Novel Automation of an Enzyme-Linked Immunosorbent Spot Assay Testing Method: Comparable Diagnostic Performance of the T-SPOT.TB Test Using Manual Density Gradient Cell Isolation versus Automated Positive Selection with the T-Cell Select Kit

The diagnosis of latent tuberculosis (TB) infection (LTBI) is critical to improve TB treatment and control, and the T-SPOT.TB test is a commercial enzyme-linked immunosorbent spot assay used for this purpose. The objective of the study was to increase automation and extend the time between blood collection and processing for the T-SPOT.TB test from 0 to 8 h to 0 to 54 h. The previous maximum time between blood collection and processing for the T-SPOT.TB test is 32 h using T-Cell Xtend. For this, we compared the T-SPOT.TB test using manual peripheral blood mononuclear cell (PBMC) isolation by density gradient separation at 0 to 8 h (reference method, control arm) to an automated PBMC isolation method using magnetic beads (T-Cell Select kit) at 0 to 55 h postcollection. A total of 620 subjects were enrolled from 4 study sites, and blood samples were collected from each volunteer, comprising 1,850 paired samples in total. Overall agreement between both methods was 96.8% (confidence interval [CI], 95.9 to 97.6%), with 95.8% (CI, 93.5 to 97.5%) positive and 97.1% negative agreement (CI, 96.1 to 97.9%). In summary, there was a strong overall agreement between the automated and manual T-SPOT.TB test processing methods. The results suggest that the T-SPOT.TB test can be processed using automated positive selection with magnetic beads using T-Cell Select to decrease hands-on time. Also, this cell isolation method allowed for the time between blood collection and processing to range from 0 to 55 h. Additional studies in larger and diverse patient populations including immunocompromised and pediatric patients are needed

Supernova 2017eaw: Molecule and Dust Formation from Infrared Observations

We present infrared (IR) photometry and spectroscopy of the Type II-P SN 2017eaw and its progenitor in the nearby galaxy NGC 6946. Progenitor observations in the Ks band in four epochs from 1 yr to 1 day before the explosion reveal no significant variability in the progenitor star greater than 6% that lasts longer than 200 days. SN 2017eaw is a typical SN II-P with near-IR and mid-IR photometric evolution similar to those of SNe 2002hh and 2004et, other normal SNe II-P in the same galaxy. Spectroscopic monitoring during the plateau phase reveals a possible high-velocity He I 1.083 μm absorption line, indicative of a shock interaction with the circumstellar medium. Spectra between 389 and 480 days postexplosion reveal a strong CO first overtone emission at 389 days, with a line profile matching that of SN 1987A from the same epoch, indicating ~10^(−3) M⊙ of CO at 1800 K. From the 389 days epoch until the most recent observation at 566 days, the first overtone feature fades while the 4.5 μm excess, likely from the CO fundamental band, remains. This behavior indicates that the CO has not been destroyed, but that the gas has cooled enough that the levels responsible for first overtone emissions are no longer populated. Finally, the evolution of Spitzer 3.6 μm photometry shows evidence for dust formation in SN 2017eaw, with a dust mass of 10^(−6) or 10^(−4) M⊙ assuming carbonaceous or silicate grains, respectively

Supernova 2017eaw: Molecule and Dust Formation from Infrared Observations

We present infrared (IR) photometry and spectroscopy of the Type II-P SN 2017eaw and its progenitor in the nearby galaxy NGC 6946. Progenitor observations in the Ks band in four epochs from 1 yr to 1 day before the explosion reveal no significant variability in the progenitor star greater than 6% that lasts longer than 200 days. SN 2017eaw is a typical SN II-P with near-IR and mid-IR photometric evolution similar to those of SNe 2002hh and 2004et, other normal SNe II-P in the same galaxy. Spectroscopic monitoring during the plateau phase reveals a possible high-velocity He I 1.083 μm absorption line, indicative of a shock interaction with the circumstellar medium. Spectra between 389 and 480 days postexplosion reveal a strong CO first overtone emission at 389 days, with a line profile matching that of SN 1987A from the same epoch, indicating ~10^(−3) M⊙ of CO at 1800 K. From the 389 days epoch until the most recent observation at 566 days, the first overtone feature fades while the 4.5 μm excess, likely from the CO fundamental band, remains. This behavior indicates that the CO has not been destroyed, but that the gas has cooled enough that the levels responsible for first overtone emissions are no longer populated. Finally, the evolution of Spitzer 3.6 μm photometry shows evidence for dust formation in SN 2017eaw, with a dust mass of 10^(−6) or 10^(−4) M⊙ assuming carbonaceous or silicate grains, respectively

Bone marrow microenvironments that contribute to patient outcomes in newly diagnosed multiple myeloma: A cohort study of patients in the Total Therapy clinical trials.

BackgroundThe tumor microenvironment (TME) is increasingly appreciated as an important determinant of cancer outcome, including in multiple myeloma (MM). However, most myeloma microenvironment studies have been based on bone marrow (BM) aspirates, which often do not fully reflect the cellular content of BM tissue itself. To address this limitation in myeloma research, we systematically characterized the whole bone marrow (WBM) microenvironment during premalignant, baseline, on treatment, and post-treatment phases.Methods and findingsBetween 2004 and 2019, 998 BM samples were taken from 436 patients with newly diagnosed MM (NDMM) at the University of Arkansas for Medical Sciences in Little Rock, Arkansas, United States of America. These patients were 61% male and 39% female, 89% White, 8% Black, and 3% other/refused, with a mean age of 58 years. Using WBM and matched cluster of differentiation (CD)138-selected tumor gene expression to control for tumor burden, we identified a subgroup of patients with an adverse TME associated with 17 fewer months of progression-free survival (PFS) (95% confidence interval [CI] 5-29, 49-69 versus 70-82 months, χ2 p = 0.001) and 15 fewer months of overall survival (OS; 95% CI -1 to 31, 92-120 versus 113-129 months, χ2 p = 0.036). Using immunohistochemistry-validated computational tools that identify distinct cell types from bulk gene expression, we showed that the adverse outcome was correlated with elevated CD8+ T cell and reduced granulocytic cell proportions. This microenvironment develops during the progression of premalignant to malignant disease and becomes less prevalent after therapy, in which it is associated with improved outcomes. In patients with quantified International Staging System (ISS) stage and 70-gene Prognostic Risk Score (GEP-70) scores, taking the microenvironment into consideration would have identified an additional 40 out of 290 patients (14%, premutation p = 0.001) with significantly worse outcomes (PFS, 95% CI 6-36, 49-73 versus 74-90 months) who were not identified by existing clinical (ISS stage III) and tumor (GEP-70) criteria as high risk. The main limitations of this study are that it relies on computationally identified cell types and that patients were treated with thalidomide rather than current therapies.ConclusionsIn this study, we observe that granulocyte signatures in the MM TME contribute to a more accurate prognosis. This implies that future researchers and clinicians treating patients should quantify TME components, in particular monocytes and granulocytes, which are often ignored in microenvironment studies

Treatment of severe COVID-19 with convalescent plasma in Bronx, NYC

Convalescent plasma with severe acute respiratory disease coronavirus 2 (SARS-CoV-2) antibodies (CCP) may hold promise as a treatment for coronavirus disease 2019 (COVID-19). We compared the mortality and clinical outcome of patients with COVID-19 who received 200 mL of CCP with a spike protein IgG titer ≥ 1:2430 (median 1:47,385) within 72 hours of admission with propensity score–matched controls cared for at a medical center in the Bronx, between April 13 and May 4, 2020. Matching criteria for controls were age, sex, body mass index, race, ethnicity, comorbidities, week of admission, oxygen requirement, D-dimer, lymphocyte counts, corticosteroid use, and anticoagulation use. There was no difference in mortality or oxygenation between CCP recipients and controls at day 28. When stratified by age, compared with matched controls, CCP recipients less than 65 years had 4-fold lower risk of mortality and 4-fold lower risk of deterioration in oxygenation or mortality at day 28. For CCP recipients, pretransfusion spike protein IgG, IgM, and IgA titers were associated with mortality at day 28 in univariate analyses. No adverse effects of CCP were observed. Our results suggest CCP may be beneficial for hospitalized patients less than 65 years, but data from controlled trials are needed to validate this finding and establish the effect of aging on CCP efficacy