Co‐expression of the protease furin in Nicotiana benthamiana leads to efficient processing of latent transforming growth factor‐β1 into a biologically active protein

Transforming growth factor beta (TGF‐β) is a signalling molecule that plays a key role in developmental and immunological processes in mammals. Three TGF‐β isoforms exist in humans, and each isoform has unique therapeutic potential. Plants offer a platform for the production of recombinant proteins, which is cheap and easy to scale up and has a low risk of contamination with human pathogens. TGF‐β3 has been produced in plants before using a chloroplast expression system. However, this strategy requires chemical refolding to obtain a biologically active protein. In this study, we investigated the possibility to transiently express active human TGF‐β1 in Nicotiana benthamiana plants. We successfully expressed mature TGF‐β1 in the absence of the latency‐associated peptide (LAP) using different strategies, but the obtained proteins were inactive. Upon expression of LAP‐TGF‐β1, we were able to show that processing of the latent complex by a furin‐like protease does not occur in planta. The use of a chitinase signal peptide enhanced the expression and secretion of LAP‐TGF‐β1, and co‐expression of human furin enabled the proteolytic processing of latent TGF‐β1. Engineering the plant post‐translational machinery by co‐expressing human furin also enhanced the accumulation of biologically active TGF‐β1. This engineering step is quite remarkable, as furin requires multiple processing steps and correct localization within the secretory pathway to become active. Our data demonstrate that plants can be a suitable platform for the production of complex proteins that rely on specific proteolytic processing.     

Does Syntactic Alignment Effectively Influence How Speakers Are Perceived by Their Conversation Partner?

The way we talk can influence how we are perceived by others. Whereas previous studies have started to explore the influence of social goals on syntactic alignment, in the current study, we additionally investigated whether syntactic alignment effectively influences conversation partners’ perception of the speaker. To this end, we developed a novel paradigm in which we can measure the effect of social goals on the strength of syntactic alignment for one participant (primed participant), while simultaneously obtaining usable social opinions about them from their conversation partner (the evaluator). In Study 1, participants’ desire to be rated favorably by their partner was manipulated by assigning pairs to a Control (i.e., primed participants did not know they were being evaluated) or Evaluation context (i.e., primed participants knew they were being evaluated). Surprisingly, results showed no significant difference in the strength with which primed participants aligned their syntactic choices with their partners’ choices. In a follow-up study, we used a Directed Evaluation context (i.e., primed participants knew they were being evaluated and were explicitly instructed to make a positive impression). However, again, there was no evidence supporting the hypothesis that participants’ desire to impress their partner influences syntactic alignment. With respect to the influence of syntactic alignment on perceived likeability by the evaluator, a negative relationship was reported in Study 1: the more primed participants aligned their syntactic choices with their partner, the more that partner decreased their likeability rating after the experiment. However, this effect was not replicated in the Directed Evaluation context of Study 2. In other words, our results do not support the conclusion that speakers’ desire to be liked affects how much they align their syntactic choices with their partner, nor is there convincing evidence that there is a reliable relationship between syntactic alignment and perceived likeability.     

Polymorphisms in the human X-linked pyruvate dehydrogenase E1α gene

Summary Pyruvate dehydrogenase E1 deficiency is an X-chromosome-linked disorder, often with fatal consequences. We have searched for genetically useful polymorphisms in or near this gene. No restriction fragment length polymorphisms were detected using a battery of 36 different restriction enzymes and probing with a fulllength cDNA fragment, or two single-copy genomic fragments located within intron 8, and 15 kb 3 of the coding region, respectively. The chemical cleavage method was then applied to the detection of base changes in or near the gene. One polymorphism was found in exon 8 of the coding region. However, no base changes were detected in intron 3 or in the part of intron 8 covered by fragment gB2. Three blocks of microsatellite DNA containing variable numbers of CA-repeats were isolated from the 5 end of the gene and characterized. Length polymorphisms in these microsatellite DNAs were analysed using the polymerase chain reaction. Although the three loci are tightly linked, the polymorphisms appear not to be in disequilibrium, making them useful markers in linkage studies of the pyruvate dehydrogenase E1 gene. Of 31 females analysed 12 (39%) were heterozygous for at least one length polymorphism of the three (CA)_n alleles.     

Zur protoplasmatischen Anatomie des Keimblattes vonSoja hispida

Ohne ZusammenfassungDas Manuskript wurde im Jänner 1945 abgeschlossen und konnte infolge von Kriegseinwirkungen bis jetzt nicht zur Drucklegung gelangen.     

Identification of the lipopolysaccharide O‐antigen biosynthesis priming enzyme and the O‐antigen ligase in Myxococcus xanthus: critical role of LPS O‐antigen in motility and development

Myxococcus xanthus is a model bacterium to study social behavior. At the cellular level, the different social behaviors of M. xanthus involve extensive cell–cell contacts. Here, we used bioinformatics, genetics, heterologous expression and biochemical experiments to identify and characterize the key enzymes in M. xanthus implicated in O‐antigen and lipopolysaccharide (LPS) biosynthesis and examined the role of LPS O‐antigen in M. xanthus social behaviors. We identified WbaP_Mx (MXAN_2922) as the polyisoprenyl‐phosphate hexose‐1‐phosphate transferase responsible for priming O‐antigen synthesis. In heterologous expression experiments, WbaP_Mx complemented a Salmonella enterica mutant lacking the endogenous WbaP that primes O‐antigen synthesis, indicating that WbaP_Mx transfers galactose‐1‐P to undecaprenyl‐phosphate. We also identified WaaL_Mx (MXAN_2919), as the O‐antigen ligase that joins O‐antigen to lipid A‐core. Our data also support the previous suggestion that Wzm_Mx (MXAN_4622) and Wzt_Mx (MXAN_4623) form the Wzm/Wzt ABC transporter. We show that mutations that block different steps in LPS O‐antigen synthesis can cause pleiotropic phenotypes. Also, using a wbaP_Mx deletion mutant, we revisited the role of LPS O‐antigen and demonstrate that it is important for gliding motility, conditionally important for type IV pili‐dependent motility and required to complete the developmental program leading to the formation of spore‐filled fruiting bodies.     

Impact of salinomycin on the intestinal microflora of broiler chickens

Background  

The ionophoric coccidiostat salinomycin is widely used in chicken feed. In the near future the use of ionophore coccidiostats may be banned as has been the case for other antimicrobial growth promoters. This study was conducted to examine the effect of salinomycin on Campylobacter jejuni infection and on the composition of the caecal microflora in broiler chickens.