A High-Throughput, High-Containment Human Primary Epithelial Airway Organ-on-Chip Platform for SARS-CoV-2 Therapeutic Screening.

COVID-19 emerged as a worldwide pandemic in early 2020, and while the rapid development of safe and efficacious vaccines stands as an extraordinary achievement, the identification of effective therapeutics has been less successful. This process has been limited in part by a lack of human-relevant preclinical models compatible with therapeutic screening on the native virus, which requires a high-containment environment. Here, we report SARS-CoV-2 infection and robust viral replication in PREDICT96-ALI, a high-throughput, human primary cell-based organ-on-chip platform. We evaluate unique infection kinetic profiles across lung tissue from three human donors by immunofluorescence, RT-qPCR, and plaque assays over a 6-day infection period. Enabled by the 96 devices/plate throughput of PREDICT96-ALI, we also investigate the efficacy of Remdesivir and MPro61 in a proof-of-concept antiviral study. Both compounds exhibit an antiviral effect against SARS-CoV-2 in the platform. This demonstration of SARS-CoV-2 infection and antiviral dosing in a high-throughput organ-on-chip platform presents a critical capability for disease modeling and therapeutic screening applications in a human physiology-relevant in vitro system.

Screening in serum-derived medium reveals differential response to compounds targeting metabolism.

A challenge for screening new anticancer drugs is that efficacy in cell culture models is not always predictive of efficacy in patients. One limitation of standard cell culture is a reliance on non-physiological nutrient levels, which can influence cell metabolism and drug sensitivity. A general assessment of how physiological nutrients affect cancer cell response to small molecule therapies is lacking. To address this, we developed a serum-derived culture medium that supports the proliferation of diverse cancer cell lines and is amenable to high-throughput screening. We screened several small molecule libraries and found that compounds targeting metabolic enzymes were differentially effective in standard compared to serum-derived medium. We exploited the differences in nutrient levels between each medium to understand why medium conditions affected the response of cells to some compounds, illustrating how this approach can be used to screen potential therapeutics and understand how their efficacy is modified by available nutrients.

Serum D-2-hydroxyglutarate and the ratio of D-2HG/L-2HG predict IDH mutation in acute myeloid leukemia.

This study investigates whether serum D-2HG (D-2-hydroxyglutarate) produced by the mutated isocitrate dehydrogenase (IDH) can predict IDH mutations in acute myeloid leukemia (AML) at diagnosis. D-2HG and L-2HG are measured by liquid chromatography-tandem mass spectrometry. D-2HG, total 2HG and the D/L ratio (D-2HG/L-2HG) are significantly higher in IDH mutated cases than in IDH wild cases. The optimal cutoff values to predict IDH mutations at 100% sensitivity (specificity 91%-94%) are >588 ng/mL for D-2HG and >2.33 for the D/L ratio. Our study indicates that elevated serum D-2HG and the D/L ratio may serve as noninvasive biomarkers of IDH mutation in AML.

Screening in serum-derived medium reveals differential response to compounds targeting metabolism.

A challenge for screening new candidate drugs to treat cancer is that efficacy in cell culture models is not always predictive of efficacy in patients. One limitation of standard cell culture is a reliance on non-physiological nutrient levels to propagate cells. Which nutrients are available can influence how cancer cells use metabolism to proliferate and impact sensitivity to some drugs, but a general assessment of how physiological nutrients affect cancer cell response to small molecule therapies is lacking. To enable screening of compounds to determine how the nutrient environment impacts drug efficacy, we developed a serum-derived culture medium that supports the proliferation of diverse cancer cell lines and is amenable to high-throughput screening. We used this system to screen several small molecule libraries and found that compounds targeting metabolic enzymes were enriched as having differential efficacy in standard compared to serum-derived medium. We exploited the differences in nutrient levels between each medium to understand why medium conditions affected the response of cells to some compounds, illustrating how this approach can be used to screen potential therapeutics and understand how their efficacy is modified by available nutrients.

Global Temporal Patterns of Age Group and Sex Distributions of COVID-19.

Since the beginning of 2020, COVID-19 has been the biggest public health crisis in the world. To help develop appropriate public health measures and deploy corresponding resources, many governments have been actively tracking COVID-19 in real time within their jurisdictions. However, one of the key unresolved issues is whether COVID-19 was distributed differently among different age groups and between the two sexes in the ongoing pandemic. The objectives of this study were to use publicly available data to investigate the relative distributions of COVID-19 cases, hospitalizations, and deaths among age groups and between the sexes throughout 2020; and to analyze temporal changes in the relative frequencies of COVID-19 for each age group and each sex. Fifteen countries reported age group and/or sex data of patients with COVID-19. Our analyses revealed that different age groups and sexes were distributed differently in COVID-19 cases, hospitalizations, and deaths. However, there were differences among countries in both their age group and sex distributions. Though there was no consistent temporal change across all countries for any age group or either sex in COVID-19 cases, hospitalizations, and deaths, several countries showed statistically significant patterns. We discuss the potential mechanisms for these observations, the limitations of this study, and the implications of our results on the management of this ongoing pandemic.

Readability and Quality of Online Patient Education Material on Websites of Breast Imaging Centers.

OBJECTIVES: The purpose of this study was to assess the readability and quality of patient information regarding mammography, tomosynthesis, and breast density on the websites of ACR-designated mammography Breast Imaging Centers of Excellence (BICOEs) in the United States. MATERIALS AND METHODS: In this institutional review board-exempt study, websites of mammography BICOEs were identified by state. Websites were assessed for presence of patient education material on screening mammography, tomosynthesis, and breast density. This material was then assessed for readability using five well-established readability metrics (Flesch-Kincaid Reading Ease Index, Flesch-Kincaid Grade Level, Gunning-Fog Score, Coleman-Liau Index, and Simple Measure of Gobbledygook Index) and for quality using the DISCERN instrument. RESULTS: We identified 1,482 BICOE centers, of which 1,451 (98%) had websites; 79% (1,156 of 1,451) of websites had information on screening mammography, 45% (657 of 1,451) on tomosynthesis, and 16% (228 of 1,451) on breast density. Readability analysis showed that the overall average grade level of patient information was 12.4. Average readability grade levels by tested indexes were Flesch-Kincaid Grade Level of 11.5, Gunning-Fog Score of 14.1, Coleman-Liau Index of 14.1, and Simple Measure of Gobbledygook Index of 10.9. The average Flesch-Kincaid Reading Ease score was 45, with 0 being the most difficult and 100 being the easiest text to read. The overall DISCERN score nationally was 2.61 of 5 (with 5 representing the highest quality). CONCLUSIONS: Although most BICOEs have patient education material on their websites regarding mammography and some have information regarding tomosynthesis and breast density, the average readability grade level of this material is more than double the nationally recommended readability grade level of 6 for health care information. Additionally, the overall quality of this material is relatively low per established quality metrics.

K-D Balance: An objective measure of balance in tandem and double leg stances.

BACKGROUND AND OBJECTIVE: Subjective grade-based scoring balance assessments tend to be lengthy and have demonstrated poor repeatability and reliability. This study examined the reliability of a mobile balance assessment tool and differences in balance measurements between individuals at risk for a balance deficit secondary to a diagnosed neurological or musculoskeletal condition and a control group of healthy individuals. METHODS: Objective balance testing was measured using K-D Balance on a compatible iPhone. Seventy-seven participants were enrolled (control group, n = 44; group at risk for balance deficits, n = 33). Mean and standard deviation of K-D Balance were recorded for each stance. Intra-rater reliability was calculated by repeating the trial. RESULTS: Overall balance scores were superior for the control group compared with the group at risk for balance deficits in double leg stance (mean (SD): 0.15 (0.12) versus 0.18 (0.13), p = 0.260), tandem stance right leg (mean (SD): 0.27 (0.17) versus 0.45 (0.49), p = 0.028), and tandem stance left leg (mean (SD): 0.26 (0.17) versus 0.35 (0.35), p = 0.136). Intra-rater reliability was good to excellent for K-D Balance double leg stance (intra-class correlation coefficient (ICC) = 0.80, 95% confidence interval (CI) 0.58-1.03), tandem stance right leg (ICC = 0.96, 95% CI 0.86-1.06) and tandem stance left leg (ICC = 0.98, 95% CI 0.95-1.0). CONCLUSIONS: K-D Balance revealed differences in balance performance between healthy individuals compared with individuals with neurological or musculoskeletal impairment. Objective balance measures may improve the accuracy and reliability of clinical balance assessment by detecting subtle differences in balance and aid in early detection of diseases that impair balance.

Lumbee Native American ancestry and the incidence of aggressive histologic subtypes of endometrial cancer.

OBJECTIVE: The Lumbee Indian tribe is the largest Native American tribe in North Carolina, with about 55,000 enrolled members who mostly reside in southeastern counties. We evaluated whether Lumbee heritage is associated with high-risk histologic subtypes of endometrial cancer. METHODS: We retrospectively analyzed the available records from IRB-approved endometrial cancer databases at two institutions of patients of Lumbee descent (year of diagnosis range 1980-2014). Each Lumbee case was matched by age, year of diagnosis, and BMI to two non-Lumbee controls. Chi-square test was used to compare categorical associations. Kaplan-Meier methods and log-rank test were used to display and compare disease-free survival (DFS) and overall survival (OS). Multivariate Cox proportional hazards regression was used to adjust for age and BMI while testing cohort as a predictor of DFS and OS. RESULTS: Among 108 subjects, 10/35 (29%) Lumbee and 19/72 (26%) non-Lumbee subjects had high-risk (serous/clear cell/carcinosarcoma) histologic types (p = 0.8). 12/35 (34%) Lumbee and 24/72 (33%) non-Lumbee subjects had grade 3 tumors (p = 0.9). 5/33 (15%) Lumbee and 13/72 (18%) non-Lumbee had advanced stage endometrial cancer at diagnosis (p = 0.7). Lumbee ancestry was not associated with worse survival outcomes. OS (p = 0.054) and DFS (p = 0.01) were both worse in Blacks compared to Lumbee and White subjects. CONCLUSION: In this retrospective cohort analysis, Lumbee Native American ancestry was not a significant independent predictor of rates of high-risk histological subtypes of endometrial cancer or poor survival outcomes.

Relationship between minimally invasive hysterectomy, pelvic cytology, and lymph vascular space invasion: a single institution study of 458 patients.

OBJECTIVE: The aim of this study is to determine whether a minimally invasive approach to hysterectomy is associated with an increased rate of lymph vascular space invasion (LVSI) and/or malignant pelvic peritoneal cytology in endometrial cancer. METHODS: We performed a single institution analysis of 458 women with endometrial cancer who underwent either total abdominal hysterectomy (TAH) or minimally invasive hysterectomy (MIH) with use of a disposable uterine manipulator. All patients had endometrial cancer diagnosed by endometrial biopsy at a single academic institution between 2002 and 2012. Exclusion criteria were pre-operative D&C and/or hysteroscopy, uterine perforation or morcellation, and conversion to laparotomy. Multivariate logistic regression models to determine if type of hysterectomy predicts either LVSI or presence of abnormal cytology were controlled for grade, stage, depth of invasion, tumor size, cervical and adnexal involvement. RESULTS: LVSI was identified in 39/214 (18%) MIH and 44/242 (18%) TAH (p=0.99). Pelvic washings were malignant in 14/203 (7%) MIH and 16/241 (7%) TAH (p=1.0). Washings were atypical or inconclusive in 16/203 (8%) MIH and 6/241 (2.5%) TAH (p=0.014). In multivariate analyses, type of hysterectomy was not a significant predictor of either LVSI (p=0.29) or presence of malignant washings (p=0.66), but was a predictor of atypical or inconclusive washings (p=0.03). CONCLUSION: Minimally invasive hysterectomy with use of a uterine manipulator for endometrial cancer is not associated with LVSI or malignant cytology. Algorithms that better determine the etiology and implications of inconclusive or atypical pelvic cytology are needed to inform the possible additional risk associated with a minimally invasive approach to endometrial cancer.

Acrolein-induced oxidative stress in NAD(P)H Oxidase Subunit gp91phox knock-out mice and its modulation of NFκB and CD36.

An essential component of NAD(P)H, gp91phox, maintains the functionality of the enzyme in producing oxygen radicals. NAD(P)H oxidase plays an important role in oxidative stress but its precise contribution in acrolein-induced toxicity was not explored. We examined the involvement of NAD(P)H oxidase and other oxidant system in acrolein toxicity using gp91phox knockout mice. Male gp91phox knockout (KO) mice (20-25 gm) or wild type (WT) controls were treated with acrolein (0.5 µg/kg; 1 week). Animals were sacrificed and the liver was used to determine biochemical parameters. Knockout mice generated low (1.43 ±.02 pg/µg protein) free radicals as evident in 8-Isoprostane compared with the WT mice (2.19 ± 0.1). Acrolein increased 8-Isoprostane in WT (P<.05) and KO (p<.05) mice. Xanthine Oxidase (XO) activity was higher (p<.05) in KO (0.56 ± 0.06 µ unit/µg protein) than WT mice. Acrolein increased XO in KO mice, but significantly increased it only in WT. Cycloxygenase (COX) activity was not different between WT and KO mice, although acroelin increased COX in WT. Knockout mice exhibited a significantly low (2.1 ± 0.2 µmol/mg protein) total antioxidant status (TAS) compared with the WT (3.5 ± 0.3). Acrolein reduced TAS in both WT and KO mice equally. Baseline NFκB was significantly higher in KO mice, although acrolein increased NFκB in WT but not in KO. CD36 was higher (p<.05) in KO mice than the WT and acrolein increased (p<.05) CD36 further in KO but not in WT mice. These data suggest that NAD(P)H oxidase contributes significantly in acrolein-induced oxidative stress. We also suggests that in the absence of NAD(P)H oxidase XO plays a definitive role together with reduced antioxidant ability to compound the toxic effects of acrolein. We propose that in absence of NAD(P)H oxidase a different signaling process may involve that utilizes CD36 besides NFκB.