Improved Combinatorial Assembly and Barcode Sequencing for Gene-Sized DNA Constructs.

Synergistic and supportive interactions among genes can be incorporated in engineering biology to enhance and stabilize the performance of biological systems, but combinatorial numerical explosion challenges the analysis of multigene interactions. The incorporation of DNA barcodes to mark genes coupled with next-generation sequencing offers a solution to this challenge. We describe improvements for a key method in this space, CombiGEM, to broaden its application to assembling typical gene-sized DNA fragments and to reduce the cost of sequencing for prevalent small-scale projects. The expanded reach of the method beyond currently targeted small RNA genes promotes the discovery and incorporation of gene synergy in natural and engineered processes such as biocontainment, the production of desired compounds, and previously uncharacterized fundamental biological mechanisms.

COVID-19 and human resources for health: analysis of planning, policy responses and actions in Latin American and Caribbean countries.

BACKGROUND: The COVID-19 pandemic led to worldwide health service disruptions, due mainly to insufficient staff availability. To gain insight into policy responses and engage with policy-makers, the World Health Organization (WHO) developed a global approach to assess and measure the impact of COVID-19 on the health workforce. As part of this, WHO, together with the Pan American Health Organization (PAHO), supported an impact analysis of COVID-19 on health workers and policy responses, through country case studies in Latin America and the Caribbean (LAC). METHODS: We sought to identify lessons learned from policies on human resources for health (HRH) during health emergencies, to improve HRH readiness. First, we performed a rapid literature review for information-gathering. Second, we used the WHO interim guidance and impact measurement framework for COVID-19 and HRH to systematically organize that information. Finally, we used the Health Labour Market Framework to guide the content analysis on COVID-19 response in eight LAC countries and identify lessons learned to improve HRH readiness. RESULTS: Planning and implementing the COVID-19 response required strengthening HRH governance and HRH data and information systems. The results suggest two main aspects for HRH governance crucial to enabling an agile response: (1) aligning objectives among ministries to define and produce regulation and policy actions; and (2) agreeing on the strategy for HRH management between the public and private sectors, and between central and local governments. We identified three areas for improvement: (a) HRH information systems; (b) methodologies to estimate HRH needs; and (c) teams to analyse information for decision-making. Three key actions were identified during countries monitored, reviewed, and updated their response stages: (i) strengthening response through primary health care; (ii); planning HRH needs to implement the vaccination plan; and (iii) securing long-term HRH availability. CONCLUSION: Countries coordinated and articulated with different stakeholders to align objectives, allocate resources, and agree on policy actions to implement the COVID-19 response. Data and information for HRH preparedness and implementation were key in enabling an agile COVID-19 response and are key areas to explore for improved pandemic preparedness.

Preoperative Computed Tomography Assessment of Risk of Right Ventricle Failure After Left Ventricular Assist Device Placement.

Identification of patients who are at a high risk for right ventricular failure (RVF) after left ventricular assist device (LVAD) implantation is of critical importance. Conventional tools for predicting RVF, including two-dimensional echocardiography, right heart catheterization (RHC), and clinical parameters, generally have limited sensitivity and specificity. We retrospectively examined the ability of computed tomography (CT) ventricular volume measures to identify patients who experienced RVF after LVAD implantation. Between September 2017 and November 2021, 92 patients underwent LVAD surgery at our institution. Preoperative CT-derived ventricular volumes were obtained in 20 patients. Patients who underwent CT evaluation had a similar demographics and rate of RVF after LVAD as patients who did not undergo cardiac CT imaging. In the study cohort, seven of 20 (35%) patients experienced RVF (2 unplanned biventricular assist device, 5 prolonged inotropic support). Computed tomography-derived right ventricular end-diastolic and end-systolic volume indices were the strongest predictors of RVF compared with demographic, echocardiographic, and RHC data with areas under the receiver operating curve of 0.79 and 0.76, respectively. Computed tomography volumetric assessment of RV size can be performed in patients evaluated for LVAD treatment. RV measures of size provide a promising means of pre-LVAD assessment for postoperative RV failure.

Pressure Volume Loop Analysis of the Right Ventricle in Heart Failure With Computed Tomography.

Right ventricular (RV) function is an important marker of mortality in chronic left-sided heart failure. Right ventricular function is particularly important for patients receiving left ventricular assist devices as it is a predictor of postoperative RV failure. RV stroke work index (RVSWI), the area enclosed by a pressure-volume (PV) loop, is prognostic of RV failure. However, clinical RVSWI approximates RVSWI as the product of thermodilution-derived stroke volume and the pulmonary pressure gradient. This ignores the energetic contribution of regurgitant flow and does not allow for advanced energetic measures, such as pressure-volume area and efficiency. Estimating RVSWI from forward flow may underestimate the underlying RV function. We created single-beat PV loops by combining data from cine computed tomography (CT) and right heart catheterization in 44 heart failure patients, tested the approximations made by clinical RVSWI and found it to underestimate PV loop RVSWI, primarily due to regurgitant flow in tricuspid regurgitation. The ability of RVSWI to predict post-operative RV failure improved when the single-beat approach was used. Further, RV pressure-volume area and efficiency measures were obtained and show broad agreement with other functional measures. Future work is needed to investigate the utility of these PV metrics in a clinical setting.

Experiencia de una niña de nueve años ante la separación parental / Effects of parental separation: listening to a nine-year-old girl / Efeitos da separação parental: ouvindo uma menina de nove anos

Resumen (analítico) En este estudio cualitativo se describen y analizan los efectos de la separación parental, a través de la mirada y voz de una niña de nueve años. Se empleó como instrumento una entrevista semiestructurada y un análisis de resultados de tipo categórico. Las repercusiones observadas se clasificaron en efectos individuales como el manejo de la información y la construcción de explicaciones, así como una mayor independencia y mejora en su autoconcepto; y efectos en su entorno familiar, tales como la modificación en los vínculos afectivos con sus padres, dinámicas familiares más consistentes y mayores momentos de bienestar. Concluyendo que los infantes son participantes activos en la construcción de sus significados y que la separación de los padres puede permitir la generación de experiencias de crecimiento.

Abstract (analytical) In this qualitative study, the effects of parental separation are described and analyzed through the voice and point of view of a nine-year-old girl. The authors used a semi-structured interview and categorical data analysis to identify these effects. The identified repercussions of the parental separation were classified into individual effects, such as information management and construction of explanations, as well as higher levels of independence and improvements in self-awareness. The authors also identified effects on the family environment such as modification of affective bonding with parents, more consistent family dynamics and increased moments of wellbeing. The study concludes that children perform an active role in the construction of their meaning and parental separation can facilitate the generation of self-growth experiences.

Resumo (analítico) Neste estudo qualitativo, são descritos e analisados os efeitos da separação parental através da visão e da voz de uma menina de nove anos. Utilizou-se como instrumento uma entrevista semiestruturada e uma análise dos resultados categóricos. As repercussões observadas foram classificadas em efeitos individuais, como gerenciamento de informações e construção de explicações, além de maior independência e melhoria de seu autoconceito; Os efeitos no ambiente familiar, como mudanças nos laços emocionais com os pais, dinâmicas familiares mais consistentes e mais momentos de bem-estar, concluindo que as crianças são participantes ativas na construção de seus significados. E, por último, que a separação dos pais pode gerar experiências de crescimento.