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Western blots are a commonly used method for protein detection and quantification in biological samples. Compensation of loading variations is achieved by housekeeping protein (HKP) normalization and/or total protein normalization (TPN). However, under infection conditions, HKP normalization, traditionally used in cell biology for quantification of western blots, can be problematic. Binding of microbes to target cells via specific receptors can induce signal transduction events resulting in drastic changes in the level of expression of HKPs. Additionally, samples collected after infection assays will include cellular and microbial proteins altering the analysis with TPN. Here we demonstrate under experimental infection conditions, how a reliable semi-quantitative analysis of proteins in western blots can be achieved using the Stain-Free technology. [ABSTRACT FROM AUTHOR]

Regulated loading of eIF3-bound 40S ribosomes on capped mRNA is generally dependent upon the translation initiation factor eIF4E; however, mRNA translation often proceeds during physiological stress, such as virus infection, when eIF4E availability and activity are limiting. It remains poorly understood how translation of virus and host mRNAs are regulated during infection stress. While initially sensitive to mTOR inhibition, which limits eIF4E-dependent translation, we show that protein synthesis in human cytomegalovirus (HCMV)-infected cells unexpectedly becomes progressively reliant upon eIF3d. Targeting eIF3d selectively inhibits HCMV replication, reduces polyribosome abundance, and interferes with expression of essential virus genes and a host gene expression signature indicative of chronic ER stress that fosters HCMV reproduction. This reveals a strategy whereby cellular eIF3d-dependent protein production is hijacked to exploit virus-induced ER stress. Moreover, it establishes how switching between eIF4E and eIF3d-responsive cap-dependent translation can differentially tune virus and host gene expression in infected cells. [Display omitted] • mRNA translation in HCMV-infected cells becomes progressively reliant upon eIF3d • eIF3d depletion selectively restricts HCMV replication and late gene expression • Chronic ER stress-induced host genes regulated by eIF3d promote HCMV replication • An eIF3d-dependent switch tunes infected cell mRNA translation to support HCMV growth Instead of eIF4E-regulated ribosome loading, Thompson et al. show capped mRNA translation in HCMV-infected cells becomes reliant upon eIF3d. Depleting eIF3d inhibits HCMV replication, reduces polyribosomes, and restricts virus late gene and host chronic ER stress-induced gene expression. Thus, switching to eIF3d-responsive translation tunes gene expression to support virus replication. [ABSTRACT FROM AUTHOR]

Abstract: Infection biology and invasion strategies of Kretzschmaria deusta were investigated through the analysis of patterns of colonization and population diversity in three case studies, comprising different host species. Molecular analysis and isolation assays performed on stem sections at different heights indicated a prevalent heart rot mode of expansion. Random amplified microsatellites – RAMs and somatic incompatibility assays on isolates allowed detection, in one case, of a genet occupying the entire decay column and, in the other cases, of several different genets in each individual tree. Hypotheses on modes of arrival and entrance of K. deusta in trees are discussed on the basis of the distribution of genets and population genetics analyses. Significant correlation (Spearman ρ = 1.0; p < 0.001) between the number of genets and the number of areas delimited by pseudosclerotial plates (PSPs) on the decayed portions of stem sections suggests PSPs are interaction zone lines between different individuals of K. deusta occupying adjacent decay columns. [Copyright &y& Elsevier]

Abstract: We investigated the impact of molecular mimicry between pathogenic microbes and their antigenic surrounding on the clinical course and outcome of pneumonia induced sepsis. Using mathematical prediction, we estimated the mimicry tendency of the identified pathogenic flora of patients with the human proteome as well as intestinal microbes. Since gut bacteria become invasive and hostile in critical illness, mimicry between these organisms and the infectious flora is expected to be rather hyperinflammatory type, in contrast to the expectedly tolerogenic self versus pathogen cross-reactions. Differential effects of these two kinds of cross-reactions were studied. The predicted similarity of the identified pathogenic flora and intestinal microbes was higher in non-survivor patients compared to survivors (P =0.019). Higher values of “pathogen versus intestinal flora/pathogen versus human proteome” mimicry ratios (inflammatory quotients) were associated with mortality at a higher extent of significance (P <0.01), and correlated with admission APACHE II disease severity scores (R =0.311; P =0.017). We also found a correlation between the previously reported sepsis mortality rates by causative agent and the corresponding inflammatory quotients of these pathogens (R =0.738; P <0.05). Gram negative species showed higher similarity to intestinal bacteria and reached higher inflammatory quotients compared to Gram positives (P =0.01 and P <0.01, respectively). The disadvantageous effect of “pathogen versus intestinal flora” mimicry – presumably due to the extension of inflammation from the infectious focus to the already injured gut – is in accordance with the gut-lymph hypothesis, assessing that the destruction of the intestinal symbiosis culminates in the formation of damageous gut origin lymph. Our results raise the idea that molecular mimicry between pathogenic microbes and their antigenic surrounding might be a contributing factor behind the clinically and experimentally observed differences in microbiologically distinct forms of sepsis syndrome. [Copyright &y& Elsevier]

Abstract: Fusarium graminearum is one of the main causal agents of Fusarium Ear Blight on wheat. How the pathogen colonises the entire ear is not known. There is controversy over whether this mycotoxin producing pathogenic fungus invades wheat floral tissue using a necrotrophic or another mode of nutrition. A detailed microscopic investigation has revealed how wild-type fungal hyphae, of the sequenced strain PH-1, colonised susceptible wheat ears and spread from spikelet to spikelet. At the advancing infection front, colonisation of the host cortex occurred ahead of any vascular colonisation and the hyphae adapted to the available intercellular space between host cells. Intercellular hyphae then became abundant and host cells lost their entire cellular contents just prior to intracellular colonisation. No host cells died ahead of the infection. However, while these deep cortex infections progressed, just below the surface the highly photosynthetic chlorenchyma cells were observed to have died prior to colonisation. Behind the infection front, hyphae were abundant in the vasculature and the cortex, often growing through the pit fields of thick walled cells. This high level of inter- and intracellular fungal colonisation resulted in the collapse of the non-lignified cell-types. In this middle zone of infection, hyphal diameters were considerably enlarged. Far behind the infection front inter- and intracellular hyphae were devoid of contents and had often collapsed. At later stages of infection, the pathogen switched from predominately vertical to lateral growth and accumulated below the surface of the rachis. Here the lignified host cell walls became heavily degraded and hyphae ruptured the epidermis and produced an aerial mycelium. [Copyright &y& Elsevier]

Abstract: The first part of the present review gives an overview on the history of infectious agents of the order Chlamydiales and the general infection biology of Chlamydophila (C.) psittaci, the causative agent of psittacosis. In the second part, the classification of C. psittaci strains, as well as issues of epidemiology of avian chlamydiosis., disease transmission routes, clinical disease, public health significance, present legislation and recommendations for prevention and control are reviewed. [Copyright &y& Elsevier]