The Association of Racism and Discrimination in Disparities of Hypertensive Disorders of Pregnancy in the United States: An Analysis of PRAMS Data.

BACKGROUND: Hypertensive disorders of pregnancy are a leading cause of maternal mortality. Racial disparities in maternal outcomes such as maternal mortality in the United States (US) are well-documented, but the relationship of racism and/or discrimination with one's risk of developing a hypertension during pregnancy (HDP) is not well-studied. METHODS: Data from 17 sites that asked questions regarding experiences with racism and/or discrimination during pregnancy via the Pregnancy Risk Assessment Monitoring System (PRAMS) Phase 8 (2016-2020) was used. Logistic regression models were used compare the potential of stress induced HDP from perceived experiences of racism and/or discrimination versus the effect of systemic racism and/or discrimination (in healthcare settings and generally) on racial disparities in HDP diagnosis. RESULTS: Among 9,907 live births, 18% of participants reported they were diagnosed with hypertension during pregnancy, with non-Hispanic Black individuals having the highest rate (21.8%). Regarding experiences of racism and/or discrimination, 76.4% of participants responded "yes", with all races/ethnicities studied here except non-Hispanic White individuals responding "yes" at rates higher than 89%. Perceived experiences of racism and/or discrimination did not statistically significantly affect one's odds of being diagnosed with HDP (OR = 0.94, CI: 0.74, 1.20). The disparity in odds of having hypertension during pregnancy between Non-Hispanic Black individuals and non-Hispanic White individuals was not statistically significant when perceived experiences of racism and/or discrimination were included in the model. CONCLUSIONS: Experiences of racism and/or discrimination drive racial disparities in hypertensive disorders in pregnancy. PUBLIC HEALTH IMPLICATIONS: It is vital to eliminate racist and discriminatory practices and behaviors to reduce maternal morbidity and mortality.

Electro-DBS: A Simple Method to Rapidly Extract Genomic DNA from Dried Blood Spots.

Dried blood spots (DBSs) have been used for more than 50 years and are used as a method of sample collection for pharmacological testing, genetic analyses, monitoring of viral infections, and more. Protocols for nucleic acid extraction from DBSs involve several steps that require specialized equipment and can be lengthy. Thus, we sought to explore ways to reduce the analytical burden of DBSs. We developed a DBS extraction method that uses the synergistic action of electrophoretic and diffusive transport mechanisms to extract genomic DNA (gDNA) through the DBS matrix. This method (which we termed "Electro-DBS") reduces the time of extraction from 40 min to 5 min and removes the need for heat, shaking, and DNA purification steps while maintaining gDNA quality and yield. We found that the electrophoretic transport speeds up gDNA elution, allowing for the diffusive transport of polymerase chain reaction (PCR) inhibitors to be minimized due to a reduced elution time. Overall, this work added mechanistic insight into the extraction of DNA from DBSs and developed a method that was ideal for point of care and automation.

Insight into Increased Recovery and Simplification of Genomic DNA Extraction Methods from Dried Blood Spots.

There is no consensus on how to perform the manual extraction of nucleic acids from dried blood spots (DBSs). Current methods typically involve agitation of the DBSs in a solution for varying amounts of time with or without heat, and then purification of the eluted nucleic acids with a purification protocol. We explored several characteristics of genomic DNA (gDNA) DBS extraction such as extraction efficiency, the role of red blood cells (RBCs) in extraction and critical kinetic factors to understand if these protocols can be simplified while maintaining sufficient gDNA recovery. We found that agitation in a RBC lysis buffer before performing a DBS gDNA extraction protocol increases yield 1.5 to 5-fold, depending upon the anticoagulant used. The use of an alkaline lysing agent along with either heat or agitation was sufficient to elute quantitative polymerase chain reaction (qPCR) amplifiable gDNA in 5 minutes. This work adds insight into the extraction of gDNA from DBSs with the intention of informing a simple, standardized manual protocol for extraction.

An investigation into simplifying total RNA extraction with minimal equipment using a low volume, electrokinetically driven microfluidic protocol.

Current methods for total RNA extraction are time-consuming and require several hands-on steps and specialized equipment. Microfluidic devices can offer the opportunity to reduce the number of hands-on steps, decrease the volumes of reagents required for purification, and make extraction high throughput. Here, we investigated the translation of a high volume magnetic bead-based total RNA extraction method (from human whole blood) onto a low input volume microfluidic device. Our results first show that RNA integrity is maintained when the reagent volumes are scaled down by a factor of 22 and the wash buffers are combined 1:1. With our microfluidic method, the number of wash steps can be reduced from four to one. Thus, the time to complete RNA extraction can be reduced from 2 h to 40 min. These manipulations to the conventional protocol yielded RNA amplifiable within 40 cycles of reverse transcription quantitative PCR (RT-qPCR) when using the microfluidic device to simplify the wash steps. To improve the purification of the RNA during the bead transport through the microchannel, we also investigated the effect of a synergetic application of the electrokinetic flow. Our results show that DNase I and other contaminants surrounding the beads get washed away more effectively via electrophoretic transport. Most notably, RNA adsorption on the beads is strong enough to counter electrophoretically-driven desorption. In all, our work opens new ways to extract high-quality total RNA rapidly and simply from a small quantity of blood, making the process of RNA extraction more accessible.

Pre-eclampsia: a Scoping Review of Risk Factors and Suggestions for Future Research Direction.

Abstract: Most of maternal deaths are preventable, and one-quarter of maternal deaths are due to pre-eclampsia and eclampsia. Prenatal screening is essential for detecting and managing pre-eclampsia. However, pre-eclampsia screening is solely based on maternal risk factors and has low (< 5% in the USA) detection rates. This review looks at pre-eclampsia from engineering, public health, and medical points of view. First, pre-eclampsia is defined clinically, and the biological basis of established risk factors is described. The multiple theories behind pre-eclampsia etiology should serve as the scientific basis behind established risk factors for pre-eclampsia; however, African American race does not have sufficient evidence as a risk factor. We then briefly describe predictive statistical models that have been created to improve screening detection rates, which use a combination of biophysical and biochemical biomarkers, as well as aspects of patient medical history as inputs. Lastly, technologies that aid in advancing pre-eclampsia screening worldwide are explored. The review concludes with suggestions for more robust pre-eclampsia research, which includes diversifying study sites, improving biomarker analytical tools, and for researchers to consider studying patients before they become pregnant to improve pre-eclampsia detection rates. Additionally, researchers must acknowledge the systemic racism involved in using race as a risk factor and include qualitative measures in study designs to capture the effects of racism on patients. Lay Summary: Pre-eclampsia is a pregnancy-specific hypertensive disorder that can affect almost every organ system and complicates 2-8% of pregnancies globally. Here, we focus on the biological basis of the risk factors that have been identified for the condition. African American race currently does not have sufficient evidence as a risk factor and has been poorly studied. Current clinical methods poorly predict a patient's likelihood of developing pre-eclampsia; thus, researchers have made statistical models that are briefly described in this review. Then, low-cost technologies that aid in advancing pre-eclampsia screening are discussed. The review ends with suggestions for research direction to improve pre-eclampsia screening in all settings.Overall, we suggest that the future of pre-eclampsia screening should aim to identify those at risk before they become pregnant. We also suggest that the clinical standard of assessing patient risk solely on patient characteristics needs to be reevaluated, that study locations of pre-eclampsia research need to be expanded beyond a few high-income countries, and that low-cost technologies should be developed to increase access to prenatal screening.

Parallel DNA Extraction From Whole Blood for Rapid Sample Generation in Genetic Epidemiological Studies.

Large-scale genetic epidemiological studies require high-quality analysis of samples such as blood or saliva from multiple patients, which is challenging at the point of care. To expand these studies' impact, minimal sample storage time and less complex extraction of a substantial quantity and good purity of DNA or RNA for downstream applications are necessary. Here, a simple microfluidics-based system that performs genomic DNA (gDNA) extraction from whole blood was developed. In this system, a mixture of blood lysate, paramagnetic beads, and binding buffer are first placed into the input well. Then, the gDNA-bound paramagnetic beads are pulled using a magnet through a central channel containing a wash buffer to the output well, which contains elution buffer. The gDNA is eluted at 55°C off the chip. The 40-minute microfluidic protocol extracts gDNA from six samples simultaneously and requires an input of 4 µL of diluted blood and a total reagent volume of 75 µL per reaction. Techniques including quantitative PCR (qPCR) and spectrofluorimetry were used to test the purity and quantity of gDNA eluted from the chip following extraction. Bead transport and molecular diffusional analysis showed that an input of less than 4 ng of gDNA (∼667 white blood cells) is optimal for on-chip extraction. There was no observable transport of inhibitors into the eluate that would greatly affect qPCR, and a sample was successfully prepared for next-generation sequencing (NGS). The microfluidics-based extraction of DNA from whole blood described here is paramount for future work in DNA-based point-of-care diagnostics and NGS library workflows.