approaches could be applied or how they might benefit public health and disease control. In this chapter, we review ecological concepts, and discuss their relevance to health, with an emphasis on emerging infectious diseases (EIDs). Despite the fact that most EIDs originate in wildlife, few studies account for the population, community, or ecosystem ecology of the host, reservoir, or vector. The dimensions of ecological approaches to public health that we propose in this chapter are, in essence, networks of population dynamics, community structure, and ecosystem matrices incorporating concepts of complexity, resilience, and biogeochemical processes.


title: Food Safety: At the Center of a One Health Approach for Combating Zoonoses

text: Food Safety is at the center of One Health. Many, if not most, of all important zoonoses relate in some way to animals in the food production chain. Therefore, the food becomes an important vehicle for many, but not all, of these zoonotic pathogens. One of the major issues in food safety over the latest decennia has been the lack of cross-sectoral collaboration across the food production chain. Major food safety events have been significantly affected by the lack of collaboration between the animal health, the food control, and the human health sector. Examples range from BSE and E. coli outbreaks over dioxin crises to intentional melamine contamination. One Health formulates clearly both the need for and the benefit of cross-sectoral collaboration. In this chapter, we will focus on the human health risk related to zoonotic microorganisms present both in food animals and food from these animals, and typically transmitted to humans through food. We focus on these issues because they are very important in relation to the human disease burden, but also because this is the area where some experience of cross-sectoral collaboration already exist. Food related zoonoses can be separated in three major classes: parasites, bacteria, and viruses. While parasites often relate to very specific animal hosts and contribute significantly to the human disease burden, virus have often been related to major, well-published global outbreaks, e.g. SARS and avian- and swine-influenza. The bacterial zoonoses on the other hand often result in sporadic, but very wide-spread disease cases, resulting in a major disease burden in all countries, e.g. Salmonella and Campylobacter. Next to these traditional zoonotic problems, the use of antimicrobials in (food) animals has also caused the emergence of antimicrobial resistant (AMR) zoonotic bacteria. It is important to realize the difference in the nature of disease epidemiology, as well as, in society’s reaction to these diseases in different socio-economic settings. Some diseases have global epidemic—or pandemic—potential, resulting in dramatic action from international organizations and national agricultural—and health authorities in most countries, for instance as was the case with avian influenza. Other diseases relate to the industrialized food production chain and have been—in some settings—dealt with efficiently through farm-to-fork preventive action in the animal sector, e.g. Salmonella. Finally, an important group of zoonotic diseases are ‘neglected diseases’ in poor settings, while they have been basically eradicated in affluent economies through vaccination and culling policies in the animal sector, e.g. Brucella.


title: Current Concepts of Severe Pneumococcal Community-acquired Pneumonia

text: Community-acquired pneumonia (CAP) is a major health problem, even in developed countries, being the leading cause of death due to infectious diseases in the USA [1]. CAP has a wide clinical spectrum of severity: up to 80% of patients are successfully managed in primary care, but 1 % of patients with CAP are classified as having severe disease, needing intensive care unit (ICU) admission, with 20–50% dying despite all available support and treatment options being utilized. Streptococcus pneumoniae is the most common cause of CAP, enclosing the subset group of patients having severe disease [2]. Moreover, bacteremia is not uncommon in pneumococcal CAP (20%) and has been associated with increased severity and mortality compared with non-bacteremic pneumonia [3].


title: The Historical, Present, and Future Role of Veterinarians in One Health

text: The renewed interest in the concept of One Health has occurred as a result of the increased emergence of zoonotic infectious diseases over the past decade. The subsequent impacts of these diseases on human, livestock, and wildlife health, as well as the economic effects, have given international health organizations and national governments a greater appreciation of the importance of collaborative efforts in solving health problems. The One Health concept is not new, but under its umbrella, a new generation of veterinarians, physicians, ecologists, biologists, and social scientists is shaping the concept in novel ways. This has led to increased support for One Health initiatives to control disease by international agencies, national governments, and nongovernmental organizations as well as a growing emphasis on One Health concepts in training the veterinary workforce. Veterinary schools are reorganizing veterinary education to better teach students the precepts of One Health. This chapter explores the evolution and application of the One Health concept from the perspective of the veterinarian. The veterinary profession is positioned to be a strong advocate and leader of One Health. Veterinarians have a long history of involvement with One Health activities, and this involvement has adjusted and shifted with the changing needs of society. A new area of work for veterinarians is ecosystem health, which is becoming more relevant as a result of the impact that the ever-increasing human population is having on the environment that supports them.


title: Airway Epithelia

text: Cystic fibrosis (CF) is a common inherited disorder affecting a variety of epithelial tissues. The disease is caused by mutations in the cystic fibrosis transmembrane conductance regulator gene (CFTR) that lead to abnormal secretions, recurrent infection and inflammation, bronchiectasis, and premature death. Because airways disease is the major cause of morbidity and mortality in cystic fibrosis, gene therapy efforts have focused on luminal delivery of vector to the airways of CF patients. Retroviruses are attractive as a gene transfer vector system since integration of the wild-type CFTR cDNA into the host genome may lead to long-term expression and perhaps, a cure. However, simple retroviruses are limited as vectors for airway gene transfer by the low rates of epithelial cell proliferation in human airways (∼0.1–0.2%) combined with the traditionally low titers. Advances in vector design and production have improved titers, and the development of human and animal lentiviruses may help overcome the requirement for cell proliferation. These developments have raised hopes for retroviral approaches for treatment of CF lung disease.


title: Measles Studies in the Macaque Model

text: Much of our current understanding of measles has come from experiments in non-human primates. In 1911, Goldberger and Anderson showed that macaques inoculated with filtered secretions from measles patients developed measles, thus demonstrating that the causative agent of this disease was a virus. Since then, different monkey species have been used for experimental measles virus infections. Moreover, infection studies in macaques demonstrated that serial passage of the virus in vivo and in vitro resulted in virus attenuation, providing the basis for all current live-attenuated measles vaccines. This chapter will review the macaque model for measles, with a focus on vaccination and immunopathogenesis studies conducted over the last 15 years. In addition, recent data are highlighted demonstrating that the application of a recombinant measles virus strain expressing enhanced green fluorescent protein dramatically increased the sensitivity of virus detection, both in living and sacrificed animals, allowing new approaches to old questions on measles vaccination and pathogenesis.


title: Mapping Protein–Protein Interactions with Phage-Displayed Combinatorial Peptide Libraries and Alanine Scanning

text: One avenue for inferring the function of a protein is to learn what proteins it may bind to in the cell. Among the various methodologies, one way for doing so is to affinity select peptide ligands from a phage-displayed combinatorial peptide library and then to examine if the proteins that carry such peptide sequences interact with the target protein in the cell. With the protocols described in this chapter, a laboratory with skills in microbiology, molecular biology, and protein biochemistry can readily identify peptides in the library that bind selectively, and with micromolar affinity, to a given target protein on the time scale of 2 months. To illustrate this approach, we use a library of bacteriophage M13 particles, which display 12-mer combinatorial peptides, to affinity select different peptide ligands for two different targets, the SH3 domain of the human Lyn protein tyrosine kinase and a segment of the yeast serine/threonine protein kinase Cbk1. The binding properties of the selected peptide ligands are then dissected by sequence alignment, Kunkel mutagenesis, and alanine scanning. Finally, the peptide ligands can be used to predict cellular interacting proteins and serve as the starting point for drug discovery.


title: Nucleocapsid Protein: A Desirable Target for Future Therapies Against HIV-1

text: The currently available anti-HIV-1 therapeutics is highly beneficial to infected patients. However, clinical failures occur as a result of the ability of HIV-1 to rapidly mutate. One approach to overcome drug resistance is to target HIV-1 proteins that are highly conserved among phylogenetically distant viral strains and currently not targeted by available therapies. In this respect, the nucleocapsid (NC) protein, a zinc finger protein, is particularly attractive, as it is highly conserved and plays a central role in virus replication, mainly by interacting with nucleic acids. The compelling rationale for considering NC as a viable drug target is illustrated by the fact that point mutants of this protein lead to noninfectious viruses and by the inability to select viruses resistant to a first generation of anti-NC drugs. In our review, we discuss the most relevant properties and functions of NC, as well as recent developments of small molecules targeting NC. Zinc ejectors show strong antiviral activity, but are endowed with a low therapeutic index due to their lack of specificity, which has resulted in toxicity. Currently, they are mainly being investigated for use as topical microbicides. Greater specificity may be achieved by using non-covalent NC inhibitors (NCIs) targeting the hydrophobic platform at the top of the zinc fingers or key nucleic acid partners of NC. Within the last few years, innovative methodologies have been developed to identify NCIs. Though the antiviral activity of the identified NCIs needs still to be improved, these compounds strongly support the druggability of NC and pave the way for future structure-based design and optimization of efficient NCIs.


title: Transgene Design

text: Transgenics are powerful mouse models to understand the biological functions of genes. This chapter gives a short overview of the requirements and considerations in designing a transgene. In addition, potential important choices that have to be made in advance for the successful designing and generating a transgenic mouse model are discussed. Methods for DNA purification for microinjection are also provided in this chapter.


title: Therapeutic Uses of Antioxidant Liposomes

text: This chapter focuses on the use of antioxidant liposomes in the general area of free radical biology and medicine. The term antioxidant liposome is relatively new and refers to liposomes containing lipid-soluble chemical antioxidants, water-soluble chemical antioxidants, enzymatic antioxidants, or combinations of these various antioxidants. The role of antioxidants in health and disease has been extensively discussed, and many excellent reviews and books are available (1–3). Antioxidant liposomes hold great promise in the treatment of many diseases in which oxidative stress plays a prominent role. Oxidative stress is a physiological condition in which the production of damaging free radicals exceeds the in vivo capacity of antioxidant protection mechanisms to prevent pathophysiology. Free radicals are molecules with unpaired electrons, often highly reactive and damaging to biological systems. The biological membranes of subcellular organelles are a major site of free radical damage but proteins and DNA are also significant targets. Moreover, free radicals can alter cellular signal transduction pathways and stimulate the synthesis of inflammatory cytokines. Oxygen radicals and other reactive oxygen species (ROS) arise from the single electron reductions of oxygen.


title: An Overview on GPCRs and Drug Discovery: Structure-Based Drug Design and Structural Biology on GPCRs

text: G protein-coupled receptors (GPCRs) represent 50–60% of the current drug targets. There is no doubt that this family of membrane proteins plays a crucial role in drug discovery today. Classically, a number of drugs based on GPCRs have been developed for such different indications as cardiovascular, metabolic, neurodegenerative, psychiatric, and oncologic diseases. Owing to the restricted structural information on GPCRs, only limited exploration of structure-based drug design has been possible. Much effort has been dedicated to structural biology on GPCRs and very recently an X-ray structure of the β2-adrenergic receptor was obtained. This breakthrough will certainly increase the efforts in structural biology on GPCRs and furthermore speed up and facilitate the drug discovery process.


title: Suicide Gene Therapy in Liver Tumors

text: Charaterization of a variety of genomic defects in malignant cells (1) has led to attempts to treat cancer by gene therapy. Gene therapy is a therapeutic approach in which therapeutic nucleic acids are transferred into the affected organs. Although the ideal concept would be the replacement of the abnormal gene by a copy of the functional gene, currently there have not been reliable and safe techniques to allow the site-specific integration of DNA into the human genome (2). Thus, almost all gene therapies are developed by simply transferring the therapeutic gene into somatic cells without replacing the abnormal gene. The goal is to identify and correct genetic abnormalities interfering with the cell cycle and to correct them in all cells. Technically, there are two methods amenable for gene transfer: reintroduction of in vitro transferred gene into the body and direct transfer of gene into the target cells in vivo.


title: Protein Microarrays

text: Protein microarrays containing nearly the entire yeast proteome have been constructed. They are typically prepared by overexpression and high-throughput purification and printing onto microscope slides. The arrays can be used to screen nearly the entire proteome in an unbiased fashion and have enormous utility for a variety of applications. These include protein–protein interactions, identification of novel lipid- and nucleic acid-binding proteins, and finding targets of small molecules, protein kinases, and other modification enzymes. Protein microarrays are thus powerful tools for individual studies as well as systematic characterization of proteins and their biochemical activities and regulation.


title: Identifying Functional Adenovirus-Host Interactions Using Tandem Mass Spectrometry

text: We describe a systematic, high-throughput approach to identify proteins involved in functional adenovirus (Ad)host interactions in vitro and in vivo. We were particularly interested in identifying cellular proteins that interact with fiber knob, which is the moiety within the Ad capsid responsible for high-affinity attachment of virus to cellular receptors. We used recombinant fiber knob domains from members of group C and B Ads to purify virus interacting proteins from cell membrane lysates and from human and mouse plasma. Using tandem mass spectrometry, we identified a number of candidate Ad-interacting proteins, including functional cellular receptors and previously unknown interacting partners such as complement component C4-binding protein and other blood proteins that presumably are involved in Ad infection after intravenous virus application. The ability of these proteins to bind to Ad was further confirmed using in vitro protein binding assays as well as infection competition assays. The approach of using a structural protein can be universally applied for a variety of viral and nonviral pathogens and can reveal host cell factors critical in viral infection, immune evasion, and tissue specificity. This information is also a prerequisite to assess in vivo safety and efficacy of Ad-based gene transfer vectors.


title: Principles of Ocular Pharmacology

text: Recently, in a poll by Research America, a significant number of individuals placed losing their eyesight as having the greatest impact on their lives more so than other conditions, such as limb loss or memory loss. When they were also asked to rank which is the worst disease that could happen to them, blindness was ranked first by African-Americans and second by Caucasians, Hispanics, and Asians. Therefore, understanding the mechanisms of disease progression in the eye is extremely important if we want to make a difference in people’s lives. In addition, developing treatment programs for these various diseases that could affect our eyesight is also critical. One of the most effective treatments we have is in the development of specific drugs that can be used to target various components of the mechanisms that lead to ocular disease. Understanding basic principles of the pharmacology of the eye is important if one seeks to develop effective treatments. As our population ages, the incidence of devastating eye diseases increases. It has been estimated that more than 65 million people suffer from glaucoma worldwide (Quigley and Broman. Br J Ophthalmol 90:262–267, 2006). Add to this the debilitating eye diseases of age-related macular degeneration, diabetic retinopathy, and cataract, the number of people effected exceeds 100 million. This chapter focuses on ocular pharmacology with specific emphasis on basic principles and outlining where in the various ocular sites are drug targets currently in use with effective drugs but also on future drug targets.


title: Display of Peptides on the Surface of Tobacco Mosaic Virus Particles

text: In this review, we focus on the potential that tobacco mosaic virus (TMV) has as a carrier for immunogenic epitopes, and the factors that must be considered in order to bring products based on this platform to the market. Large Scale Biology Corporation developed facile and scaleable methods for manufacture of candidate peptide display vaccines based on TMV. We describe how rational design of peptide vaccines can improve the manufacturability of particular TMV products. We also discuss downstream processing and purification of the vaccine products, with particular attention to the metrics that a product must attain in order to meet criteria for regulatory approval as injectable biologics.


title: Rotavirus Assembly: An Alternative Model That Utilizes an Atypical Trafficking Pathway

text: We review here recent advances in our knowledge on trafficking and assembly of rotavirus and rotaviral proteins in intestinal cells. Assembly of rotavirus has been extensively studied in nonpolarized kidney epithelial MA104 cells, where several data indicate that most if not all the steps of rotavirus assembly take place within the endoplasmic reticulum (ER) and that rotavirus is release upon cell lysis. We focus here on data obtained in intestinal cells that argue for another scheme of rotavirus assembly, where the final steps seem to take place outside the ER with an apically polarized release of rotavirus without significant cell lysis. One of the key observations made by different groups is that VP4 and other structural proteins interact substantially with specialized membrane microdomains enriched in cholesterol and sphingolipids termed rafts. In addition, recent data point to the fact that VP4 does not localize within the ER or the Golgi apparatus in infected intestinal cells. The mechanisms by which VP4, a cytosolic protein, may be targeted to the apical membrane in these cells and assembles with the other structural proteins are discussed. The identification of cellular proteins such as Hsp70, flotillin, rab5, PRA1 and cytoskeletal components that interact with VP4 may help to define an atypical polarized trafficking pathway to the apical membrane of intestinal cells that will be raft-dependent and by-pass the classical exocytic route.


title: Arenaviruses

text: The Arenaviridae family contains 22 recognized virus species, each of them strongly associated with a rodent species (except Tacaribe virus which is associated with a species of bat), suggesting an ancient co-evolutionary process. Although the concept of co-evolution between rodents and arenaviruses is now largely accepted, little has been uncovered in terms of dating the phenomenon and the mechanisms of evolution, including speciation and pathogenicity. These questions are targeted in the present chapter. Old World arenaviruses are associated with the Eurasian rodents in the family Muridae. New World arenaviruses are associated with American rodents in the subfamily Sigmodontinae. The correlation between the rodent host phylogeny and the viruses suggests a long association and a co-evolutionary process. Furthermore, three distinct New World arenaviruses share a common ancestor, demonstrating a unique recombination event that probably occurred in that ancestor. This shows that recombination among arenaviruses of different lineages might occur in nature. Recombination and co-evolutionary adaptation appear as the main mechanisms of arenavirus evolution, generating a high degree of diversity. The diversity among rodent host reservoir and virus species and the potential to exchange genomic material provide a basis for the emergence of new viruses and the risk of these becoming pathogenic for humans.


title: Peripheral Mechanisms II: The Pharmacology of Peripherally Active Antitussive Drugs

text: Cough is an indispensable defensive reflex. Although generally beneficial, it is also a common symptom of diseases such as asthma, chronic obstructive pulmonary disease, upper respiratory tract infections, idiopathic pulmonary fibrosis and lung cancer. Cough remains a major unmet medical need and although the centrally acting opioids have remained the antitussive of choice for decades, they have many unwanted side effects. However, new research into the behaviour of airway sensory nerves has provided greater insight into the mechanisms of cough and new avenues for the discovery of novel non-opioid antitussive drugs. In this review, the pathophysiological mechanisms of cough and the development of novel antitussive drugs are reviewed.


Efficacy of an Inactivated PRRSV Vaccine: Induction of Virus-Neutralizing Antibodies and Partial Virological Protection upon Challenge


title: BRD4 inhibition exerts anti-viral activity through DNA damage-dependent innate immune responses

text: Chromatin dynamics regulated by epigenetic modification is crucial in genome stability and gene expression. Various epigenetic mechanisms have been identified in the pathogenesis of human diseases. Here, we examined the effects of ten epigenetic agents on pseudorabies virus (PRV) infection by using GFP-reporter assays. Inhibitors of bromodomain protein 4 (BRD4), which receives much more attention in cancer than viral infection, was found to exhibit substantial anti-viral activity against PRV as well as a range of DNA and RNA viruses. We further demonstrated that BRD4 inhibition boosted a robust innate immune response. BRD4 inhibition also de-compacted chromatin structure and induced the DNA damage response, thereby triggering the activation of cGAS-mediated innate immunity and increasing host resistance to viral infection both in vitro and in vivo. Mechanistically, the inhibitory effect of BRD4 inhibition on viral infection was mainly attributed to the attenuation of viral attachment. Our findings reveal a unique mechanism through which BRD4 inhibition restrains viral infection and points to its potent therapeutic value for viral infectious diseases.


title: Ceramide in Stress Response

text: Evidence has consistently indicated that activation of sphingomyelinases and/or ceramide synthases and the resulting accumulation of ceramide mediate cellular responses to stressors such as lipopolysaccharide, interleukin 1ß, tumor necrosis factor a, serum deprivation, irradiation and various antitumor treatments. Recent studies had identified the genes encoding most of the enzymes responsible for the generation of ceramide and ongoing research is aimed at characterizing their individual functions in cellular response to stress. This chapter discusses the seminal and more recent discoveries in regards to the pathways responsible for the accumulation of ceramide during stress and the mechanisms by which ceramide affects cell functions. The former group includes the roles of neutral sphingomyelinase 2, serine palmitoyltransferase, ceramide synthases, as well as the secretory and endosomal/lysosomal forms of acid sphingomyelinase. The latter summarizes the mechanisms by which ceramide activate its direct targets, PKC?, PP2A and cathepsin D. The ability of ceramide to affect membrane organization is discussed in the light of its relevance to cell signaling. Emerging evidence to support the previously assumed notion that ceramide acts in a strictly structure-specific manner are also included. These findings are described in the context of several physiological and pathophysiological conditions, namely septic shock, obesity-induced insulin resistance, aging and apoptosis of tumor cells in response to radiation and chemotherapy.


title: Electrochemical Biosensors Combined with Isothermal Amplification for Quantitative Detection of Nucleic Acids

text: In recent years, various isothermal amplification techniques have been developed as alternatives to polymerase chain reaction (PCR). The integration of isothermal amplification with electrical or electrochemical devices has enabled high-throughput nucleic acid-based assays with high sensitivity. We performed solid-phase rolling circle amplification (RCA) on the surface of a Au electrode, and detected RCA products in situ using chronocoulometry (CC) with [Ru (NH(3))(6)](3+) as the signaling molecule. Detection sensitivity for DNA and a microRNA (miR-143) was 100 fM and 1 pM, respectively. Furthermore, we conducted potentiometric DNA detection using an ethidium ion (Et(+))-selective electrode (Et(+)ISE) for real-time monitoring of isothermal DNA amplification by primer-generation RCA (PG-RCA). The Et(+)ISE potential enabled real-time monitoring of the PG-RCA reaction in the range of 10 nM–1 μM of initial target DNA. Devices based on these electrochemical techniques represent a new strategy for replacing conventional PCR for on-site detection of nucleic acids of viruses or microorganisms.


title: Viral Pathogenesis, Modulation of Immune Receptor Signaling and Treatment

text: During the co-evolution of viruses and their hosts, the latter have equipped themselves with an elaborate immune system to defend themselves from the invading viruses. In order to establish a successful infection, replicate and persist in the host, viruses have evolved numerous strategies to counter and evade host antiviral immune responses as well as exploit them for productive viral replication. These strategies include those that target immune receptor transmembrane signaling. Uncovering the exact molecular mechanisms underlying these critical points in viral pathogenesis will not only help us understand strategies used by viruses to escape from the host immune surveillance but also reveal new therapeutic targets for antiviral as well as immunomodulatory therapy. In this chapter, based on our current understanding of transmembrane signal transduction mediated by multichain immune recognition receptors (MIRRs) and the results of sequence analysis, we discuss the MIRR-targeting viral strategies of immune evasion and suggest their possible mechanisms that, in turn, reveal new points of antiviral intervention. We also show how two unrelated enveloped viruses, human immunodeficiency virus and human cytomegalovirus, use a similar mechanism to modulate the host immune response mediated by two functionally different MIRRs—T-cell antigen receptor and natural killer cell receptor, NKp30. This suggests that it is very likely that similar general mechanisms can be or are used by other viral and possibly nonviral pathogens.


Molecular Dissection of Porcine Reproductive and Respiratory Virus Putative Nonstructural Protein 2


title: Epithelial Sodium Channels in the Adult Lung – Important Modulators of Pulmonary Health and Disease

text: Absorption of excess fluid from the airways and alveolar lumen requires active vectorial transepithelial transport of sodium ions (Na(+)) by alveolar type II and possibly type I cells. The rate-limiting step in this process is the activity of the heterotrimeric apical membrane epithelial Na+ channel (ENaC). Pharmacologic inhibitors and genetic manipulations that disrupt Na+ transport result in fluid accumulation within the lung and failure of gas exchange. The importance of Na(+) transport in the lung is also demonstrated in conditions such as ARDS, where abnormal absorption of Na(+) contributes to the pathophysiology of pulmonary disease. ENaC expression and function is influenced by diverse factors, such as oxygen tension, glucocorticoids, and cytoskeletal proteins. In addition, ENaC dysfunction has been shown to be induced by purinergic nucleotide activation of P2Y receptors (in paramyxoviral bronchiolitis) and reactive species (in acute lung injury). Finally, βadrenergic agonists have been shown experimentally to reverse defects in ENaC function, and improve hypoxemia and pulmonary edema, and may provide a novel therapeutic modality for ARDS, although some viral lung pathogens appear to induce insensitivity to their actions.


title: The Use of Integrated and Intelligent Testing Strategies in the Prediction of Toxic Hazard and in Risk Assessment

text: There is increasing concern that insurmountable differences between humans and laboratory animals limit the relevance and reliability for hazard identification and risk assessment purposes of animal data produced by traditional toxicity test procedures. A way forward is offered by the emerging new technologies, which can be directly applied to human material or even to human beings themselves. This promises to revolutionise the evaluation of the safety of chemicals and chemical products of various kinds and, in particular, pharmaceuticals. The available and developing technologies are summarised and it is emphasised that they will need to be used selectively, in integrated and intelligent testing strategies, which, in addition to being scientifically sound, must be manageable and affordable. Examples are given of proposed testing strategies for general chemicals, cosmetic ingredients, candidate pharmaceuticals, inhaled substances, nanoparticles and neurotoxicity.


title: Searching the Tritryp Genomes for Drug Targets

text: The recent publication of the complete genome sequences of Leishmania major, Trypanosoma brucei and Trypanosoma cruzi revealed that each genome contains 8300-12,000 protein-coding genes, of which -6500 are common to all three genomes, and ushers in a new, post-genomic, era for trypanosomatid drug discovery. This vast amount of new information makes possible more comprehensive and accurate target identification using several new computational approaches, including identification of metabolic “choke-points”, searching the parasite proteomes for orthologues of known drug targets, and identification of parasite proteins likely to interact with known drugs and drug-like small molecules. In this chapter, we describe several databases (such as GENEDB, BRENDA, KEGG, METACYC, the THERAPEUTIC TARGET DATABASE, and CHEMBANK) and algorithms (including PATHOLOGIC, PATHWAY HUNTER TOOL, AND AUTODOCK) which have been developed to facilitate the bioinformatic analyses underlying these approaches. While target identification is only the first step in the drug development pipeline, these new approaches give rise to renewed optimism for the discovery of new drugs to combat the devastating diseases caused by these parasites. Traditionally, drug discovery in the trypanosomatids (and other organisms) has proceeded from two different starting points: screening large numbers of existing compounds for activity against whole parasites or more focused screening of compounds for activity against defined molecular targets. Most existing anti-trypanosomatids drugs were developed using the former approach, although the latter has gained much attention in the last twenty years under the rubric of “rational drug design”. Until recently, one of the major bottlenecks in anti-trypanosomatid drug development has been our ability to identify good targets, since only a very small percentage of the total number of trypanosomatid genes were known. That has now changed forever, with the recent (July, 2005) publication of the “Tritryp” (Trypanosonm brucei, Trypanosoma cruzi and Leishmania major) genome sequences.(1-4) This vast amount of information now makes possible several new approaches for target identffication and ushers in a post-genomic era for trypanosomatid drug discovery.


title: The Antigenome: From Protein Subunit Vaccines to Antibody Treatments of Bacterial Infections?

text: New strategies are needed to master infectious diseases. The so-called “passive vaccination”, i.e., prevention and treatment with specific antibodies, has a proven record and potential in the management of infections and entered the medical arena more than 100 years ago. Progress in the identification of specific antigens has become the hallmark in the development of novel subunit vaccines that often contain only a single immunogen, frequently proteins, derived from the microbe in order to induce protective immunity. On the other hand, the monoclonal antibody technology has enabled biotechnology to produce antibody species in unlimited quantities and at reasonable costs that are more or less identical to their human counterparts and bind with high affinity to only one specific site of a given antigen. Although, this technology has provided a robust platform for launching novel and successful treatments against a variety of devastating diseases, it is up till now only exceptionally employed in therapy of infectious diseases. Monoclonal antibodies engaged in the treatment of specific cancers seem to work by a dual mode; they mark the cancerous cells for decontamination by the immune system, but also block a function that intervenes with cell growth. The availability of the entire genome sequence of pathogens has strongly facilitated the identification of highly specific protein antigens that are suitable targets for neutralizing antibodies, but also often seem to play an important role in the microbe’s life cycle. Thus, the growing repertoire of well-characterized protein antigens will open the perspective to develop monoclonal antibodies against bacterial infections, at least as last resort treatment, when vaccination and antibiotics are no options for prevention or therapy. In the following chapter we describe and compare various technologies regarding the identification of suitable target antigens and the foundation of cognate monoclonal antibodies and discuss their possible applications in the treatment of bacterial infections together with an overview of current efforts.


Gamma-Interferon Involvement in the Pathogenesis of Lactate Dehydrogenase–Elevating Virus Infection


title: Die neue Grippe (porzine Influenza/Schweinegrippe): Eine Zusammenfassung aktueller Informationen zu Influenza A/H1N1

text: Porcine influenza is a viral disease. Along with other viral diseases such as avian influenza or SARS it may lead to serious consequences for the population. The disease occurs after infection with the influenza virus A/H1N1. Reassortment of RNA genome sequences of porcine and human influenza viruses can lead to new antigenic variants (antigen shift) with human pathogenic features. The present articles supplies recent information with relevance for occupational health.


title: Die humane Hantavirus-Infektion: Eine Literaturübersicht

text: Hantaviruses belong to the Bunyaviridae family. A large number of different subtypes are known worldwide that show a host-specific geographical distribution. Humans may become infected through contact with rodent urine, saliva or feces. In humans, hantavirus infections are an important and worldwide emerging zoonotic disease that may result in a variety of potentially life-threatening diseases, e. g. hemorrhagic fever with renal syndrome (HFRS) and hantavirus cardiopulmonary syndrome (HCPS). There are various aspects of major occupational importance, particularly for agriculturists, veterinarians and forestry workers. This article reviews the current knowledge about epidemiology, pathology, diagnostics, therapy and prevention of human hantavirus infections worldwide.


title: The Mannose-Binding Lectin: An Infection Susceptibility Gene

text: A critical but unanswered question is what defines each individual’s pre-morbid susceptibility to infection? w e propose that individuals must have an “immune haplotype” that shapes their response to infectious agents. Infection is a balance between the intrinsic virulence of the infectious agent and the host defenses. Recent viral outbreaks of SARS and influenza serve to illustrate this point as these viruses cause severe disease in certain individuals, yet there are others in whom the same infectious challenge results in minimal symptoms. On the other hand it might be that those self same people who are resistance to one particular viral infection might be susceptible to other infection challenges. Similar rules can apply to susceptibility to bacterial infections.


title: Immunoregulation by Naturally Occurring and Disease-Associated Autoantibodies: Binding to Cytokines and Their Role in Regulation of T-Cell Responses

text: The role of naturally occurring autoantibodies (NAbs) in homeostasis and in disease manifestations is poorly understood. In the present chapter, we review how NAbs may interfere with the cytokine network and how NAbs, through formation of complement-activating immune complexes with soluble self-antigens, may promote the uptake and presentation of self-molecules by antigen-presenting cells. Both naturally occurring and disease-associated autoantibodies against a variety of cytokines have been reported, including NAbs against interleukin (IL)-1α, IL-6, IL-8, IL-10, granulocyte-macrophage colony-stimulating factor, interferon (IFN)-α, IFN-β, IFN-γ, macrophage chemotactic protein-1 and IL-21. NAbs against a variety of other self-antigens have also been reported, and using thyroglobulin as an example we discuss how NAbs are capable of promoting uptake of immune complexes via complement receptors and Fc-receptors on antigen-presenting cells and thereby regulate T-cell activity. Knowledge of the influence of NAbs against cytokines on immune homeostasis is likely to have wide-ranging implications both in understanding pathogenesis and in treatment of many immunoinflammatory disorders, including a number of autoimmune and autoinflammatory diseases.


title: Influenza Pandemics

text: The recent H1N1 pandemic that emerged in 2009 has illustrated how swiftly a new influenza virus can circulate the globe. Here we explain the origins of the 2009 pandemic virus, and other twentieth century pandemics. We also consider the impact of the 2009 pandemic in the human population and the use of vaccines and antiviral drugs. Thankfully this outbreak was much less severe than that associated with Spanish flu in 1918. We describe the viral factors that affect virulence of influenza and speculate on the future course of this virus in humans and animals.


title: New Therapeutics Targets in Chronic Viral Cardiomyopathy

text: Dilated cardiomyopathy (DCM) is a prevalent heart muscle disease characterized by impaired contractility and dilation of the ventricles. Recent clinical research suggests that cardiotropic viruses are important environmental pathogenic factors in human DCM, which may therefore be considered as a chronic viral cardiomyopathy. All virus-positive DCM patients thus come into the focus of virological research and should be considered for antiviral strategies. Interferon-β therapy has been shown to mediate virus elimination in patients with adenovirus or coxsackievirus persistence.We discuss here several possible new molecular targets for patients infected with cardiotropic viruses in (1) the cellular virus uptake system, (2) virus-induced cellular signaling pathways, and (3) interactions between virus-encoded proteins with important cellular target proteins. The potential of these approaches in the setting of a chronic viral infection is significantly different from that in an acute viral infection. Specific problems encountered in a chronic situation and possible solutions are discussed.


title: Management of Severe Malaria and Severe Dengue in Resource-Limited Settings

text: This chapter summarizes recommendations on important aspects of the management of patients with severe malaria and severe dengue. Severe falciparum malaria requires rapid parasitological diagnosis by microscopy or rapid diagnostic test (RCT) and prompt initiation of parenteral artesunate. Fluid bolus therapy should be avoided in patients without hypotensive shock, and we suggest initial (24 h) crystalloid fluid therapy of 2–4 mL/kg/h, which may subsequently be reduced to 1 mL/kg/h in patients receiving additional fluids, e.g., through enteral tube feeding. In the minority of those patients presenting with hypotensive shock, we suggest fluid bolus therapy (30 mL/kg) with an isotonic crystalloid and early initiation of vasopressor support. Enteral feeding in non-intubated adult patients with cerebral malaria can start after 60 h, to avoid aspiration pneumonia. There are insufficient data to suggest this in pediatric cerebral malaria. The diagnosis of severe dengue is commonly with a combined dengue antigen (NS1) and antibody RDT. No antiviral treatment is currently available. Dengue shock results from capillary leakage, although hemorrhage or depression of myocardial contractility can contribute. The World Health Organization guidelines recommend restoration of the circulation guided by pulse pressure, capillary refill time, hematocrit, and urine output. Large (>15 mL/kg) rapid (<30 min) fluid boluses should be avoided, but prompt fluid administration with crystalloids is essential and should be restricted as soon as the critical phase is over to avoid pulmonary edema. Corticosteroids are not recommended, neither is platelet transfusion for thrombocytopenia in the absence of active bleeding or other risk factors.


Recognition and Management of Encephalitis in Children


title: Early and Acute Complications and the Principles of HSCT Nursing Care

text: Haematopoietic stem cell transplantation (HSCT) generally includes preparative or conditioning regimes containing chemotherapy and/or radiotherapy in high doses. These regimens, as well as other treatments before and after HSCT such as immunosuppressive drugs to prevent graft versus host disease (GvHD) (see Chap. 11), may affect the patient’s organs and tissues and may cause both acute and long-term complications. In the evolving field of stem cell therapies, some complications that traditionally have been regarded as early complications are now, due to changes in preparative regimens and choice of stem cell source, sometimes seen later in the post-transplant out-patient setting. The complications covered in this chapter generally occur within 100 days post HSCT and are thus classified as early complications. Two of the most common early complications are oral complications/mucositis and sepsis. Some other relatively rare complications are also covered here: haemorrhagic cystitis (HC), endothelial damage (ED) syndromes including engraftment syndrome (ES), idiopathic pneumonia syndrome (IPS), diffuse alveolar haemorrhage (DAH), transplant-associated microangiopathy (TAM) and sinusoidal obstruction syndrome/veno-occlusive disease (SOS/VOD). For all complications, recommendations for prevention and principles for nursing care are presented since careful nursing monitoring, prompt intervention and care may have an influence on patients’ morbidity and mortality.


Read all about it: The Lancet's Paper of the Year, 2003


title: GPU Acceleration of Dock6’s Amber Scoring Computation

text: Dressing the problem of virtual screening is a long-term goal in the drug discovery field, which if properly solved, can significantly shorten new drugs’ R&D cycle. The scoring functionality that evaluates the fitness of the docking result is one of the major challenges in virtual screening. In general, scoring functionality in docking requires a large amount of floating-point calculations, which usually takes several weeks or even months to be finished. This time-consuming procedure is unacceptable, especially when highly fatal and infectious virus arises such as SARS and H1N1, which forces the scoring task to be done in a limited time. This paper presents how to leverage the computational power of GPU to accelerate Dock6’s (http://dock.compbio.ucsf.edu/DOCK_6/) Amber (J. Comput. Chem. 25: 1157–1174, 2004) scoring with NVIDIA CUDA (NVIDIA Corporation Technical Staff, Compute Unified Device Architecture – Programming Guide, NVIDIA Corporation, 2008) (Compute Unified Device Architecture) platform. We also discuss many factors that will greatly influence the performance after porting the Amber scoring to GPU, including thread management, data transfer, and divergence hidden. Our experiments show that the GPU-accelerated Amber scoring achieves a 6.5× speedup with respect to the original version running on AMD dual-core CPU for the same problem size. This acceleration makes the Amber scoring more competitive and efficient for large-scale virtual screening problems.


title: Noninfectious Pulmonary Complications

text: Lung injury occurs frequently following HSCT and significantly contributes to morbidity and mortality in the immediate post transplant period and in the months and years that follow.


title: Influenza Virus Entry

text: As all the enveloped viruses, the entry of influenza viruses includes a number of steps in host cell infection. This chapter summarizes the current knowledge of the entry pathway and the role of the fusion protein of influenza virus, hemagglutinin, in this process. Hemagglutinin (HA) is a trimeric glycoprotein that is present in multiple copies in the membrane envelope of influenza virus. HA contains a fusion peptide, a receptor binding site, a metastable structural motif, and the transmembrane domain. The first step of influenza virus entry is the recognition of the host cell receptor molecule, terminal α-sialic acid, by HA. This multivalent attachment by multiple copies of trimetric HA triggers endocytosis of influenza virus that is contained in the endosome. The endosome-trapped virus traffics via a unidirectional pathway to near the nucleus. At this location, the interior pH of the endosome becomes acidic that induces a dramatic conformational change in HA to insert the fusion peptide into the host membrane, induce juxtaposition of the two membranes, and form a fusion pore that allows the release of the genome segments of influenza virus. HA plays a key role in the entire entry pathway. Inhibitors of virus entry are potentially effective antiviral drugs of influenza viruses.


title: Inflammasome in the Pathogenesis of Pulmonary Diseases

text: Lung diseases are common and significant causes of illness and death around the world. Inflammasomes have emerged as an important regulator of lung diseases. The important role of IL-1 beta and IL-18 in the inflammatory response of many lung diseases has been elucidated. The cleavage to turn IL-1 beta and IL-18 from their precursors into the active forms is tightly regulated by inflammasomes. In this chapter, we structurally review current evidence of inflammasome-related components in the pathogenesis of acute and chronic lung diseases, focusing on the “inflammasome-caspase-1-IL-1 beta/IL-18” axis.


HCoV-229E Infects and Activates Monocytes


title: Cell Cycle-Regulated Transcription: Effectively Using a Genomics Toolbox

text: The cell cycle comprises a series of temporally ordered events that occur sequentially, including DNA replication, centrosome duplication, mitosis, and cytokinesis. What are the regulatory mechanisms that ensure proper timing and coordination of events during the cell cycle? Biochemical and genetic screens have identified a number of cell-cycle regulators, and it was recognized early on that many of the genes encoding cell-cycle regulators, including cyclins, were transcribed only in distinct phases of the cell cycle. Thus, “just in time” expression is likely an important part of the mechanism that maintains the proper temporal order of cell cycle events. New high-throughput technologies for measuring transcript levels have revealed that a large percentage of the Saccharomyces cerevisiae transcriptome (~20 %) is cell cycle regulated. Similarly, a substantial fraction of the mammalian transcriptome is cell cycle-regulated. Over the past 25 years, many studies have been undertaken to determine how gene expression is regulated during the cell cycle. In this review, we discuss contemporary models for the control of cell cycle-regulated transcription, and how this transcription program is coordinated with other cell cycle events in S. cerevisiae. In addition, we address the genomic approaches and analytical methods that enabled contemporary models of cell cycle transcription. Finally, we address current and future technologies that will aid in further understanding the role of periodic transcription during cell cycle progression.


title: The Immune Response to Viral Lower Respiratory Tract Infection

text: Viruses are responsible for the majority of respiratory infections in childhood,causing considerable morbidity and mortality. It is estimated that in the United States approximately $ 652 million per year is spent on medical costs for respiratory syncytial virus (RSV) related disease alone (Paramore et al., 2004). Viruses cause a variety of respiratory diseases in children from the common cold to life-threatening pneumonia and bronchiolitis. The host reacts to a viral infection with a combination of innate and adaptive immune mechanisms, usually resulting in the clearance of the virus and clinical recovery. However, there is an accumulating evidence for a number of viral infections that the host immune response actually enhances disease in the course of clearing virus from the infected organs. Interestingly, the effectiveness of the immune response seems to be dependent on the age and probably genetic background of the child. This has important implications for treatment as well as vaccine development.


title: Imaging of Pulmonary Infection

text: The spectrum of organisms known to cause respiratory infections is broad and constantly increasing as new pathogens are identified, and an increasing number of patients have impaired immunity due to disease or medications. The radiographic manifestations of a given organism may be variable depending on the immunologic status of the patient and the presence of pre- or coexisting lung disease. Moreover, the clinical data and radiographic findings often fail to lead to a definitive diagnosis of pneumonia because there are an extensive number of noninfectious processes associated with febrile pneumonitis. This chapter describes and illustrates the characteristic imaging manifestations of the most common community- acquired pneumonias, nosocomial pneumonias, and the various infections seen in both immunocompetent and immunocompromised patients.


title: Drugs and pharmaceuticals: management of intoxication and antidotes

text: The treatment of patients poisoned with drugs and pharmaceuticals can be quite challenging. Diverse exposure circumstances, varied clinical presentations, unique patient-specific factors, and inconsistent diagnostic and therapeutic infrastructure support, coupled with relatively few definitive antidotes, may complicate evaluation and management. The historical approach to poisoned patients (patient arousal, toxin elimination, and toxin identification) has given way to rigorous attention to the fundamental aspects of basic life suppport — airway management, oxygenation and ventilation, circulatory competence, thermoregulation, and substrate availability. Selected patients may benefit from methods to alter toxin pharmacokinetics to minimize systemic, target organ, or tissue compartment exposure (either by decreasing absorption or increasing elimination). These may include syrup of ipecac, orogastric lavage, activated single- or multi-dose charcoal, whole bowel irrigation, endoscopy and surgery, urinary alkalinization, saline diuresis, or extracorporeal methods (hemodialysis, charcoal hemoperfusion, continuous venovenous hemofiltration, and exchange transfusion). Pharmaceutical adjuncts and antidotes may be useful in toxicant-induced hyperthermias. In the context of analgesic, anti-inflammatory, anticholinergic, anticonvulsant, antihyperglycemic, antimicrobial, antineoplastic, cardiovascular, opioid, or sedative-hypnotic agents overdose, N-acetylcysteine, physostigmine, l-carnitine, dextrose, octreotide, pyridoxine, dexrazoxane, leucovorin, glucarpidase, atropine, calcium, digoxin-specific antibody fragments, glucagon, high-dose insulin euglycemia therapy, lipid emulsion, magnesium, sodium bicarbonate, naloxone, and flumazenil are specifically reviewed. In summary, patients generally benefit from aggressive support of vital functions, careful history and physical examination, specific laboratory analyses, a thoughtful consideration of the risks and benefits of decontamination and enhanced elimination, and the use of specific antidotes where warranted. Data supporting antidotes effectiveness vary considerably. Clinicians are encouraged to utilize consultation with regional poison centers or those with toxicology training to assist with diagnosis, management, and administration of antidotes, particularly in unfamiliar cases.


title: Anti-endotoxin Properties of Polymyxin B-immobilized Fibers

text: Polymyxin B is an antibiotic that shows strong bactericidal activity against Gram-negative bacteria, by binding to and inactivating endotoxin. Systemic administration of polymyxin B in humans is restricted because of its nephrotoxicity and neurotoxicity, and this compound was therefore considered a strong candidate ligand for the extracorporeal selective adsorption of circulating endotoxin in the blood. Toraymyxin® is a direct hemoperfusion column that uses polymyxin B attached to an insoluble carrier to bind endotoxin in the blood. In 1994, the Japanese National Health Insurance system approved the use of Toraymyxin for the treatment of endotoxemia and septic shock. In this chapter, we will review the development, clinical use, and efficacy of Toraymyxin, examine the structure of the Toraymyxin column, and comment on the current position of Toraymyxin in the treatment of severe sepsis and septic shock. We will also highlight some potential new applications of Toraymyxin for pulmonary diseases.


title: Clinical Aspects of Sepsis: An Overview

text: Sepsis is one of the oldest and most elusive syndromes in medicine. With the confirmation of germ theory by Semmelweis, Pasteur, and others, sepsis was considered as a systemic infection by a pathogenic organism. Although the germ is probably the beginning of the syndrome and one of the major enemies to be identified and fought, sepsis is something wider and more elusive. In this chapter clinically relevant themes of sepsis will be approached to provide an insight of everyday clinical practice for healthcare workers often not directly involved in the patient’s management.


title: The Damage-Response Framework of Microbial Pathogenesis and Infectious Diseases

text: Historical and most currently held views of microbial pathogenesis and virulence are plagued by confusing and imprecise terminology and definitions that require revision and exceptions to accommodate new basic science and clinical information about microbes and infectious diseases. These views are also inherently unable to account for the ability of some microbes to cause disease in certain, but not other hosts, because they are grounded in singular, either microbe-or host-centric views. The damage-response framework is an integrated theory of microbial pathogenesis that puts forth the view that microbial pathogenesis reflects the outcome of an interaction between a host and a microbe, with each entiry contributing to the nature of the outcome, which in turn depends on the amount of host damage that results from the host-microbe interaction. This view is able to accommodate new information and explain why infection with the same microbe can have different outcomes in different hosts. This chapter describes the origins and conceptual underpinnings of and the outcomes of infection put forth in, the damage-response framework.


title: How Immune Complexes from Certain IgG NAbs and Any F(ab′)(2) Can Mediate Excessive Complement Activation

text: In sepsis death follows an excessive inflammatory response involving cytokines and complement that is activated primarily via the amplifying C3/C5 convertase. Excessive stimulation of complement amplification requires IgG-containing or F(ab′)(2)-containing immune complexes (IC) that capture dimeric C3b on one of their heavy chains or heavy chain fragments. The ability of IgG-IC to capture dimeric C3b by the Fab portion is dependent on an affinity for C3 within the Fab portion, but outside the antigen-binding region. This property is rare among IgG NAbs. In contrast to this, the lack of the Fc portion renders the Fab regions of any F(ab′)(2)-IC accessible to nascent C3b, but dimeric C3b deposits only if F(ab′)(2)-IC form secondary IC with anti-hinge NAbs that rigidify the complex and thereby promote deposition of dimeric C3b. Both types of complexes, C3b(2)-IgG-IC and C3b(2)-F(ab′)(2)-IC/anti-hinge NAbs, are potent precursors of alternative C3 convertases and stimulate complement amplification along with properdin up to 750 times more effectively than C3b and properdin. F(ab′)(2) fragments are not normally generated, but are formed from NAbs by enzymes from pathogens and neutrophils in sepsis. Unlike IgG-IC F(ab′)(2)-IC are not cleared by Fc-receptor dependent processes and circulate long enough to form secondary IC with anti-hinge NAbs that rigidify the complexes such that they capture dimeric C3b and gain the potency to stimulate complement amplification.


Febrile Neutropenia in Children with Cancer


title: Macrophage Pattern Recognition Receptors in Immunity, Homeostasis and Self Tolerance

text: Macrophages, a major component of innate immune defence, express a large repertoire of different classes of pattern recognition receptors and other surface antigens which determine the immunologic and homeostatic potential of these versatile cells. In the light of present knowledge of macrophage surface antigens, we discuss self versus nonself recognition, microbicidal effector functions and self tolerance in the innate immune system.


Construction of a Full-Length cDNA Infectious Clone of a European-Like Type 1 PRRSV Isolated in the U.S.


title: The Role of OX40 (CD134) in T-Cell Memory Generation

text: Memory T-cell generation is limited by activation-induced cell death during the effector T-cell stage. Cell surface proteins are known to transmit signals that either accentuate or limit T-cell death after activation. This chapter will focus on the TNF-receptor family member OX40, which is expressed on effector T cells and when engaged greatly enhances survival of T cells leading to increased memory T-cell generation. Targeting OX40 in vivo can alter the fate of T-cell survival. Enhancing OX40 signaling during Ag priming through agonists increases memory T-cell development, while blocking OX40 signaling decreases the memory T-cell pool. These two opposing outcomes provide therapeutic tools for blocking inflammation in autoimmune conditions and enhancing immunity in hosts harboring cancer or chronic pathogens. OX40 agonists and antagonists are in the first stages of human clinical trials and their therapeutic potential will soon be realized.


title: Modelling Environmentally-Mediated Infectious Diseases of Humans: Transmission Dynamics of Schistosomiasis in China

text: Macroparasites of humans are sensitive to a variety of environmental variables, including temperature, rainfall and hydrology, yet current comprehension of these relationships is limited. Given the incomplete mechanistic understanding of environment-disease interactions, mathematical models that describe them have seldom included the effects of time-varying environmental processes on transmission dynamics and where they have been included, simple generic, periodic functions are usually used. Few examples exist where seasonal forcing functions describe the actual processes underlying the environmental drivers of disease dynamics. Transmission of human schistosomes, which involves multiple environmental stages, offers a model for applying our understanding of the environmental determinants of the viability, longevity, infectivity and mobility of these stages to controlling disease in diverse environments. Here, a mathematical model of schistosomiasis transmission is presented which incorporates the effects of environmental variables on transmission. Model dynamics are explored and several key extensions to the model are proposed.


title: What the Paediatrician Needs to Know When Pandemic Influenza Arrives in Clinical Practice

text: Avian (H5N1) influenza or “bird ‘flu” has received considerable attention in both the medical literature and the mass media in the last few years. Despite the tabloids’ portrayal of an imminent threat, to date there have been relatively few cases in humans in spite of large numbers of infected poultry (Hien et al. 2004). However, this may be falsely reassuring. Most indications suggest that it is just a matter of time until the next influenza pandemic occurs (Osterholm 2005). In the words of the UK Chief Medical Officer: “most experts believe that it is not a question of whether there will be another severe influenza pandemic but when” (Department of Health 2005). Although experts are agreed that a future influenza pandemic is almost inevitable, its timing is unpredictable and it is uncertain whether the virus responsible will be H5N1 or another, novel, influenza strain (Osterholm 2005). A recent editorial described avian influenza as a “predicament of extraordinary proportions” (Anonymous 2006). The next influenza pandemic will have a dramatic impact on all levels of health care including the everyday work of doctors. This chapter focuses on the clinical aspects of pandemic influenza about which paediatricians need to be familiar.


Porcine Arterivirus Entry in Macrophages: Heparan Sulfate – Mediated Attachment, Sialoadhesin-Mediated Internalization, and a Cell-Specific Factor Mediating Virus Disassembly and Genome Release


Equine Viral Arteritis


title: An Overview of Brevinin Superfamily: Structure, Function and Clinical Perspectives

text: Antimicrobial peptides are the backbone of first-line defense against various microorganisms in the animal kingdom. Thus, not surprisingly, they are gaining attention in the science and medical fields as a rich repository of new pro-drugs. Below, we focus our attention on the Brevinin family of anuran peptides. While most of them show strong antibacterial activities, some, e.g. Brevinin-2R, appear to be promising anticancer molecules, exhibiting better a therapeutic window than widely-use anticancer drugs like doxorubicin. We briefly introduce the field, followed by highlighting the promising therapeutic properties of Brevinins. Next, we provide information about the cloning and phylogenetic aspects of Brevinin genes. In the final paragraphs of this chapter, we discuss possible large-scale production methods of Brevinins, giving examples of some systems that are already in use. Towards the end, we discuss various means of modification of biologic properties of Brevinins, either by chemical modifications or by amino acid substitution and sequence rearrangements. In this context, also other unique properties of Brevinins are briefly mentioned. Finally, we discuss the future of the Brevinin field, particularly highlighting yet to be answered biologic questions, like for example presumed anti-viral and antitumor activities of Brevinin family members.


title: Vaccine Containing Natural TLR Ligands Protects from Salmonella typhimurium Infection in Mice and Acute Respiratory Infections in Children

text: It has been shown that a single parenteral administration of vaccine containing bacterial ligands for TLR1, TLR2, TLR4, TLR6, and TLR9 in mice induced rapid (24 h after administration) and effective (100%), but short-term (96 h) protection against lethal challenge with Salmonella typhimurium. Repeated mucosal applications of this vaccine stimulated long-term (up to 9 months) protection against acute respiratory infections in children of preschool age.


NKG2D Signaling and Host Defense after Mouse Hepatitis Virus Infection of the Central Nervous System


title: Nuclear Morphometry, Epigenetic Changes, and Clinical Relevance in Prostate Cancer

text: Nuclear structure alterations in cancer involve global genetic (mutations, amplifications, copy number variations, translocations, etc.) and epigenetic (DNA methylation and histone modifications) events that dramatically and dynamically spatially change chromatin, nuclear body, and chromosome organization. In prostate cancer (CaP) there appears to be early (<50 years) versus late (>60 years) onset clinically significant cancers, and we have yet to clearly understand the hereditary and somatic-based molecular pathways involved. We do know that once cancer is initiated, dedifferentiation of the prostate gland occurs with significant changes in nuclear structure driven by numerous genetic and epigenetic processes. This review focuses upon the nuclear architecture and epigenetic dynamics with potential translational clinically relevant applications to CaP. Further, the review correlates changes in the cancer-driven epigenetic process at the molecular level and correlates these alterations to nuclear morphological quantitative measurements. Finally, we address how we can best utilize this knowledge to improve the efficacy of personalized treatment of cancer.


Human Coronavirus NL63 Infection is Associated with Croup


The Nuclear Localization Signal of the Prrs Virus Nucleocapsid Protein Modulates Viral Replication in vitro and Antibody Response in vivo


title: Viral Infections

text: CMV can cause symptoms from almost any organ as well as unspecific symptoms such as fever, malaise, and bone marrow suppression in stem cell transplant patients. However, the most important clinical entities in allo-HSCT patients are pneumonia, gastroenteritis, and retinitis.


title: Infrastructure and Organization of Adult Intensive Care Units in Resource-Limited Settings

text: In this chapter, we provide guidance on some basic structural requirements, focusing on organization, staffing, and infrastructure. We suggest a closed-format intensive care unit (ICU) with dedicated physicians and nurses, specifically trained in intensive care medicine whenever feasible. Regarding infrastructural components, a reliable electricity supply is essential, with adequate backup systems. Facilities for oxygen therapy are crucial, and the choice between oxygen concentrators, cylinders, and a centralized system depends on the setting. For use in mechanical ventilators, a centralized piped system is preferred. Facilities for proper hand hygiene are essential. Alcohol-based solutions are preferred, except in the context of Ebola virus disease (chloride-based solutions) and Clostridium difficile infection (soap and water). Availability of disposable gloves is important for self-protection; for invasive procedures masks, caps, sterile gowns, sterile drapes, and sterile gloves are recommended. Caring for patients with highly contagious infectious diseases requires access to personal protective equipment. Basic ICU equipment should include vital signs monitors and mechanical ventilators, which should also deliver noninvasive ventilator modes. We suggest that ICUs providing invasive ventilatory support have the ability to measure end-tidal carbon dioxide and if possible can perform blood gas analysis. We recommend availability of glucometers and capabilities for measuring blood lactate. We suggest implementation of bedside ultrasound as diagnostic tool. Finally, we recommend proper administration of patient data; suggest development of locally applicable bundles, protocols, and checklists for the management of sepsis; and implement systematic collection of quality and performance indicators to guide improvements in ICU performance.


Use of a PRRSV Infectious Clone to Evaluate in Vitro Quasispecies Evolution


Want to Improve Public Health Access? Let's Start with the Basics: Measuring Efficiency Correctly.


title: Virus-Like Particles as a Vaccine Delivery System: Myths and Facts

text: Vaccines against viral disease have traditionally relied on attenuated virus strains or inactivation of infectious virus. Subunit vaccines based on viral proteins expressed in heterologous systems have been effective for some pathogens, but have often suffered from poor immunogenicity due to incorrect protein folding or modification. In this chapter we focus on a specific class of viral subunit vaccine that mimics the overall structure of virus particles and thus preserves the native antigenic conformation of the immunogenic proteins. These virus-like particles (VLPs) have been produced for a wide range of taxonomically and structurally distinct viruses, and have unique advantages in terms of safety and immunogenicity over previous approaches. With new VLP vaccines for papillomavirus beginning to reach the market place we argue that this technology has now ‘come-of-age’ and must be considered a viable vaccine strategy.


The Missing Antibody: The Pitfalls of ANCA Testing


Fatal outcome of SARS in a patient with reactivation of chronic hepatitis B


title: The membrane-bound basic carboxypeptidase from hog intestinal mucosa

text: The carboxypeptidase activity occurring in hog intestinal mucosa is apparently due to two distinct enzymes which may be responsible for the release of basic COOH-terminal amino acids from short peptides. The plasma membrane-bound carboxypeptidase activity which occurs at neutral optimum pH levels was found to be enhanced by CoCl(2) and inhibited by guanidinoethylmercaptosuccinic acid, o-phenanthroline, ethylenediamine tetraacetic acid and cadmium acetate; whereas the soluble carboxypeptidase activity which occurs at an optimum pH level of 5.0 was not activated by CoCl(2) and only slightly inhibited by o-phenanthroline, ethylenediamine tetraacetic acid, NiCl(2) and CdCl(2). The latter activity was presumably due to lysosomal cathepsin B, which is known to be present in the soluble fraction of hog intestinal mucosa. Although the membrane-bound enzyme was evenly distributed along the small intestine, it was not anchored in the phospholipidic bilayer via a glycosyl-phosphatidylinositol moiety, as carboxypeptidase M from human placenta is. The enzyme was not solubilized by phosphatidylinositol-specific phospholipase C, but was solubilized to practically the same extent by several detergents. The purified trypsin-solubilized form is a glycoprotein with a molecular mass of 200 kDa, as determined by performing SDS–PAGE and gel filtration, which differs considerably from the molecular mass of human placental carboxypeptidase M (62 kDa). It was found to cleave lysyl bonds more rapidly than arginyl bonds, which is not so in the case of carboxypeptidase M, and immunoblotting analysis provided further evidence that hog intestinal and human placental membrane-bound carboxypeptidases do not bear much resemblance to each other. Since the latter enzyme has been called carboxypeptidase M, it is suggested that the former might be carboxypeptidase D, the recently described new member of the carboxypeptide B-type family.


Introduction to the Special Issue ‘Viral Membrane Proteins — Channels for Cellular Networking’


A Fall in Ghana


title: Applications of Luminex® xMAP™ technology for rapid, high-throughput multiplexed nucleic acid detection

text: BACKGROUND: As we enter the post-genome sequencing era and begin to sift through the enormous amount of genetic information now available, the need for technologies that allow rapid, cost-effective, high-throughput detection of specific nucleic acid sequences becomes apparent. Multiplexing technologies, which allow for simultaneous detection of multiple nucleic acid sequences in a single reaction, can greatly reduce the time, cost and labor associated with single reaction detection technologies. METHODS: The Luminex® xMAP™ system is a multiplexed microsphere-based suspension array platform capable of analyzing and reporting up to 100 different reactions in a single reaction vessel. This technology provides a new platform for high-throughput nucleic acid detection and is being utilized with increasing frequency. Here we review specific applications of xMAP technology for nucleic acid detection in the areas of single nucleotide polymorphism (SNP) genotyping, genetic disease screening, gene expression profiling, HLA DNA typing and microbial detection. CONCLUSIONS: These studies demonstrate the speed, efficiency and utility of xMAP technology for simultaneous, rapid, sensitive and specific nucleic acid detection, and its capability to meet the current and future requirements of the molecular laboratory for high-throughput nucleic acid detection.


Middle East respiratory syndrome coronavirus infection control: The missing piece?


Combined infection training – should we be concerned about its impact on infection prevention and control training of microbiologists in the UK?


title: Recent advances in avian virology

text: Selected, recent research on the following avian diseases, and their causative viruses, has been reviewed: chicken anaemia, infectious bursal disease, turkey rhinotracheitis, avian nephritis, fowlpox, influenza, infectious bronchitis and turkey enteritis.


Facing an outbreak of highly transmissible disease: problems in emergency department response


Middle East Respiratory Syndrome Interpreted: Human Herpesvirus 6B Pneumonia


Pomalidomide-Induced Pulmonary Toxicity in Multiple Myeloma


What are the main drivers for public policies to prevent healthcare-associated infections?


The open-air factor and infection control


Transmission of severe acute respiratory syndrome in an emergency department


Incidental Administration of Corticosteroid Can Mask the Diagnosis of Tuberculosis


Career options in aerospace and aviation medicine()


Stamping out SARS


DEPARTMENT OF ERROR


Novel coronavirus and severe acute respiratory syndrome


The truth about PHEICs


Zoonotic Viruses: The Mysterious Leap From Animals to Man


Personal view of SARS: confusing definition, confusing diagnoses


End of an era as Lee confirmed to succeed Brundtland


WHO to strengthen commitment to polio eradication


title: ROTAVIRUS ENTERITIS IN THE WEST MIDLANDS DURING 1974

text: During 1974 in the West Midlands of England, 38% of children less than 6 years old with enteritis were excreting rotaviruses. Children aged from 6 months to 3 years were those most commonly infected. Rotavirus infections were most common during winter with only a few sporadic cases during summer. A possible pathogen was detected in 59% of patients examined.


AIDS campaign signals new WHO priorities and approach


Treatment of severe acute respiratory syndrome in health-care workers


title: Standardization and kinetics of in vitro bovine blood lymphocyte stimulation with bovine rotavirus

text: Two groups of 3-month old calves were immunized intramuscularly with attenuated bovine rotavirus and boosted 21 and 42 days later. The first group of three calves were vaccinated with live virus emulsified with incomplete Freund's adjuvant (IFA) and the second group was immunized with live virus suspended in phosphate buffered saline (PBS). Three other calves, serving as controls, were inoculated with PBS emulsified with IFA. The specific cell-mediated and antibody responses of the animals were studied. Preliminary analysis of in vitro peripheral blood lymphocyte transformation to bovine rotavirus determined optimal conditions as: 96 h culture period, 5 × 10(5) cells per culture in RPMI 1640 medium containing 10% heat-inactivated bovine fetal serum and the use of inactivated virus in the cell culture at a concentration of 5 × 10(6) median tissue culture infective dose before inactivation. Specific blastic stimulation was observed on calves immunized with the rotavirus emulsified with IFA after the second and third vaccine inoculation with stimulation index values varying from 2.00 to 5.73. Serum neutralizing antibody titers of 125,600 were also induced in the same calves. Calves immunized with rotavirus-PBS suspension developed a mean antibody titer of 11,600, but showed no specific lymphocyte stimulation. No increase in specific immune responses was detected in the control animals.


title: Streamlined selection of cancer antigens for vaccine development through integrative multi-omics and high-content cell imaging

text: Identification of tumor antigens that induce cytotoxic T lymphocytes (CTLs) is crucial for cancer-vaccine development. Despite their predictive ability, current algorithmic approaches and human leukocyte antigen (HLA)-peptidomic analysis allow limited selectivity. Here, we optimized a method to rapidly screen and identify highly immunogenic epitopes that trigger CTL responses. We used a combined application of this method involving immune-specific signature analysis and HLA-associated peptidomics using samples from six patients with triple-negative breast cancer (TNBC) in order to select immunogenic HLA epitopes for in vitro testing. Additionally, we applied high-throughput imaging at the single-cell level in order to confirm the immunoreactivity of the selected peptides. The results indicated that this method enabled identification of promising CTL peptides capable of inducing antitumor immunity. This platform combining high-resolution computational analysis, HLA-peptidomics, and high-throughput immunogenicity testing allowed rapid and robust identification of highly immunogenic epitopes and represents a powerful technique for cancer-vaccine development.


title: Epidemics with mutating infectivity on small-world networks

text: Epidemics and evolution of many pathogens occur on similar timescales so that their dynamics are often entangled. Here, in a first step to study this problem theoretically, we analyze mutating pathogens spreading on simple SIR networks with grid-like connectivity. We have in mind the spatial aspect of epidemics, which often advance on transport links between hosts or groups of hosts such as cities or countries. We focus on the case of mutations that enhance an agent’s infection rate. We uncover that the small-world property, i.e., the presence of long-range connections, makes the network very vulnerable, supporting frequent supercritical mutations and bringing the network from disease extinction to full blown epidemic. For very large numbers of long-range links, however, the effect reverses and we find a reduced chance for large outbreaks. We study two cases, one with discrete number of mutational steps and one with a continuous genetic variable, and we analyze various scaling regimes. For the continuous case we derive a Fokker-Planck-like equation for the probability density and solve it for small numbers of shortcuts using the WKB approximation. Our analysis supports the claims that a potentiating mutation in the transmissibility might occur during an epidemic wave and not necessarily before its initiation.


title: 3D-quantitative structure–activity relationship and antiviral effects of curcumin derivatives as potent inhibitors of influenza H1N1 neuraminidase

text: Curcumin derivatives have been shown to inhibit replication of human influenza A viruses (IAVs). However, it is not clear whether curcumin and its derivatives can inhibit neuraminidase (NA) of influenza virus. In this study, a meaningful 3D quantitative structure–activity relationship model (comparative molecular field analysis R(2) = 0.997, q(2) = 0.527, s = 0.064, F = 282.663) was built to understand the chemical–biological interactions between their activities and neuraminidase. Molecular docking was used to predict binding models between curcumin derivatives and neuraminidase. Real-time polymerase chain reactions showed that the five active curcumin derivatives might have direct effects on viral particle infectivity in H1N1-infected lung epithelial (MDCK) cells. Neuraminidase activation assay showed that five active curcumin derivatives decreased H1N1-induced neuraminidase activation in MDCK cells. Indirect immunofluorescence assay indicated that two active curcumin derivatives (tetramethylcurcumin and curcumin) down-regulated the nucleoprotein expression. Curcumin inhibited IAV in vivo. The therapeutic mechanism of curcumin in the treatment of influenza viral pneumonia is related to improving the immune function of infected mice and regulating secretion of tumor necrosis-α, interleukin-6, and interferon-γ. These results indicate that curcumin derivatives inhibit IAV by blocking neuraminidase in the cellular model and curcumin also has anti-IAV activity in the animal model.


title: Innate immune activation as a broad-spectrum biodefense strategy: Prospects and research challenges

text: Biodefense strategies require protection against a broad and largely unforeseen spectrum of pathogens—the forte of innate immune system defenses—that have evolved over millennia to function within moments of encountering either ancient or newly emerging pathogens. Although constitutive, the innate immune system is activated by the presence of microbes or their products, providing a rationale for a potential biodefense strategy. Both prophylactic and postexposure strategies involving innate immune stimulation have been shown to be plausible to prevent or ameliorate infections in animal models. Innate immune-activating compounds based on conserved microbial components recognized by toll-like molecules and other receptors could be synthesized and delivered like drugs by using an entirely different strategy from conventional vaccination. However, important theoretic and practical questions emerge about developing and deploying innate immune protective strategies for biodefense. This rostrum discusses prospects and problems in the overall approach itself. Important topics include microbe-specific issues about innate immune system effectiveness against highly virulent pathogens and general questions, such as whether innate immune responses will be safe and effective if used in a diverse human population of different age groups and with different genetic makeups.


title: Possible immunoenhancement of persistent viremia by feline leukemia virus envelope glycoprotein vaccines in challenge-exposure situations where whole inactivated virus vaccines were protective

text: Kittens immunized with purified native FeLV-gp70 or -gp85 envelope proteins developed ELISA, but not virus neutralizing, antibodies in their serum to both whole FeLV and FeLV-gp70. Kittens vaccinated with envelope proteins and infected with feline sarcoma virus (FeSV) developed smaller tumors than nonvaccinates, but a greater incidence of persistent retroviremia. Similarly, FeLV-gp70 and -gp85 vaccinated kittens were more apt to become persistently retroviremic following virulent FeLV challenge exposure than nonvaccinates. Kittens vaccinated with inactivated whole FeLV developed smaller tumors after FeSV inoculation and had a lower incidence of persistent retroviremia than nonvaccinates. The protective effect of inactivated whole FeLV vaccine against persistent retroviremia was also seen with FeLV challenge-exposed cats. Protection afforded by inactivated whole FeLV vaccine was not associated with virus neutralizing antibodies, although ELISA antibodies to both whole FeLV and FeLV-gp70 were induced by vaccination.


The Regulation and Maturation of Antiviral Immune Responses


title: Pulmonary high-resolution computed tomography findings in nephropathia epidemica

text: PURPOSE: To evaluate lung high-resolution computed tomography (HRCT) findings in patients with Puumala hantavirus-induced nephropathia epidemica (NE), and to determine if these findings correspond to chest radiograph findings. MATERIALS AND METHODS: HRCT findings and clinical course were studied in 13 hospital-treated NE patients. Chest radiograph findings were studied in 12 of them. RESULTS: Twelve patients (92%) showed lung parenchymal abnormalities in HRCT, while only 8 had changes in their chest radiography. Atelectasis, pleural effusion, intralobular and interlobular septal thickening were the most common HRCT findings. Ground-glass opacification (GGO) was seen in 4 and hilar and mediastinal lymphadenopathy in 3 patients. Atelectasis and pleural effusion were also mostly seen in chest radiographs, other findings only in HRCT. CONCLUSION: Almost every NE patient showed lung parenchymal abnormalities in HRCT. The most common findings of lung involvement in NE can be defined as accumulation of pleural fluid and atelectasis and intralobular and interlobular septal thickening, most profusely in the lower parts of the lung. As a novel finding, lymphadenopathy was seen in a minority, probably related to capillary leakage and overall fluid overload. Pleural effusion is not the prominent feature in other viral pneumonias, whereas intralobular and interlobular septal thickening are characteristic of other viral pulmonary infections as well. Lung parenchymal findings in HRCT can thus be taken not to be disease-specific in NE and HRCT is useful only for scientific purposes.


title: Attenuation of acute lung injury in mice by oxymatrine is associated with inhibition of phosphorylated p38 mitogen-activated protein kinase

text: Oxymatrine is one of the alkaloids extracted from Chinese herb Sophora japonica (Sophora flavescens Ait.) with activities of anti-inflammation, inhibiting immune reaction, antivirus, protecting hepatocytes and antihepatic fibrosis. However, the effect of oxymatrine on acute lung injury (ALI) has not been known yet. In this study, the effect of oxymatrine on ALI was investigated using an oleic acid-induced ALI mouse model. Morphological findings showed that the oleic acid group demonstrated a marked lung injury represented by prominent atelectasis, intraalveolar and interstitial patchy hemorrhage, edema, thickened alveolar septum, formation of hyaline membranes and the existence of inflammatory cells in alveolar spaces. While in the oxymatrine/dexamethasone group, these changes were less severe and in the vicinity of the control group. Furthermore, pretreatment with oxymatrine significantly alleviated oleic acid-induced lung injury accompanied by reduction of lung index and wet-to-dry weight ratio, decreases in serum TNF-α level and inhibition of phosphorylated p38 MAPK. These findings suggest that oxymatrine has a beneficial effect on acute lung injury induced by oleic acid in mice and may inhibit the production of proinflammatory cytokine, TNF-α, by means of the inhibition of p38 MAPK.


Inhibitors of protein glycosylation and glycoprotein processing in viral systems


title: The main neutral aminopeptidase activity of human lymphoid tumour cell lines does not originate from the aminopeptidase N-(APN; CD13) gene

text: Lymphocytes and related cell lines are predominantly CD13-negative, however, there are reports describing neutral aminopeptidase activity in or on these cells. The aim of this study was to answer the question, whether this activity originates from APN-gene expression. The total cellular activities (Ala-pNA hydrolysis) of lymphoid cell lines are up to 15 times higher than that of normal lymphocytes. Despite weak or lacking CD13 surface expression all lymphoid cell lines tested contain APNmRNA as quantified by competitive RT-PCR as well as low enzymatic activity in their particulate fractions. By isoelectric focusing two enzyme species with isoelectric points of 5.4 or between 3.5 to 4.8, respectively, were detected. To investigate whether these activities result from APN-gene we established transfectants lacking cellular APN expression of the CD13-positive histiocytic cell line U937 and the CD13-negative T cell line H9. Studies on these transfectants proved (I) that the main neutral aminopeptidase activity expressed in lymphoid cells is definitively not related to APN and (II) that APN is also expressed in lymphoid cells, although on a low level only.


On the Interface of Infectious Diseases and Critical Care Medicine


SELECTED EPIDEMICS & EMERGING PATHOGENS


Virus-Induced Demyelination: From Animal Models to Human Diseases


Dealing With Threat of Drug-Resistant Tuberculosis: Background Information for Interpreting the Andrew Speaker and Related Cases


title: Patients Would Prefer Ward to Emergency Department Boarding While Awaiting an Inpatient Bed

text: Boarding of admitted patients in the Emergency Department (ED), rather than in inpatient care areas, is widespread. We surveyed boarded patients, patients without a disposition, and visitors at a county hospital ED serving a mixed urban and rural population. Subjects were asked “If you needed to be admitted to the hospital but no inpatient bed is available, would you prefer to be kept in an ER hallway or a hallway on an inpatient ward?” Boarded patients said they would prefer ward to ED boarding, 117/213 (54.9%; 95% confidence interval [CI] 48.0%–61.7%). Patients without a disposition 314/477 (65.8%; 95% CI 61.4%–70.0%) and visitors 370/532 (69.5%; 95% CI 65.4%–73.4%) stated a preference for ward boarding in 314/477 (65.8%; 95% CI 61.4%–70.0%) and in 370/532 (69.5%; 95% CI 65.4%–73.4%), respectively. Common reasons for preferring inpatient ward boarding were privacy concerns and reduced noise levels. Those preferring ED boarding valued easy access to a doctor.


Autophagy: Nobel Prize 2016 and allergy and asthma research


Safe, high-throughput screening of natural compounds of MERS-CoV entry inhibitors using a pseudovirus expressing MERS-CoV spike protein


Comparación entre la técnica shell vial y una PCR múltiple en el diagnóstico de las infecciones respiratorias víricas


Serum Procalcitonin and Infective Exacerbations of Asthma


Emerging and Reemerging Infectious Diseases: Global Overview