bluuebunny/crossref_metadata_2025_split huggingface dataset:
Evaluation of Bio-Rad D100 for HbA1c testing of dried blood spot samples
Authors: Wei Ruhan , Kroner Grace , Reineks Edmunds 
Published: November 2022

Abstract
Hemoglobin A1c (HbA1c) testing is critical for diabetes diagnosis and monitoring. Although many studies have demonstrated the feasibility of measuring HbA1c from dried blood spots (DBS) using immunoassays, enzymatic assays, and certain high-performance liquid chromatography (HPLC) assays, there is limited evidence supporting the use of the Bio-Rad D100 HPLC method. We evaluated the feasibility of testing HbA1c from DBS using Bio-Rad D100 to support the assessment of glycemic control for the growing field of telehealth.
Deidentified residual EDTA whole blood samples were processed, and HbA1c was measured on the Bio-Rad D100 on day zero. Concurrently, DBS were prepared for each sample by spotting 50 µL of blood onto a Whatman® 903 filter paper. The samples were dried overnight and stored at room temperature for two days. On day 2, a DBS disc with a diameter of 3.2 mm was eluted with 4.5 mL of Bio-Rad diluent for 20 minutes. The supernatants of the DBS samples were then processed on the platform. HbA1c values from the DBS samples were compared to the EDTA whole blood results.
Compared to the EDTA samples, the HbA1c concentrations of the corresponding DBS samples were elevated, especially for samples with original results less than 7.0%. Furthermore, the chromatograms of the DBS samples were significantly altered; we observed baseline elevation and the appearance of unknown minor peaks. This issue was investigated. First, blank DBS samples were created by directly spotting filter paper with diluent or adding diluent into an empty EDTA tube and then spotting the filter paper with the mixture of diluent and EDTA. The blank DBS samples were tested, and the lack of peaks suggested that the interference was not from the diluent, EDTA, or the filter paper. DBS samples were also prepared with sodium heparin whole blood, finger-stick capillary blood, and multiple brands of filter papers and tested using a different HPLC method. However, the interference was still present in those samples, indicating the interference is most likely due to the time-dependent degradation of the samples themselves. Of note, the alternate HPLC method showed less chromatogram alteration. To prevent possible adduct formation, DBS samples were eluted in the presence of various concentrations of cysteine (Jeppsson et al. 1986). However, the pretreatment did not eliminate the chromatogram changes. Finally, a multiple regression model was created using the results of day two A1c, A0, and F peaks, and a correction factor was generated. The corrected HbA1c concentrations were well within +/-6% of the initial HbA1c concentrations. In conclusion, the Bio-Rad D100 HPLC method appears particularly susceptible to stability changes in DBS samples, and further analysis is needed before employing a correction factor.
Incidence of Heparin Induced Thrombocytopenia (HIT) in patients with severe COVID-19
Authors: Barouqa Mohammad , Arackal Anita , Reyes-Gil Morayma 
Published: November 2022

Abstract

Introduction
To prevent and treat thrombotic complications in patients hospitalized with severe COVID-19 infection, anticoagulation treatments primarily with heparin and low molecular weight heparin have been recommended. Heparin-induced thrombocytopenia (HIT) is a rare but conceivably fatal reaction to heparin that is characterized by a sudden drop in platelet count accompanied by new onset of thrombosis 4-10 days after heparin exposure. The purpose of this retrospective study was to investigate the prevalence of thrombocytopenia and HIT in hospitalized COVID-19 patients, as well as their association with mortality.

Methods
3,672 plasma samples were collected from patients admitted to the first wave of COVID-19 in our institution at New York City (March to May 2020). All patients admitted with a platelet count of less than 150 k/ul were assigned to the thrombocytopenic group. In addition, two groups with similar demographics and normal platelet counts were randomly selected based on discharge outcome: alive vs. deceased (n= 88 per group). PF4 IgG Elisa and heparin neutralization were carried out in accordance with the manufacturer's instructions. A positive HIT result required an optical density (OD) greater than 0.4 and heparin neutralization greater than 50%. Statistical analysis was done in R studio (V.1.4.1717) to analyze demographics (age, gender, ethnicity), initial laboratory data, anticoagulation on admission, and thrombosis.

Results
Only 86 of the 3,672 (2.3%) patients admitted had thrombocytopenia. Only 1 of the 86 patients tested positive for HIT (1.1% ). 4 cases of the non-survivors (4.5%) tested positive for HIT compared to none of the survivors in the two groups with normal platelet counts. One of these 4 cases had a history of thrombosis (DVT). Interestingly, the PF4 Elisa ODs in non-survivors were significantly higher than in survivors (0.09 vs. 0.06, p-value&lt; 0.001). Although the platelet count did not differ significantly between the two groups, the mean platelet volume (MPV) on admission and its maximum peak during hospitalization were significantly higher in non-survivors than in survivors.

Conclusions
We only found HIT positive cases among non-survivors, implying that HIT is associated with COVID severity. The incidence of HIT in severe COVID-19 patients appears to be higher than the pre-COVID-19 historical rates of HIT in hospitalized patients (&lt;1%). Although thrombocytopenia is relatively uncommon in COVID-19 patients, the MPV was significantly higher in non-survivors, suggesting that platelet activation and destruction may explain the higher rate of HIT in COVID-19.
On-Cell Stability for Therapeutic Drug Monitoring
Authors: Nelson Heather , Pearson Lauren , Rudolf Joseph , Doyle Kelly 
Published: November 2022

Abstract

Introduction
A common problem in many clinical laboratories and outpatient clinics is processing specimens rapidly to comply with specimen stability criteria. Labs receive thousands of samples a day and may face challenges with staffing, equipment malfunction, or workflow, which can prevent immediate processing of samples. Currently, for therapeutic drug monitoring, the lab is required to centrifuge specimens and remove the serum/plasma from cells within 2-hours (CLSI-GP44-A4). Many of these drugs are monitored in the outpatient setting, which may require longer transportation times to get specimens from the collection location to the laboratory. This creates challenges for the lab in processing samples within 2-hours and may result in unnecessary rejection of otherwise acceptable specimens. Not all clinical sites have the staffing or space to process the specimens on site. Our objective was to determine if the on-cell stability of therapeutic drugs can be extended to beyond two hours.

Methods
We performed spiking studies to determine the on-cell stability of phenytoin in whole blood that was collected from healthy donors in barrier-free red top tubes. Phenytoin was spiked into the whole blood at a final concentration of about 10 (therapeutic) or 40 (toxic) µg/mL. The spiked whole blood samples were mixed by inverting six times and then were left at ambient temperature for 30 minutes, 2-, 4-, 6-, or 12-hours, after which they were centrifuged, and the serum was removed for analysis of total phenytoin concentration on the Abbott Architect and free and total phenytoin on the Beckman AU (n=3 per time point and concentration). In vivo studies are ongoing for an extended list of therapeutic drugs, in which two extra red top tubes are collected from patients being monitored for phenytoin, valproic acid, digoxin, vancomycin, lithium, or carbamazepine therapy. The two study tubes are kept at ambient temperature for 6- or 12-hours after collection before being centrifuged and separated from cells. The 6- and 12-hour drug concentrations were compared to the patient’s standard of care tube that was processed within 2-hours, after clotting for 30 minutes.

Results
Spiking studies demonstrated phenytoin was stable on cells for at least 12-hours at both 10 and 40 µg/mL. The percent total phenytoin remaining at 12-hours relative to the 30-minute control was 104 ± 2% at 10 µg/mL and 99 ± 16% at 40 µg/mL. Furthermore, the fraction of free phenytoin did not change over the 12 hours (10 µg/mL: 8 ± 1% free; 40 µg/mL: 10 ± 1% free). Our preliminary in vivo studies have shown no change in vancomycin or lithium concentration in samples processed 6- and 12-hours post-collection.

Conclusion
We have demonstrated that phenytoin, vancomycin, and lithium collected in red-top tubes are stable on cells at ambient temperature for at least 12-hours.
Levels and in-unit stability of 11-nor-9-carboxy-9-tetrahydrocannabinol (9-carboxy-THC) in donor platelets and whole blood: is enough present to have an impact on recipients?
Authors: Potterveld Susan , Annen Kyle , DomBourian Melkon , Pasko Bryce , Helander Louise 
Published: November 2022

Abstract

Objective
The safety of the United States donor blood supply has been questioned in the context of recreational marijuana legalization in Colorado and other states nationwide. It is unclear whether cannabis use by donors presents a danger to recipients, either through direct toxicity or socioeconomic ramifications. Our objectives are to both determine if the active metabolite of tetrahydrocannabinol (THC), 11-nor-9-carboxy-9-tetrahydrocannabinol (9-carboxy-THC), is present in donated units in a dosage high enough to impact the recipient and to assess for in-unit metabolism or degradation.

Methods
We will quantify the amount of 9-carboxy-THC in platelet units donated by self-reported cannabis users through liquid chromatography with tandem mass spectrometry (LC-MS/MS) testing and correlate the assay data with donor survey responses. To gain insight into in-unit metabolite stability, we also will test whole blood units at several time points throughout storage, collected from participants who self-report taking a known oral dose of THC prior to donation.

Results
Institutional review board approval for a study evaluating marijuana at a children’s hospital presented a challenge, requiring the addition of an organizational risk review and an update to the hospital transfusion consent for approval. Thus, the projected timeline of this study has been extended. Study recruitment flyers and an email screening survey have generated interest from volunteer donors. The email survey was sent to 9,208 donors who had previously donated at least once, from which we received 372 responses (4.04% response rate). Of the respondents, 49 individuals reported daily marijuana use, 33 reported marijuana usage 4-6 times per week, 32 reported usage 1-3 times/week, 30 reported usage 1-3 times/month, 52 reported rare usage, and 167 reported never using marijuana. Overall, out of the survey respondents, over 50% reported at least some marijuana usage, and 31% reported at least weekly marijuana usage. Although this number is small, our prior study found that many were repeat donors. Research visits are currently being scheduled for interested participants.

Conclusions
This study highlights the challenges of performing marijuana research at a children’s hospital and illustrates why there is a lack of published data evaluating the effects of recreational marijuana legalization. Although completion of this project investigating this controversial topic has been delayed, persistent advocation for this area of interest is opening doors for future studies and will hopefully pave the way to fill our knowledge gaps. The results of this research study will inform decisions regarding donor eligibility and safety of the national blood supply as well as address associated socioeconomic implications.
Utilization of system-wide urine drug testing to safely confront opioid misuse.
Authors: Bava Ejas Palathingal , Arora Kanika , Totten Amanda , Copeland Jacqueline , Crow Joseph , Shah Vinay , Frank Cathrine , Sibai Nabil , Winston-Mcpherson Gabrielle 
Published: November 2022

Abstract
Abuse and addiction of opioids leading to overdose deaths has increased significantly in Michigan since 2014. Moreover, a recent study showed that a majority (92%) of clinicians at Henry Ford Health System (HFHS) had reported prescribing narcotics in a way that was out of their comfort zone. To tackle this menace, we needed to equip our physicians with the appropriate tools to monitor patient compliance. In 2017, our laboratory implemented a reflexive urine drug testing (UDT) strategy called the directed pain panel (DPP). This panel included testing for natural/semi-synthetic /synthetic opiates, benzodiazepines, and cocaine metabolites. In 2019, the DPP was enhanced by adding buprenorphine, fentanyl, and tramadol. It is beneficial to order the DPP as opposed to UDSC (Urine drug screen) because UDSC does not detect the synthetic narcotics methadone, fentanyl, tramadol, and buprenorphine. DPP also has better sensitivity for detection of oxycodone. The primary aim of the current study was to evaluate UDT orders for chronic pain patients and to focus our efforts on improving the lab testing available to our providers to manage patients safely and effectively. Therefore, we evaluated the utilization of our testing strategy by doing a survey among chronic pain physicians in addition to retrospectively analyzing total orders for DPP (chronic pain panel, directed, urine) and UDSC (drug screen, urine) for chronic pain patients at five locations in Michigan over a one-year period (8/2020-8/2021). We collaborated with key providers and accessed their test ordering history to assess compliance with ordering the preferred UDT panel. The DPP represented 68% of all UDTs ordered for this population, however there were still a significant number of UDSC orders (32%). When order location was evaluated, the majority of UDSC orders originated from two locations (80%). When provider ordering patterns were evaluated, most providers (65%) ordered the DPP &gt; 50% of the time. In addition, 70% of all UDSC orders came from just two providers. This data showed that HFHS pain management clinicians favored ordering the DPP, however a small minority of ordering providers may benefit from targeted education regarding the advantages of the DPP. Hence, we educated the physicians who were ordering UDSC. They were willing to modify their practice to order DPP, instead of UDSC. This may pave the way for better patient compliance as well as reduced overdose and abuse of opioids.
Development of a generalizable UMAP-based approach for comparing clinical flow cytometry data with application to NPM1-mutated AML cohorts
Authors: van den Akker Tayler , Patel Sanjay , Simonson Paul 
Published: November 2022

Abstract

Introduction
AML with mutated NPM1 is associated with heterogeneous clinicopathologic features. We sought to study the association between phenotype, genetics, and clinical behavior using treatment-naïve bone marrow samples of NPM1-mutated AML. Prior phenotypic studies using flow cytometry data have primarily focused on the blast population in isolation or used less comprehensive analysis techniques, such as simple visual histogram assessment. We applied a dimensionality-reduction algorithm (UMAP) to analyze retrospective clinical flow cytometry data of tumor and non-tumor cells in an initial small cohort of NPM1-mutated samples.

Methods
Custom software was developed using python, FlowKit, and umap-learn to create a dictionary of the various antibody panels, detect the antibody panels that were used in each raw data (FCS) file, and determine the flow cytometer channels that should be disregarded. Subsamples from each FCS file for a given antibody panel were combined and analyzed using UMAP to create an embedding that could then be applied to all FCS files of the given antibody panel. FCS files were subsequently prepared for analysis in FlowJo, including using UMAP coordinates. The initial pilot phase included analysis of the EuroFlow AML1 panel of 11 cases, which included 3 primary refractory cases, 3 early relapsed cases, and 5 cases which achieved clinical remission without relapse. FlowJo was used to gate and examine the clusters identified by UMAP with respect to phenotypic parameters. These same UMAP gates were applied to all 11 cases for direct comparison.

Results
The blast count ranged from 50 to 88 in these cases. The blast phenotype was determined to be myeloid (n=3), monocytic (n=4), or other (CD34-/HLA-DR-)(n=4). Although standard CD45 by side scatter gating delineates four major cell types (lymphocytes, monocytes, granulocytes, blasts), gating using the UMAP algorithm with input data from eight phenotypic markers in conjunction with scattering parameters, produced at least 10 distinct UMAP clusters of variable cellular composition. Interestingly, applying those gates to side scatter (SSC-A) by CD45 histoplots revealed 5 distinct gates falling into the traditional “blast” region of the histoplot. The UMAP gates thus identified provided quantitative values for further statistical analysis.

Conclusion
We have developed a useful tool to automatically identify the antibody panels used to generate prior flow cytometry data, preprocess the data, and apply the UMAP algorithm for creating embeddings that can be applied to additional cases. Our preliminary analyses revealed significant phenotypic heterogeneity among a small cohort of NPM1-mutated cases. Ongoing work includes expansion of the cohort and number of antibody panels incorporated into the analyses to elucidate prognostic and predictive features of tumor and non-tumor populations in treatment naïve samples of NPM1-mutated AML.
A retrospective study of the eGFR differences between the 2021 CKD-EPI Equation and the MDRD equation in Black and White Patients
Authors: Chang Jeffrey , Redden David , Lima Jose , Reddy Vishnu , Cao Liyun 
Published: November 2022

Abstract

Objectives
In implementing the 2021 CKD-EPI without race equation in our institution, we sought to investigate the differences between the new CKD-EPI equation and the current MDRD equation to assist clinicians in transitioning to the new equation.

Methods
A retrospective study was performed in patients with eGFR&lt;60 ml/min/1.73 m2 calculated by MDRD. Stata was used to analyze the eGFR calculated by the two equations and measure the differences according to respective categories.

Results
eGFRs calculated by the 2021 CKD-EPI without race equation were well correlated to eGFRs calculated by the MDRD. However, notable differences were observed. Compared to the MDRD, the CKD-EPI w/o race equation for Black males with a serum creatinine less than 1.5 yielded decreased eGFR results with mean negative differences of -4.49 (18-60 years old), -6.43 (60-80 years old), and -8.87 (&gt;80 years old). For serum creatinine cohorts between 1.5 to 2.0 and greater than 2.0, the differences were -3.35, -5.46, -7.48 and -1.86, -3.40, -4.14, respectively. A similar trend was noted in Black females. For those with a serum creatinine less than 1.5, the CKD-EPI equation yielded mean negative differences that increased with advancing age: -2.56, -4.74 and -7.27. For serum creatinine between 1.5 to 2.0 and greater than 2.0, the negative differences were -2.31, -3.73 and -5.28 and -1.30, -2.06 and -2.70, respectively. Therefore, in Black patients, the mean eGFR difference between the two equations increased as age increased and serum creatinine level decreased. In contrast, the CKD-EPI equation for White males with a serum creatinine less than 1.5 yielded increased eGFR results with mean positive difference of 7.22 (18-60 years old), 4.09 (60-80 years old), and 1.26 (&gt;80 years old). For serum creatinine cohorts between 1.5 to 2.0 and greater than 2.0, the differences were 5.15, 2.54 and 0.18 and 2.01, 0.69 and -0.15, respectively. Similarly, White females had mean positive differences of 8.09 (18-60 years old), 4.61 (60-80 years old) and 2.00 (&gt;80 years old). For cohorts with serum creatinine between 1.5 to 2.0 and greater than 2.0, the positive differences were 4.21, 2.24, 0.51 and 2.02, 0.82, 0.10. Therefore, in White patients, the difference of eGFR between the two equations increased as age decreased and creatinine level decreased.

Conclusions
Although the 2021 CKD-EPI equation strongly correlated with the MDRD, distinct differences exist. In our study, Black males and females exhibited a negative difference of around -5.0 and -4.0, respectively. The difference in eGFR was greater in those of older age and serum creatinine less than 1.5. Meanwhile, White males and females exhibited a positive difference of around 2.5 and 2.7. The difference in eGFR was greater in those of younger age and serum creatinine less than 1.5.
Synovial Sarcoma Isn’t Supposed to To This!
Authors: Scholl A , Chen J , Cardona D 
Published: November 2022

Abstract

Introduction/Objective
NA.

Methods/Case Report
A 35 year old male construction worker with no significant medical history presented with left posterior knee pain, weakness and numbness. Onset of symptoms coincided with an acute gout attack three months prior. Imaging revealed a 6.3 x 6.4 x 6.2 cm intraarticular, multicystic mass arising from the anterior cruciate ligament and extending posteriorly to displace the popliteal artery and vein. It was heterogenous, partially enhancing and aggressive appearing with associated bone erosion of the femoral condyle. Needle core biopsy showed a biphasic synovial sarcoma (SS), confirmed by SS18 gene rearrangement by fluorescence in situ hybridization. The patient underwent an above the knee amputation. Histologic examination of the resected tumor revealed a heterogenous histology. There were areas characteristic of a biphasic SS, hyperchromatic spindle cells, intermixed epithelial clusters and calcifications, and solid areas of round cells with necrosis and staghorn vessels, consistent with poorly differentiated SS. Additionally, a third histologic appearance with large pleomorphic tumor cells, markedly atypical mitotic figures and what appears to be matrix production was noted. These latter findings are not well described in SS but is suggestive of dedifferentiation. Next generation sequencing was performed, focused on the dedifferentiated appearing area, which confirmed the presence of SS18-SSX1 fusion. Additional findings included PDGFRA amplification and p53 mutation. We present this interesting case in order to demonstrate a previously undescribed variant of synovial sarcoma which could be a potential pitfall, especially had this been the area initially sampled in the biopsy.

Results (if a Case Study enter NA)
NA.

Conclusion
NA.