An in vitro translation system, based on cell components of the hyperthermophilic archaeon, Thermococcus kodakaraensis, has previously been developed. The system has been optimized and applied for protein production at high temperatures (60-65 degrees C). In this paper, we have examined the possibilities to utilize this system at a lower temperature range using green fluorescence protein (GFP) as the reporter protein. By optimizing the composition of the reaction mixture, and adding chaperonins from the mesophilic Escherichia coli, the yield of protein production at 40 degrees C was increased by fivefold. For liposome encapsulation of the optimized system, water-in-oil cell-sized emulsions were prepared by adding the translation system/GFP mRNA mixture to mineral oil supplemented with 1,2-dioleoyl-sn -glycero-3-phosphatidylcholine (DOPC). Giant liposomes were formed when these emulsions passed across a water/oil interface occupied with DOPC. The liposomes were incubated at 40 degrees C for 90 min, and fluorescence was examined by laser confocal microscopy. A significant increase in average fluorescence intensity was observed in liposomes with GFP mRNA, but not in those without mRNA. Our results indicate that the T. kodakaraensis in vitro translation system is applicable for protein production within giant liposomes, and these artificial cell models should provide the methodology to reconstitute various cell functions from a constitutional biology approach.
Background Cardiopulmonary arrest (CPA) is the most serious presentation of acute myocardial infarction (AMI). However, the frequency and prognostic impact of CPA in young patients with AMI have been still unclear. Objectives This study aimed to characterize AMI in young patients who underwent primary percutaneous coronary intervention using large-scale nationwide all-comer registry data in Japan (J-PCI registry). Methods Data on risk factor profiles, clinical features, post-procedural complications, and in-hospital outcomes were reviewed within the J-PCI registry between 2014 and 2018. Results Among 213,297 patients with AMI, 23,985 (11.2%) were young (age, 20–49 years). Compared with the older group (age, 50–79 years; n=189,312), the young group included a higher number of men, smokers, patients with dyslipidemia, and patients with single-vessel disease, and a lower number of patients with hypertension and diabetes. Despite favorable clinical profiles, younger age was associated with a higher rate of presentation with CPA (Figure 1). Further, concomitant CPA was strongly associated with in-hospital mortality in the young group (Table 1). Conclusions Young patients with AMI presented a higher risk of CPA than older patients, which was strongly associated with in-hospital mortality. Funding Acknowledgement Type of funding sources: Public grant(s) – National budget only. Main funding source(s): JSPS KAKENHI
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
hi@scite.ai
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.