Tumor Markers Research Articles

7426 Tamoxifen-Induced Ovarian Hyperstimulation in a Pre-Menopausal Patient with Breast Cancer History: Spontaneous Resolution with Conservative Management

Abstract Disclosure: S. Gondi: None. M. Contento: None. A. Leiter: None. Background: Tamoxifen is a selective estrogen receptor modulator (SERM) that competitively binds to estrogen receptors (ER) on tumors and inhibits the effect of estrogen, which is used in the treatment and prevention of ER-positive breast cancer. Tamoxifen therapy has been reported to increase serum estrogens and cause ovarian hyperstimulation in premenopausal women, but knowledge on how to manage these conditions is limited. Clinical Case: A 36-year-old pre-menopausal female with history of malignant neoplasm of the right breast (ER positive) who completed neoadjuvant chemotherapy, lumpectomy, and radiation, on tamoxifen for seven years presented with new-onset oligomenorrhea. Laboratory testing done by the patient’s gynecologist revealed total estrogen 2,723 pg/mL (reference range [RR] 34-501), testosterone 132 ng/dL (RR 8-60), anti-Mullerian hormone 12.7 ng/mL (RR for age 0.66-8.75), ovulation-phase follicle stimulating hormone 11.5 mIU/mL (RR 4.7-21.5) and luteinizing hormone (LH) 63.3 mIU/mL (RR 14-95.6). Further workup revealed that ovarian tumor markers alpha-fetoprotein and serum human chorionic gonadotropin and other labs including dehydroepiandrosterone sulfate, progesterone 17-OH, lactate dehydrogenase, prolactin, and thyroid stimulating hormone were normal. Transvaginal ultrasound (TVUS) showed a new 2.8 x 1.9 x 2.3 cm complex left ovarian cyst, which, along with the elevated estrogen level, was concerning for ovarian hyperstimulation. Treatment with LH-releasing hormone (LHRH) agonists, surgical consultation, and discontinuation of tamoxifen were offered as potential management strategies, but the patient opted for observation and continued tamoxifen. Two months later, estradiol was 989 pg/mL (RR 85-498), and then three months later decreased to 152 pg/mL with repeat TVUS showing resolution of the left ovarian cyst. The patient’s oligomenorrhea resolved with resumption of regular menstrual cycles. Conclusion: While it is well-known that tamoxifen causes elevated serum estrogen levels and can lead to ovarian hyperstimulation, there is limited research on the management of these side effects. Treatment strategies in past cases have included oophorectomy or medical management with LHRH agonists or gonadotropin-RH agonists. However, this case demonstrates that a tamoxifen-induced ovarian cyst and high estrogen levels can resolve on its own with continued tamoxifen treatment. Conservative management with monitoring via imaging and repeat estrogen levels may be sufficient in management of pre-menopausal women with tamoxifen-induced ovarian hyperstimulation. Presentation: 6/1/2024

Carbohydrate antigens Lewis a and Lewis b act as tumor markers cooperating with CA19.9 in the management of PDAC patients

BackgroundCA19.9 is the unique marker recommended for the preoperative staging and the follow-up of patients suffering from pancreatic ductal adenocarcinoma (PDAC) but up to 30% of PDAC patients maintain normal CA19.9 values and cannot be monitored in this way. Lewis a (Lea Galβ1,3[Fucα1,4]GlcNAc) and b (Leb, Fucα1,2Galβ1,3[Fucα1,4]GlcNAc) are antigens which are structurally similar to sialyl-Lewis a (Siaα2,3Galβ1,3[Fucα1,4]GlcNAc), the epitope of CA19.9. MethodsWe set an ELISA procedure determining the levels of Lea, Leb, and CA19.9 in the blood of healthy individuals or PDAC patients. Moreover, such antigens were also detected in cancer resections by immunofluorescence microscopy, and the levels of glycosyltransferase transcripts involved in Lewis antigen biosynthesis were determined by RT-qPCR. ResultsIn our cohort of 116 healthy individuals, the distribution of circulating Lea and Leb was similar to that of CA19.9, allowing us to set putative cutoff values for both antigens. In a cohort of 115 PDAC patients, the differential distribution with respect to the controls was statistically significant for both antigens (p < 0.001). Out of 37 patients presenting normal CA19.9 values, 15 patients presented Lea or Leb above the cutoffs. By immunofluorescence, Lea, Leb and CA19.9 were all detected in cancer resections and expression levels were heterogeneous among patients in terms of intensity, localization and diffusion. The levels of relevant glycosyltransferase transcripts were found to be heterogeneous between cancers of different patients and no association was detectable with the levels of any circulating antigen. ConclusionsThe concurrent quantification of Lea and Leb together with CA19.9 improves the management of PDAC patients.

B-320 Understanding CA 72-4: Exploring False Positives in Clinical Context

Abstract Background CA 72-4, initially identified in 1981 as an antigen reactive to murine antibodies, is a glycoprotein expressed on the cell surface of certain carcinoma cells. Its production is elevated in various cancers, including ovarian, pancreatic, colorectal, and gastric cancer. However, CA 72-4 serum concentration may rise in non-cancerous conditions, leading to potential misdiagnosis and a low positive predictive value (PPV), particularly in asymptomatic patients. Our aim in this study was to describe CA 72-4 false positive values and assess the pathological conditions and pharmacological treatments associated with this. We also intended to compare other tumour markers status in false and true positive results of CA 72-4. Methods A retrospective analysis form pseudo-anonymized database was conducted using data from 1290 laboratory analysis from different patients taking a blood test at our laboratory hospital from 2017 to 2019. CA 72-4 and other tumour markers were analysed by ECLIA (Roche Diagnostics Gmbh. Mannheim, Germany). Results Among 1290 CA 72-4 laboratory tests in different patients, 204 were considered positive because they were above the reference interval (&amp;lt;7 U/mL). Within this subgroup, 44 of the 204 were clearly classified as false positives, as patients were not diagnosed with any form of neoplastic process. 45% of cases were between 7-14 U/mL (group 1), 30% were between 14-28 U/mL (group 2), and finally 25% of patients were above 28 U/mL ranging from 35.1 to 240.4 U/mL (group 3). Interestingly, CA19-9, CA 125 and CEA were measured in 37 of the false positive cases and it was found that 59.5% had at least another tumour marker above the reference range. In 103 true positive cases it was found that 88% of patients had at least one tumour marker above the normal range. Patients exhibiting false positive results for CA 72-4 demonstrated a wide spectrum of pathological conditions, with pulmonary disease, inflammation, infection, and heart and kidney disease emerging as the most prevalent. Group 1 patients most common underlying pathologies were pulmonary disease in 25%, heart disease in 25% and either viral or bacterial infection in 60% of patients. Group 2 patients most common pathologies were heart disease in 42%, pulmonary disease in 33% and infection in 25% of patients. In group 3 patients, renal disease was present in 45% of patients, heart disease was found in 45% of patients and inflammation/sepsis was found in 36% of patients. When looking into pharmacological treatment, 86.3% of patients were taking at least one of previously described treatments increasing CA 72-4 in serum; proton pump inhibitors, corticosteroids, antiplatelet drugs and colchicine. Drug treatment proportion was very similar across the different groups. Conclusions CA72-4 false positive cases are sometimes in conjunction with other false positive tumour markers but not in the same proportion as true positives. In addition, most false positive patients can be explained by pharmacological treatments. This research may help in the understanding and identification of potential factors influencing tumor marker levels and thus contribute to the interpretation of results.

Nucleic Acid-Based Biological Nanopore Sensing Strategies for Tumor Marker Detection.

Cancer, which is characterized by high mortality rates, poses a significant threat to global human health. Early diagnosis is of paramount importance in managing cancer, and tumor markers have emerged as crucial indicators for achieving this goal. The advent of precision medicine has further emphasized the need for the effective detection of these markers. However, traditional detection methods are hampered by numerous limitations. In recent years, nanopore technology has emerged as a promising alternative, due to its unique physical and chemical properties, which facilitate rapid, label-free, and amplification-free detection. This Review focuses on the direct detection of tumor markers through nucleic acid analysis and indirect detection mediated by nucleic acids and facilitated by biological nanopores. Furthermore, it also discusses the challenges and prospects of applying biological nanopore sensing technology in early cancer diagnosis, underscoring its potential to revolutionize tumor marker detection.

B-141 Analysis of risk factors and construction of prediction model for pneumonia in patients with pneumoconiosis based on the clinical laboratory indexes

Abstract Background Pneumoconiosis has been the leading industrial disease in China. Pneumonia is one of the common comorbidities for patients with pneumoconiosis, which may lead to the worse condition and poor prognosis. Timely and accurate diagnosis is necessary to retard the pneumonia progression in pneumoconiosis patients. Due to the similarity of clinical features between pneumoconiosis and pneumonia, clinical laboratory tests are needed for assessing the hazard of pneumonia occurrence in patients with pneumoconiosis. Therefore, we aim to explore the risk factors of pneumonia from laboratory test indexes for patients with pneumoconiosis, construct prediction model, and evaluate its feasibility. Methods A total of 243 patients with pneumoconiosis from the General Hospital of Xuzhou Mining Group from September 2021 to August 2023 were selected as the modeling set, which was further separated as the non-pneumonia group and the pneumonia group. Nineteen laboratory test indexes related to inflammation, tumor markers, renal function, and blood routine examination were compared between the two groups. The univariate and multivariate Logistic analysis was used to screen out risk factors and construct the prediction model. Two hundred and twenty-eight patients with pneumoconiosis from General Hospital of Xuzhou Mining Group (Yi'an Branch) were enrolled as the validation set. The predictive efficiency of the model on pneumonia for patients with pneumoconiosis was evaluated through Hosmer-Lemeshow(H-L)test and receiver operating characteristic (ROC) curve. Results C-reactive protein (CRP), interleukin-6 (IL-6), interleukin-8 (IL-8), interleukin-10 (IL-10), β2 microglobulin, creatinine, urea, neutrophil percentage, lymphocyte percentage, and lymphocyte count in pneumonia group were significant different from those in non-pneumonia group (&amp;lt;0.05). Logistic regression analysis suggested that CRP (OR=3.883, &amp;lt;0.001), IL-6 (OR=1.217, =0.003), IL-8 (OR=1.215, =0.002) and β2-microglobulin (OR=2.838, =0.002) were independent risk factors for pneumonia in patients with pneumoconiosis. The prediction model was constructed as Logit (P) =1.357×CRP+0.196×IL-6+0.195×IL-8+1.043×β2-microglobulin-9.772. The H-L test demonstrated that there was no significantly difference between predicted values and the true values. The areas under ROC curves of the model in the modeling set and the validation set were 0.909 and 0.848. The sensitivity was 86.8% and 75.6%, and accuracy 82.1% and 79.7%, respectively. Conclusions CRP, IL-6, IL-8, and β2-microglobulin were independent risk factors of pneumonia for pneumoconiosis patients. The risk model constructed by laboratory test indexes showed a good predictive performance, which might provide laboratory evidence for the pneumonia diagnosis in patients with pneumoconiosis.

B-313 Assessment of β2-microglobulin concentration in pleural effusions

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 &amp;gt;60mL/min β2-m median concentration was 4,12mg/L and if GFR &amp;lt;60 mL/min, β2-m was 8,62g/L significant difference have found (p&amp;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&amp;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&amp;gt;60 mL/min), the area under the ROC curve was 0.98 (95% CI: 0.88-0.98) (p&amp;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&amp;gt;60 mL/min and tuberculosis. Using 5,15 mg/L cut-off point, 100% of NPV was obtained.

B-314 Utility of he4 in differential diagnosis of pleural effusions

Abstract Background The usefulness of tumor markers in the differential diagnosis of cancer in patients with pleural effusions (PE) remains controversial. Few studies have reported the measurement of human epididymal protein 4 (HE4) in pleural fluid. HE4 is a tumor marker associated with different types of cancer. An elevated serum concentration has been observed in ovarian and lung cancers. Other benign pathologies can increase HE4 levels, which can be detect with different laboratory tests identified with adenosine deaminase (ADA), C-reactive protein (CRP), % polynuclear cell count (%PMN) and glomerular filtration rate (GFR) can be used in order to detect possible false positives. The objective of this study was to assess the diagnostic usefulness of HE4 in malignant pleural effusions by comparing HE4 concentrations between benign and malignant samples, while considering the causes of HE4 false positives. Methods HE4, ADA, differential leukocyte count, CRP and GFR concentrations were determined in 238 pleural effusion samples. Diagnostic efficacy analysis using receiver operating curves (ROC) identified the ability of HE4 to detect malignancy, searching for cut-off points with 100% specificity. Results HE4 concentrations showed significant differences (P&amp;lt;0.01) between malignant (median 1065pmol/L) and benign (median 699pmol/L) pleural effusions. Significant differences (P&amp;lt;0.01) were found in HE4 concentrations for gynecological and lung cancer origin (median 7397 pmol/L and 5150 pmol/L respectively) and other malignant pleural effusions (median 1065 pmol/L). Patients with benign diseases with renal failure showed a higher HE4 concentration (median 964pmol/L) than in patients without these pathologies (median 696pmol/L) identified with GFR&amp;lt;30 mL/min. No differences were demonstrated in HE4 concentration levels using ADA, differential leukocyte count or CRP, so only glomerular filtration rate was considered cause of false positives. A sensitivity of 23% was obtained for HE4, with a cut-off point of 3050pmol/L and a specificity of 100% for the whole group. In the potential false positive group, the sensitivity of HE4 was 28% with a cutoff point of 3050 pmol/L with a specificity of 100%. In the group without potential false positives, we obtained a sensitivity of 30% with a specificity of 100% for a cutoff point of 1992pmol/L. When we used a specific cut-off point for each group, we obtained a sensitivity of 38% with a specificity of 100%. Conclusions HE4 levels are higher in malignant pleural effusions compared to other pathologies. HE4 levels allow the identification of malignant pleural effusions with moderate sensitivity and maximum specificity. HE4 levels can differentiate between tumors of gynecological or lung origin and others. Glomerular filtration rate (GFR&amp;lt;30 mL/min) was considered the main cause of false positives. If we identify possible false positives, we can use specific cut-offs and improve sensitivity up to 38% with maximum specificity compared to using a single cut-off for all effusions.

B-240 Multi-omic blood test for early detection of cancer and pre-cancer

Abstract Background The earlier diagnosis of cancer is vital to improve patient survival. Many liquid biopsy technologies are focused on single tumor-derived biomarkers, which limits test sensitivity, especially for early-stage cancers that do not shed sufficient genetic material into the bloodstream. The Dxcover liquid biopsy employs Multi-Omic Spectral Analysis (MOSA) that interrogates a blood sample with Infrared (IR) radiation and produces a distinctive signature that represents the whole biomolecular profile of the sample. This multi-omic blood test analyzes the full complement of tumor and immune-derived markers present within blood derivatives and could facilitate the earlier detection of cancer. The technology can be tuned for high sensitivity (rule out) or high specificity (rule in) depending on the desired application, clinical priorities and international healthcare markets. Methods Three clinical studies have been completed, assessing the technology for the detection of: (1) brain cancer, (2) multiple cancers, and (3) colorectal cancer (CRC) and pre-cancerous lesions. In the first study, we recruited 988 patients prospectively with non-specific symptoms associated with a brain tumor. Secondly, we carried out a large-scale multi-cancer evaluation (n = 2092 patients), targeting eight different cancers. Moreover, the CREATE (ColoREctal cancer &amp; Adenoma Test Evaluation) feasibility study (3) examined 100 CRC, 92 adenoma samples and 104 colonoscopy screening control patients. Results In study 1, the brain cancer algorithm detected 96% of the patients with brain cancer, and 100% of glioblastomas, the most aggressive primary brain tumor. In the multi-cancer study (2), area under the receiver operating characteristic curve values were calculated for the eight cancer types versus symptomatic non-cancer controls: brain (0.90), breast (0.76), colorectal (0.91), kidney (0.91), lung (0.91), ovarian (0.86), pancreatic (0.84) and prostate (0.86). We also assessed the test performance when all eight cancer types were pooled to classify ‘any cancer’ against non-cancer patients. The cancer versus asymptomatic non-cancer classification detected 64% of stage I cancers when specificity was 99% (overall sensitivity 57%). When tuned for higher sensitivity, this model identified 99% of Stage I cancers (with specificity 59%). In the CREATE study, the test reported promising results for pre-cancer detection; at 90% specificity, the test reported sensitivities of 80% for CRC and 59% for advanced adenomas. Conclusions A rapid liquid biopsy that is sensitive to pre-cancerous lesions and early-stage cancer could substantially improve patient outcomes. There is a low barrier to integrating this blood test into existing diagnostic pathways as the technology is rapid, simple to use, and sample preparation is minimal. In addition, the spectroscopic liquid biopsy has the potential to be combined with other orthogonal tests, such as cell-free DNA, which could provide an efficient route to diagnosis. Cancer treatment can be more effective when given earlier, and this low-cost strategy has the potential to improve patient prognosis.

B-316 Evaluation of an FDA-cleared Chromogranin A Assay on an Automated Analyzer

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 (&amp;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 &amp;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 &amp;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.

Significance of apparent diffusion coefficient in diagnosis of rectal carcinoma

IntroductionThe apparent diffusion coefficient (ADC) is a quantitative parameter that facilitates the detection and reliable differentiation of rectal cancer. MR differentiation between rectal carcinoma, post-radiation proctitis, and normal rectal wall with the ADC values and their comparison depending on the level of tumor markers and pathohistological characteristics of rectal carcinoma.MethodsThe retrospective study performed at the Oncology Institute of Vojvodina included 300 patients, 100 each with rectal cancer, post-radiation proctitis, and normal rectum. Mean ADC values were obtained by measuring the region of interest (ROI) of the rectal wall.ResultsRectal cancer showed lower ADC values (0.665 ± 0.086 x 10-3mm2/s) compared to both post-radiation proctitis (1.648 ± 0.268 x 10-3mm2/s) and normal rectum (1.180 ± 0.110 x 10-3mm2/s) (p&amp;lt;0.001). No significant differences in ADC values were observed between different grades of rectal cancer (p=0.874; p&amp;gt;0.05), depending on the presence of metastases in the lymph nodes (p=0.357; p&amp;gt;0.05), different TN stage (p=0.196; p&amp;gt;0.05), local spread of the tumor (p=0.312; p&amp;gt;0.05), the presence of RAS mutation (p=0.829; p&amp;gt;0.05) and the value of tumor markers (p=0.923; p&amp;gt;0.05). ADC values below 1.013 x 10-3mm2/s with 100% sensitivity and 96% specificity indicate the presence of rectal cancer in relation to normal wall, with a positive predictive value of 96.1% and a negative of 100%. ADC values below 1.255 x 10-3mm2/s with 100% sensitivity and 95% specificity indicate rectal cancer in relation to post-radiation proctitis. ADC values above 1.339 x 10-3mm2/s with 87% sensitivity and 89% specificity indicate post-radiation proctitis in relation to normal wall.DiscussionThe ADC is a useful marker in differentiating between rectal cancer, post-radiation proctitis, and normal rectal wall with high sensitivity and specificity, but it cannot be used to distinguish the histological grades of rectal cancer, nor other pathohistological parameters.

A-074 A Laboratory Developed Serum Anti-Mullerian Hormone Electrochemiluminescent Immunoassay: Performance and Retrospective Analysis of Over 2.2 Million Tests in Females and Males Across Lifespan

Abstract Background The quantification of serum Anti-Mullerian hormone (AMH), a glycoprotein dimer produced by granulosa cells of the ovarian follicle in females and by Sertoli cells of the testes in males, has clinical utility across lifespan in both females and males. AMH is most frequently ordered to assess fertility in females of reproductive age, where it correlates with ovarian reserve. The clinical applications of serum AMH beyond ovarian reserve assessment, especially in males, pediatric, and postmenopausal populations, include the investigation of peri-menopause transition, delayed or precocious onset of puberty, differential diagnosis of disorders of sexual development, the evaluation of male gonadal function, and as an ovarian tumor marker. The objective was to establish and validate a highly sensitive assay to quantify AMH and to use our database of more than 2 million tests to understand the clinical utility of measuring AMH across sex and lifespan. Methods A validated two-step electrochemiluminescence assay was used for the quantification of serum AMH. The analytical performance of the assay was assessed, including accuracy, precision, and dilutional linearity. ICD-10 codes ordered between 2012 and 2023 were retrospectively analyzed for each subgroup based on age and sex to determine test usage. The analysis includes results from 2,229,845 female and 10,362 male patients across lifespan. Results The AMH analytical measurement range (AMR) was determined to be 0.015 - 15.0 ng/mL with a maximum reportable concentration of 960 ng/mL when run on dilution. Both inter- and intra-assay standard accuracy and precision were ≤ ±15.4% bias and ≤ 11.0% CV for all levels. Inter- and intra-assay sample precision was ≤ 9.6% CV across the AMR. Samples diluted linearly and recovered within 80 - 120% of the expected values when diluted up to 64X. Analytical interference caused by icterus, lipemia, or hemolysis was not observed. The most common ICD-10 associated with AMH orders was for fertility testing in females of reproductive age. AMH was also ordered in females for evaluation of ovarian cancer, menstrual disorders, and PCOS. In males, common ICD-10 codes associated with AMH include delayed puberty, hypoplasia of the penis, fertility testing, hypogonadism, and childhood short stature. Conclusions This study demonstrates a highly sensitive assay for the quantification of serum AMH with clinical utility across sex and lifespan. Using retrospective data, we determined that in addition to being used primarily in fertility testing, AMH is also commonly used to evaluate and manage other endocrine disorders and malignancies, including disorders of sexual development, disorders of puberty, PCOS, male gonadal dysfunction, and granulosa cell tumors. This assay can aid in defining the clinically relevant age and sex-specific levels of AHM in non-fertility-related conditions.

B-149 Diverse glycosylation of circulating alpha fetoprotein in chronic liver diseases profiled by intact mass high resolution mass spectrometry

Abstract Background Alpha fetoprotein (AFP) is a circulating tumor marker utilized clinically to detect liver cirrhosis, liver cancer and testicular seminomas. AFP molecule contains one single N-glycosylation site, with highly variable glycan structures, resulting in diverse AFP glycoforms in circulating blood. The core fucosylation of AFP (AFP-L3) is a clinical biomarker to monitor and predict risks of liver cancer progression and recurrence. Clinical testing for AFP-L3 utilizes lectin-binding and gel-shift electrophoresis. We developed a novel method to measure AFP glycoforms, based on immune-enrichment and liquid chromatography with high resolution mass spectrometry (LC-HRMS). Methods Immune enrichment of AFP was achieved by anti-AFP antibody, immobilized to magnetic beads. After binding and enrichment, the magnetic beads were washed, AFP was eluted, and the samples were analyzed by LC-HRMS (Exploris 480 MS and Vanquish UHPLC, Thermo Fisher Scientific), using full scan, positive polarity acquisition mode. Chromatographic separation was performed on PLRP column (Agilent); flow rate 200 µL/min. The HRMS data were processed and deconvoluted with intact mass analysis workflow in Biopharma Finder 5.1 (Thermo Fisher Scientific) to derive the intact AFP masses. The method’s sensitivity and imprecision were assessed by replicate analysis of serial diluted patient serum samples. AFP and its associated glycoforms were matched and identified with an in-house developed bioinformatics workflow. The deconvoluted intact masses and the sum intensity of detected charged species for each AFP glycoform were used to quantify relative abundance of AFP glycoforms. Results The method specificity for AFP was confirmed by bottom-up analysis after immune-enrichment with observed peptide sequence coverage over 35%. The recovery of the immune enrichment was shown to be independent of the AFP glycoform compositions. The identified AFP glycoforms and their relative abundance were consistent among replicates between AFP concentration 100 ng/mL and 5000 ng/mL. Analysis of authentic serum specimens from patients with different chronic liver diseases and liver cancer (n=102), revealed multiple complex biantennary branched AFP glycan modifications, with different numbers of terminal sialic acid and core fucose. Four AFP glycoforms were identified in over 60% patient samples with elevated AFP concentrations. Although occurring consistently, relative abundance of these AFP glycoforms varied significantly among patients. Assessment of the AFP fucosylation and sialylation features suggested that the two are orthogonal processes. Clinical correlations suggested that fewer terminal sialic acids were associated with impaired liver functions, while the fucosylation was linked to inflammatory responses and presence of liver malignancy. Strong linear correlation (Spearman’s rs=0.952, N=35) was observed between AFP-L3% quantitated by the LC-HRMS method, and AFP-L3% determined by the lectin-binding based isotachophoresis assay (Wako Diagnostics). Discrepancies between the two methods were observed in specimens containing &amp;lt; 400 ng/mL of AFP, in which the LC-HRMS method detected only a single predominant glycoform. Conclusions We developed and evaluated the LC-HRMS method for profiling circulating AFP glycoforms in patients with chronic liver diseases and liver cancer. The detected AFP glycoforms and their relative abundance were used to establish glycan features, which can be further linked to cancer development, impaired liver function, and inflammation.

Chronic granulomatous intestinal disease: case report

Chronic intestinal granulomatous disease is a disorder that directly compromises the entire immune system. The estimated incidence is one in a million inhabitants. Among the associated risk factors, the presence of chronic and recurrent infections stands out. In the present case, it is a 39-year-old female patient with a history of intestinal obstruction and endometriosis, who presents abdominal pain in the left abdomen and recurrent abdominal pain without an identifiable cause. Laboratory tests revealed leukocytosis and mild neutrophilia. The tomography showed intestinal dilation at the jejunoileal level without transition zones, and the serological tests were negative. Exploratory laparoscopy was performed and the presence of tumor implants was found in the omentum, extending to the abdominal, uterine, intestinal, and peritoneal regions, as well as sallow fluid at the abdominal level. Tumor markers are normal. Histopathology details fungal colonization with eosinophilic necrosis and presence of granulomas, negative immunohistochemistry for Aspergillus and negative Lowestein-jensen culture. Antifungal and antituberculous treatment was started, with a decrease in symptoms and improvement in the clinical picture.

Differentiating Primary from Metastatic Ovarian Tumors of Gastrointestinal Origin by CT

PurposeTo determine differentiating CT imaging features of primary ovarian cancers from ovarian metastases of gastrointestinal origin. MethodsRetrospective study of 50 patients with new ovarian lesions on CT, half were primary ovarian cancers and half gastrointestinal metastases. Two blinded independent readers described tumor characteristics on CT (size, laterality, margin, etc.) and ancillary features (ascites, peritoneal seeding, lymphadenopathy, etc.). Patient age, sex, cancer history, and tumor marker levels for CA-125 and CEA were collected. Wilcoxon test and Pearson's chi-squared test were used for statistical analysis. Results50 patients with mean age of 62.1 years were included. Ovarian metastases were more likely to be cystic/mainly cystic (p=0.013), have smooth margins (p=0.011), and have no/mild enhancement (p<0.001). Primary ovarian lesions were associated with moderate to large volume of ascites (p=0.047) and more commonly seen with lymphadenopathy (p=0.008). Laterality was not significantly different between the two groups. CA-125 level was more commonly elevated in primary ovarian lesions (87% vs 50%, p=0.018), and with much higher values (1076.5 vs 155.1, p=0.013). CEA level was more commonly elevated in metastatic ovarian lesions (83.3% vs 15.4%, p<0.001), and with higher values (72.4 vs 2.1, p<0.001). ConclusionOvarian metastases were more frequently smooth-margined and cystic with little enhancement. Primary ovarian lesions were more commonly associated with lymphadenopathy and larger volume of ascites. Tumor markers CEA and CA-125 were more frequently elevated in metastatic and primary lesions, respectively. Cancer history was the only variable that increased the odds of metastasis and therefore it is important to always correlate with history of cancer.

Modified reconstruction approach after pancreaticoduodenectomy optimizes postoperative outcomes: Results from a multivariate cohort analysis.

Despite technical advances in recent decades and a decrease in hospital mortality (<5%), pancreaticoduodenectomy (PD) is still associated with major postoperative complications, even in high-volume centers. The present study aimed to assess the effect of a modified reconstruction technique on postoperative morbidity and mortality. A cohort study of all patients (n=218) undergoing PD between January 2010 and December 2019 was performed at Attikon University Hospital (Athens, Greece). Several variables were studied, including demographic data, past medical history, perioperative parameters, tumor markers and pathology, duration of hospitalization, postoperative complications, 30-day-survival, postoperative mortality and overall survival using multivariate logistic regression and survival analysis techniques. In this cohort, 123 patients [modified PD (mPD) group] underwent a modified reconstruction after a pylorus-preserving pancreaticoduodenectomy, which consisted of gastrojejunostomy and pancreaticojejunostomy on the same loop and an isolated hepaticojejunostomy on another loop. In the standard PD (StPD) group, 95 patients underwent standard reconstruction. The median age was 67 years, ranging from 25 to 89 years. Compared with in the StPD group, the mPD group had significantly lower rates of grade B and C pancreatic fistula (4.9% vs. 28.4%), delayed gastric emptying (7.3% vs. 42.1%), postoperative hemorrhage (3.3% vs. 20%), intensive care unit admission (8.1% vs. 18.9%), overall morbidity (Clavien-Dindo grade III-V: 14.7% vs. 42.0%), perioperative mortality (4.1% vs. 14.7%), and shorter hospitalization stay (11 days vs. 20 days). However, no difference was noted regarding median survival (35 months vs. 30 months). In this single-center series, a modified reconstruction after PD appears to be associated with improved postoperative outcomes. However, further evaluation in larger multi-center trials is required.

Application of a single-cell-RNA-based biological-inspired graph neural network in diagnosis of primary liver tumors

Single-cell technology depicts integrated tumor profiles including both tumor cells and tumor microenvironments, which theoretically enables more robust diagnosis than traditional diagnostic standards based on only pathology. However, the inherent challenges of single-cell RNA sequencing (scRNA-seq) data, such as high dimensionality, low signal-to-noise ratio (SNR), sparse and non-Euclidean nature, pose significant obstacles for traditional diagnostic approaches. The diagnostic value of single-cell technology has been largely unexplored despite the potential advantages. Here, we present a graph neural network-based framework tailored for molecular diagnosis of primary liver tumors using scRNA-seq data. Our approach capitalizes on the biological plausibility inherent in the intercellular communication networks within tumor samples. By integrating pathway activation features within cell clusters and modeling unidirectional inter-cellular communication, we achieve robust discrimination between malignant tumors (including hepatocellular carcinoma, HCC, and intrahepatic cholangiocarcinoma, iCCA) and benign tumors (focal nodular hyperplasia, FNH) by scRNA data of all tissue cells and immunocytes only. The efficacy to distinguish iCCA from HCC was further validated on public datasets. Through extending the application of high-throughput scRNA-seq data into diagnosis approaches focusing on integrated tumor microenvironment profiles rather than a few tumor markers, this framework also sheds light on minimal-invasive diagnostic methods based on migrating/circulating immunocytes.

Strategies for Non-Covalent Attachment of Antibodies to PEGylated Nanoparticles for Targeted Drug Delivery.

Polyethylene glycol (PEG)-modified nanoparticles (NPs) often struggle with reduced effectiveness against metastasis and liquid tumors due to limited tumor cell uptake and therapeutic efficacy. To address this, actively targeted liposomes with enhanced tumor selectivity and internalization are being developed to improve uptake and treatment outcomes. Using bi-functional proteins to functionalize PEGylated NPs and enhance targeted drug delivery through non-covalent attachment methods has emerged as a promising approach. Among these, the one-step and two-step targeting strategies stand out for their simplicity, efficiency, and versatility. The one-step strategy integrates streptavidin-tagged antibodies or bispecific antibodies (bsAbs: PEG/DIG × marker) directly into PEGylated NPs. This method uses the natural interactions between antibodies and PEG for stable, specific binding, allowing the modification of biotin/Fc-binding molecules like protein A, G, or anti-Fc peptide. Simply mixing bsAbs with PEGylated NPs improves tumor targeting and internalization. The two-step strategy involves first accumulating bsAbs (PEG/biotin × tumor marker) on the tumor cell surface, triggering an initial attack via antibody-dependent and complement-dependent cytotoxicity. These bsAbs then capture PEGylated NPs, initiating a second wave of internalization and cytotoxicity. Both strategies aim to enhance the targeting capabilities of PEGylated NPs by enabling specific recognition and binding to disease-specific markers or receptors. This review provides potential pathways for accelerating clinical translation in the development of targeted nanomedicine.

The relationship between serum tumor markers and immune response in patients with lung cancer.

The relationship between serum tumor markers and immune response in patients with lung cancer.

7510 Use of Tumor Markers and Surgery in the Management of Medullary Thyroid Cancer During Pregnancy

7510 Use of Tumor Markers and Surgery in the Management of Medullary Thyroid Cancer During Pregnancy

Development of transition metal oxide platforms for aptasensing of PSA in cell cultures.

In this study, a novel aptasensor based on a transition metal oxide-modified pencil graphite electrode (PGE) was developed for the diagnosis of early-stage prostate cancer (PCa) via monitoring the prostate-specific antigen (PSA), which is the main biomarker for PCa. Single-use PGEs modified with pulsed deposited manganese oxide (MnOx) film were used to attach the amino-terminated aptamer specific to the PSA via carbodiimide chemistry. The designed aptasensor was placed in an electrochemical cell containing ferri/ferrocyanide ions as a redox probe to measure the charge transfer resistances (Rct) of the electrode surface by electrochemical impedance spectroscopy (EIS) to follow the response of each modification step. The effect of the medium pH on the ionic structure of the aptamer molecule according to its pI value and, thus, the reversing of the direction of the response (ΔRct) by the pH change was also discussed. The level of PSA secreted from PCa cells was investigated using impedimetric transduction. The specificity of the aptasensor was validated through selectivity studies against non-specific tumor markers like VEGF and different cancer cell lines including breast cancer and androgen-insensitive prostate cancer. The developed system showcases a label-free, fast, specific, and cost-effective approach for PSA detection, highlighting the importance of medium pH and the electrostatic environment on the aptamer's response. Our work emphasizes the potential for such aptasensors in clinical diagnostics and paves the way for further exploration into using transition metal oxides in biosensing applications.