Optimization of DNA Extraction from Human Milk

Background: Techniques are emerging for determining the best and most cost-effective way to test for human milk adulteration. Currently, the literature is focused on the use of qPCR testing, a technique used to isolate and amplify pieces of DNA for analysis. However, no recommendation currently exists on the best DNA extraction kit to use to achieve optimal DNA yield or purity from human milk samples for downstream qPCR use. Thus, the objective of this study is to assess and compare two DNA extraction kits for use with human milk samples for future DNA-based analysis in the testing for bovine milk adulteration in human milk. Methods: Forty mothers pumped human milk samples under the observation of a researcher using a brand-new hand pump. Eight unadulterated samples were then randomly chosen for DNA-extraction. The eight samples were thawed, pooled, and DNA was isolated using the Omega Bio-Tek\u27s E.Z.N.A.® Blood DNA Mini Spin Kit and the Norgen Biotek Corporation Plasma/Serum Circulating DNA Purification Mini Kit (Slurry Format) per the protocols included in the kits on arrival. An overnight incubation modification was also added to both kits to try to obtain optimal yield and purity. UV/VIS spectroscopy was used to determine DNA yield and purity using the ThermoFisher Scientific NanoDrop 2000TM 260/280 ratio, and a cost comparison was done between kits. Results: The Norgen kit with no modification provided 143% more DNA than the E.Z.N.A kit with no modifications. Similarly, the average nucleic acid yield was 134% greater when comparing the Norgen and E.Z.N.A kit with an overnight incubation. The Norgen kit provided a 17.0% greater 260/280 ratio and an 11.4% greater 260/280 ratio than the iii E.Z.N.A. kit, with and without modifications, respectively. The Norgen kit costs $2.37 more per extraction than the E.Z.N.A kit. Modifying both DNA extraction kits with an overnight incubation decreased the average nucleic acid yield and purity of the resulting DNA. Conclusion: From these results, the Norgen kit without overnight incubation is a better extraction kit for DNA extraction from raw human milk for both nucleic acid yield and purity. However, the EZNA kit costs less per extraction at$1.45 vs. $3.82. For extraction purposes, purity should be prioritized over nucleic acid yield because contaminants can compromise results and shorten the shelf-life of samples

Building theatres/theatre buildings: reinventing Mull Theatre

Mull Theatre is a professional touring theatre company based on a small island off the west coast of Scotland. In 2008 the company relocated from a small converted cow byre which seated 42 people to a new purpose-built venue –Druimfin - on a different part of the island. The move was made possible through a grant from the Scottish Arts Council in 2006, which was awarded on the expectation that the new building would be a ‘production centre’ as opposed to a theatre. That is to say the emphasis in the design of the new space was to be placed on the production rather than the reception of the theatrical event. This stands in contrast to the expectation of many theatre attendees that the new space would continue as it had been – as a place to go and see a theatre production - but that it would do so out of a much larger, more comfortable and better equipped venue. Building Theatres/Theatre Buildings stems from a three year Collaborative Doctoral Award between Mull Theatre and the University of Glasgow, which was funded by the Arts and Humanities Research Council (AHRC). Using the partnership that emerged from this award, the thesis explores what was potentially lost and gained in the move in order to draw conclusions about the wider relationship between spaces of performance and the creation of theatrical meaning in relation to small and medium scale touring theatre. It also uses the company’s dual identity as a touring company with its own permanent building to extend the discussion and to examine the wide range of venues which currently form the rural touring circuit in Scotland. By bringing together primary fieldwork from a pivotal moment in the company’s identity alongside current dialogues regarding theatre space and touring theatre, this research provides new knowledge about this often overlooked theatre company, its buildings and its role within contemporary Scottish theatre and small scale rural touring

Design manual for excavation support using deep mixing technology

Deep mixing (DM) is the modification of in situ soil to increase strength, control deformation, and reduce permeability. Multiaxis augers and mixing paddles are used to construct overlapping columns strengthened by mixing cement with in situ soils. This method has been used for excavation support to increase bearing capacity, reduce movements, prevent sliding failure, control seepage by acting as a cutoff barrier, and as a measure against base heave. DM is effectively used in excavations both in conjunction with and in substitution of traditional techniques, where it results in more economical and convenient solutions for the stability of the system and the prevention of seepage. Although DM is currently used for excavation control in numerous projects, no standard procedure has been developed and the different applications have not been evaluated. As this technique emerges as a more economical and effective alternative to traditional excavation shoring, there is a need for guidelines describing proven procedures for evaluation of design, analysis and construction. The main objective of this research is to develop a methodology to design retaining systems using deep mixing technology. The method will be evaluated using numerical analysis of one selected case history

Development of a Multi-directional Direct Simple Shear Testing Device for Characterization of the Cyclic Shear Response of Marine Clays

This dissertation describes the development of a new multi-directional direct simple shear testing device, the Texas A&M Multi-directional Direct Simple Shear (TAMU-MDSS), for testing marine soil samples under conditions, which simulate, at the element level, the state of stress acting within a submarine slope under dynamic loading. Prototype testing and an experimental program to characterize the response of marine clays to complex loading conditions are presented. The work is divided into four major components: 1) Equipment Development: Design and construction of a prototype multi-directional direct simple shear testing device (TAMU-MDSS) that addresses the limitations of previous devices. 2) Support systems: selection of control software, development of data acquisition system and design of back pressure systems for direct pore pressure measurements. 3) Prototype Testing: performance of the TAMU MDSS system and testing of strain-control and stress-control capabilities. 4) Experimental Testing: characterize the response of marine clays to monotonic, dynamic and random loads. The two-directional monotonic, cyclic, circular and figure-8 tests demonstrated the undrained shear strength increases with increasing initial shear stress, (i.e, slope), for shearing in the same direction (equivalent to downhill). The strength decreases for shearing in the direction opposite to the initial stress (shearing uphill). The response is as brittle for shearing in the same direction as the shear stress applied during consolidation initial shear stress and ductile for shearing opposite to initial shear stress. These findings have important implications for the stability of the slope, predicting that forces acting downward in the slope direction will need to mobilize less strain to reach peak strength and initiate failure. This information provides insight into the behavior of marine soils under complex loading conditions, and provides high quality laboratory data for use in constitutive and finite element model development for analysis of submarine slopes

Conceptualizing a Theory of Ethical Behavior in Engineering

Traditional engineering courses typically approach teaching and problem solving by focusing on the physical dimensions of those problems without consideration of dynamic social and ethical dimensions. As such, projects can fail to consider human rights, community questions and concerns, broader impacts upon society, or otherwise result in inequitable outcomes. And, despite the fact that students in engineering receive training on the Professional Code of Ethics for Engineers, to which they are expected to adhere in practice, many students are unable to recognize and analyze real-life ethical challenges as they arise. Indeed, research has found that students are typically less engaged with ethics—defined as the sensitivity and judgment of microethics and macroethics, sensitivity to diversity, and interest in promoting organizational ethical culture—at the end of their engineering studies than they were at the beginning. As such, many studies have focused on developing and improving the curriculum surrounding ethics through, for instance, exposing students to ethics case studies. However, such ethics courses often present a narrow and simplified view of ethics that students may struggle to integrate with their broader experience as engineers. Thus, there is a critical need to unpack the complexity of ethical behavior amongst engineering students in order to determine how to better foster ethical judgment and behavior. Promoting ethical behavior among engineering students and developing a culture of ethical behavior within institutions have become goals of many engineering programs. Towards this goal, we would like to present an overview of the current scholarship of engineering ethics and propose a theoretical framework of ethical behavior using a review of articles related to engineering ethics from 1997-2020. The review engages in theories across disciplines including philosophy, education, and psychology. In this work-in-progress paper, we present a subset of initial results based on a review of the first 50 articles out of the systematically selected 409 articles from Springer, Engineering Village, and EBSCO-Education Full Text. Preliminary results identify two major kinds of drivers of ethical behavior, namely individual level ethical behavior drivers (sensitivity to microethics, sensitivity to macroethics, implicit understanding, and explicit understanding) and institutional drivers (sensitivity to diversity and institutional ethical culture). Our preliminary results indicate that a sensitivity to both microethics and macroethics as well as the implicit and explicit understanding of ethics are essential in promoting ethical behavior amongst students. Furthermore, while drivers of ethical behavior at the individual level is important, one should not ignore the roles of the drivers of ethical behavior at the institutional level in promoting a collective ethical culture within organizations. The review also points to a need to focus on increasing students’ macroethical sensitivity to topics such as sustainability and protection of human rights. This research thus addresses the need, driven by existing scholarship, 2 to identify a conceptual framework for explaining how ethical judgment and behavior in engineering can be further promote

NFL and BMI

Over the years, the position of quarterback in the National Football League has undergone various transformations. Of those transformations, the need for taller and heavier players has risen. As player stature has changed, so has the style of play in the NFL. This study is designed to determine if there is a correlation between quarterback body mass index (BMI) and total yards from scrimmage (TY). The study uses the stratified sampling method to look at the quarterback from each team who started the most games during the 2017-2018 football season. Data on age, height, weight, and game performance were collected from the NFL website. We hypothesize that quarterback BMI will have no correlation with total yards, but will have an affect on quarterback rushing yards. The study will also look to examine if age and family size may also have impact on game performance

A semi-empirical relationship for the small-strain shear modulus of soft clays

The small-strain shear modulus (Gmax) is a soil property that has many practical applications. The authors compiled a database of Gmax measurements for 40 normally consolidated to slightly overconsolidated low to high plasticity clays. Using these data, the authors propose a semi-empirical relationship between Gmax, effective stress (σ'v or σ'c), preconsolidation stress (σ'p) and in-situ void ratio (e0) for four ranges of plasticity index (Ip): Ip < 30%, 30% ≤ Ip < 50%, 50% ≤ Ip < 80% and 80% ≤ Ip < 120%. With results from bender element tests on a Gulf of Mexico clay subjected to multiple load-unload consolidation loops, the authors were able to validate the proposed relationships for 30% ≤ Ip < 50% and 50% ≤ Ip < 80%. The proposed relationship for 30% ≤ Ip < 50% and 50% ≤ Ip < 80% captures changes in laboratory Gmax resulting from variations in effective stress (σ'c), maximum past stress (σ'v,max), and void ratio. The proposed relationships are a simple and efficient tool that can provide independent insight on Gmax if the stress history of a clay is known, or on stress history if Gmax is known

Static and Dynamic Response of Micropiles Used for Reinforcing Slopes

To study the static and dynamic response of micropile-reinforced slopes, static model tests and shaking table tests were performed. The failure modes, the pile-slope interaction, the displacement, and the static/dynamic earth pressure distributions were analyzed based on static and dynamic model tests with a prescribed sliding surface. The test results indicated: (1) The micropile failure mode is mainly bending failure under both loading conditions. As far as the damage to the pile body is concerned, under static loading, the rear row piles showed more damage than the middle row piles followed by the front row piles. Under dynamic loading, the damage of the rear row piles was approximately the same as the middle row piles, which was greater than the front row piles; (2) The earth pressures in front of and behind each row of micropiles and the axial force of the pile body distributed triangularly for both loading conditions, with the bending moment of the pile body distributed in an “S” shape; (3) The landslide thrust experienced by the micropiles has a relatively large group effect. The group effect or shear ratio parameters are recommended for each loading case; (4) The interaction between the micropiles and the soil landslide presents evident progressive failure and load transfer between the rows

Improving the Effectiveness of Saturated Riparian Buffers for Removing Nitrate from Subsurface Drainage

A saturated riparian buffer (SRB) is an edge‐of‐field conservation practice that reduces nitrate export from agricultural lands by redistributing tile drainage as shallow groundwater and allowing for denitrification and plant uptake. We propose an approach to improve the design of SRBs by analyzing a tradeoff in choosing the SRB width, and we apply the approach to six sites with SRBs in central Iowa. A larger width allows for more residence time, which increases the opportunity for removing nitrate that enters the buffer. However, because the SRBs considered here treat only a portion of the tile flow when it is large, for the same difference in hydraulic head, a smaller width allows more of the total tile flow to enter the buffer and therefore treats more of the drainage. By maximizing the effectiveness of nitrate removal—defined as the ratio of total nitrate removed by the SRB to total nitrate leaving the field in tile drainage, an equation for the optimal width was derived in terms of soil properties, denitrification rates, and head difference. All six sites with existing SRBs considered here have optimal widths smaller than the current width, and two are below the minimum width listed in current design standards. In terms of uncertainty, the main challenges in computing the optimal width for a site are estimating the removal coefficient for nitrate and determining the saturated hydraulic conductivity. Nevertheless, including a width that accounts for site conditions in the design standards would improve water quality, locally and regionally.This is the peer reviewed version of the following article: McEachran, AR, Dickey, LC, Rehmann, CR, et al. Improving the effectiveness of saturated riparian buffers for removing nitrate from subsurface drainage. J. Environ. Qual. 2020; 49: 1624–1632, which has been published in final form at doi: 10.1002/jeq2.20160. This article may be used for non-commercial purposes in accordance with Wiley Terms and Conditions for Use of Self-Archived Versions.</p

Macroethics in Undergraduate Engineering: an Institutional View

Engineering programs tend to focus on technical knowledge rather than developing ethical understanding. Those programs that do teach ethics typically focus on microethics, i.e. issues that arise in particular contexts and interactions between individuals, rather than macroethics, i.e. issues that address societal concerns more broadly. We conducted a systematic literature review of previous work assessing the inclusion of ethics education in undergraduate engineering programs. We used an institutional framework to understand where, in the context of their university experiences, undergraduate students are exposed to ethics. Through this analysis, we found that the most effective way to help students develop an understanding of macroethics is programming outside of the classroom. However, while equity and inclusion are key aspects of macroethics in the engineering profession, exposure to this topic is not accessible to all groups of students due to financial and time constraints that may preclude their participation.This proceeding is published as LaPatin, Michaela, Kasey Faust, C. P. Poleacovschi, Kate Padgett-Walsh, Scott Feinstein, Cassandra Rutherford, and Luan Nguyen. "Macroethics in Undergraduate Engineering: an Institutional View." In Engineering Project Organization Conference (EPOC). 2020. Copyright 2020 The Authors. Posted with permission