Search results for: tandem repeats

Authors: Aryati Yashadhana, Ted Fields Jnr., Wendy Fernando, Kelvin Brown, Godfrey Blitner, Francis Hayes, Ruby Stanley, Brian Donnelly, Bridgette Jerrard, Anthea Burnett, Anthony B. Zwi

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Indigenous Australians experience some of the worst health outcomes globally, with life expectancy being significantly poorer than those of non-Indigenous Australians. This is largely attributed to preventable diseases such as diabetes (prevalence 39% in Indigenous Australian adults > 55 years), which is attributed to a raised risk of diabetic visual impairment and cataract among Indigenous adults. Our study aims to explore the interface between structural and sociocultural determinants and human agency, in order to understand how they impact (1) accessibility of eye health and chronic disease services and (2) the potential for Indigenous patients to achieve positive clinical eye health outcomes. We used Participatory Action Research methods, and aimed to privilege the voices of Indigenous people through community collaboration. Semi-structured interviews (n=82) and patient focus groups (n=8) were conducted by Indigenous Community-Based Researchers (CBRs) with diabetic Indigenous adults (> 40 years) in four remote communities in Australia. Interviews (n=25) and focus groups (n=4) with primary health care clinicians in each community were also conducted. Data were audio recorded, transcribed verbatim, and analysed thematically using grounded theory, comparative analysis and Nvivo 10. Preliminary analysis occurred in tandem with data collection to determine theoretical saturation. The principal investigator (AY) led analysis sessions with CBRs, fostering cultural and contextual appropriateness to interpreting responses, knowledge exchange and capacity building. Identified themes were conceptualised into three spheres of influence: structural (health services, government), sociocultural (Indigenous cultural values, distrust of the health system, ongoing effects of colonialism and dispossession) and individual (health beliefs/perceptions, patient phenomenology). Permeating these spheres of influence were three core determinants: economic disadvantage, health literacy/education, and cultural marginalisation. These core determinants affected accessibility of services, and the potential for patients to achieve positive clinical outcomes at every level of care (primary, secondary, tertiary). Our findings highlight the clinical realities of institutionalised and structural inequities, illustrated through the lived experiences of Indigenous patients and primary care clinicians in the four sampled communities. The complex determinants surrounding inequity in health for Indigenous Australians, are entrenched through a longstanding experience of cultural discrimination and ostracism. Secure and long term funding of Aboriginal Community Controlled Health Services will be valuable, but are insufficient to address issues of inequity. Rather, working collaboratively with communities to build trust, and identify needs and solutions at the grassroots level, while leveraging community voices to drive change at the systemic/policy level are recommended.

Keywords: indigenous, Australia, culture, public health, eye health, diabetes, social determinants of health, sociology, anthropology, health equity, aboriginal and Torres strait islander, primary care

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Authors: Adriana Muñoz-Talavera, Ana Rosa Rincón-Sánchez, Abraham Escobedo-Moratilla, María Cristina Islas-Carbajal, Miguel Ángel Gómez-Lim

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The highest rates of diabetes incidence and prevalence worldwide will increase the number of diabetic patients requiring insulin or insulin analogues. Then, current production systems would not be sufficient to meet the future market demands. Therefore, developing efficient expression systems for insulin and insulin analogues are needed. In addition, insulin analogues with better pharmacokinetics and pharmacodynamics properties and without mitogenic potential will be required. SCI-57 (single chain insulin-57) is an insulin analogue having 10 times greater affinity to the insulin receptor, higher resistance to thermal degradation than insulin, native mitogenicity and biological effect. Plants as expression platforms have been used to produce recombinant proteins because of their advantages such as cost-effectiveness, posttranslational modifications, absence of human pathogens and high quality. Immunoglobulin production with a yield of 50% has been achieved by transient expression in Nicotiana benthamiana (Nb). The aim of this study is to produce SCI-57 by transient expression in Nb. Methodology: DNA sequence encoding SCI-57 was cloned in pICH31070. This construction was introduced into Agrobacterium tumefaciens by electroporation. The resulting strain was used to infiltrate leaves of Nb. In order to isolate SCI-57, leaves from transformed plants were incubated 3 hours with the extraction buffer therefore filtrated to remove solid material. The resultant protein solution was subjected to anion exchange chromatography on an FPLC system and ultrafiltration to purify SCI-57. Detection of SCI-57 was made by electrophoresis pattern (SDS-PAGE). Protein band was digested with trypsin and the peptides were analyzed by Liquid chromatography tandem-mass spectrometry (LC-MS/MS). A purified protein sample (20µM) was analyzed by ESI-Q-TOF-MS to obtain the ionization pattern and the exact molecular weight determination. Chromatography pattern and impurities detection were performed using RP-HPLC using recombinant insulin as standard. The identity of the SCI-57 was confirmed by anti-insulin ELISA. The total soluble protein concentration was quantified by Bradford assay. Results: The expression cassette was verified by restriction mapping (5393 bp fragment). The SDS-PAGE of crude leaf extract (CLE) of transformed plants, revealed a protein of about 6.4 kDa, non-present in CLE of untransformed plants. The LC-MS/MS results displayed one peptide with a high score that matches SCI-57 amino acid sequence in the sample, confirming the identity of SCI-57. From the purified SCI-57 sample (PSCI-57) the most intense charge state was 1069 m/z (+6) on the displayed ionization pattern corresponding to the molecular weight of SCI-57 (6412.6554 Da). The RP-HPLC of the PSCI-57 shows the presence of a peak with similar retention time (rt) and UV spectroscopic profile to the insulin standard (SCI-57 rt=12.96 and insulin rt=12.70 min). The collected SCI-57 peak had ELISA signal. The total protein amount in CLE from transformed plants was higher compared to untransformed plants. Conclusions: Our results suggest the feasibility to produce insulin analogue SCI-57 by transient expression in Nicotiana benthamiana. Further work is being undertaken to evaluate the biological activity by glucose uptake by insulin-sensitive and insulin-resistant murine and human cultured adipocytes.

Keywords: insulin analogue, mass spectrometry, Nicotiana benthamiana, transient expression

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Authors: Boris Nemzer, Nebiyu Abshiru, Z. B. Pietrzkowski

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Coffee cherry is one of the most ubiquitous agricultural commodities which possess nutritional and human health beneficial properties. Between the two most widely used coffee cherries Coffea arabica (Arabica) and Coffea canephora (Robusta), Coffea arabica remains superior due to its sensory properties and, therefore, remains in great demand in the global coffee market. In this study, the phytochemical contents and pharmacokinetics of Coffeeberry® Energy (CBE), a commercially available Arabica whole coffee fruit caffeine extract, are investigated. For phytochemical screening, 20 mg of CBE was dissolved in an aqueous methanol solution for analysis by mass spectrometry (MS). Quantification of caffeine and chlorogenic acids (CGAs) contents of CBE was performed using HPLC. For the bioavailability study, serum samples were collected from human subjects before and after 1, 2 and 3 h post-ingestion of 150mg CBE extract. Protein precipitation and extraction were carried out using methanol. Identification of compounds was performed using an untargeted metabolomic approach on Q-Exactive Orbitrap MS coupled to reversed-phase chromatography. Data processing was performed using Thermo Scientific Compound Discover 3.3 software. Phytochemical screening identified a total of 170 compounds, including organic acids, phenolic acids, CGAs, diterpenoids and hydroxytryptamine. Caffeine & CGAs make up more than, respectively, 70% & 9% of the total CBE composition. For serum samples, a total of 82 metabolites representing 32 caffeine- and 50 phenolic-derived metabolites were identified. Volcano plot analysis revealed 32 differential metabolites (24 caffeine- and 8 phenolic-derived) that showed an increase in serum level post-CBE dosing. Caffeine, uric acid, and trimethyluric acid isomers exhibited 4- to 10-fold increase in serum abundance post-dosing. 7-Methyluric acid, 1,7-dimethyluric acid, paraxanthine and theophylline exhibited a minimum of 1.5-fold increase in serum level. Among the phenolic-derived metabolites, iso-feruloyl quinic acid isomers (3-, 4- and 5-iFQA) showed the highest increase in serum level. These compounds were essentially absent in serum collected before dosage. More interestingly, the iFQA isomers were not originally present in the CBE extract, as our phytochemical screen did not identify these compounds. This suggests the potential formation of the isomers during the digestion and absorption processes. Pharmacokinetics parameters (Cmax, Tmax and AUC0-3h) of caffeine- and phenolic-derived metabolites were also investigated. Caffeine was rapidly absorbed, reaching a maximum concentration (Cmax) of 10.95 µg/ml in just 1 hour. Thereafter, caffeine level steadily dropped from the peak level, although it did not return to baseline within the 3-hour dosing period. The disappearance of caffeine from circulation was mirrored by the rise in the concentration of its methylxanthine metabolites. Similarly, serum concentration of iFQA isomers steadily increased, reaching maximum (Cmax: 3-iFQA, 1.54 ng/ml; 4-iFQA, 2.47 ng/ml; 5-iFQA, 2.91 ng/ml) at tmax of 1.5 hours. The isomers remained well above the baseline during the 3-hour dosing period, allowing them to remain in circulation long enough for absorption into the body. Overall, the current study provides evidence of the potential health benefits of a uniquely formulated whole coffee fruit product. Consumption of this product resulted in a distinct serum profile of bioactive compounds, as demonstrated by the more than 32 metabolites that exhibited a significant change in systemic exposure.

Keywords: phytochemicals, mass spectrometry, pharmacokinetics, differential metabolites, chlorogenic acids

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Authors: Suresh Salla

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In this paper, author explores for most sustainable last mile transport mode addressing present problems of traffic congestion, jams, pollution and travel stress. Development of energy-efficient sustainable integrated transport system(s) is/are must to make our cities more livable. Emphasis on autonomous, connected, electric, sharing system for effective utilization of systems (vehicles and public infrastructure) is on the rise. Many surface mobility innovations like PBS, Ride hailing, ride sharing, etc. are, although workable but if we analyze holistically, add to the already congested roads, difficult to ride in hostile weather, causes pollution and poses commuter stress. Sustainability of transportation is evaluated with respect to public adoption, average speed, energy consumption, and pollution. Why public prefer certain mode over others? How commute time plays a role in mode selection or shift? What are the factors play-ing role in energy consumption and pollution? Based on the study, it is clear that public prefer a transport mode which is exhaustive (i.e., less need for interchange – network is widespread) and intensive (i.e., less waiting time - vehicles are available at frequent intervals) and convenient with latest technologies. Average speed is dependent on stops, number of intersections, signals, clear route availability, etc. It is clear from Physics that higher the kerb weight of a vehicle; higher is the operational energy consumption. Higher kerb weight also demands heavier infrastructure. Pollution is dependent on source of energy, efficiency of vehicle, average speed. Mode can be made exhaustive when the unit infrastructure cost is less and can be offered intensively when the vehicle cost is less. Reliable and seamless integrated mobility till last ¼ mile (Five Minute Walk-FMW) is a must to encourage sustainable public transportation. Study shows that average speed and reliability of dedicated modes (like Metro, PRT, BRT, etc.) is high compared to road vehicles. Electric vehicles and more so battery-less or 3rd rail vehicles reduce pollution. One potential mode can be Cycleloop PRT, where commuter rides e-cycle in a dedicated path – elevated, at grade or underground. e-Bike with kerb weight per rider at 15 kg being 1/50th of car or 1/10th of other PRT systems makes it sustainable mode. Cycleloop tube will be light, sleek and scalable and can be modular erected, either on modified street lamp-posts or can be hanged/suspended between the two stations. Embarking and dis-embarking points or offline stations can be at an interval which suits FMW to mass public transit. In terms of convenience, guided e-Bike can be made self-balancing thus encouraging driverless on-demand vehicles. e-Bike equipped with smart electronics and drive controls can intelligently respond to field sensors and autonomously move reacting to Central Controller. Smart switching allows travel from origin to destination without interchange of cycles. DC Powered Batteryless e-cycle with voluntary manual pedaling makes it sustainable and provides health benefits. Tandem e-bike, smart switching and Platoon operations algorithm options provide superior through-put of the Cycleloop. Thus Cycleloop PRT will be exhaustive, intensive, convenient, reliable, speedy, sustainable, safe, pollution-free and healthy alternative mode for last mile connectivity in cities.

Keywords: cycleloop PRT, five-minute walk, lean modular infrastructure, self-balanced intelligent e-cycle

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Authors: Lauren B. Birney

Abstract:

The envisioned long-term outcome of this three-year research, and implementation plan is for 1) teachers and students to design and build their own computational models of real-world environmental-human health phenomena occurring within the context of the “Human Harbor” and 2) project researchers to evaluate the degree to which these integrated Computer Science (CS) education experiences in New York City (NYC) public school classrooms (PreK-12) impact students’ computational-technical skill development, job readiness, career motivations, and measurable abilities to understand, articulate, and solve the underlying phenomena at the center of their models. This effort builds on the partnership’s successes over the past eight years in developing a benchmark Model of restoration-based Science, Technology, Engineering, and Math (STEM) education for urban public schools and achieving relatively broad-based implementation in the nation’s largest public school system. The Billion Oyster Project Curriculum and Community Enterprise for Restoration Science (BOP-CCERS STEM + Computing) curriculum, teacher professional developments, and community engagement programs have reached more than 200 educators and 11,000 students at 124 schools, with 84 waterfront locations and Out of School of Time (OST) programs. The BOP-CCERS Partnership is poised to develop a more refined focus on integrating computer science across the STEM domains; teaching industry-aligned computational methods and tools; and explicitly preparing students from the city’s most under-resourced and underrepresented communities for upwardly mobile careers in NYC’s ever-expanding “digital economy,” in which jobs require computational thinking and an increasing percentage require discreet computer science technical skills. Project Objectives include the following: 1. Computational Thinking (CT) Integration: Integrate computational thinking core practices across existing middle/high school BOP-CCERS STEM curriculum as a means of scaffolding toward long term computer science and computational modeling outcomes. 2. Data Science and Data Analytics: Enabling Researchers to perform interviews with Teachers, students, community members, partners, stakeholders, and Science, Technology, Engineering, and Mathematics (STEM) industry Professionals. Collaborative analysis and data collection were also performed. As a centerpiece, the BOP-CCERS partnership will expand to include a dedicated computer science education partner. New York City Department of Education (NYCDOE), Computer Science for All (CS4ALL) NYC will serve as the dedicated Computer Science (CS) lead, advising the consortium on integration and curriculum development, working in tandem. The BOP-CCERS Model™ also validates that with appropriate application of technical infrastructure, intensive teacher professional developments, and curricular scaffolding, socially connected science learning can be mainstreamed in the nation’s largest urban public school system. This is evidenced and substantiated in the initial phases of BOP-CCERS™. The BOP-CCERS™ student curriculum and teacher professional development have been implemented in approximately 24% of NYC public middle schools, reaching more than 250 educators and 11,000 students directly. BOP-CCERS™ is a fully scalable and transferable educational model, adaptable to all American school districts. In all settings of the proposed Phase IV initiative, the primary beneficiary group will be underrepresented NYC public school students who live in high-poverty neighborhoods and are traditionally underrepresented in the STEM fields, including African Americans, Latinos, English language learners, and children from economically disadvantaged households. In particular, BOP-CCERS Phase IV will explicitly prepare underrepresented students for skilled positions within New York City’s expanding digital economy, computer science, computational information systems, and innovative technology sectors.

Keywords: computer science, data science, equity, diversity and inclusion, STEM education

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