bluuebunny/crossref_metadata_2025_split huggingface dataset:
B-299 Comparison of a Recently FDA-Cleared Urine Benzodiazepine Screening Assay with β-Glucuronidase Hydrolysis to Liquid Chromatography-Mass Spectrometry
Authors: Zakrajsek J T , Zakrajsek J A , Spiesman A M , McShane A J 
Published: October 2024

Abstract

Background
Benzodiazepines are a class of prescribed psychoactive drugs that act as a depressant on the central nervous system. Over the last decade, the fast-acting sedative effects of these drugs have led to misuse, therefore making detection important for monitoring proper drug compliance. Immunoassays on urine samples are commonly used as a screening method and the results are confirmed using high-performance liquid chromatography/tandem mass spectrometry (LC-MS/MS). The metabolites of benzodiazepine in urine are predominantly excreted as glucuronides which may lead to false negative results when utilizing immunoassay-based methods. The purpose of this study was to compare a new generation of benzodiazepine reagent, that includes a β-glucuronidase enzyme to increase sensitivity, to LC-MS/MS. Secondly, the positivity rate was compared before and after the increased sensitivity screening assay was implemented.

Methods
Validation of benzodiazepine II assay was performed on an automated chemistry analyzer (Roche Diagnostics, cobas c502), utilizing a kinetic interaction of microparticles in a solution (KIMS) method. A total of thirty urine samples that screened positive on the benzodiazepine II assay and thirty samples that screened negative, were compared with LC-MS/MS. The LC-MS/MS panel included α- hydroxytriazolam, 7- aminoclonazepam, nordiazepam, temazepam, oxazepam, lorazepam, and α-hydroxyalprazolam. The lower limit of quantitation was 40 ng/mL for all compounds except α-hydroxyalprazolam which was 60 ng/mL. The cutoff concentration for the benzodiazepine II assay was 200 ng/mL. Three months of data was also collected before and after converting to the new generation of benzodiazepine assay.

Results

Conclusions
The addition of β-glucuronidase to the benzodiazepine II assay allowed for high sensitivity when detecting urine benzodiazepine metabolites. With the increased sensitivity, almost a doubling of positivity rate was observed. The overall high diagnostic accuracy of the test makes it a reliable screening method for urine benzodiazepine detection.
B-300 Fentanyl and Xylazine Positivity Patterns at a Northeast U.S. Children’s Hospital
Authors: Poventud-Fuentes I , Unsihuay Vila D M , Osterhoudt K , Patel K 
Published: October 2024

Abstract

Background
In the U.S., fentanyl mixed with xylazine has been designated as an emerging health threat. Information on whether xylazine is present with fentanyl poisoning could help better navigate the management of children exposed to fentanyl mixed with xylazine. The detection of xylazine can be a marker used to differentiate medical vs non-medical fentanyl exposure. We aimed to explore the trends in positivity rates of fentanyl and/or xylazine at an urban pediatric tertiary care hospital.

Methods
We conducted a retrospective analysis from October 2021 - January 2024 of fentanyl positive urine drug screen (UDS) results by immunoassay performed in-house, and the reflex confirmatory testing by liquid chromatography-tandem mass spectrometry (LC-MS/MS), performed by an external laboratory. When available, xylazine detection results by gas chromatography mass spectrometry (GC-MS) were also included in the analysis. A chart review of fentanyl positive UDS cases was conducted to determine if fentanyl was medically administered prior to the UDS. We evaluated the positivity rate of fentanyl, the co-detection of xylazine, and the positivity patterns based on age, sex, and collection department.

Results
Fentanyl positive tests encompassed 3.4% (227/6632) of all UDS performed at our institution and 11.0% (227/2058) of all positive UDS tests, with medical fentanyl administration prior to UDS documented in 76.2% (173/227) of cases. 217/227 samples had confirmatory testing with 96.8% (210/217) confirmed by LC-MS/MS. The median age of fentanyl positives was 7 years (41.9% ≤ 3 years old) and 57.3% were male. Fentanyl-only positive UDS were seen in 33.0% (75/227) of all fentanyl positives and 45.6% (108/227) were also positive for benzodiazepines. 68.7% (156/227) were collected in the intensive care units, and 19.8% (45/227) were from the emergency department. 46.7% (106/227) of fentanyl positive cases had concurrent GC-MS results available. Xylazine was detected in 13.2% (14/106) of these cases, with 13 cases reported as unintentional exposure on patients' chart, and 12 cases having a fentanyl-only positive UDS. The median age of xylazine positives was 2 years (71.4% ≤ 3 years old, age ranged from 19 days to 14 years) and 78.6% were male. 7/14 fentanyl mixed with xylazine cases were collected in the emergency department and 7/14 were collected in intensive care. 2/14 of the xylazine positive cases had medical fentanyl administration prior to UDS.

Conclusions
Detection and management of fentanyl poisoning, especially when mixed with xylazine, remains a challenge in children. Presence of xylazine is a distinct marker of exposure to non-medical fentanyl. Continued surveillance of these cases could shed light on patterns that could aid further the management of affected children and contribute to harm reduction efforts.
B-309 Evaluation of the Analytical Performance of Breast and Ovarian Cancer Antigen Assays on the Atellica CI Analyzer
Authors: Quintanilla M , Rybin A , Frenna J , Arrode-Bruses G , Leipold H 
Published: October 2024

Abstract

Background
The Atellica® CI Analyzer is an automated, high-throughput integrated chemistry and immunoassay analyzer utilizing both Atellica® CH and Atellica® IM Assays. This study evaluated the analytical performance of the Atellica IM breast cancer antigen CA 15-3™ (CA 15-3) and the ovarian cancer antigen CA 125II™ (CA 125II) Assays on the Atellica CI Analyzer.

Methods
Precision studies were performed according to CLSI EP05-A3 using native and contrived human serum samples as well as contrived horse serum samples for CA 15-3. One aliquot of each sample pool was tested in duplicate in two runs per day ≥2 hours apart on each analyzer for ≥20 days. Precision studies were evaluated with one reagent lot on two systems. Method comparison studies were performed using three reagent lots according to CLSI EP09c. Individual native and contrived human serum samples were analyzed using the Atellica IM CA 15-3 and CA 125II Assays on the Atellica IM Analyzer and the Atellica CI Analyzer. LoB, LoD, and LoQ were determined as described in CLSI EP17-A2.

Results
Representative precision and method comparison results for each assay are listed in the table. For the two assays tested, repeatability and within-lab %CVs were &lt;2.4% and &lt;6.9%, respectively. Slopes determined by the weighted Deming linear regression model were approximately 1. Detection capability for the Atellica IM CA 15-3 Assay on the Atellica CI Analyzer was estimated to be 1.0 U/mL, 2.0 U/mL and 3.0 U/mL for LoB, LoD, and LoQ, respectively. When using the Atellica IM CA 125II Assay, detection capability was estimated to be 2.0 U/mL, 3.0 U/mL and 3.0 U/mL for LoB, LoD, and LoQ, respectively.

Conclusions
Evaluation of the Atellica IM CA 15-3 and CA 125II Assays using the Atellica CI Analyzer demonstrated good precision and equivalent performance compared to the same assays on the Atellica IM Analyzer.
B-313 Assessment of β2-microglobulin concentration in pleural effusions
Authors: Bérgamo Vázquez S , Trapé Pujol J , Rives Jimenez J , González Fernández C , González García L 
Published: October 2024

Abstract

Background
β2-microglobulin (β2-m) is a biomarker used in serum for different types of pathologies. It has been described that lymphocyte activation processes us tuberculosis infections, AIDS virus or cytomegalovirus infections, can produce an increase in the concentration of this marker. As a tumor marker is useful in lymphomas diagnosis. In the bibliography has reported that processes involving renal failure lead to a significant increase in its concentration. In patients with renal failure and treated with hemodialysis, have been described medians twenty times the upper reference limit. The objective is to assess whether the concentration of (β2-m) in pleural fluid is affected by the same factors as in serum.

Methods
We analyzed 303 pleural effusions (PE) samples from undiagnosed patients with pleural effusion. β2-microglobulin levels were prospectively measured by electrochemiluminescence (ECLIA) assays (Cobas c702, Roche Diagnostics®). Glomerular filtration rate (GFR) was calculated by CKD-EPI equation. The diagnostic performance of β2-microglobulin was evaluated by receiver operating characteristic (ROC) curve analysis. Statistical analyses were performed using SPSS software.

Results
Pleural fluid of 303 patients (104 female, 204 male) were collected. The median age was 72 ± 10,6 years. For all samples β2-m median concentration was 5,85 mg/L (IQR:3.4 -6.45).When we compared all results according to kidney failure values; GFR &gt;60mL/min β2-m median concentration was 4,12mg/L and if GFR &lt;60 mL/min, β2-m was 8,62g/L significant difference have found (p&lt;0,001). Comparing results between malignant pleural effusions (MPE) (n=96) and benign pleural effusions (BPE) (n=207), MPE median concentration was 5.38 mg/L and 6.07mg/L significant difference (p&lt;0,001). Benign pathologies associated with pleural effusions, tuberculosis samples had the highest concentration of β2-m. Tuberculosis samples (n=6) had a normal distribution, obtaining a value [mean (SD)] of 7.28 mg/mL (2,13) mg/mL. For tuberculosis samples and patients without renal failure (n=5) (GFR&gt;60 mL/min), the area under the ROC curve was 0.98 (95% CI: 0.88-0.98) (p&lt;0.005). For β2-m cut-off point was 5.150 mg/L, a sensitivity of 100%, specificity of 16%, positive predictive value (PPV) of 14% and negative predictive value (NPV) of 100% were obtained. Patients with lymphomas present the highest concentration in the group of malignant pleural effusions, but further investigations could not be performed due to the small sample size.

Conclusions
Based on the results obtained from our present study, it can be concluded that β2-microglobulin marker in pleural effusions has significant association with glomerular filtration rate, as well as occurs in serum. Higher concentrations were found in patients with GFR&gt;60 mL/min and tuberculosis. Using 5,15 mg/L cut-off point, 100% of NPV was obtained.
B-316 Evaluation of an FDA-cleared Chromogranin A Assay on an Automated Analyzer
Authors: Boyert N , Schroeder G , Dee M , Colon-Franco J M 
Published: October 2024

Abstract

Background
Chromogranin A (CgA) is a hydrophilic and acidic protein, present in the chromaffin granules of neuroendocrine cells. It is the preferred tumor marker for the monitoring of neuroendocrine tumors (NETS), which are often found in the lungs, appendix, pancreas, and gastrointestinal tract. Previous CgA assays were performed by manual ELISA testing, and were not FDA-approved, leading to a longer, more rigorous validation process and longer run times. The introduction of an automated, FDA-cleared CgA assay allows the laboratory to process patient samples more quickly and reliably, shortening a multiple hour process to minutes. The objective of this study was to evaluate the performance of an automated CgA assay recently cleared by the FDA.

Methods
We validated the the B·R·A·H·M·S™ Chromogranin A II (CGAII) assay on the automated B·R·A·H·M·S KRYPTOR Compact PLUS (Thermo Fisher Scientific Inc.). The CGAII assay is an immunofluorescent assay, which uses Time-Resolved Amplified Cryptate Emission (TRACE) to quantitate CGA in human serum. Assay performance characteristics were evaluated including measuring range (MR) and maximum dilution, precision, method comparison, stability, reference interval, stability, and carryover.

Results
The MR for CGAII was established with triplicate analysis of serum at 8 levels, ranging from 25.8 - 2,700 ng/mL. Linearity was demonstrated with a slope of 0.907, intercept of 1.948 and observed error of 2.54 ng/mL. Recovery at each point ranged from 83.3-110.3%. Carryover (&lt;5.0 ng/mL) was not observed at a concentration approximately 2 times the upper limit of the MR. A 1:25 or 1:500 automatic dilution was determined to be acceptable (% bias &lt;16%). Intraday precision coefficient of variation (CV) ranged from 4.1-1.7% (SDs from 2.32-6.64 ng/mL) at concentrations of 56.7 and 393.0 ng/mL. The interday CVs ranged from 6.0-7.1% (SDs from 3.43 - 28.84 ng/mL) at concentrations of 57.5 and 403.8 ng/mL. Twenty samples with concentrations ranging from 56.4-2,907 ng/mL were compared to a reference laboratory utilizing a similar methodology. The correlation coefficient was 0.9934, with a slope of 1.014, and intercept of -32.95. An additional twenty samples with concentrations ranging from 67.0-2,446 ng/mL were compared to a CgA ELISA kit (CGA-ELISA-NG, CISBIO) to determine the bias between the two methodologies. The correlation coefficient was 0.9902, with a slope of 0.845, and intercept of -15.31, and a bias of -19.9%. The serum samples were stable for 3 days when stored at 2-8°C. Recovered CgA concentrations fell outside of the 95% confidence interval after day 3 at 2-8°C. The manufacturer provided reference interval of &lt;187 ng/mL was verified by testing 20 healthy adult samples, all of which fell within this range.

Conclusions
This assay has been validated and shown to be an accurate and precise method for monitoring CgA. Further, the method was validated on an automated analyzer to allow expeditious turnaround time in a core laboratory.
B-317 Novel prognostic marker TGFBI affects the migration and invasion function of ovarian cancer cells via integrin αvβ3-PI3K-Akt signaling pathway
Authors: Guo Y , Wang H , Xu Y 
Published: October 2024

Abstract

Background
Patients with ovarian cancer show remarkably different prognosis. Present prognostic models for ovarian cancer mainly focus on clinico-pathological parameters, so quantifiable prognostic markers at molecular level are urgently needed. Platelets contribute to ovarian cancer progression, but have not been considered as biomarkers likely due to their instability. Therefore, we aimed to screened out stable prognostic markers from platelet-treated ovarian cancer cells, and explored mechanisms by which they promote ovarian cancer metastasis.

Methods
Microarrays analysis was done with platelet-treated SKOV-3 ovarian cancer cells, and similar studies were searched in the Gene Expression Omnibus (GEO) database. The candidate genes were determined by analyzing differentially expressed genes (DEGs), protein-protein interaction (PPI) network, Cox proportional hazard models and Kaplan-Meier curves. The expression of TGFBI in the tissue microarray was assessed by immunehistochemical staining (IHC), and its prognostic efficiency was evaluated by univariate and multivariate analysis. The functions of TGFBI were predicted using data from TCGA, and validated by in vitro and in vivo experiments. The mechanism exploration was performed based on proteomic analysis, molecular docking and intervention study.

Results
TGFBI was significantly higher expressed in the platelet-treated ovarian cancer cells. Tissue microarray showed that increased expression of TGFBI led to significant decrease of overall survival (OS), progression-free survival (PFS), and TGFBI was proved to be an independent prognostic factor for ovarian cancer. Functionally, TGFBI affected the migration and invasion of ovarian cancer cells by regulation of epithelial mesenchymal transition (EMT) markers (CDH1 and CDH2) and extracellular matrix (ECM) degradation proteins (MMP-2). Mechanistically, TGFBI phosphorylated PI3K and Akt by combining integrin αvβ3.

Conclusions
We found out TGFBI as a novel prognostic indicator for ovarian cancer patients. TGFBI could promote metastasis in ovarian cancer by EMT induction and ECM remodeling, which might be associated with the activation of integrin αvβ3-PI3K-Akt signaling pathway.
B-319 Kappa and Lambda Free Light Chains Quantification in Serum Samples with 3 or 4 Monoclonal Components Detected by Serum Immunofixation
Authors: Loureiro G , Rocha L , Siciliano S , Benini M , Martins M , Souza G , Pelegrini A , Facchin M , de Sá V , Sumita N 
Published: October 2024

Abstract

Background
Serum free light chain (FLC) quantification is a diagnostic criterion for monoclonal gammopathy. Oligoclonal expansion is defined when two or more small bands are detected in the gamma region of serum protein electrophoresis (SPEP) or immunofixation (SIFE) within the context of infectious diseases, autoimmune diseases, or some lymphoproliferative processes. This study aims to investigate the frequency of FLC disturbances in samples with oligoclonal pattern of 3 or 4 monoclonal bands observable in SIFE.

Methods
We have analyzed 7553 consecutive samples during the period of January to December 2023 in which both determination of FLC and SIFE were ordered in a large central laboratory at São Paulo, Brazil. FLC quantification and SIFE analysis were performed with FreeLite assay and Optilite system (The Binding Site, UK) and SPIFE 3000 (Helena Laboratories, USA), respectively.

Results
In our analysis, 119 samples (50 males, 69 females, median age 69 years (range 37-90)), with 3 or 4 monoclonal bands were identified, constituting 1.57% of the overall study population. These samples were then categorized into two groups. The first group had normal FLC Ratio (range 0.26 to 1.65) and the second group had altered FLC Ratio (outside 0.26-1.65 range). Seventy-seven samples (65%) had altered FLC ratio, with kappa elevation being the most common alteration (51 samples). Within these samples, median FLC ratio was 2.99 (range 1.72-719.05). Lambda elevation was present in 26 samples, with median FLC ratio 0.04 (range 0.005-0.16).

Conclusions
Presence of 3 or more oligoclonal bands in SIFE analysis may be indicative of various pathological processes (benign and malignant) and also be observed in post-transplantation. In this study, we have observed that most of these samples present altered FLC ratio, which indicate monoclonal disturbances in the production of free light chains. Those disturbances may be associated with a lymphoproliferative disease (e.g. multiple myeloma) or may be transient. Clinicians should be aware of the need for follow-up of these cases.
B-321 Enhancing Early Detection of Prostate Cancer Using PSA-Guided Screening in High-Risk Men Under 50
Authors: Chen L , Meng Q 
Published: October 2024

Abstract

Background
Prostate Cancer remains a significant global public health concern. It is the most common cancer and the second leading cause of cancer death among men in the United States. Current guidelines, including those by the USPSTF, ACS and ASCO offer varying recommendations on PSA screening. Present clinical practice, primarily targeting men aged 50-69. Our study aimed to assess the feasibility and validity of using PSA for targeted screening in men under 50 who are at high risk, enhancing early detection of prostate cancer.

Methods
This retrospective analysis utilized data from our hospital's prostate cancer screening program collected from 2022 to 2023, including104 individuals under 50. Participants were categorized based on PSA levels 4-10 ng/mL (n=71) and over10 ng/mL (n=33). We analyzed the number of cases diagnosed with prostate cancer and the correlation with PSA levels, Gleason Scores (GS), family history, and ethnicity/race.

Results
In the104 participants, 88 of them were diagnosed with prostate cancer, highlighting an 84.6% detection rate. Among PSA levels 4-10 ng/mL group, 36 out of 43 White or Caucasian and 13 out of 13 Black or African American men were identified as GS 6-10. In the group PSA over 10 ng/mL, 18 White or Caucasian and 6 Black or African American individuals with more aggressive disease (GS 6-10). Importantly, a substantial fraction of these cases had a first-degree relative with prostate cancer, emphasizing the influence of family history on PSA screening decision.

Conclusions
Our findings suggest that PSA-guided screening in men under 50 at high risk can enhance the early detection of prostate cancer. The significant presence of intermediate-risk prostate cancer (GS 6 or 7) in populations at higher risk due to ethnicity/race and family history. Our study highlights the potential benefits of revisiting current screening guidelines to lower the age threshold for high-risk populations.
B-323 Measurement of PIVIKA II Blood Level in Patients with Liver Diseases by Immunoassays
Authors: TAMIMI W , AlBagaei N , Alhamad A , Altwaijry A , Alkhidhr A , Alhuwayshil S , Alasam H A , Almaiman S , Althubiti F M , AlAjlan A 
Published: October 2024

Abstract

Background
Liver diseases are the most common diseases globally and can be complicated to cirrhosis and cancer and lead to death if not managed. It is crucial to prevent patients with liver diseases to reach the level of complications in particular liver cancer or hepatocellular Carcinoma (HCC). There are many screening lab tests to monitor and evaluate patients with liver disease such as alpha fetoprotein (AFP), however, it lacks optimal sensitivity and specificity. Another promising screening test is Prothrombin Induced by Vitamin K deficiency or antagonist- II (PIVKA-II) to complements AFP in screening of HCC. The aim of this study was to evaluate two PIVKA-II immunoassays and compare it to AFP in blood samples of patients with liver diseases.

Methods
Blood samples were collected from 56 patients with liver diseases, chronic hepatitis B (n=14), chronic hepatitis C (n=18), liver cirrhosis (n=14), and HCC (n=10) at King Abdulaziz medical city, Riyadh, SA. Measurement of PIVKA-II levels were performed on all samples by Snibe and Abbott analyzers. Clinical data was collected from the HIS BestCare. AFP was performed on Abbott analyzer. Statistical analysis was carried by Excel sheet Microsoft and p value was set at 0.05.

Results
There was an excellent correlation (R2= 0.9327) between the PIVKA-II two immunoassays from Snibe and Abbott among patients with HCC (p=0.1384). The diagnostic sensitivity PIVKA-II for Abbott and Snibe were calculated to be 78% and 90% respectively among HCC patients at cutoff 32 mAU/mL with specificity of 16% and 44.4% respectively. This was corresponding to sensitivity of 90% and specificity of 67% on AFP at cutoff 10 ng/mL.

Conclusions
The two PIVKA-II two immunoassays were perfectly correlated among the samples of HCC patients. The sensitivity of both assays was high but with lower specificity. Snibe assay has superior sensitivity and specificity among patients with HCC.