Discrepant Cases Research Articles

Implementation of calculations of normal sections of reinforced concrete structures in software packages

Introduction. In previous publications, the authors examined the features of calculating the strength of normal sections of reinforced concrete structures associated with the possibility of using two approaches – the nonlinear deformation model (NDM) and the method of limiting forces (MLF). Using specific examples, the results obtained with both approaches are compared. Cases of significant discrepancies in the consumption of selected reinforcement have been identified, trends have been explained, and the physical essence of the phenomenon has been analyzed. Calculations were performed according to the OM SNiP Reinforced Concrete program.
 
 Aim. To analyze the implementation of features when using various computer programs for calculations of reinforced concrete structures, identify differences in results, establish their physical cause, evaluate economic aspects and problems, and make the necessary recommendations.
 
 Materials and methods. The studies were performed for three types of structures. The first type included four sections of elements under two stress states: off-center compression (rectangular and annular) and oblique offcenter compression (rectangular and I-beam). The second type of structures included columns and pylons of a projected and constructed reinforced concrete building. The third type was an illustrative example – the T-section of the bent element at different operating moments and percentages of reinforcement. Calculations of the first type of structures were carried out by the authors by the OM SNiP Reinforced Concrete program, as well as the program developers of the Ing+, Arbat, Lira-SAPR, Lira 10, STARK ES programs. The calculation of structures of the second type was carried out by the authors by the OM SNiP Reinforced Concrete program and the designers by the Lira-SAPR 2022 program. The test example was calculated by the authors according to the OM SNiP Reinforced Concrete program.
 
 Results. When calculating according to the NDM, a significant unjustified overspending of the required reinforcement was revealed compared to the calculation according to the MLF. In an illustrative example, the overspend is 220 %.
 
 Conclusions. The general state of calculations of reinforced concrete structures should be recognized as requiring adjustments. To obtain a multibillion-dollar economic effect, it is proposed to provide in SP 63.13330.2018 two equal approaches to the calculation of normal sections of reinforced concrete structures - NDM and MLF. It is advisable to develop a document with a set of reference examples that allows you to get the same results for different programs with the same source data.

Expect the unexpected: investigating discordant prostate MRI and biopsy results

ObjectivesTo evaluate discrepant radio-pathological outcomes in biopsy-naïve patients undergoing prostate MRI and to provide insights into the underlying causes.Materials and methodsA retrospective analysis was conducted on 2780 biopsy-naïve patients undergoing prostate MRI at a tertiary referral centre between October 2015 and June 2022. Exclusion criteria were biopsy not performed, indeterminate MRI findings (PI-RADS 3), and clinically insignificant PCa (Gleason score 3 + 3). Patients with discrepant findings between MRI and biopsy results were categorised into two groups: MRI-negative/Biopsy-positive and MRI-positive/Biopsy-negative (biopsy-positive defined as Gleason score ≥ 3 + 4). An expert uroradiologist reviewed discrepant cases, retrospectively re-assigning PI-RADS scores, identifying any missed MRI targets, and evaluating the quality of MRI scans. Potential explanations for discrepancies included MRI overcalls (including known pitfalls), benign pathology findings, and biopsy targeting errors.ResultsPatients who did not undergo biopsy (n = 1258) or who had indeterminate MRI findings (n = 204), as well as those with clinically insignificant PCa (n = 216), were excluded, with a total of 1102 patients analysed. Of these, 32/1,102 (3%) were classified as MRI-negative/biopsy-positive and 117/1102 (11%) as MRI-positive/biopsy-negative. In the MRI-negative/Biopsy-positive group, 44% of studies were considered non-diagnostic quality. Upon retrospective image review, target lesions were identified in 28% of cases. In the MRI-positive/Biopsy-negative group, 42% of cases were considered to be MRI overcalls, and 32% had an explanatory benign pathological finding, with biopsy targeting errors accounting for 11% of cases.ConclusionProstate MRI demonstrated a high diagnostic accuracy, with low occurrences of discrepant findings as defined. Common reasons for MRI-positive/Biopsy-negative cases included explanatory benign findings and MRI overcalls.Clinical relevance statementThis study highlights the importance of optimal prostate MRI image quality and expertise in reducing diagnostic errors, improving patient outcomes, and guiding appropriate management decisions in the prostate cancer diagnostic pathway.Key Points• Discrepancies between prostate MRI and biopsy results can occur, with higher numbers of MRI-positive/biopsy-negative relative to MRI-negative/biopsy-positive cases.• MRI-positive/biopsy-negative cases were mostly overcalls or explainable by benign biopsy findings.• In about one-third of MRI-negative/biopsy-positive cases, a target lesion was retrospectively identified.

Reappraisal of p16 for Determining HPV Status of Head and Neck Carcinomas Arising in HPV Hotspots.

In an era of head and neck oncology where HPV status will soon dictate patient management, reliable HPV detection is critical. P16 immunohistochemistry (IHC) is currently recommended as the test of choice for oropharyngeal squamous cell carcinomas (OPSCCs). The purpose of this study was to determine the performance characteristics of p16 IHC based on a large clinical experience of squamous cell carcinomas (SCC) arising from HPV hot-spot regions of the head and neck. Consecutive OPSCCs, sinonasal SCCs, and metastatic SCCs of unknown primary sites were evaluated for the presence of HPV by p16 IHC and PCR-based HPV DNA testing as part of clinical care. For discrepant cases, high-risk HPV E6/E7 mRNA in situ hybridization (ISH) and, when possible, matrix-assisted laser desorption/ionization-time of flight (MALDI-TOF) mass spectrometry (MassArray) genotyping were performed. 746 cancers underwent HPV testing by p16 IHC and DNA PCR genotyping. There was a 95.6% concordance between the 2 assays. Of the 33 discrepant cases, 32 cases (4.3%) were p16 positive but HPV DNA negative. In these cases, 68% were positive for mRNA ISH, invariably related to a non-16 HPV genotype. P16 IHC had an overall accuracy of 98.8%, a sensitivity of 99.8%, and a specificity of 92.1%. P16 IHC is a sensitive and specific assay for determining HPV status. HPV DNA PCR appears vulnerable to HPV genotype diversity and is prone to missing rare non-16 genotypes. HPV mRNA ISH is a practical and reliable direct measure of HPV that may help eliminate the small number of false-positive p16 cases and avoid potential patient harm related to erroneous HPV classification.

Loss of Blood Group Antigens in Haematolymphoid Malignancy: A Case Series from a Cancer Institute from North East India

Introduction: Altered expression of blood group antigens has been reported to be associated with both solid as well as haematological malignancies. This usually results from genetic mutation leading to incomplete as well as abnormal synthesis of antigens. Transitional loss of RBC antigens especially from ABO system is most commonly found in haematological malignancies causing blood group discrepancies and followed by reversal to its historical blood group during remission. Loss of A, B antigens has also been associated to play a role in lung, bladder and colon cancers. However not much data is available regarding haematological malignancies.Methods and Methodology: Retrospective review of blood group discrepancy (Forward group) in patients admitted for haematolymphoid malignancy at our institute was done during a period of 3.5 years (13/3/2020 till 12/9/2023). Blood grouping done by Column agglutination test were repeated by conventional tube method. Further confirmation of the group was done by saliva test and adsorption elution test as indicated. The detail history of the patient was collected from the hospital records and analysed. Results: A total of 7 patients presented with either loss or decrease in antigen expression. Most common diagnosis was Acute Myeloid Leukaemia (AML) with FLT3 mutation and most common antigen affected was A. Conclusion: Haematolymphoid malignancies has been associated with loss of blood group antigens causing group discrepancy. Blood transfusion is an integral part of the supportive care in these patients. Hence, proper work up of blood group discrepancy cases should be done and recorded so as to prevent delay in blood transfusion.

PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay: A highly sensitive method for detection of MYD88 L265P mutation.

Agarose gel-based conventional and real-time allele-specific polymerase chain reaction (AS-PCR) assays are currently used for sensitive detection and quantification of MYD88 L265P mutation. Visual inspection of an agarose gel can often be ambiguous. We propose a new allele-specific quantification PCR (AS-qPCR) assay, PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay, that uses Intercalating Nucleic Acid (INA®) technology for increased affinity and specificity. This study compares PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay with conventional AS-PCR. We included a total of 102 peripheral and bone marrow blood samples from 94 patients with a lymphoproliferative disorder. Droplet digital PCR (ddPCR) was used as a third method in case of discrepancy. A positive percent agreement of 100% (95% CI 0.92-1.0) and a negative percent agreement of 98% (95% CI 0.90-1.0) were found between the conventional AS-PCR and the AS-qPCR methods. Including the ddPCR results to validate the discrepant cases, the sensitivity and specificity of PlentiPlex™ MYD88 Waldenström lymphoma qPCR Assay were 1.0 (95% CI 0.97-1.0) and 1.0 (95% CI 0.96-1.0), respectively. Our data demonstrate that PlentiPlex™ MYD88 Waldenström lymphoma qPCR assay is a fast, highly sensitive, and specific method for the detection of MYD88 L265P compared with conventional AS-PCR.

Retrospective analysis of discordant results between histology and other clinical diagnostic tests on helicobacter pylori infection.

A reliable test is essential for diagnosing Helicobacter pylori (H. pylori) infection, and crucial for managing H. pylori-related diseases. Serving as an excellent method for detecting H. pylori infection, histologic examination is a test that clinicians heavily rely on, especially when complemented with immunohistochemistry (IHC). Additionally, other diagnostic tests for H. pylori, such as the rapid urease test (CLO test) and stool antigen test (SA), are also highly sensitive and specific. Typically, the results of histology and other tests align with each other. However, on rare occasions, discrepancy between histopathology and other H. pylori diagnostic tests occurs. To investigate the discordance between histology and other H. pylori tests, the underlying causes, and the impact on clinical management. Pathology reports of gastric biopsies were retrieved spanning August 2013 and July 2018. Reports were included in the study only if there were other H. pylori tests within seven days of the biopsy. These additional tests include CLO test, SA, and H. pylori culture. Concordance between histopathology and other tests was determined based on the consistency of results. In instances where histology results were negative while other tests were positive, the slides were retrieved for re-assessment, and the clinical chart was reviewed. Of 1396 pathology reports were identified, each accompanied by one additional H. pylori test. The concordance rates in detecting H. pylori infection between biopsy and other tests did not exhibit significant differences based on the number of biopsy fragments. 117 discrepant cases were identified. Only 20 cases (9 with CLO test and 11 with SA) had negative biopsy but positive results in other tests. Four cases initially stained with Warthin-Starry turned out to be positive for H. pylori with subsequent IHC staining. Among the remaining 16 true discrepant cases, 10 patients were on proton pump inhibitors before the biopsy and/or other tests. Most patients underwent treatment, except for two who were untreated, and two patients who were lost to follow-up. There are rare discrepant cases with negative biopsy but positive in SA or CLO test. Various factors may contribute to this inconsistency. Most patients in such cases had undergone treatment.

Discrepancies Between Morphological and Immunohistochemical Classifications Are Associated With Prognoses and Subtypes of Lung Cancer.

Immunohistochemical (IHC) staining has been routinely used to distinguish adenocarcinoma (ADC) and squamous cell carcinoma (SCC) of the lungs; however, it is challenging to obtain an accurate diagnosis, especially for cases with discrepancies between IHC and hematoxylin and eosin (H&E) staining results. This study aimed to clarify the clinicopathological characteristics of these discrepant cases. Tissue microarray specimens from 321 patients with ADC and SCC were used for H&E and IHC staining of thyroid transcription factor 1 (TTF-1), Napsin A, cytokeratin 5/6 (CK5/6), p40, and p63. The pathological diagnosis was made based on (1) H&E, (2) IHC, and (3) both H&E and IHC results. Discrepant cases were defined as those with different diagnoses based on the H&E and IHC results. A total of 32 (10%) discrepant cases were identified. ADC (3.9%) showed fewer discrepant cases than SCC (51%). Discrepant cases of ADC had a significantly higher proportion of poorly differentiated tumors and subtypes of solid and invasive mucinous ADC, and they also had shorter overall and disease-free survival than concordant cases. Solid and invasive mucinous ADC cases showed low positivity for TTF-1 (84% and 40%, respectively) and Napsin A (88% and 80%, respectively), and invasive mucinous ADC cases showed high positivity for CK5/6 (80%). The sensitivity and specificity of TTF-1+Napsin A for ADC were 91% and 83%, respectively, whereas those of CK5/6+p40 for SCC cases were 90% and 96%, respectively. Discrepant cases of ADC are associated with solid and invasive mucinous subtypes and shorter survival.

Comparison of tissue based FGFR mutation detection by Therascreen FGFR with UroTyper FGFR and ADC test and relevance for potential co-targeting with TROP2 and NECTIN4: Preview of Bladder BRIDGister.

679 Background: In view of the efficacy of FGFR targeting in early and advanced bladder cancer, as has been shown for erdafitinib in the THOR and NORSE trial, molecular testing of FGFR mutations and fusions will soon become clinical routine worldwide. Therascreen FGFR testing has been approved as companion diagnostic to select patients for erdafitinib. However, technical problems and shortages as well as a limited number of detectable mutations indicate the need of FGFR test systems that can be performed easily & fast worldwide. The objective was to evaluate concordance of UroTyper FGFR with Therascreen FGFR mutation testing and unravel molecular characteristics of FGFR mutated tumors to assess potential co-targeting options for erdafitinib to justify subsequent adopted clinical trials with the Bladder BRIDGister framework. Methods: FFPE tumor tissues from 100 TURB samples were prospectively collected as part of the Bladder BRIDGister. RNA from FFPE tissues were extracted by commercial kits to assess Therascreen FGFR & UroTyper FGFR MODAPLEX (BIOTYPE GmbH, Dresden) in conjunction with relative gene expression of subtyping markers (i.a.. PPARG), ADC targets (TROP2, NECTIN4) and CPI targets by RT-qPCR systems. Hierarchical clustering, Spearman correlation tests were done by JMP 9.0.0 (SAS software). Results: After exclusion of FGFR fusion samples, a total of 94 samples was available for comparative mutation analysis. Both PCR based FGFR tests concordantly identified S249C (n=15), R248C (n=2) and G370C (n=1) mutations, while the results for Y373C were in part discrepant with Therascreen FGFR detecting 9 mutations, while UroTyper FGFR did detect 6 mutations. Further examination of the 3 discrepant Y373C cases by SNaPshot and Uromonitor FGFR test revealed that the Therascreen FGFR test resulted in 3 false positive results. Moreover, one sample was invalid from Therascreen FGFR test. Overall, the concordance was very good and reached a kappa value of 0.92. Interestingly, FGFR3 mutations by UroTyper FGFR test were positively associated not only with FGFR3 mRNA overexpression (r=0.6699, p<0.0001), but also with TROP2 and NECTIN4 (r=0.5026 and r=0.4406, p<0.0001) indicating, that combinations of FGFR targeting with ADC targeting of TROP2 or NECTIN4 could have strong synergistic effects. Hierarchical clustering revealed a homogenous FGFR3 mutated group of tumors simultaneously co-expressing the target genes TROP2, NECTIN4 and PPARG. Conclusions: PCR-based FGFR assessment by Therascreen and UroTyper is highly concordant and enables fast, local FGFR assessment within few hours. FGFR3 mutations are associated with increased TROP2 & NECTIN4 expression indicating potential synergistic options which warrants further exploration as part of molecularly stratified clinical trials.

Comparison of Immunohistochemistry, Next-generation Sequencing and Fluorescence In Situ Hybridization for Detection of MTAP Loss in Pleural Mesothelioma

9p21 deletions involving MTAP/CDKN2A genes are detected in diffuse pleural mesotheliomas (DPM) but are absent in benign mesothelial proliferations. Loss of MTAP expression by immunohistochemistry (IHC) is well accepted as a surrogate for 9p21 deletion to support a diagnosis of DPM. Accurate interpretation can be critical in the diagnosis of DPM, but variations in antibody performance may impact interpretation. The objectives of this study were to compare the performance of MTAP monoclonal antibodies (mAbs) EPR6893 and 1813 and to compare MTAP expression by IHC with 9p21 copy number status in DPM. Cytoplasmic expression of MTAP IHC with mAbs EPR6893 (ab126770; Abcam) and 1813 (NBP2-75730, Novus Biologicals) was evaluated in 56 DPM (47 epithelioid, 7 biphasic, and 2 sarcomatoid) profiled by targeted next-generation sequencing. 9p21 Copy number status was assessed by Fraction and Allele-Specific Copy Number Estimates from Tumor Sequencing (FACETS) analysis and also by CDKN2A fluorescence in situ hybridization in discrepant cases when material was available. MTAP mAb 1813 showed stronger immunoreactivity, more specific staining, and no equivocal interpretations compared to mAb EPR6893 which showed equivocal staining in 19 (34%) of cases due to weak or heterogenous immunoreactivity, lack of definitive internal positive control, and/or nonspecific background staining. MTAP expression with mAb 1813 showed near perfect agreement with 9p21 copy number by combined FACETS/fluorescence in situ hybridization calls (κ = 0.85; 95% CI, 0.71-0.99; P < .001). MTAP IHC with mAb 1813 was 96% sensitive, 86% specific, and 93% accurate for 9p21 homozygous deletion. The findings of this study suggest that interpretation of MTAP IHC is improved with mAb 1813 because mAb EPR6893 was often limited by equivocal interpretations. We show that MTAP IHC and molecular assays are complementary in detecting 9p21 homozygous deletion. MTAP IHC may be particularly useful for low tumor purity samples and in low-resource settings.

Оценка расстояний между российскими регионами с учетом транспортной инфраструктуры

This study is devoted to assessing the weighted average distances between the regions of Russia according to different metrics – the shortest distances on the sphere, as well as distances by railways and highways. Such aggregated estimates are necessary for researchers of interregional processes (from migration to cargo transportation), but these estimates are not publicly available. The article fills this gap. We describe the methods for estimating distances using various metrics (relying on the Russian Railways data on ‘tariff distances’ between railway stations, information on cities’ coordinates and population, and web-services for calculating inter-city distances). The proposed metrics can be considered as an economic distance metric developed in the context of gravitational interaction studies. We estimate the distances between Russian regions using recent and highly detailed data (the distances between 2840 railway stations, the population data gathered for about 10,000 settlements). For scientific and practical purposes, the open access to resulting estimates is provided. We discuss and interpret the cases of significant discrepancies in the estimates of distances between Russian regions according to different metrics. We demonstrate that, for a number of regions, location of transport infrastructure determines the need to choose the metric of interregional distance carefully, depending on the task. We estimate the weighted distance between all regions of Russia with an account for changes in regional structure of population; on this basis, we show that since 1990s migration has been directed to regions with greater relative transport connectivity

UTILITY OF THE 2023 BETHESDA SYSTEM AUS SUBCATEGORIZATION ON THYROID ASPIRATES.

The Bethesda System for Reporting Thyroid Cytopathology (TBSRTC) is widely used for the standardized reporting of thyroid fine-needle aspiration (FNA) cytology. The 2023 revision of TBSRTC introduced specific subcategories for the classification of atypia of undetermined significance (AUS). This study tests the association between AUS subtypes and malignant diagnoses, namely AUS-nuclear atypia and AUS-other, in archived thyroid - FNA specimens with atypia from 2018-2022 at King Abdulaziz University Hospital. A total of 104 thyroid - FNA specimens with AUS were re-evaluated cytologically and correlated with subsequent surgical outcomes, along with a discussion of discrepant cases.

Development of a deep learning model for automated detection of calcium pyrophosphate deposition in hand radiographs.

Calcium pyrophosphate deposition (CPPD) disease is a leading cause of arthritis, which can mimic or strongly interfere with other rheumatic diseases such as gout, osteoarthritis (OA) or rheumatoid arthritis (RA). In the recently established ACR/EULAR CPPD classification criteria, calcification and OA features of the wrist and hand joints are substantial features. To develop and test a deep-learning algorithm for automatically and reliably detecting CPPD features in hand radiographs, focusing on calcification of the triangular fibrocartilage complex (TFCC) and metacarpophalangeal (MCP)-2 and -3 joints, in separate or combined models. Two radiologists independently labeled a dataset of 926 hand radiographs, yielding 319 CPPD positive and 607 CPPD negative cases across the three sites of interest after adjudicating discrepant cases. CPPD presence was then predicted using a convolutional neural network. We tested seven CPPD models, each with a different combination of sites out of TFCC, MCP-2 and MCP-3. The model performance was assessed using the area under the receiver operating characteristic (AUROC) and area under the precision-recall (AUPR) curves, with heatmaps (Grad-CAM) aiding in case discrimination. All models trialed gave good class separation, with the combined TFCC, MCP-2 and MCP-3 model showing the most robust performance with a mean AUROC of 0.86, mean AUPR of 0.77, sensitivity of 0.77, specificity of 0.80, and precision of 0.67. The TFCC-alone model had a slightly lower mean AUROC of 0.85 with a mean AUPR of 0.73. The MCP-2-alone and MCP-3-alone models exhibited mean AUROCs of 0.78-0.87, but lower mean AUPRs of 0.29-0.47. Heatmap analysis revealed activation in the regions of interest for positive cases (true and false positives), but unexpected highlights were encountered possibly due to correlated features in different hand regions. A combined deep-learning model detecting CPPD at the TFCC and MCP-2/3 joints in hand radiographs provides the highest diagnostic performance. The algorithm could be used to screen larger OA or RA databases or electronic medical records for CPPD cases. Future work includes dataset expansion and validation with external datasets.

Specialist learning curves and clinical feasibility of introducing a new MRI grading system for skeletal maturity.

MRI is an emerging imaging modality to assess skeletal maturity. This study aimed to chart the learning curves of paediatric radiologists when using an unfamiliar MRI grading system of skeletal maturity and to assess the clinical feasibility of implementing said system. 958 healthy paediatric volunteers were prospectively included in a dual-facility study. Each subject underwent a conventional MRI scan at 1.5 T. To perform the image reading, the participants were grouped into five subsets (subsets 1-5) of equal size (n∼192) in chronological order for scan acquisition. Two paediatric radiologists (R1-2) with different levels of MRI experience, both of whom were previously unfamiliar with the study's MRI grading system, independently evaluated the subsets to assess skeletal maturity in five different growth plate locations. Congruent cases at blinded reading established the consensus reading. For discrepant cases, the consensus reading was obtained through an unblinded reading by a third paediatric radiologist (R3), also unfamiliar with the MRI grading system. Further, R1 performed a second blinded image reading for all included subjects with a memory wash-out of 180 days. Weighted Cohen kappa was used to assess interreader reliability (R1 vs consensus; R2 vs consensus) at non-cumulative and cumulative time points, as well as interreader (R1 vs R2) and intrareader (R1 vs R1) reliability at non-cumulative time points. Mean weighted Cohen kappa values for each pair of blinded readers compared to consensus reading (interreader reliability, R1-2 vs consensus) were ≥0.85, showing a strong to almost perfect interreader agreement at both non-cumulative and cumulative time points and in all growth plate locations. Weighted Cohen kappa values for interreader (R1 vs R2) and intrareader reliability (R1 vs R1) were ≥0.72 at non-cumulative time points, with values ≥0.82 at subset 5. Paediatric radiologists' clinical confidence when introduced to a new MRI grading system for skeletal maturity was high from the outset of their learning curve, despite the radiologists' varying levels of work experience with MRI assessment. The MRI grading system for skeletal maturity investigated in this study is a robust clinical method when used by paediatric radiologists and can be used in clinical practice. Radiologists with fellowship training in paediatric radiology experienced no learning curve progress when introduced to a new MRI grading system for skeletal maturity and achieved desirable agreement from the first time point of the learning curve. The robustness of the investigated MRI grading system was not affected by the earlier different levels of MRI experience among the readers.

Evaluation of solid-phase panreactivity with negative direct antiglobulin testing.

Solid-phase red cell adherence (SPRCA) is a sensitive platform for antibody detection, but nonspecific reactions may occur. One pattern of apparent nonspecific reactivity is a panagglutinin with a negative direct antiglobulin test (DAT). The purpose of this study was to define the clinical characteristics of patients with these nonspecific reactions and their associated serologic findings. Twenty patients with panreactive SPRCA testing results were identified between November 2022 and May 2023. In addition to panagglutinins, these patients had (1) a negative polyethylene glycol (PEG) antibody detection test, (2) a negative PEG autocontrol, and (3) a negative DAT. The strength of SPRCA panreactivity and the results of eluate testing (by tube and SPRCA) were studied. Clinical characteristics of patients included age, sex, and primary diagnosis. Each patient was also assessed for evidence of hemolysis. Fourteen female and six male patients were evaluated (average age 44 years). Primary diagnoses included pregnancy (n = 10), acute bleeding (n = 4), orthopedic (n = 3), and other (n = 3). There was no clinical or laboratory evidence of hemolysis. The predominant strength of SPRCA panreactivity was evenly distributed across reaction grades (1+ to 3+). Fifty-five percent of the eluates tested in PEG showed panreactivity, consistent with warm-reactive autoantibodies, while 85 percent of eluates tested by SPRCA were panreactive. Six discrepant cases, in which PEG eluate testing was negative and solid-phase eluate testing showed panreactivity, were associated with weak solid-phase plasma panreactivity (1+). In addition, the reactivity strengths of the eluates tested by SPRCA were invariably more strongly reactive than those eluates tested in PEG. Panagglutination is a distinct SPRCA-only plasma reactivity pattern. Despite a negative PEG tube and DAT, most panagglutinins are warm-reactive autoantibodies. Fortunately, these "interfering" panagglutinins do not appear to be clinically significant and are easily managed by an alternative testing method such as PEG.

HER2 copy number determination in breast cancer using the highly sensitive droplet digital PCR method

Human epidermal growth factor receptor 2 (HER)-positive breast cancer (BC) is characterized by an aggressive clinical course. In the case of HER2 overexpression/amplification, patients benefit from HER2-targeting therapies. Standardized diagnostic HER2 assessment includes immunohistochemistry (IHC) and/or in situ hybridization (ISH). The aim of this study was to compare this “gold standard” with the Droplet Digital™ polymerase chain reaction (ddPCR), a method that allows sensitive and precise detection of copy number variations (CNV) in FFPE (formalin-fixed, paraffin-embedded) DNA samples. Partitioning of the PCR reaction into 20,000 droplets enables a precise quantitative “CN” discrimination also in heterogeneous samples. FFPE breast cancer samples (n = 170) with routinely assessed HER2 status by IHC/ISH were retrospectively analyzed using the ddPCR CNV ERBB2 assay. Comparison of HER2 status assessment by the two methods revealed concordant results in 92.9% (158/170) of the cases. Discrepant cases were verified and interpreted. For ddPCR, a cut off value of 3 HER2 copies was set to distinguish between HER2-negative and HER2-positive BC. Results obtained with the ddPCR CNV ERBB2 assay were consistent and reproducible, and serial dilutions demonstrated a high stability and sensitivity of the method. The ddPCR CNV ERBB2 assay may be a specific and convenient tool to quantify HER2 copy numbers in BC samples. In our study, this method showed high reproducibility in accuracy of HER2 assessment compared to IHC/ISH analysis.

EVALUATING THE UTILITY OF SPECT/CT FOR THE IDENTIFICATION OF CARTILAGE LESIONS: A SYSTEMATIC REVIEW

AbstractObjectiveA common orthopaedic pain found in a wide spectrum of individuals, from young and active to the elderly is anterior knee pain (AKP). It is a multifactorial disorder which is thought to occur through muscular imbalance, overuse, trauma, and structural malalignment. Over time, this can result in cartilage damage and subsequent chondral lesions. Whilst the current gold standard for chondral lesion detection is MRI, it is not a highly sensitive tool, with around 20% of lesions thought to be mis-diagnosed by MRI. Single-photon emission computerised tomography with conventional computer tomography (SPECT/CT) is an emerging technology, which may hold clinical value for the detection of chondral lesions. SPECT/CT may provide valuable diagnostic information for AKP patients who demonstrate absence of structural change on other imaging modalities. This review systematically assessed the value of SPECT/CT as an imaging modality for knee pain, and its ability to diagnose chondral lesions for patients who present with knee pain.MethodsUsing PRISMA guidelines, a systematic search was carried out in PubMed, Science Direct, and Web of Knowledge, CINAHL, AMED, Ovid Emcare and Embase. Inclusion criteria consisted of any English language article focusing on the diagnostic value of SPECT/CT for knee chondral lesions and knee pain. Furthermore, animal or cadaver studies, comparator technique other than SPECT/CT or patients with a pathology other than knee chondral lesions were excluded from the study. Relevant articles underwent QUADAS-2 bias assessment.Results11,982 manuscripts were identified, and the titles were screened for relevance. Seven studies were selected as being appropriate and were subjected to QUADAS-2 assessment. All 7 articles scored low for bias. Two papers deemed that the ICRS score of chondral lesions at intraoperative assessment correlated with SPECT/CT tracer uptake. Two studies concluded that MRI significantly correlated with SPECT/CT tracer uptake, with some instances whereby SPECT/CT identified more chondral lesions than MRI. Two papers compared bone scintigraphy (BS) to SPECT/CT and concluded that SPECT/CT was not only able to identify more chondral lesions than BS, but also localise and characterise the lesions.ConclusionEvidence implies that SPECT/CT may be a useful imaging modality for the detection and localisation of cartilage lesions, particularly in discrepant cases whereby there is an absence of lesions on other imaging modalities, or a lack of correspondence with patients’ symptoms. More studies would be of value to confirm the conclusions of this review.Declaration of Interest(b) declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research reported:I declare that there is no conflict of interest that could be perceived as prejudicing the impartiality of the research project.

Parallel Genomic Analysis from Paired Bone Marrow and Peripheral Blood Samples of 200 Cytopenic Patients

Background: Analysis of bone marrow is the gold standard in patients with unexplainable cytopenia for diagnosing (or excluding) myelodysplastic neoplasm (MDS). Besides morphology also molecular genetics and cytogenetics are currently assessed from BM as both are required for diagnosis and risk stratification. Substitution of BM analysis with peripheral blood (PB) analysis would be a substantial progress especially in the elderly to allow for a less invasive strategy. Aims: Validate, whether PB reliably reflects molecular and cytogenetic aberrations of BM by evaluating somatic mutations and copy number variations (CNVs) in concurrent PB and BM samples. Methods: We analyzed paired BM and PB samples (time difference of sampling: &amp;lt;15 days) of 200 cytopenic patients. All samples were subjected to targeted panel sequencing analyzing mutations (assessed as positive at variant allelic frequencies/VAFs &amp;gt;1%) in 40 genes associated with myeloid neoplasms (coverage: 1500x). If available, BM chromosome analysis (CA) and BM FISH analysis were compared to NGS CNV analysis from PB using a human CNV backbone hybridization panel (coverage: 50x). Results: In 69 patients (35%) no mutations (MUTs) were detected in either BM or PB (Figure 1A). For the remaining 131 patients at least one MUT was found in BM (median MUTs/patient: 2; range 1-10). In 130/131 patients MUTs were detected in both BM and PB (same number of MUTs: n=121; higher number of MUTs in BM than in PB: n=9) while in only one patient harboring a DNMT3A MUT as the sole MUT in BM (VAF: 9%) no MUT was detected in PB. This resulted in a negative predictive value of 98.6% (69/70). Overall, we detected 364 MUTs in 29 of the analyzed genes (BM MUTs: n=362; PB MUTs: n=349) and a concordance between BM and PB of 95% (347/364; Figure 1A). The median VAF was lower in PB compared to BM across all MUTs (median VAF: 20% vs. 12%; p&amp;lt;0.001). All MUTs detected in BM at VAF ≥13% (n=229) were also detected in corresponding PB. Next, we compared CNVs detected by PB NGS to BM karyotype (KT). For 119/200 patients PB NGS including a CNV-backbone panel was available. BM CA revealed a normal KT in 71 and an aberrant KT in 48 patients. In total, 126 aberrations were detected in BM CA including 7 balanced aberrations that can technically not be detected using the CNV backbone panel. CNV analysis from PB revealed 68 aberrations (of 119 detectable BM CNVs; 54%) with 64 being concordant with BM CA. In total, in 88/119 patients (74%) concordant results were found in BM and PB (70/71 with normal and 18/48 with aberrant BM KT). In 3 patients (2.5%) different CNVs were detected (1/3: no CNV in BM CA), while in 28 (23.5%) either fewer CNVs (n=8; all complex KTs) or no CNVs (n=20) were detected in PB compared to BM. The majority (76%) of BM CNVs not detected by PB NGS with available BM FISH data were small BM clones (clone size ≤30%) and therefore not reliably captured by NGS PB with a technical sensitivity of 30%. Interestingly, 4 of the discordant patients with aberrant BM KT did not harbor any somatic MUT in BM and PB. Regarding the IPSS-R cytogenetic risk group, 95 (80%) patients fell into the same category, while 22 (18%; all with aberrant BM KT) were assigned to a better risk category by NGS PB than based on BM CA data and 2 patients (2%) to a worse risk category by NGS PB (Figure 1B). Conclusion: In cytopenic patients mutations can be reliably detected from PB showing a concordance with BM of individual mutations of 95% and a consistency regarding overall mutation detection between PB and BM in 99.5% of patients. Cytogenetic risk group assignment based on NGS CNV analysis from PB compared to chromosome analysis from BM resulted in the same risk group in 80% of patients. However, to further decrease the number of discrepant cases, the technical limitations (sensitivity, inability to detect balanced CNVs) of the CNV backbone method might be overcome by increasing the sequencing depth and/or using CD34-enriched PB samples and adding probe sets. Alternatively, FISH on PB smears could be used for CNV detection. Overall, our data indicate a high degree of overlap between PB and BM regarding mutational and cytogenetic analysis in patients with unexplained cytopenia and suggest PB as reliable surrogate especially in patients without any genetic aberrations.

214 Angiolymphoid hyperplasia with eosinophilia and kimura disease: a case of clinical and histopathological discrepancy and response to dupilumab

214 Angiolymphoid hyperplasia with eosinophilia and kimura disease: a case of clinical and histopathological discrepancy and response to dupilumab

The initial results of ankylosing spondylitis treatment by total hip arthroplasty

Objective: To evaluate the initial results of ankylosing spondylitis treatment by total hip arthroplasty. Subject and method: This is a clinical descriptive study, with vertical follow-up and no control group study. 10 male patients with ankylosing spondylitis including 13 hips were operated with non-cemented total hip arthroplasty at the Joint Surgery Department of 108 Military Central Hospital from May 2018 to May 2022. The age range was 21 to 58 (average 35.6). The mean follow-up period was 33 months (ranged from 6 to 56 months). Result and conclusion: Complications include: 1 case of femoral neck fracture, 8/13 cases of thigh pain 12 months after operation, 3 cases of leg-length discrepancy, 2 cases of heterotopic ossification, 1 case of rapid subsidence. The average score according to the Merle d`Aubigné-Postel scale improved significantly (17.55 ± 0.76 points after 12 months compared with 2.33 ± 1.15 points before surgery, p&lt;0.05).

Comprehensive assessment of mismatch repair and microsatellite instability status in molecular classification of endometrial carcinoma

Objective: To explore the concordance and causes of different mismatch repair (MMR) and microsatellite instability (MSI) detection results in endometrial carcinoma (EC) molecular typing. Methods: A total of 214 EC patients diagnosed from January 2021 to April 2023 were selected at the Department of Pathology, Peking University Third Hospital. The immunohistochemistry (IHC) results of MMR protein were reviewed. Tumor specific somatic mutations, MMR germline mutations, microsatellite scores and tumor mutation burden (TMB) were detected by next-generation sequencing (NGS) with multi-gene panel. Methylation-specific PCR was used to detect the methylation status of MLH1 gene promoter in cases with deficient MLH1 protein expression. In cases with discrepant results between MMR-IHC and MSI-NGS, the MSI status was detected again by PCR (MSI-PCR), and the molecular typing was determined by combining the results of TMB and MLH1 gene promoter methylation. Results: (1) In this study, there were 22 cases of POLE gene mutation subtype, 55 cases of mismatch repair deficient (MMR-d) subtype, 29 cases of p53 abnormal subtype, and 108 cases of no specific molecular profile (NSMP). The median age at diagnosis of MMR-d subtype (54 years old) and the proportion of aggressive histological types (40.0%, 22/55) were higher than those of NSMP subtype [50 years old and 12.0% (13/108) respectively; all P<0.05]. (2) Among 214 patients, MMR-IHC test showed that 153 patients were mismatch repair proficient (MMR-p), 49 patients were MMR-d, and 12 patients were difficult to evaluate directly. MSI-NGS showed that 164 patients were microsatellite stable (MSS; equal to MMR-p), 48 patients were high microsatellite instability (MSI-H; equal to MMR-d), and 2 patients had no MSI-NGS results because the effective sequencing depth did not meet the quality control. The overall concordance between MMR-IHC and MSI-NGS was 94.3% (200/212). All the 12 discrepant cases were MMR-d or subclonal loss of MMR protein by IHC, but MSS by NGS. Among them, 10 cases were loss or subclonal loss of MLH1 and (or) PMS2 protein. Three discrepant cases were classified as POLE gene mutation subtype. In the remaining 9 cases, 5 cases and 3 cases were confirmed as MSI-H and low microsatellite instability (MSI-L) respectively by MSI-PCR, 6 cases were detected as MLH1 gene promoter methylation and 7 cases demonstrated high TMB (>10 mutations/Mb). These 9 cases were classified as MMR-d EC. (3) Lynch syndrome was diagnosed in 27.3% (15/55) of all 55 MMR-d EC cases, and the TMB of EC with MSH2 and (or) MSH6 protein loss or associated with Lynch syndrome [(71.0±26.2) and (71.5±20.1) mutations/Mb respectively] were significantly higher than those of EC with MLH1 and (or) PMS2 loss or sporadic MMR-d EC [(38.2±19.1) and (41.9±24.3) mutations/Mb respectively, all P<0.01]. The top 10 most frequently mutated genes in MMR-d EC were PTEN (85.5%, 47/55), ARID1A (80.0%, 44/55), PIK3CA (69.1%, 38/55), KMT2B (60.0%, 33/55), CTCF (45.5%, 25/55), RNF43 (40.0%, 22/55), KRAS (36.4%, 20/55), CREBBP (34.5%, 19/55), LRP1B (32.7%, 18/55) and BRCA2 (32.7%, 18/55). Concurrent PTEN, ARID1A and PIK3CA gene mutations were found in 50.9% (28/55) of MMR-d EC patients. Conclusions: The concordance of MMR-IHC and MSI-NGS in EC is relatively high.The discordance in a few MMR-d EC are mostly found in cases with MLH1 and (or) PMS2 protein loss or MMR protein subclonal staining caused by MLH1 gene promoter hypermethylation. In order to provide accurate molecular typing for EC patients, MLH1 gene methylation, MSI-PCR, MMR gene germline mutation and TMB should be combined to comprehensively evaluate MMR and MSI status.