A bioinformatics approach to the development of immunoassays for specified risk material in canned meat products

A bioinformatics approach to developing antibodies to specific proteins has been evaluated for the production of antibodies to heat-processed specified risk tissues from ruminants (brain and eye tissue). The approach involved the identification of proteins specific to ruminant tissues by interrogation of the annotation fields within the Swissprot database. These protein sequences were then interrogated for peptide sequences that were unique to the protein. Peptides were selected that met these criteria as close as possible and that were also theoretically resistant to either pepsin or trypsin. The selected peptides were synthesised and used as immunogens to raise monoclonal antibodies. Antibodies specific for the synthetic peptides were raised to half of the selected peptides. These antibodies have each been incorporated into a competitive enzyme-linked immunosorbent assay (ELISA) and shown to be able to detect the heat-processed parent protein after digestion with either pepsin or trypsin. One antibody, specific for alpha crystallin peptide (from bovine eye tissue), was able to detect the peptide in canned meat products spiked with 10% eye tissue. These results, although preliminary in nature, show that bioinformatics in conjunction with enzyme digestion can be used to develop ELISA for proteins in high-temperature processed foods and demonstrate that the approach is worth further stud

(Il)Legitimisation of the role of the nation state: Understanding of and reactions to Internet censorship in Turkey

This study aims to explore Turkish citizen-consumers' understanding of and reactions to censorship of websites in Turkey by using in-depth interviews and online ethnography. In an environment where sites such as YouTube and others are increasingly being banned, the citizen-consumers' macro-level understanding is that such censorship is part of a wider ideological plan and their micro-level understanding is that their relationship with the wider global network is reduced, in the sense that they have trouble accessing full information on products, services and experiences. The study revealed that citizen-consumers engage in two types of resistance strategies against such domination by the state: using irony as passive resistance, and using the very same technology used by the state to resist its domination

Pacing with restoration of respiratory sinus arrhythmia improved cardiac contractility and the left ventricular output: a translational study

Introduction: Respiratory sinus arrhythmia (RSA) is a prognostic value for patients with heart failure and is defined as a beat-to-beat variation of the timing between the heart beats. Patients with heart failure or patients with permanent cardiac pacing might benefit from restoration of RSA. The aim of this translational, proof-of-principle study was to evaluate the effect of pacing with or without restored RSAon parameters of LV cardiac contractility and the cardiac output

Long-distance endosome trafficking drives fungal effector production during plant infection

To cause plant disease, pathogenic fungi can secrete effector proteins into plant cells to suppress plant immunity and facilitate fungal infection. Most fungal pathogens infect plants using very long strand-like cells, called hyphae, that secrete effectors from their tips into host tissue. How fungi undergo long-distance cell signalling to regulate effector production during infection is not known. Here we show that long-distance retrograde motility of early endosomes (EEs) is necessary to trigger transcription of effector-encoding genes during plant infection by the pathogenic fungus Ustilago maydis. We demonstrate that motor-dependent retrograde EE motility is necessary for regulation of effector production and secretion during host cell invasion. We further show that retrograde signalling involves the mitogen-activated kinase Crk1 that travels on EEs and participates in control of effector production. Fungal pathogens therefore undergo long-range signalling to orchestrate host invasion

The rise of inconspicuous consumption

Ever since Veblen and Simmel, luxury has been synonymous with conspicuous consumption. In this conceptual paper we demonstrate the rise of inconspicuous consumption via a wide-ranging synthesis of the literature. We attribute this rise to the signalling ability of traditional luxury goods being diluted, a preference for not standing out as ostentatious during times of economic hardship, and an increased desire for sophistication and subtlety in design in order to further distinguish oneself for a narrow group of peers. We decouple the constructs of luxury and conspicuousness, which allows us to reconceptualise the signalling quality of brands and the construct of luxury. This also has implications for understanding consumer behaviour practices such as counterfeiting and suggests that consumption trends in emerging markets may take a different path from the past

Untangling knowledge creation and knowledge integration in enterprise wikis

A central challenge organizations face is how to build, store, and maintain knowledge over time. Enterprise wikis are community-based knowledge systems situated in an organizational context. These systems have the potential to play an important role in managing knowledge within organizations, but the motivating factors that drive individuals to contribute their knowledge to these systems is not very well understood. We theorize that enterprise wiki initiatives require two separate and distinct types of knowledge-sharing behaviors to succeed: knowledge creation (KC) and knowledge integration (KI). We examine a Wiki initiative at a major German bank to untangle the motivating factors behind KC and KI. Our results suggest KC and KI are indeed two distinct behaviors, reconcile inconsistent findings from past studies on the role of motivational factors for knowledge sharing to establish shared electronic knowledge resources in organizations, and identify factors that can be leveraged to tilt behaviors in favor of KC or KI

The Ustilago maydis Effector Pep1 Suppresses Plant Immunity by Inhibition of Host Peroxidase Activity

The corn smut Ustilago maydis establishes a biotrophic interaction with its host plant maize. This interaction requires efficient suppression of plant immune responses, which is attributed to secreted effector proteins. Previously we identified Pep1 (Protein essential during penetration-1) as a secreted effector with an essential role for U. maydis virulence. pep1 deletion mutants induce strong defense responses leading to an early block in pathogenic development of the fungus. Using cytological and functional assays we show that Pep1 functions as an inhibitor of plant peroxidases. At sites of Δpep1 mutant penetrations, H2O2 strongly accumulated in the cell walls, coinciding with a transcriptional induction of the secreted maize peroxidase POX12. Pep1 protein effectively inhibited the peroxidase driven oxidative burst and thereby suppresses the early immune responses of maize. Moreover, Pep1 directly inhibits peroxidases in vitro in a concentration-dependent manner. Using fluorescence complementation assays, we observed a direct interaction of Pep1 and the maize peroxidase POX12 in vivo. Functional relevance of this interaction was demonstrated by partial complementation of the Δpep1 mutant defect by virus induced gene silencing of maize POX12. We conclude that Pep1 acts as a potent suppressor of early plant defenses by inhibition of peroxidase activity. Thus, it represents a novel strategy for establishing a biotrophic interaction

Regulation of proteinaceous effector expression in phytopathogenic fungi

Effectors are molecules used by microbial pathogens to facilitate infection via effector-triggered susceptibility or tissue necrosis in their host. Much research has been focussed on the identification and elucidating the function of fungal effectors during plant pathogenesis. By comparison, knowledge of how phytopathogenic fungi regulate the expression of effector genes has been lagging. Several recent studies have illustrated the role of various transcription factors, chromosome-based control, effector epistasis, and mobilisation of endosomes within the fungal hyphae in regulating effector expression and virulence on the host plant. Improved knowledge of effector regulation is likely to assist in improving novel crop protection strategies

The effect of low temperature and low light intensity on nutrient removal from municipal wastewater by purple phototrophic bacteria (PPB)

There has been increased interest in alternative wastewater treatment systems to improve nutrient recovery while achieving acceptable TCOD, TN, and TP discharge limits. Purple phototrophic bacteria (PPB) have a high potential for simultaneous nutrient removal and recovery from wastewater. This study evaluated the PPB performance and its growth at different operating conditions with a focus on HRT and light optimization using a continuous-flow membrane photobioreactor (PHB). Furthermore, the effect of low temperature on PPB performance was assessed to evaluate the PPB’s application in cold-climate regions. In order to evaluate PPB performance, TCOD, TN, and TP removal efficiencies and Monod kinetic parameters were analyzed at different HRTs (36, 18, and 9 h), at temperatures of 22°C and 11°C and infrared (IR) light intensities of 50, 3, and 1.4 Wm-2. The results indicated that low temperature had no detrimental impact on PPB’s performance. The photobioreactor (PHB) with cold-enriched PPB has a high potential to treat municipal wastewater with effluent concentrations below target limits (TCOD˂ 50mgL-1, TN˂10 mgL-1, and TP˂1 mgL-1). Monod kinetic parameters Ks, K, Y, and Kd were estimated at 20-29 mgCODL-1, 1.6-1.9 mgCOD(mgVSS.d)-1, 0.47 mgVSS mgCOD-1, and 0.07-0.08 d-1 at temperatures of 11°C-22°C respectively. The results of the steady-state mass balances showed TCOD, TN, and TP recoveries of 80%-86%, which reflected PPB’s substrate and nutrient assimilation. Previous studies utilized high light intensities (˃ 50 Wm-2) to provide PPB with the maximum energy required for its growth. In order to enable the PPB technology as a practical approach in municipal wastewater treatment, light intensity must be optimized. Based on the literature, there is no study on PPB performance at low light intensities using a continuous-flow membrane photobioreactor. The effect of low light intensities of 3, and 1.4 Wm-2 on PPB performance was addressed in this study. The results indicated that PPB at a light intensity as low as 1.4 Wm-2 were able to treat municipal wastewater with effluent concentrations below above-mentioned target limits. Light intensity (1-50 Wm-2) had no detrimental impact on PPB performance and Monod kinetic parameters. This study showed that the optimized light intensity required for municipal wastewater treatment with PPB is significantly lower than previously indicated in the literature. The energy consumptions attributed to PHB’s illumination of 3, and 1.4 Wm-2 were determined to be 1.44, and 0.67 kWh/m3 which is significantly lower than previous studies (˃ 24 kWh/m3)

A fungal substrate mimicking molecule suppresses plant immunity via an inter-kingdom conserved motif

Ustilago maydis is a biotrophic fungus causing corn smut disease in maize. The secreted effector protein Pit2 is an inhibitor of papain-like cysteine proteases (PLCPs) essential for virulence. Pit2 inhibitory function relies on a conserved 14 amino acids motif (PID14). Here we show that synthetic PID14 peptides act more efficiently as PLCP inhibitors than the full-length Pit2 effector. Mass spectrometry shows processing of Pit2 by maize PLCPs, which releases an inhibitory core motif from the PID14 sequence. Mutational analysis demonstrates that two conserved residues are essential for Pit2 function. We propose that the Pit2 effector functions as a substrate mimicking molecule: Pit2 is a suitable substrate for apoplastic PLCPs and its processing releases the embedded inhibitor peptide, which in turn blocks PLCPs to modulate host immunity. Remarkably, the PID14 core motif is present in several plant associated fungi and bacteria, indicating the existence of a conserved microbial inhibitor of proteases (cMIP)