The role of Exon Junction Complex (EJC) and SOLE in oskar mRNA transport

Asymmetric localization of specific RNAs is essential in early embryo development. During Drosophila melanogaster oogenesis, oskar mRNA is transcribed in nurse cells, transported to the transcriptional silent oocyte and subsequently localized at the posterior pole. The first transport step from the nurse cells into the oocyte is mediated by dynein transport, whereas the second step to the posterior pole is dependent on kinesin. At the posterior pole, oskar mRNA is required for the formation of germ plasm and abdominal segments. The transport of oskar mRNA by kinesin requires the exon junction complex (EJC) core components eIF4AIII, Mago nashi, Y14 and Barentsz as well as the spliced oskar localization element (SOLE). The exact mechanism by which the EJC and SOLE are involved in kinesin transport has remained unclear. I utilized RNA-protein interaction footprinting to analyze how the protein occupancy on the RNA changes upon disrupting the SOLE secondary structure. Through RNA immunoprecipitation against Mago nashi, I discovered that the presence of the SOLE led to a more stable binding of the EJC to oskar mRNA. This demonstrates that the SOLE has a stabilizing effect on EJC binding to oskar mRNA. Although the EJC and SOLE were found to be dispensable for kinesin recruitment to oskar mRNA, they were essential for oskar mRNA transport, suggesting that the EJC-SOLE interactome plays a role in activating kinesin transport to the posterior pole. I optimized a previously described transcript-specific RNA binding protein (RBP) capture protocol and utilized it to analyze the EJC-SOLE interactome. Flies expressing transgenic lacZ RNA containing the oskar 3’UTR and either wild-type or mutant SOLE were created and used to determine the difference in RBP composition upon disrupting the SOLE secondary structure. I identified RBPs that were specifically bound to the transgenic lacZ RNA with the wild-type SOLE, which localizes to the posterior pole and RBPs bound to the lacZ RNA with a disrupted SOLE, which does not localize. Further investigations will be carried out to explore which of the RBPs are involved in the kinesin-mediated transport of oskar mRNA

Impact of Brand Label on Purchase Intention of a Customer

Main target of this research is examining the relationship between the purchase intention of cosmetics and brand label. The image of the store mediates the relationship between purchase intention and brand label in the cosmetics sector of Pakistan. To check the direct or indirect effect of brand label on purchase intention, data was collected from 200 customers of different stores. Snow ball sampling technique is used for the collection of data from respondents, and PLS-SEM 3 was used to test the hypothesis. The findings indicate that brand label positively and significantly impact customer purchase intention of cosmetics and store image is also positively linked with brand label and mediating the relationship between purchase intention and brand label. Findings will help the retail service sector to understand the importance of store image and how to enhance the consumers purchase intentions. Over long term, customer retention is better and business profitability is greater

How Social Studies Teachers Conceptualize Civic Teaching and Learning in 2020: Insights from a Research-Practice Partnership

High-quality civic learning opportunities remain the exception, rather than the norm, in public schools across the country. The health and future of the American democracy is dependent on all its public schools to foster democratic classrooms and prepare informed citizens—a reality far from realized in this third decade of the 21st century. Through an inductive analysis of in-depth interviews, this article makes visible how educators conceptualize civic teaching and learning in the political moment of the year 2020. Because of the known link between teachers’ conceptualizations, instructional visions, and their practice, it is necessary when engaged in any district change effort to first understand how teachers understand the existing phenomenon—in this case, civic teaching and learning. In shedding light on teachers’ contemporary conceptualizations of civic teaching and learning, this article contributes to the necessary and timely conversation on how to support civic teaching, even in politically contentious times, so that all students can experience high-quality civic learning on a routine basis

The Crime of Sedition: At the Crossroads of Reform and Resurgence

The offense of “sedition” — often characterized as criminalizing the incitement of rebellion against the government — is an archaic crime that is frequently used to target political speech. Introduced in the sixteenth century in England specifically to suppress dissent, sedition laws spread through the British colonies. These laws still persist in some legal systems, and while there are reforms underway in some of those jurisdictions, in a few outliers, the offense continues to be prosecuted — and in some there has been a resurgence in cases. Sedition laws have been criticized by the United Nations (“U.N.”), human rights experts, courts, legislatures, advocates, and others for being a weapon used by governments to violate the right to freedom of expression. Moreover, the significant criminal penalties that usually accompany sedition laws have a chilling effect on political debate and can undermine democratic processes. This report examines the movement to repeal or reform sedition laws, the reasons for these efforts, and abuses that take place where the charge of sedition continues to be used. It proceeds in three parts: first, a brief overview of sedition laws and the criticisms they have faced at the international and regional levels; second, an update on the substantial progress Commonwealth States have made to reform these laws; and third, an overview of several examples of States where prosecutors have weaponized sedition laws to stifle dissent, including examples arising from TrialWatch’s monitoring experience

Free Trade Agreements and Environmental Nexus in Pakistan

This study attempts to examine the relationship between Free Trade Agreements (FTAs) and Carbon Dioxide (CO2) emissions in Pakistan. Panel unit root, panel co-integration, and Fully Modified OLS (FMOLS) estimators were employed to investigate the existence of long-run relationship between Gross Domestic Product (GDP), Trade openness (TO) and energy consumption (EC) with CO2 emissions in Pakistan from 1980 to 2014. The results show that GDP per capita have a significant positive effect on CO2 emissions. This implies that an increase in GDP per capita increases CO2 emissions and show a scale effect. The results also indicate the existence of long run positive relationship of energy consumption on carbon emission and negative relationship with trade openness and FTA. This research is helpful for policy makers to eliminate the negative impact by adopting appropriate policy instruments and promote Pakistan's trade in the international market. </p

A computer-aided polyp detection system in screening and surveillance colonoscopy:an international, multicentre, randomised, tandem trial

Background: Studies on the effect of computer-aided detection (CAD) in a daily clinical screening and surveillance colonoscopy population practice are scarce. The aim of this study was to evaluate a novel CAD system in a screening and surveillance colonoscopy population. Methods: This multicentre, randomised, controlled trial was done in ten hospitals in Europe, the USA, and Israel by 31 endoscopists. Patients referred for non-immunochemical faecal occult blood test (iFOBT) screening or surveillance colonoscopy were included. Patients were randomomly assigned to CAD-assisted colonoscopy or conventional colonoscopy; a subset was further randomly assigned to undergo tandem colonoscopy: CAD followed by conventional colonoscopy or conventional colonoscopy followed by CAD. Primary objectives included adenoma per colonoscopy (APC) and adenoma per extraction (APE). Secondary objectives included adenoma miss rate (AMR) in the tandem colonoscopies. The study was registered at ClinicalTrials.gov, NCT04640792. Findings: A total of 916 patients were included in the modified intention-to-treat analysis: 449 in the CAD group and 467 in the conventional colonoscopy group. APC was higher with CAD compared with conventional colonoscopy (0·70 vs 0·51, p=0·015; 314 adenomas per 449 colonoscopies vs 238 adenomas per 467 colonoscopies; poisson effect ratio 1·372 [95% CI 1·068–1·769]), while showing non-inferiority of APE compared with conventional colonoscopy (0·59 vs 0·66; p&lt;0·001 for non-inferiority; 314 of 536 extractions vs 238 of 360 extractions). AMR in the 127 (61 with CAD first, 66 with conventional colonoscopy first) patients completing tandem colonoscopy was 19% (11 of 59 detected during the second pass) in the CAD first group and 36% (16 of 45 detected during the second pass) in the conventional colonoscopy first group (p=0·024). Interpretation: CAD increased adenoma detection in non-iFOBT screening and surveillance colonoscopies and reduced adenoma miss rates compared with conventional colonoscopy, without an increase in the resection of non-adenomatous lesions. Funding: Magentiq Eye.</p

Autoreactivity to Glucose Regulated Protein 78 Links Emphysema and Osteoporosis in Smokers

Rationale: Emphysema and osteoporosis are epidemiologically associated diseases of cigarette smokers. The causal mechanism(s) linking these illnesses is unknown. We hypothesized autoimmune responses may be involved in both disorders. Objectives: To discover an antigen-specific autoimmune response associated with both emphysema and osteoporosis among smokers. Methods: Replicate nonbiased discovery assays indicated that autoimmunity to glucose regulated protein 78 (GRP78), an endoplasmic reticulum chaperone and cell surface signaling receptor, is present in many smokers. Subject assessments included spirometry, chest CT scans, dual x-ray absorptiometry, and immunoblots for anti-GRP78 IgG. Anti-GRP78 autoantibodies were isolated from patient plasma by affinity chromatography, leukocyte functions assessed by flow cytometry, and soluble metabolites and mediators measured by immunoassays. Measurements and Main Results Circulating anti-GRP78 IgG autoantibodies were detected in plasma specimens from 86 (32%) of the 265 smoking subjects. Anti-GRP78 autoantibodies were singularly prevalent among subjects with radiographic emphysema (OR 3.1, 95%CI 1.7–5.7, p = 0.003). Anti-GRP78 autoantibodies were also associated with osteoporosis (OR 4.7, 95%CI 1.7–13.3, p = 0.002), and increased circulating bone metabolites (p = 0.006). Among emphysematous subjects, GRP78 protein was an autoantigen of CD4 T-cells, stimulating lymphocyte proliferation (p = 0.0002) and IFN-gamma production (p = 0.03). Patient-derived anti-GRP78 autoantibodies had avidities for osteoclasts and macrophages, and increased macrophage NFkB phosphorylation (p = 0.005) and productions of IL-8, CCL-2, and MMP9 (p = 0.005, 0.007, 0.03, respectively). Conclusions: Humoral and cellular GRP78 autoimmune responses in smokers have numerous biologically-relevant pro-inflammatory and other deleterious actions, and are associated with emphysema and osteoporosis. These findings may have relevance for the pathogenesis of smoking-associated diseases, and development of biomarker immunoassays and/or novel treatments for these disorders

Spontaneous Sinus Conversion of Permanent Atrial Fibrillation During Treatment of Hyperkalemia

Hyperkalemia is a common adverse effect of treatment for heart failure and is associated with high mortality and morbidity. The cardiac manifestations of hyperkalemia include various electrocardiogram changes. We describe a case of a 74-year-old woman with heart failure and permanent atrial fibrillation who reverted to normal sinus rhythm during recovery from hyperkalemia

Early Treatment with Fumagillin, an Inhibitor of Methionine Aminopeptidase-2, Prevents Pulmonary Hypertension in Monocrotaline-Injured Rats

Pulmonary Hypertension (PH) is a pathophysiologic condition characterized by hypoxemia and right ventricular strain. Proliferation of fibroblasts, smooth muscle cells, and endothelial cells is central to the pathology of PH in animal models and in humans. Methionine aminopeptidase-2 (MetAP2) regulates proliferation in a variety of cell types including endothelial cells, smooth muscle cells, and fibroblasts. MetAP2 is inhibited irreversibly by the angiogenesis inhibitor fumagillin. We have previously found that inhibition of MetAP2 with fumagillin in bleomycin-injured mice decreased pulmonary fibrosis by selectively decreasing the proliferation of lung myofibroblasts. In this study, we investigated the role of fumagillin as a potential therapy in experimental PH. In vivo, treatment of rats with fumagillin early after monocrotaline injury prevented PH and right ventricular remodeling by decreasing the thickness of the medial layer of the pulmonary arteries. Treatment with fumagillin beginning two weeks after monocrotaline injury did not prevent PH but was associated with decreased right ventricular mass and decreased cardiomyocyte hypertrophy, suggesting a direct effect of fumagillin on right ventricular remodeling. Incubation of rat pulmonary artery smooth muscle cells (RPASMC) with fumagillin and MetAP2-targeting siRNA inhibited proliferation of RPASMC in vitro. Platelet-derived growth factor, a growth factor that is important in the pathogenesis of PH and stimulates proliferation of fibroblasts and smooth muscle cells, strongly increased expression of MetP2. By immunohistochemistry, we found that MetAP2 was expressed in the lesions of human pulmonary arterial hypertension. We propose that fumagillin may be an effective adjunctive therapy for treating PH in patients