Synthesis of bacteriophage lytic proteins against Streptococcus pneumoniae in the chloroplast of Chlamydomonas reinhardtii.

There is a pressing need to develop novel antibacterial agents given the widespread antibiotic resistance among pathogenic bacteria and the low specificity of the drugs available. Endolysins are antibacterial proteins that are produced by bacteriophage-infected cells to digest the bacterial cell wall for phage progeny release at the end of the lytic cycle. These highly efficient enzymes show a considerable degree of specificity for the target bacterium of the phage. Furthermore, the emergence of resistance against endolysins appears to be rare as the enzymes have evolved to target molecules in the cell wall that are essential for bacterial viability. Taken together, these factors make recombinant endolysins promising novel antibacterial agents. The chloroplast of the green unicellular alga Chlamydomonas reinhardtii represents an attractive platform for production of therapeutic proteins in general, not least due to the availability of established techniques for foreign gene expression, a lack of endotoxins or potentially infectious agents in the algal host, and low cost of cultivation. The chloroplast is particularly well suited to the production of endolysins as it mimics the native bacterial expression environment of these proteins while being devoid of their cell wall target. In this study the endolysins Cpl-1 and Pal, specific to the major human pathogen Streptococcus pneumoniae, were produced in the C. reinhardtii chloroplast. The antibacterial activity of cell lysates and the isolated endolysins was demonstrated against different serotypes of S. pneumoniae, including clinical isolates and total recombinant protein yield was quantified at ~1.3 mg/g algal dry weight. This article is protected by copyright. All rights reserved

Gate stability of GaN-Based HEMTs with P-Type Gate

status: publishe

When the going gets tough, the tough get going: Social identification and individual effort in intergroup competition.

Based on social identity theory, the authors predicted that in ongoing intergroup competition, people’s strength of social identification will have a positive impact on their behavioral efforts on behalf of an ingroup when its current status is low, whereas this will not be the case when its current status is high. In a first experiment, male participants showed the expected pattern of behavior. Female participants, however, tended to display opposite reactions. As a possible explanation, it was argued that the experimental procedure may have inadvertently evoked a gender-based stereotype threat for female participants. In an attempt to obtain more consistent support for their hypothesis, the authors therefore replicated the experiment with modifications to avoid such a threat. These changes proved to be effective in the sense that this time the predicted interaction effect between ingroup identification and current group status was obtained for both male and female participants

Microalgae as a novel production platform for antibacterial proteins

Widespread antibiotic resistance among pathogenic bacteria and the low specificity of these drugs cause a pressing need for the development of novel antibiotics. Endolysins are proteins that are produced by bacteriophage infected cells and digest the bacterial cell wall for phage progeny release. These efficient enzymes are highly specific for the target bacteria without affecting other species. Development of resistance against endolysins is rare, because they evolved to target molecules that are essential for bacterial viability. Taken together, this makes them promising novel antibiotics. The development of recombinant endolysins as antibacterial agents requires an inexpensive and safe production platform. Microalgae emerged as an alternative expression platform in the last years. This study investigated therefore the production of endolysins in two distinct microalgal systems, the eukaryotic green microalga Chlamydomonas reinhardtii and the cyanobacterium Synechocystis sp. PCC 6803. C. reinhardtii is an attractive production platform for therapeutic proteins, due to the lack of endotoxins and infectious agents and its GRAS status (Generally Recognised as Safe). Synechocystis is a prokaryotic system and is natural transformable. Both offer established techniques for the expression of foreign genes and can be cultivated in full containment in simple photobioreactors. Transgenic lines of C. reinhardtii and Synechocystis expressing endolysins specific to the human pathogens Streptococcus pneumoniae and Staphylococcus aureus were created. The S. pneumoniae-specific endolysin Pal has been purified in an active form and its specificity and efficiency in killing the target bacterium demonstrated in vitro. Another endolysin specific to Propionibacterium acnes, a bacterium involved in the skin condition acne vulgaris, was synthesised in Synechocystis. The synthesis of the endolysins in the microalgae was analysed under the influence of different expression elements and at different growth stages, and the yields of recombinant protein were quantified to evaluate microalgae as a production platform for antibacterial and other protein therapeutics. !

Cytokine secretion in breast cancer cells – MILLIPLEX assay data

© 2019 The Author(s) Metastatic breast cancer is the most advanced stage of breast cancer and the leading cause of breast cancer mortality. Although understanding of the cancer progression and metastasis process has improved, the bi-directional communication between the tumor cell and the tumor microenvironment is still not well understood. Breast cancer cells are highly secretory, and their secretory activity is modulated by a variety of inflammatory stimuli present in the tumor microenvironment. Here, we characterized the cytokine expression in human breast cancer cells (MDA-MB-231, MCF-7, T-47D, and BT-474) in vitro using 41 cytokine MILLIPLEX assay. Further, we compared cytokine expression in breast cancer cells to those in non-tumorigenic human breast epithelial MCF-10A cells

Green biologics: the algal chloroplast as a platform for making biopharmaceuticals

Most commercial production of recombinant pharmaceutical proteins involves the use of mammalian cell lines, E. coli or yeast as the expression host. However, recent work has demonstrated the potential of eukaryotic microalgae as platforms for light-driven synthesis of such proteins. Expression in the algal chloroplast is particularly attractive since this organelle contains a minimal genome suitable for rapid engineering using synthetic biology approaches; with transgenes precisely targeted to specific genomic loci and amenable to high-level, regulated and stable expression. Furthermore, proteins can be tightly contained and bio-encapsulated in the chloroplast allowing accumulation of proteins otherwise toxic to the host, and opening up possibilities for low-cost, oral delivery of biologics. In this commentary we illustrate the technology with recent examples of hormones, protein antibiotics and immunotoxins successfully produced in the algal chloroplast, and highlight possible future applications

Danger- and pathogen-associated molecular patterns recognition by pattern-recognition receptors and ion channels of the transient receptor potential family triggers the inflammasome activation in immune cells and sensory neurons.

An increasing number of studies show that the activation of the innate immune system and inflammatory mechanisms play an important role in the pathogenesis of numerous diseases. The innate immune system is present in almost all multicellular organisms and its activation occurs in response to pathogens or tissue injury via pattern-recognition receptors (PRRs) that recognize pathogen-associated molecular patterns (PAMPs) or danger-associated molecular patterns (DAMPs). Intracellular pathways, linking immune and inflammatory response to ion channel expression and function, have been recently identified. Among ion channels, the transient receptor potential (TRP) channels are a major family of non-selective cation-permeable channels that function as polymodal cellular sensors involved in many physiological and pathological processes.In this review, we summarize current knowledge of interactions between immune cells and PRRs and ion channels of TRP families with PAMPs and DAMPs to provide new insights into the pathogenesis of inflammatory diseases. TRP channels have been found to interfere with innate immunity via both nuclear factor-kB and procaspase-1 activation to generate the mature caspase-1 that cleaves pro-interleukin-1ß cytokine into the mature interleukin-1ß.Sensory neurons are also adapted to recognize dangers by virtue of their sensitivity to intense mechanical, thermal and irritant chemical stimuli. As immune cells, they possess many of the same molecular recognition pathways for danger. Thus, they express PRRs including Toll-like receptors 3, 4, 7, and 9, and stimulation by Toll-like receptor ligands leads to induction of inward currents and sensitization in TRPs. In addition, the expression of inflammasomes in neurons and the involvement of TRPs in central nervous system diseases strongly support a role of TRPs in inflammasome-mediated neurodegenerative pathologies. This field is still at its beginning and further studies may be required.Overall, these studies highlight the therapeutic potential of targeting the inflammasomes in proinflammatory, autoinflammatory and metabolic disorders associated with undesirable activation of the inflammasome by using specific TRP antagonists, anti-human TRP monoclonal antibody or different molecules able to abrogate the TRP channel-mediated inflammatory signals

Atmospheric pressure plasma analysis by modulated molecular beam mass spectrometry

Fractional no. d. measurements for a radiofrequency plasma needle operating at atm. pressure were obtained using a mol. beam mass spectrometer (MBMS) system designed for diagnostics of atm. plasmas. The MBMS system comprises three differentially pumped stages and a mass/energy analyzer and includes an automated beam-to-background measurement facility as a software-controlled chopper mechanism. The automation of the beam modulation allows the neutral components in the plasma to be rapidly and accurately measured using the mass spectrometer by threshold ionization techniques. Data are reported for plasma generated by a needle plasma source operated using a He/air mixt. In particular, data for the conversion of atm. O and N into nitric oxide are discussed with ref. to its significance for medical applications such as disinfecting wounds and dental cavities and for microsurgery. [on SciFinder (R)