Studies on the palmitoyl-CoA hydrolase activity in carrot

Carrot homogenate contains a palmitoyl-CoA hydrolase. The enzyme was detected by spectrophotometric and radiochemical methods. The activation energy (Ea) for the reaction was determined to be 16700±2000 cal mol^?1 with a Q10 of 1.8. The rate of reaction reaches its maximum at 45°C and ceases completely at 55°C. The inactivation energy was found to be 67000±11000 cal mol^?1. A purification factor of 60 was obtained when polyvinylpyrolidone, polyethylene glycol, dithiothreitol and glycerol were used as stabilising agents, and the purification of palmitoyl-CoA hydrolase was carried out without delay. The method used successively, polyethylene glycol fractionation and DEAE-cellulose, hydroxylapatite and Ultrogel AcA 34 chromatography. Palmitoyl-CoA hydrolase has a molecular weight of 42000 daltons and a pI of 4.6. The role of this enzyme in the development of off-flavour in carrot has been discussed.     

Makerspaces on Social Media: Shaping Access to Open Design

Open Design is an emerging area of research that seeks to connect and extend the culture of making, social innovation, open-source software, and open-source hardware. A cornerstone for Open Design is to broaden participation in technology innovation by allowing people to use and contribute to publicly shared resources. Makerspaces are important access points to share and contribute to these resources. However, prior to entering the physical door of a makerspace, makerspaces’ social media presence serve as the ‘front door’ for Open Design activities. In this paper, we examine different ‘front doors’ of Open Design, asking: What are the characteristics that produce makerspaces’ social media presences, and how do these representations shape potential access to Open Design activities? We manually collected and qualitatively analyzed 500 public posts on the Facebook sites of five makerspaces in Copenhagen, Denmark. By choosing the same geographical area, we were able to explore the characteristics of makerspaces’ social media presences for the same potential population of Open Design participants. Our analysis identifies three main characteristics of makerspaces’ social media presence, which together shape access to Open Design activities, namely reach, transparency, and discourse. The display of these characteristics produce openness and availability in unique ways, and is constitutive for how Open Design activities are produced online. In this paper, we do not argue for or against specific social media representations. Instead, we argue that the specificities and differences between makerspaces’ social media presences in the same geographical region have the strength of producing different identities across spaces, allowing for a broader definition and potential participation in Open Design.     

3- Instead of 4-helix formation in a de novo designed protein in solution revealed by small-angle X-ray scattering

De novo design and chemical synthesis of proteins and their mimics are central approaches for understanding protein folding and accessing proteins with novel functions. We have previously described carbohydrates as templates for the assembly of artificial proteins, so-called carboproteins. Here, we describe the preparation and structural studies of three alpha-helical bundle carboproteins, which were assembled from three different carbohydrate templates and one amphiphilic hexadecapeptide sequence. This heptad repeat peptide sequence has been reported to lead to 4-alpha-helix formation. The low resolution solution structures of the three carboproteins were analyzed by means of small-angle X-ray scattering (SAXS) and synchrotron radiation circular dichroism (SRCD). The ab initio SAXS data analysis revealed that all three carboproteins adopted an unexpected 3+1-helix folding topology in solution, while the free peptide formed a 3-helix bundle. This finding is consistent with the calculated alpha-helicities based on the SRCD data, which are 72 and 68 % for two of the carboproteins. The choice of template did not affect the overall folding topology (that is for the 3+1 helix bundle) the template did have a noticeable impact on the solution structure. This was particularly evident when comparing 4-helix carboprotein monomers with the 2x2-helix carboprotein dimer as the latter adopted a more compact conformation. Furthermore, the clear conformational differences observed between the two 4-helix (3+1) carboproteins based on D-altropyranoside and D-galactopyranoside support the notion that folding is affected by the template, and subtle variations in template distance-geometry design may be exploited to control the solution fold. In addition, the SRCD data show that template assembly significantly increases thermostability.     

Decellularised Human Umbilical Artery as a Vascular Graft Elicits Minimal Pro-Inflammatory Host Response Ex Vivo and In Vivo

Small diameter (<6 mm) vessel grafts still pose a challenge for scientists worldwide. Decellularised umbilical artery (dUA) remains promising as small diameter tissue engineered vascular graft (TEVG), yet their immunogenicity remains unknown. Herein, we evaluated the host immune responses, with a focus on the innate part, towards human dUA implantation in mice, and confirmed our findings in an ex vivo allogeneic human setup. Overall, we did not observe any differences in the number of circulating white blood cells nor the number of monocytes among three groups of mice (1) dUA patch; (2) Sham; and (3) Mock throughout the study (day −7 to 28). Likewise, we found no difference in systemic inflammatory and anti-inflammatory cytokine levels between groups. However, a massive local remodelling response with M2 macrophages were observed in the dUA at day 28, whereas M1 macrophages were less frequent. Moreover, human monocytes from allogeneic individuals were differentiated into macrophages and exposed to lyophilised dUA to maximize an eventual M1 response. Yet, dUA did not elicit any immediate M1 response as determined by the absence of CCR7 and CXCL10. Together this suggests that human dUA elicits a minimal pro-inflammatory response further supporting its use as a TEVG in an allogeneic setup.     

Sulfite Action in Glycolytic Inhibition: In Vivo Real‐Time Observation by Hyperpolarized 13C NMR Spectroscopy

Detecting the molecular targets of xenobiotic substances in vivo poses a considerable analytical challenge. Here, we describe the use of an NMR‐based tracer methodology for the instantaneous in vivo observation of sulfur(IV) action on cellular metabolism. Specifically, we find that glycolytic flux is directed towards sulfite adducts of dihydroxyacetone phosphate and pyruvate as off‐pathway intermediates that obstruct glycolytic flux. In particular, the pyruvate–sulfite association hinders the formation of downstream metabolites. The apparent in vivo association constant of pyruvate and sulfite agrees with the apparent inhibition constant of CO₂ formation, thus supporting the importance of pyruvate interception in disturbing central metabolism and inhibiting NAD regeneration.     

Managerial Competences for ERP Journeys

This paper investigates the managerial competences required for the complex interactions required to successfully integrate an Enterprise Resource Planning system into an organization—the ERP journey. A feature of the study (a five year longitudinal study of a Danish production company implementing SAP R/3) was the rise and fall of the ERP project managers. The study showed that different stages of the ERP journey required different competences from the managers. A manager with a certain competence mix might successfully oversee part of the ERP journey, but a different blend of competences was required to manage other parts. The paper concludes that a wide range of competences are required: personal, business and technical. The competence mix should be expected to vary through the journey, and is often too much to expect of one person.     

Accountability in Brazilian Governmental Software Project: How Chat Technology Enables Social Translucence in Bug Report Activities

Fixing software bug is part of the daily work routine in software engineering which requires collaboration and thus has been explored as a core CSCW domain, since the early inception of the research field. In this paper, we explore the use of chat technology in software engineering by analyzing the coordination between client and vendor in a large government software project in Brazil (Gov-IT). We collected our empirical material through face-to-face and online interviews, site and chat forums observations. Looking closely at the bug fixing activities within Gov-IT, we find that the client and the vendor use chat technology to coordinate their cooperative work by enabling the participants to monitor the availability of developers and the urgency of detecting bugs synchronously. This way, the chat technology made it possible for the client to report bugs and developers to resolve bugs in a timely manner. Moreover, the chat technology enabled the participants to request and share artefacts synchronously, making it possible to analyze and understand the contextual nature surrounding bugs faster than using the bug tracking system. Finally, the chat technology enabled participants in enacting commitment and interdependence across vendor and client, creating cooperative situations of mutual dependence. Our results suggest that we, as CSCW designers, must rethink the design of bug tracking systems and find new ways to re-configure systems, so they support the coordinative practices involved in detecting, analyzing, and resolving critical and severe software bugs synchronously.     

Triage Drift: A Workplace Study in a Pediatric Emergency Department

This paper presents a workplace study of triage work practices within an emergency department (ED). We examine the practices, procedures, and organization in which ED staff uses tools and technologies when coordinating the essential activity of assessing and sorting patients arriving at the ED. The paper provides in-depth empirical observations describing the situated work practices of triage work, and the complex collaborative nature of the triage process. We identify and conceptualize triage work practices as comprising patient trajectories, triage nurse activities, coordinative artefacts and exception handling; we also articulate how these four features of triage practices constitute and connect workflows, organize and re-organize time and space during the triage process. Finally we conceptualize these connections as an assessing and sorting mechanism in collaborative work. We argue that the complexities involved in this mechanism are a necessary asset of triage work, which calls for a reassessment of the concept of triage drift.     

A technique for segmental rectal and colonic perfusion in humans

To enable a better characterization of pathophysiologic processes in colon and rectum, we have developed a perfusion technique for collection of soluble substances and cells from standardized intestinal segments. A tube with balloons attached to its outer wall was endoscopically introduced into the rectum and sigmoid colon, and its position ascertained fluoroscopically. The balloons delimited two segments, one in the sigmoid colon and one in the rectum. The segments were simultaneously perfused by a buffer at 37 degrees C. After a 30-min rinsing period, perfusate and cells were collected at 20-min intervals. Of 51 attempted perfusions, 45 were successfully completed. Recovered volumes equaled those infused. Leakage from the proximal intestine to the segments was negligible. In 18 healthy volunteers, the mean perfusate concentration from the rectal segment was 57.5 (27.5-120.2) mg/L for albumin, 1.3 (1.0-1.7) micrograms/L for eosinophil cationic protein, 5.1 (2.8-9.5) ng/L for prostaglandin E2, and 61.7 (41.7-89.1) micrograms/L for hyaluronan, and all values were lower in the sigmoid segment. Steady state conditions were achieved from the second perfusion period. The perfusate contained 4-80 x 10(4) cells, more than 95% of which were epithelial cells. The technique is safe, has a good subject compliance, and seems to be a valuable tool in investigations on quantitative release of soluble substances and cells in, e.g., colorectal inflammation.