Archive for June, 2010

Antimicrobials: treasures from the oceans

Thursday, June 17th, 2010

Actinomycete The ocean is teaming with sunken treasure and for some treasure hunters the sand itself is the target. In this article in Microbiology Today, Jem Stach tells how novel actinomycetes on the seabed could be a source of much-needed novel antimicrobial drugs:

For most, the search for sunken treasure evokes images of glistening gold discovered when a diver’s hand wafts away the sand. However, for another kind of treasure hunter, the bioprospector, the sand itself is the target, brought from the depths of the sea, to the laboratory bench. In this case, the bioprospector is interested in marine microbes and their potential to produce antimicrobial compounds. With global sales of these life-saving products set to exceed $100 billion by 2015, such micro-organisms could be worth more than their weight in gold.

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Virus tricks to grid-lock the type I interferon system

Wednesday, June 16th, 2010

Interferon signalling The innate immune system forms the first line of defence against invading micro-organisms such as viruses. It dampens initial virus replication and ensures survival of the host until specialized adaptive responses are developed. Type I interferons (IFNs) are secreted key cytokines on the innate immune axis that protect uninfected cells and stimulate leukocytes residing at the interface of innate and adaptive immunity, such as macrophages and dendritic cells. These cells prod the adaptive immune system to mount a full, specialized response against the invading microbe.

The ability to outrun innate immunity before adaptive immune responses are mounted is crucial for the survival of virtually all the mammalian viruses, regardless of their genome type and complexity. Relatively simple viruses such as RNA viruses from the Picornavirus family, as well as DNA viruses with large genomes, such as members from the Poxvirus family, have been shown to inhibit the IFN system. This review covers the latest insights into how virus-encoded antagonists sidetrack the IFN machinery and how this knowledge is currently used to generate second generation live vaccines and antiviral compounds.

Viral tricks to grid-lock the type I interferon system. Curr Opin Microbiol. Jun 9 2010
Type I interferons (IFNs) play a crucial role in the innate immune avant-garde against viral infections. Virtually all viruses have developed means to counteract the induction, signaling, or antiviral actions of the IFN circuit. Over 170 different virus-encoded IFN antagonists from 93 distinct viruses have been described up to now, indicating that most viruses interfere with multiple stages of the IFN response. Although every viral IFN antagonist is unique in its own right, four main mechanisms are employed to circumvent innate immune responses: (i) general inhibition of cellular gene expression, (ii) sequestration of molecules in the IFN circuit, (iii) proteolytic cleavage, and (iv) proteasomal degradation of key components of the IFN system. The increasing understanding of how different viral IFN antagonists function has been translated to the generation of viruses with mutant IFN antagonists as potential live vaccine candidates. Moreover, IFN antagonists are attractive targets for inhibition by small-molecule compounds.

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Statins and Candida

Tuesday, June 15th, 2010

Statins Candida infections are the fourth most common cause of nosocomial blood stream infections and are associated with a significant mortality. Delays in antifungal therapy have been associated with increased hospital costs of over US$6,000 per patient and overall mortality. The role of HMG CoA reductase inhibitors (statins) in improving outcomes bacteremic sepsis is currently being debated with recent papers showing significantly improved survival in patients with systemic inflammatory response syndrome in the intensive care unit, in patients with chronic kidney renal disease and patients with community acquired pneumonia and influenza. One explanation of this effect is that statins in animal models have shown to reduce inflammatory markers, in particular the release of cytokines and cytotoxic effects of neutrophils. The reduction in inflammatory cytokines has also been demonstrated in patients in a prospective randomized study comparing simvastatin to placebo where there was a significant reduction in tumor necrosis factor alpha (TNF-α) and interleukin-6 (IL-6) in the statin group. Yeasts use the same HMG CoA reductase as humans, however their end-product is ergosterol rather than cholesterol. In vitro studies have demonstrated that simvastatin greatly inhibits the growth of Candida species. This review suggests there is a clinical benefit of statin therapy throughout antifungal therapy in intensive care unit patients with confirmed candidemia.

Statins in Candidemia: clinical outcomes from a matched cohort study. 2010 BMC Infectious Diseases 10: 152 doi:10.1186/1471-2334-10-15210
HMG CoA reductase inhibitors (statins) in patients with bacteremic sepsis have shown significant survival benefits in several studies. There is no data on the effect of statins in candidemic patients, however in-vitro models suggest that statins interfere with ergesterol formation in the wall of yeasts.
This retrospective matched- cohort study from 1/2003 to 12/2006 evaluated the effects of statins on patients with candidemia within intensive care units. Statin-users had candidemia as a cause of their systemic inflammatory response and were on statins throughout their antifungal therapy, while non-statin-users were matched based on age +/- 5 years and co-morbid factors. Primary analysis was 30-day survival or discharge using bivariable comparisons. Multivariable comparisons were completed using conditional logistic regression. All variables with a p-value less than 0.10 in the bivariable comparisons were considered for inclusion in the conditional logistic model.
There were 15 statin-users and 30 non-statin users that met inclusion criteria, all with similar demographics and co-morbid conditions except the statin-users had significantly more coronary artery disease (P<0.01), peripheral vascular disease (P=0.03) and lower median APCAHE II scores (p=0.03). There were no differences in duration of candidemia , antifungal therapy or Candida species between the groups. Statins were associated with lower mortality on bivariable and multivariable analyses compared to controls; although, in the latter the protective effect lacked statistical signficance.
In our small, single-center matched cohort study, statins appear may provide a survival benefit in candidemia, however further studies are warranted to validate and further explore this association.

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Moonlighting bacteriophage proteins

Monday, June 14th, 2010

Genomic island Pathogenicity islands have a major role in spreading virulence genes among bacterial populations. A notable example are the phage-related pathogenicity islands of staphylococci, the SaPIs, which are responsible for the inter- as well as intrageneric spread of toxins – such as TSST-1 (toxic shock syndrome toxin) and other superantigens – through the exploitation of specific staphylococcal helper phages for high-frequency transfer within phage-encoded particles. Toxic shock syndrome is a rare, potentially fatal illness that can be caused by the release of toxins from Staphylococcus. The toxic particles are encoded by discrete genetic units called pathogenicity islands, which reside passively in the host chromosome, under the control of the global repressor Stl, unless activated by a helper phage. This paper shows that a non-essential and specific protein from the helper phage 80α is responsible for de-repression of the pathogenicity island, providing the mechanism for the first step of its mobilization. The proteins involved are ‘moonlighters’, because they have two different and genetically distinct activities. Through a remarkable evolutionary adaptation, various related pathogenicity islands co-opt entirely unrelated phage proteins to aid in their mobilization.

Moonlighting bacteriophage proteins derepress staphylococcal pathogenicity islands. 2010 Nature. 465(7299): 779-782
Staphylococcal superantigen-carrying pathogenicity islands (SaPIs) are discrete, chromosomally integrated units of ~15 kilobases that are induced by helper phages to excise and replicate. SaPI DNA is then efficiently encapsidated in phage-like infectious particles, leading to extremely high frequencies of intra- as well as intergeneric transfer. In the absence of helper phage lytic growth, the island is maintained in a quiescent prophage-like state by a global repressor, Stl, which controls expression of most of the SaPI genes4. Here we show that SaPI derepression is effected by a specific, non-essential phage protein that binds to Stl, disrupting the Stl–DNA complex and thereby initiating the excision-replication-packaging cycle of the island. Because SaPIs require phage proteins to be packaged, this strategy assures that SaPIs will be transferred once induced. Several different SaPIs are induced by helper phage 80α and, in each case, the SaPI commandeers a different non-essential phage protein for its derepression. The highly specific interactions between different SaPI repressors and helper-phage-encoded antirepressors represent a remarkable evolutionary adaptation involved in pathogenicity island mobilization.

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When you’re hot, you’re hot

Friday, June 11th, 2010

Guardian University Guide 2011 Biosciences
Guardian University Guide 2011: Biosciences

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Virus ecology

Thursday, June 10th, 2010

Virus ecology Amazingly, all the phages on Earth, if placed end to end, would probably extend a distance equivalent to that of the nearest 60 galaxies. In this article in Microbiology Today, Eric Wommack explains how metagenomics is gradually revealing the amazing diversity and abundance of viruses in the biosphere:

Although the throughput and accuracy of methods for viral direct counting has improved since the 1989 report based on transmission electron microscopy, the ‘factor of 10’ ratio of virus to bacterial abundance within aquatic environments has remained a surprisingly common observation. Extrapolating the ‘factor of 10’ rule to the biosphere has lead to estimates that global viral abundance is in the order of 1031 individuals. Assuming an average length dimension of 100 nm, Curtis Suttle has proposed that, lined end-to-end, all the phages on earth would extend a distance equivalent to that of the nearest 60 galaxies (10 million light years, 1024m).

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Bacteria from Hot Springs to Reveal Clues to Evolution of Early Life

Wednesday, June 9th, 2010

Bacterial mat, Yellowstone A bacteria that lives in hot springs in Japan may help solve one of the mysteries of the early evolution of complex organisms. It may also be the key to 21st century biofuel production. Group II introns are bacterial mobile elements thought to be ancestors of introns and retroelements in higher organisms. They comprise a catalytically active intron RNA and an intron-encoded reverse transcriptase, which promotes splicing of the intron from precursor RNA and integration of the excised intron into new genomic sites. While most bacteria have small numbers of group II introns, in the thermophilic cyanobacterium Thermosynechococcus elongatus, a single intron has proliferated and constitutes 1.3% of the genome. Researchers investigated how the T. elongatus introns proliferated to such high copy numbers. They found divergence of DNA target specificity, evolution of reverse transcriptases that splice and mobilize multiple degenerate introns, and preferential insertion into other mobile introns or insertion elements, which provide new integration sites in non-essential regions of the genome. Unlike mesophilic group II introns, the thermophilic T. elongatus introns rely on higher temperatures to help promote DNA strand separation, facilitating access to DNA target sites. They discuss how these mechanisms, including elevated temperature, might have contributed to intron proliferation in early eukaryotes.

Mechanisms Used for Genomic Proliferation by Thermophilic Group II Introns. 2010 PLoS Biol 8(6): e1000391. doi:10.1371/journal.pbio.1000391
Mobile group II introns, which are found in bacterial and organellar genomes, are site-specific retroelments hypothesized to be evolutionary ancestors of spliceosomal introns and retrotransposons in higher organisms. Most bacteria, however, contain no more than one or a few group II introns, making it unclear how introns could have proliferated to higher copy numbers in eukaryotic genomes. An exception is the thermophilic cyanobacterium Thermosynechococcus elongatus, which contains 28 closely related copies of a group II intron, constituting ,1.3% of the genome. Here, by using a combination of bioinformatics and mobility assays at different temperatures, we identified mechanisms that contribute to the proliferation of T. elongatus group II introns. These mechanisms include divergence of DNA target specificity to avoid target site saturation; adaptation of some intron-encoded reverse transcriptases to splice and mobilize multiple degenerate introns that do not encode reverse transcriptases, leading to a common splicing apparatus; and preferential insertion within other mobile introns or insertion elements, which provide new unoccupied sites in expanding non-essential DNA regions. Additionally, unlike mesophilic group II introns, the thermophilic T. elongatus introns rely on elevated temperatures to help promote DNA strand separation, enabling access to a larger number of DNA target sites by base pairing of the intron RNA, with minimal constraint from the reverse transcriptase. Our results provide insight into group II intron proliferation mechanisms and show that higher temperatures, which are thought to have prevailed on Earth during the emergence of eukaryotes, favor intron proliferation by increasing the accessibility of DNA target sites. We also identify actively mobile thermophilic introns, which may be useful for structural studies, gene targeting in thermophiles, and as a source of thermostable reverse transcriptases.

Lipids and Bluetongue Virus

Tuesday, June 8th, 2010

BTV The replication cycle of viruses involves entry into host cells, synthesis of virus genes and proteins, assembly of progeny virus particles and their subsequent release. Along with the plasma membrane, viruses also have to interact with the endosomal and vesicular membranes during their replication in host cells. All cellular membranes are composed of lipids and proteins that are usually arranged in various micro domains. During infection of cells by enveloped viruses, the lipids present in both viral and cellular membranes mediate fusion and fission reactions to facilitate virus entry and release. Since non-enveloped viruses do not have a lipid envelope, it is generally believed that their entry mechanism does not involve membrane fusion activity and that these viruses are mainly released by cell lysis. Usually, non-enveloped viruses enter the cells by penetrating the membrane barrier, either via the endocytic pathway using clathrin-coated vesicles, or by the formation of a pore at the cell surface. Recent data obtained from biochemical and structural studies indicate that the overall mechanisms of both entry (Reoviridae) and release of certain non-enveloped viruses (e.g., members of the Picornaviridae and Reoviridae) are analogous to that of enveloped viruses, and that the capsid proteins can function in these activities in a similar manner to the membrane viral proteins. This review discusses the role of lipids in the entry, maturation and release of non-enveloped viruses, focusing mainly on Bluetongue virus (BTV).

Role of Lipids on Entry and Exit of Bluetongue Virus, a Complex Non-Enveloped Virus. Viruses 2010 2 (5) 1218-1235. doi:10.3390/v2051218

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Innate immunity against Helicobacter pylori

Monday, June 7th, 2010

The spiral, microaerophilic, Gram-negative bacterium Helicobacter pylori (H. pylori) induces chronic gastritis and is a well known risk factor for peptic ulcer and gastric cancer. Although H. pylori infection can persist for decades, only a fraction of colonized individuals ever develop clinical diseases. Clinical outcome is influenced by a balance between H. pylori virulence factors and the host immune response. However, the mechanisms by which bacterial and/or host factors cause disease remain unclear. Identification of immune response genes that regulate the H. pylori-host interactions will not only have diagnostic and therapeutic implications, but may also provide insights into other inflammation related cancer.

Olfactomedin 4 down-regulates innate immunity against Helicobacter pylori infection. PNAS USA June 1 2010. doi: 10.1073/pnas.100126910
Olfactomedin 4 (OLFM4) is a glycoprotein that has been found to be up-regulated in inflammatory bowel diseases and Helicobacter pylori infected patients. However, its role in biological processes such as inflammation or other immune response is not known. In this study, we generated OLFM4 KO mice to investigate potential role(s) of OLFM4 in gastric mucosal responses to H. pylori infection. H. pylori colonization in the gastric mucosa of OLFM4 KO mice was significantly lower compared with WT littermates. The reduced bacterial load was associated with enhanced infiltration of inflammatory cells in gastric mucosa. Production and expression of proinflammatory cytokines/chemokines such as IL-1β, IL-5, IL-12 p70, and MIP-1α was increased in OLFM4 KO mice compared with infected controls. Furthermore, we found that OLFM4 is a target gene of NF-κB pathway and has a negative feedback effect on NF-κB activation induced by H. pylori infection through a direct association with nucleotide oligomerization domain-1 (NOD1) and -2 (NOD2). Together these observations indicate that OLFM4 exerts considerable influence on the host defense against H. pylori infection acting through NOD1 and NOD2 mediated NF-κB activation and subsequent cytokines and chemokines production, which in turn inhibit host immune response and contribute to persistence of H. pylori colonization.

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