microRNAs (miRNAs) hold promise as biomarkers for a variety of disease processes and for determining cell differentiation. These short RNA species are robust, survive harsh treatment and storage conditions and may be extracted from blood and tissue. Pre-analytical variables are critical confounders in the analysis of miRNAs: we elucidate these and identify best practices for minimizing sample variation in blood and tissue specimens. Preanalytical variables addressed include patient-intrinsic variation, time and temperature from sample collection to storage or processing, processing methods, contamination by cells and blood components, RNA extraction method, normalization, and storage time/conditions. For circulating miRNAs, hemolysis and blood cell contamination significantly affect profiles; samples should be processed within 2 h of collection; ethylene diamine tetraacetic acid (EDTA) is preferred while heparin should be avoided; samples should be "double spun" or filtered; room temperature or 4 °C storage for up to 24 h is preferred; miRNAs are stable for at least 1 year at -20 °C or -80 °C. For tissuebased analysis, warm ischemic time should be < 1 h; cold ischemic time (4 °C) < 24 h; common fixative used for all specimens; formalin fix up to 72 h prior to processing; enrich for cells of interest; validate candidate biomarkers with in situ visualization. Most importantly, all specimen types should have standard and common workflows with careful documentation of relevant pre-analytical variables.

BackgroundIn an ongoing study of racial/ethnic disparities in breast cancer stage at diagnosis, we consented patients to allow us to review their mammogram images, in order to examine the potential role of mammogram image quality on this disparity.MethodsIn a population-based study of urban breast cancer patients, a single breast imaging specialist (EC) performed a blinded review of the index mammogram that prompted diagnostic follow-up, as well as recent prior mammograms performed approximately one or two years prior to the index mammogram. Seven indicators of image quality were assessed on a five-point Likert scale, where 4 and 5 represented good and excellent quality. These included 3 technologist-associated image quality (TAIQ) indicators (positioning, compression, sharpness), and 4 machine associated image quality (MAIQ) indicators (contrast, exposure, noise and artifacts). Results are based on 494 images examined for 268 patients, including 225 prior images.ResultsWhereas MAIQ was generally high, TAIQ was more variable. In multivariable models of sociodemographic predictors of TAIQ, less income was associated with lower TAIQ (p < 0.05). Among prior mammograms, lower TAIQ was subsequently associated with later stage at diagnosis, even after adjusting for multiple patient and practice factors (OR = 0.80, 95% CI: 0.65, 0.99).ConclusionsConsiderable gains could be made in terms of increasing image quality through better positioning, compression and sharpness, gains that could impact subsequent stage at diagnosis.

BACKGROUND Transgender women are female individuals who were recorded men at birth based on natal sex. Supporting a person's gender identity improves their psychological health, and gender-affirming hormones reduce gender dysphoria and benefit mental health. For transgender women, estrogen administration has clinically significant benefits. Previous reviews have reported conflicting literature on the thrombotic risk of estrogen therapy in transgender women and have highlighted the need for more high-quality research. CONTENT To help address the gap in understanding thrombotic risk in transgender women receiving estrogen therapy, we performed a systematic literature review and metaanalysis. Two evaluators independently assessed quality using the Ottawa Scale for Cohort Studies. The Poisson normal model was used to estimate the study-specific incidence rates and the pooled incidence rate. Heterogeneity was measured using Higgins I2 statistic. The overall estimate of the incidence rate was 2.3 per 1000 person-years (95% CI, 0.8–6.9). The heterogeneity was significant (I2 = 74%; P = 0.0039). SUMMARY Our study estimated the incidence rate of venous thromboembolism in transgender women prescribed estrogen to be 2.3 per 1000 person-years, but because of heterogeneity this estimate cannot be reliably applied to transgender women as a group. There are insufficient data in the literature to partition by subgroup for subgroup prohibiting the analysis to control for tobacco use, age, and obesity, which is a major limitation. Additional studies of current estrogen formulations, modes of administration, and combination therapies, as well as studies in the aging transgender population, are needed to confirm thrombotic risk and clarify optimal therapy regimens.

Background This study investigated the effect on mortality of transfusing ABO‐incompatible plasma from all sources during trauma resuscitation. Methods Demographic, transfusion, and survival data were retrospectively extracted on civilian trauma patients. Patients were divided by receipt of any quantity of ABO‐incompatible plasma from any blood product (incompatible group) or receipt of solely ABO‐compatible plasma (compatible group). The primary outcome was 30‐day mortality, while other outcomes included 6‐ and 24‐hour mortality. Mixed‐effects logistic regression was used to model the effect of various predictor variables, including receipt of incompatible plasma, on mortality outcomes. Results Nine hospitals contributed data on a total of 2618 trauma patients. There were 1282 patients in the incompatible group and 1336 patients in the compatible group. In both the unadjusted and adjusted models, the 6‐hour, 24‐hour, and 30‐day mortality rates were not significantly different between these groups. The patients in the incompatible group were then divided into high volume (>342 mL) and low volume (≤342 mL) incompatible plasma recipients. In the adjusted model, the high‐volume group had higher 24‐hour mortality when the Trauma Injury Severity Score survival prediction was >50%. Mortality at 6 hours and 30 days was not higher in this model. The low‐volume group did not have increased mortality at any of the time points in this adjusted model. Conclusion The transfusion of incompatible plasma in civilian trauma resuscitation does not lead to higher 30‐day mortality. The finding of higher mortality in a select group of recipients in the secondary analysis warrants further study.

ABO incompatibility (ABOi), the most common cause of hemolytic disease of the newborn (HDN), is nearly always mild and treatable with phototherapy. Reports of ABOi HDN requiring neonatal exchange transfusion are extremely rare since the inception of modern guidelines. Here, a case of ABOi HDN clearly met criteria for exchange transfusion. An O-positive African American mother delivered a B-positive neonate that quickly developed hyperbilirubinemia. The neonatal DAT was positive from anti-B and anti-A,B, and maternal IgG titer was 1024. Double volume exchange transfusion resulted in a favorable outcome. Given early discharge of newborns, further understanding of factors predicting severe disease is needed.

PURPOSE We examined whether quality of mammography interpretation as performed by the original reading radiologist varied by patient sociodemographic characteristics. METHODS For 149 patients residing in Chicago and diagnosed in 2005-2008, we obtained the original index mammogram that detected the breast cancer and at least one prior mammogram that did not detect the cancer performed within 2 years of the index mammogram. A single breast imaging specialist performed a blinded review of the prior mammogram. Potentially missed detection was defined as an actionable lesion seen during a blinded review of the prior mammogram that was in the same quadrant as the cancer on the index mammogram. RESULTS Of 149 prior mammograms originally read as non-malignant, 46% (N=68) had a potentially detectable lesion. In unadjusted analyses, potentially missed detection was greater among minority patients (54% vs. 39%, p=0.07), for patients with incomes below $30,000 (65% vs. 36%, p<0.01), with less education (58% vs. 39%, p=0.02), and lacking private health insurance (63% vs. 40%, p=0.02). Likelihood ratio tests for the inclusion of socioeconomic variables in multivariable logistic regression models were highly significant (p<=0.02). CONCLUSIONS Disadvantaged socioeconomic status appears to be associated with potentially missed detection of breast cancer at mammography screening.

One of the most important and persistent complications of blood transfusion is red blood cell (RBC) alloimmunization. When a patient is exposed to RBC antigens that differ from their own they can form alloantibodies to these foreign antigens. Blood group antigens are highly conserved and follow ancestral patterns of inheritance that may demonstrate population restriction. Minority populations who require chronic transfusion are at particularly high risk of alloimmunization when the blood donor population does not share the same ancestral background, resulting in exposure to non-self RBC antigens. It is incumbent on blood collectors to support patients with risk factors for alloimmunization as well as patients who have already formed alloantibodies. Increasing utilization of RBC genotyping may represent an opportunity to improve access to RBC units from donors that match the extended RBC phenotype of all possible patients.

Background Severe acute respiratory syndrome coronavirus 2 (SARS-CoV-2) is a novel member of the coronavirus family that caused the global coronavirus 2019 (COVID-19) pandemic. The prevalence remains largely unknown because of early testing supply shortages. Although it cannot currently be used to determine level of immunity, antibody testing can contribute to epidemiological studies, identify convalescent plasma donors, or satisfy curiosity about previous exposure to the virus. Methods 407 samples collected from hospitalized inpatients with and without a confirmed SARS-CoV-2 infection, 170 remnant clinical specimens collected and frozen prior to the COVID-19 outbreak, and paired serum and plasma samples from 23 convalescent plasma donors were used to determine performance characteristics of the Abbott SARS-CoV-2 IgG and Roche Elecsys Anti-SARS-CoV-2 assays. The sensitivity, specificity, imprecision, interferences, and sample stability were determined. These assays were then used to characterize the antibody response in serial samples from 20 SARS-CoV-2 positive inpatients. Results Both assays exhibited 100% specificity (95% CI; 99.05 – 100.00), giving no positive results in 170 specimens collected before July 2019 and 215 specimens from patients without a confirmed SARS-CoV-2 infection. Differences between platforms were most notable in SARS-CoV-2 positive samples. Roche offered higher sensitivity in convalescent plasma donors at 95.7% (95% CI; 78.1 – 99.9) versus 91.3% (95% CI; 72.0 – 98.9) but Abbott detected antibodies in two immunocompromised patients whereas Roche did not. The Roche and Abbott platforms also exhibited different trends in antibody signal for a subset of patients. Conclusions Both the Abbott and Roche platforms offer excellent specificity but different trends in antibody signal may reflect qualitative differences in the types of antibodies recognized by the two assays. Negative serologic results do not exclude previous SARS-CoV-2 infection.