Miracle-dz/UniFAR-FTUs huggingface dataset:
Zinc Finger Structures in the Human Immunodeficiency Virus Type 1 Nucleocapsid Protein Facilitate Efficient Minus- and Plus-Strand Transfer

ABSTRACT The nucleocapsid protein (NC) of human immunodeficiency virus type 1 (HIV-1) has two zinc fingers, each containing the invariant metal ion binding residues CCHC. Recent reports indicate that mutations in the CCHC motifs are deleterious for reverse transcription in vivo. To identify reverse transcriptase (RT) reactions affected by such changes, we have probed zinc finger functions in NC-dependent RT-catalyzed HIV-1 minus- and plus-strand transfer model systems. Our approach was to examine the activities of wild-type NC and a mutant in which all six cysteine residues were replaced by serine (SSHS NC); this mutation severely disrupts zinc coordination. We find that the zinc fingers contribute to the role of NC in complete tRNA primer removal from minus-strand DNA during plus-strand transfer. Annealing of the primer binding site sequences in plus-strand strong-stop DNA [(+) SSDNA] to its complement in minus-strand acceptor DNA is not dependent on NC zinc fingers. In contrast, the rate of annealing of the complementary R regions in (−) SSDNA and 3′ viral RNA during minus-strand transfer is approximately eightfold lower when SSHS NC is used in place of wild-type NC. Moreover, unlike wild-type NC, SSHS NC has only a small stimulatory effect on minus-strand transfer and is essentially unable to block TAR-induced self-priming from (−) SSDNA. Our results strongly suggest that NC zinc finger structures are needed to unfold highly structured RNA and DNA strand transfer intermediates. Thus, it appears that in these cases, zinc finger interactions are important components of NC nucleic acid chaperone activity.
Rethinking city economic resilience: exploring deglomeration of firms in inner-city Harare

ABSTRACT City resilience is a concern for the majority of African cities especially those facing acute economic challenges. Harare city is not an exception as it shows signs of decline in the face of continual national economic crisis. Thus, this article seeks to examine how the economic shock has affected the economic well-being of the central business district (CBD) in Harare and to recommend policy strategies that can be adopted to improve the city’s competitiveness and economic resilience. This article’s originality lies in conceptualising economic resilience within a city sub-region, rather than the city as a whole. Through analysing the firms operating in the CBD, this article seeks to explain the operational challenges that have caused winding up, downsizing and/or relocation of firms highlighting their effects on the competitiveness and the economic dominance of the CBD. In closing, strategies are explored that can be adopted to avoid the general decline currently prevailing in inner-city Harare.
The Interferon-Inducible Protein Tetherin Inhibits Hepatitis B Virus Virion Secretion

ABSTRACT Interferon alpha (IFN-α) is an approved medication for chronic hepatitis B therapy. Besides acting as an immunomodulator, IFN-α elicits a pleiotropic antiviral state in hepatitis B virus (HBV)-infected hepatocytes, but whether or not IFN-α impedes the late steps of the HBV life cycle, such as HBV secretion, remains elusive. Here we report that IFN-α treatment of HepAD38 cells with established HBV replication selectively reduced HBV virion release without altering intracellular viral replication or the secretion of HBV subviral particles and nonenveloped capsids. In search of the interferon-stimulated gene(s) that is responsible for the reduction of HBV virion release, we found that tetherin, a broad-spectrum antiviral transmembrane protein that inhibits the egress of a variety of enveloped viruses, was highly induced by IFN-α in HepAD38 cells and in primary human hepatocytes. We further demonstrated that the expression of full-length tetherin, but not the C-terminal glycosylphosphatidylinositol (GPI) anchor-truncated form, inhibited HBV virion egress from HepAD38 cells. In addition, GPI anchor-truncated tetherin exhibited a dominant-negative effect and was incorporated into the liberated virions. We also found colocalization of tetherin and HBV L protein at the intracellular multivesicular body, where the budding of HBV virions takes place. In line with this, electron microscopy demonstrated that HBV virions were tethered in the lumen of the cisterna membrane under tetherin expression. Finally, knockdown of tetherin or overexpression of dominant negative tetherin attenuated the IFN-α-mediated reduction of HBV virion release. Taken together, our study suggests that IFN-α inhibits HBV virion egress from hepatocytes through the induction of tetherin. IMPORTANCE Tetherin is a host restriction factor that blocks the egress of a variety of enveloped viruses through tethering the budding virions on the cell surface with its membrane anchor domains. Here we report that interferon directly and selectively inhibits the secretion of HBV virions, but not subviral particles or nonenveloped capsids, through the induction of tetherin in hepatocyte-derived cells. The antiviral function of tetherin requires the carboxyl-terminal GPI anchor, while the GPI anchor deletion mutant exhibits dominant negative activity and attaches to liberated HBV virions. Consistent with the fact that HBV is an intracellular budding virus, microscopy analyses demonstrated that the tethering of HBV virions occurs in the intracellular cisterna and that tetherin colocalizes with HBV virions on the multivesicular body, which is the HBV virion budding site. Our study not only expands the antiviral spectrum of tetherin but also sheds light on the mechanisms of interferon-elicited anti-HBV responses.
The role of HDL and its modulators in the development of diabetes

Reduced plasma HDL cholesterol (HDL-C) level is a key feature of the metabolic syndrome, a clinical entity known to be a predisposing factor for atherosclerotic heart disease and type 2 diabetes (T2D). In recent years, compelling evidence has emerged to reveal the direct effects of HDL and its major modulators, namely, ATP-binding cassette transporter A1 (ABCA1), apolipoprotein A-I (ApoA-I), and lecithin cholesterol acyltransferase (LCAT) on the development of T2D. This review will highlight some of the most recent studies addressing the roles for HDL and their modulators in the regulation of beta-cell secretory function and insulin sensitivity.
Tumor pretargeting: role of avidin/streptavidin on monoclonal antibody internalization.

UNLABELLED Radioimmunodetection of tumor can be improved by introducing a two-step system in which radiolabeled streptavidin is administrated after the injection of a biotinylated monoclonal antibody (MAb) (two-step) or radiolabeled biotin is injected after biotinylated MAb and avidin (three-step). The anti-carcinoembryonic antigen (CEA) MAb FO23C5 has been recently exploited in a three-step protocol based on the avidin-biotin system. The anti-folate receptor (FR) MAb MOv18 has proven suitable for radioimmunodetection of ovarian cancer using directly radiolabeled MAb or in a two-step method. In this study, we analyzed the suitability of MOv18 in a three-step protocol in ovarian carcinoma patients and the internalization events after formation of the MOv18-avidin complex. METHODS Selected patients with documented metastatic lesions were enrolled in a three-step radioimaging analysis with biotinylated MOv18 and FO23C5, avidin and (111)In-labeled biotin. Two-step internalization experiments were conducted in vitro with MOv18 and MOv19 MAbs on the FR-overexpressing IGROV1 cell line and with the anti-CEA MAb FO23C5 on the LS174T cell line. Cells were incubated sequentially with biotinylated MAb and 125I-labeled streptavidin or with 125I-biotinylated MAb and cold streptavidin. RESULTS In the in vivo study, SPECT revealed the majority of metastatic lesions in patients injected with biotinylated MOv18; however, the tumor-to-background ratio was relatively low. In the in vitro study, a consistent internalization was induced by antigen-biotinylated MAb-streptavidin complex formation at the cell surface in both antigenic systems analyzed. However, the extent of internalization was lower in the CEA model. CONCLUSION The internalization ability of avidin suggests its potential clinical application for delivering toxic agents in a two-step approach (biotinylated MAb + avidin conjugate). The suitability of a given MAb for three-step clinical applications (biotinylated MAb + avidin + biotin) should be previously investigated by using appropriate in vitro experiments.
Consumer Attitudes toward Online Behavioural Advertising: The Social Media Involvement

In digital environment nowadays, data about Internet usage can be used to tailor content and advertisements to users’ interests. Interactive advertising allows customers to become more involved because they initiate most of the action. Social media allows customers and prospects to communicate directly to the brand representative or about the brand with their friends. Experiences during this interaction will drive brand attitudes. Since online behavioural advertising uses a targeted approach and the advertisement of the retailer is directed to the most appropriate and interested consumer and online retailers interact with online consumers for their transactions, online behavioural advertising becomes a very important tool especially for online retailers. However, online behavioural advertising programs through social media, which target consumers based on their interests and preferences, still raises debate concerning consumer privacy interests in connection with business data collection, transfer and use. This article aims to explain the influence of online behavioural advertising (OBA) on attitude and their purchase intention. This type of research is explanatory with quantitative approach. Facebook as one of the popular social media was chosen as the research site. Population in this research are Facebook users in Indonesia who have seen impression of OBA aged above 18 years old and ever conducted online shopping in past six months. The sample used in this research was 145 people chosen with purposive sampling technique. Self-administered questionnaire via online was employed as data collection methods in this research. The result of path analysis shows that OBA significantly influences the attitude, OBA significantly influences the purchase intention, and attitude significantly influences the purchase intention. Based on the research’s result it was suggested that users’ privacy should be considered carefully especially related to the tracking and profiling for OBA purpose, the display of OBA must be improved to attract attention and reach high click-through-rate.
Formation of effective multi-stakeholder Platforms: Lessons from coffee innovation platforms in Uganda

Agricultural innovation systems’ thinking has been adopted in agricultural research and development to harness the actor innovative capabilities, knowledge enhancement and social transformation. Hence there is growing recognition of the viability of Innovation platforms (IPs) as an approach to stimulating development of agricultural enterprises due to their emphasis on value chain development, networking and partnerships. However, despite the key role of the formation process in realization of desired outcomes, there is insufficient knowledge on how to form vibrant agricultural innovation platforms under diverse contexts. This study sought to examine the gaps and draw lessons for effective IP formation using the case of the coffee IPs in the four districts of Luwero, Ntungamo, Bushenyi and Rakai in Uganda. Data were collected through key informant interviews with 32 actors of the coffee steering committees and from document review. Qualitative data analysis was conducted using content and thematic analysis. Gaps identified in the IP formation processes included exclusion of some key stakeholders, unclear management and financing mechanisms, and weak governance structures at lower levels. To enhance legitimacy of innovation platforms, the study recommends a formation operational model with four main components of actor identification and mobilization, visioning and strategy formulation process, IP culture and development of governance structures.
Effects of land use on soil erosion and nitrogen loss in the hilly area of the Loess Plateau, China

This study examines the effects of land use and slope angle on runoff, soil loss and nitrogen loss from hillslopes of the Loess Plateau in China. Farmland, wasteland and four forest treatments (sea buckthorn+poplar, immature sea buckthorn, mature sea buckthorn, and immature Chinese pine) were the types of land use that were compared. The results showed that July was the critical period for runoff, soil loss and nitrogen loss from farmland. Farmland was the most susceptible land use. Sea buckthorn+poplar, immature sea buckthorn, and mature sea buckthorn limited the runoff, soil loss and nitrogen loss. Farmland on slopes over 15 degrees should be abandoned because of the high erosion rate and nitrogen loss. Copyright © 2003 John Wiley & Sons, Ltd.
Improving undergraduate medical student involvement in research

php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php). Advances in Medical Education and Practice 2017:8 731–733 Advances in Medical Education and Practice Dovepress
Meeting the needs of the Millennials and Generation Z: gamification in tourism through geocaching

Purpose The purpose of this paper is to present a conceptual framework based on an understanding of the principles of popular mobile-enabled games, indicating how organisations in the tourism sector could meet the needs of Millennials and Generation Z through engaging with the existing gamified location-based practice of geocaching as an information and communication technology enabled gamified enhancement to the destination experience. Design/methodology/approach As a primarily conceptual paper, the authors take an inductive qualitative approach to theory building based on the understanding of an existing practice (geocaching) that is undertaken among a community of practitioners (geocachers), which results in the presentation of a conceptual framework, which is the theory itself that the authors have constructed from the understanding of what is going on and which principles can then be applied across other tourism practices. Findings Findings indicate that through engaging with geocaching, smaller entrepreneurial businesses even in non-urban destinations that fall outside of the remit of smart city developments, and in tourism destinations on the less technologically enabled or resource-rich side of the digital divide, can reap the benefits associated with employing the principles and practices associated with smart tourism to meet the needs of this new generation of tourism consumers who seek richer digital and often gamified tourism experiences. Originality/value This paper fills a gap in the literature regarding the way many different types of tourism destinations could meet the needs of Millennials and Generation Z tourists.
1,2,3‐Triazole as a Peptide Surrogate in the Rapid Synthesis of HIV‐1 Protease Inhibitors

Given the ubiquitous nature of the peptide linkage in biological molecules, replacement of the amide bond with isosteres in potential drug candidates has been a continual goal of many laboratories. Successful replacements will provide improved stability, lipophilicity, and absorption. Many surrogates have been introduced already, yet the synthesis of many of these isosteres in a combinatorial way is difficult and requires several steps. Thus, the discovery of new peptide surrogates with easier syntheses is an important achievement that could open new opportunities for the study of amide-containing molecules and the development of inhibitors with novel physicochemical properties. We have used the copper(i)-catalyzed azide–alkyne [3+2] cycloaddition as a straightforward reaction for the preparation of inhibitor libraries. Over 100 compounds were synthesized in microtiter plates and screened in situ. Two of these compounds—AB2 (pdb-1zp8) and AB3 (pdb-1zpA)—showed the best activity against wild type and mutant HIV-1 proteases (Table 1). AB2 and AB3, were then computationally docked by using AutoDock3. The docking simulation produced two conformations of approximately equal energy. One conformation placed the triazole in the position normally adopted by the peptide unit—between P2’ and P1’—in peptidomimetic compounds. Furthermore, the central nitrogen of the triazole was perfectly positioned to form a hydrogen bond with the water molecule normally found under the protease flaps. This water molecule also formed a hydrogen bond with the sulfonamide as seen in the crystallographic structure of amprenavir when bound to HIV-1 protease. The other conformation positioned the compounds in a similar place, but with the triazole rotated by 180 8. This allowed for a slightly better fit of the triazole substituent but sacrificed the hydrogen bond with the water molecule. In this work we have solved the ambiguity in binding conformation by solving the crystal structure of two inhibitors derived from a library of triazole compounds with HIV-1 protease. Interestingly, the two structures show that the triazole ring is an effective amide surrogate that retains all hydrogen bonds in the active site (Figure 1). HIV-1 protease (3 mgmL 1 in 0.025m sodium acetate pH 5.4, 10 mm dithiothreitol, 1 mm EDTA) was combined with inhibitor (32 mm in 50% (v/v) dimethylsulfoxide and 2-methylpentane2,4-diol) at 4 8C to give a 2:1 molar ratio of inhibitor to protein, and the mixture was centrifuged to remove the precipitate. The complex was crystallized by the hanging-drop vapor-diffusion method by mixing 9.6 mL of protease solution with 4 mL of crystallization buffer (1.34m ammonium sulfate, 0.1m sodium acetate, pH 4.8–5.4). Plates were sealed at 20 8C for one to two weeks. Data were collected from frozen crystals at the Argonne National Laboratory SER-CAT beamline 22-ID and with a Rigaku Table 1. Binding constants of 1,2,3-triazole compounds to HIV-1 protease.
Cardioprotective effects of fish omega-3 fatty acids on doxorubicin-induced cardiotoxicity in rats

The aim of this study was to investigate the protective effects of fish omega-3 (n-3) fatty acids on doxorubicin (DOX)-induced acute cardiotoxicity. A total of 24 rats were divided into three groups: control, DOX-treated, and DOX treated with fish n-3 fatty acids. Control group received 0.4 ml/kg/day of saline intragastrically. The rats in the fish n-3 fatty acid-pretreated group were given 400 mg/kg/day fish n-3 fatty acids for 30 days by intragastric intubation. To induce acute cardiotoxicity, DOX (30 mg/kg) was injected intraperitoneally by a single dose and the rats were killed after 48 h. DOX treatment caused severe damage in heart tissues. Disorganization of myocardial muscle fibers, myofibrillar loss, and cardiotoxic myocardial fibers with cytoplasmic vacuoles were seen. Fish n-3 fatty acid-treated rats showed an improved histological appearance in the DOX-treated group. Our data indicate a significant reduction in the activity of terminal deoxynucleotidyl transferase-mediated deoxyuridine triphosphate nick end labeling in cardiomyocytes of the DOX-treated group with fish n-3 fatty acids therapy. The DOX-treated with fish n-3 fatty acids group showed a significant decrease in malondialdehyde levels, and an increase in superoxide dismutase and glutathione peroxidase activities in comparison with the DOX-treated group. This study showed that fish n-3 fatty acids may be a suitable cardioprotector against acute toxic effects of DOX.
Serial measurements of acute phase proteins in patients with acute coronary syndrome.

INTRODUCTION Levels of inflammatory markers increase in patients with acute coronary syndromes (ACS) and the magnitude of the inflammatory response has been related to clinical outcomes. The release patterns and, thereby, the time point of maximal increase for multiple inflammatory markers following an ACS are not fully defined. Our purpose was to serially measure three acute phase proteins (APPs) in patients with ACS. METHODS We prospectively enrolled 74 consecutive patients (54 men, age 62.2 ± 9.8 years) with ACS: 38 with ST-elevation acute myocardial infarction (STEMI), and 36 with non-ST-elevation acute myocardial infarction (NSTEMI) or unstable angina (UA). Peripheral levels of alpha-1 antitrypsin (A1AT), alpha-1 acid glycoprotein (A1GP) and haptoglobin (HPT) were measured on admission, and 6, 12, 24, 48, 72 hours, 7 days and 6 months after the initial evaluation. RESULTS Baseline levels of the APPs did not differ between the two groups. A1AT, A1GP and HPT all exhibited a similar time course among NSTEMI/UA patients, all reaching maximal values at 7 days. The markers showed an earlier increase and A1AT showed earlier peaking (at 72 hours) in STEMI patients. Peak levels were higher in patients with STEMI vs. NSTEMI/UA for all three APPs (p<0.01 for A1AT and HPT, p<0.05 for A1GP). CONCLUSIONS A1AT, A1GP and HPT levels increase significantly in patients with ACS and display different release curves in those with STEMI versus NSTEMI or UA. Knowledge of the release patterns of APPs may determine the optimal time point of measurement and thereby enhance their potential prognostic value in the setting of ACS.
Cartilage Collagen Fibril Network in Newborn Transgenic Mice Analyzed by Electron Microscopic Stereology

The objective was to investigate by electron microscopic stereology the properties of the cartilage collagen fibril network in newborn transgenic mice. The mice harbored transgenes targeted to affect the structure or assembly of the collagen fibrils. The mouse lines investigated here harbored either (i) one or (ii) two human COL2A1 genes with Arg519Cys mutation in addition to one or (iii) no active allele(s) of the murine Col2a1 gene, (iv) two inactive alleles of the procollagen N-proteinase genes, or (v) a human COL2A1 gene with deleted exons 16–27. In all newborn mice carrying the COL2A1 transgene with Arg519Cys mutation, the growth plate collagen fibrils were thinner than in the wild-type (wt) mice and showed clearly reduced volume fraction of the fibril network. In mice with the inactive procollagen N-proteinase genes, the fibril thickness and the volume fraction of collagen did not differ from the wt mice. In mice harboring the transgene of human COL2A1 gene with internally deleted exons 16–27, the collagen fibril diameter remained the same, but the volume density of collagen network was reduced. Using the indirect stereology, the differences in the collagen fibril stereological estimates could be reliably detected in newborn mice harboring mutations that affect the structure and assembly of collagen fibrils. The EM stereology permitted early detection of altered phenotype of the collagen fibril network in newborn transgenic mice. It is recommended that the indirect model-based stereological technique is utilized instead of the direct design-based technique for the estimation of collagen volume, surface, and length densities.