Atmospheric Plasma Solutions (United States)

Targeted insertion (TIN) of transgenic trait cassettes has the potential to reduce timeline and cost for GM product development and commercialization. Precise genome engineering has made remarkable progress over the last several decades, particularly with the use of site-directed nucleases as core editing machinery. However, there are still many critical factors that can impact TIN efficiency including insertion site selection, nuclease optimization and expression, donor vector design, gene delivery, and stable event regeneration. Here, we develop workflows for target site sequence identification and gRNA screening for CRISPR-Cas12a system and demonstrate its successful application for TIN in maize with donor sequences up to 10 kilobase pairs (kb) in size. We first prioritize genomic regions for inserting transgenes in silico using bioinformatics tools and then test gRNA performance using a leaf protoplast transient assay. Despite its known low efficiency, we identify homology-directed repair (HDR) as the preferential pathway for directing targeted insertions of large sequences in immature embryos and demonstrate double-junction integrations at a rate of up to 4%. We further apply a molecular analysis workflow using large amplicon TaqMan assays and nanopore sequencing for streamlined identification and characterization of high-quality insertion events with intact large inserts. Analysis of TIN events across generations suggests that efficiency bottlenecks are not limited to donor targeted integration; attrition in efficiency also results from partial or additional donor insertion, chimerism, and close linkage with undesired sequence insertions such as those encoding the editing machinery. This work represents a major step forward in realizing the potential of precise genome engineering in maize for basic research and biotech trait development applications.

ABSTRACT The Canadian Federal Government promulgated new and lower NO2 Ambient Air Quality Standards (CAAQS) that went into effect in 2020 with additional decreases scheduled for 2025. The new hourly and annual NO2 CAAQS are 60 and 17 ppb, respectively, and the 2025 hourly and annual CAAQS are 42 and 12 ppb, respectively. The province of Alberta has also promulgated Ambient Air Quality Objectives (AAAQO) for NO2 currently set to 159 and 24 ppb on an hourly and annual basis, respectively. The Wood Buffalo Environmental Association (WBEA) in northeastern Alberta, Canada monitors NO2 at 21 community and industrial sites throughout the Athabasca Oil Sands Region (AOSR), for regulatory compliance using Thermo-Environmental (TEI) Model 42i Federal Reference Method (FRM) designated NO-NO2-NOx analyzers. The 42i measures NO directly via NO-O3 chemiluminescence, and NOx following the reduction of oxidized nitrogen to NO by a heated internal molybdenum converter. The difference between the NOx and NO channels is reported as NO2. This study presents the results of a three-year (2018–2021) WBEA comparison of four continuous NO2 analyzers: TEI 42i FRM; the API Model T500U cavity attenuated phase shift (CAPS) Federal Equivalent Method (FEM); a total reactive odd nitrogen analyzer (TEI Model 42i-Y); and a TEI 42i equipped with an external photolytic converter. The study showed that NO2 data from all analyzers were highly correlated and in general agreement, with r2 values (vs. the CAPS) ranging from 0.990–0.997 and slopes ranging from 0.933–0.992. Mean NO2 concentrations over the study period ranged from 7.2–7.5 ppb. Differences between the TEI 42i, TEI 42i-Y, and PhoNO, relative to the CAPS were all positive and highly significant (p < 0.0001), based upon nonparametric tests. The potential impact from the selection of different FRM/FEM measurement methods on current and future Canadian 2025 regulatory compliance in the region is evaluated. Implications: The study objective was to compare/evaluate different regulatory NO2 measurement techniques from a regional monitoring authority in a routine network operational context. Relatively small NO2 differences resulted in significant differences with respect to regulatory compliance triggers, particularly hourly standards based on daily extreme value statistics (e.g., 99th percentiles). For example, mean hourly NO2 △ differences ranged from 0.02–0.26 ppb over the study period but resulted in 2–3 ppb differences in the 3-year hourly CAAQS metrics. These differences could affect regulatory CAAQS and LARP compliance (management level) at monitoring sites observed during 2019 annual and 2020 hourly LARP trigger exceedances.

em que registamos recuos ou objeções à educação para a cidadania nas escolas, e em que vemos decisões de tribunais que promovem a despenalização de agressores que infligem maus tratos e abusos sexuais a mulheres e meninas.Os desafios feministas, teóricos e metodológicos,

A presente dissertação de mestrado baseia-se no estudo e análise do mais recente tratado do sistema interamericano de direitos humanos, a Convenção Interamericana sobre a Proteção dos Direitos Humanos das Pessoas Idosas e alguns dos possíveis impactos, caso venha a ser ratificada em território nacional, na normativa protetora da pessoa idosa vigente no Brasil. Através de uma retrospectiva das regras internacionais (sistemas universal e regional interamericano) responsáveis pela consolidação do reconhecimento da pessoa idosa como sujeito de direitos e não somente objeto de proteção estatal, foram apresentados os principais avanços trazidos pela Convenção, com destaque para a participação do Brasil durante o procedimento de elaboração e aprovação do texto final na Organização dos Estados Americanos Diante de um cenário de envelhecimento acelerado e das inúmeras velhices particularizadas pelas diversas realidades sociais, econômicas e culturais da região, o tratado tem o mérito de jogar luz, através das lentes principiológicas dos direitos humanos, no fato de que ações de promoção da igualdade e enfrentamento da discriminação em razão da idade não podem ser feitas sem considerar as tantas situações de vulnerabilidade a que estão sujeitas as pessoas idosas. Enquanto aguarda-se a finalização do processo de ratificação da Convenção perante as instâncias brasileiras, foram analisadas nesta dissertação situações que poderão conduzir a uma ampliação dos direitos da pessoa idosa em solo nacional, em consonância com as diretrizes e objetivos propostos pelo instrumento interamericano de direitos humanos, já em vigor na região desde janeiro de 2017.

Abstract The Atmospheric Plasma Coating Removal (APCR) process provides a unique new way to quickly and safely remove the toughest industrial coatings. As with any new process a thorough examination of the possible risks to the end users and the environment is critical. APS hired an environmental testing facility to conduct a series of experiments and provide an assessment of noise, dust, and select chemicals to determine the safety of the APCR process. The project examined two operational modes of the APCR process: the in-use case, in which the device is being used to actively remove coatings from steel and the static case, in which the device was fired continuously into air without treating a surface. Direct-Reading tests for gases (Ozone, Carbon Monoxide, Carbon Dioxide, and Nitrogen Oxides) were performed over the 4 hour testing period. Area noise levels, both inside and outside the testing area, were recorded and chemical samples were collected for laboratory analysis for Aluminum, Copper, Hydrogen Cyanide, Total Hydrocarbons, Respirable Dust, Total Dust, and Sulfur Dioxide. On December 18, 2018 and March 4th and 5th, 2019, BSI EHS Services (BSI) and Atmospheric Plasma Solutions (APS) conducted a series of tests to evaluate environmental health and safety aspects of the use of APS's APCR technology. These tests were conducted within a testing enclosure located within the APS facility in Cary, North Carolina and included an assessment of noise, dust, and select chemicals to determine the safety of the APCR technology. During testing, samples were collected for a variety of media including: Direct-Reading tests for gases (Ozone, Carbon Monoxide, Carbon Dioxide, and Nitrogen Oxides)Area noise levels (both inside and outside the testing shed)Chemical samples with laboratory analysis (Aluminum, Copper, Hydrogen Cyanide, Total Hydrocarbons, Respirable Dust, Total Dust, and Sulfur Dioxide)

Requirements Engineering is the set of activities involved in managing, surveying, documenting, and maintaining a set of requirements for a product.Engineering involves the use of systematic repeatability techniques to ensure that the Software Requirements are complete, consistent, valid and verifiable.Software Requirements Specification is an organized process oriented towards defining, documenting and maintaining Software Requirements during the development life cycle.Many authors suggest that requirements should always focus their statements on what the software product needs to address without specifying how to implement them.However, the detail of the Software Requirements is influenced by several factors such as: organizational thoughts; existing specification standards; and regulatory needs.This work fits exactly with regulatory needs, where the characteristics of Software Requirements Specification in Regulated Environments such as aeronautics, rail and medical.This article briefly presents the three standards used in these regulated environments (RTCA DO-178C, IEC 62279 and IEC 62304) and analyzes their similarities from a Requirements Specification perspective.