Accurate Treatment Research Articles

Modeling and Analysis of Cancer Electrothermic Therapy Technique Based on a Digital Arm

Background: Intramuscular hemangioma (IMH) is a diffuse growth hemangioma located in the striated muscle, which is often overlooked due to its rarity. background: Intramuscular hemangioma (IMH) is a diffuse growth hemangioma located in the striated muscle, which is often overlooked due to its rarity. Objective: This patent pertains to the integration of electroacupuncture with electrothermotherapy. By introducing electrical signals into the electroacupuncture system, electromagnetic heat is produced. This heat leads to the electrolysis and thermal destruction of tumor cells, enabling targeted and precise cancer treatment. Furthermore, the patent offers a theoretical foundation for exploring the distribution of electrical signals and the associated heat in arm muscles, ensuring accurate treatment. Material and Method: To enable subsequent experimental validation, this patent integrates human anatomy and histological structure theory. The arm's geometric structure was derived from segmentation, reconstruction, and substantiation based on a digital human image dataset. Using the COMSOL Multiphysics 5.5 software, a semi-detailed finite element model was developed for the numerical simulation of electrothermotherapy. Within a time domain setting, a carrier signal of 1 MHz and 22 V was introduced to assess the distribution of electrical signals and the associated heat in the arm muscle. Results: Electrical signals, electromagnetic heat, and tissue necrosis primarily concentrate in a spherical region within 10 mm of the exposed electroacupuncture tip, with the maximum coupling temperature reaching 250 °C at the tip. Time domain analysis revealed that the coupling temperature can rise within 1 min, sufficiently to cause complete tissue damage, with the tissue necrosis ratio reaching 100% in the same timeframe. While the coupling temperature continues to rise over time, the increment is modest. After 5 min, there is negligible temperature change, and once the tissue necrosis ratio reaches 100%, it remains consistent. Conclusion: In the precision tumor treatment system utilizing electrothermotherapy, factors, such as the magnitude of the injected electrical signal, placement of the electroacupuncture tip, and treatment duration play a crucial role in the treatment's accuracy. This model delves into the treatment of intramuscular hemangiomas using electroacupuncture electrothermotherapy from two perspectives: spatial and temporal domains. It provides a theoretical foundation for precise electrothermotherapy in cancer treatment. conclusion: In the precise tumor treatment system with electrothermotherapy, the size of the injected electrical signal, the position of the electroacupuncture tip and the treatment duration all have a significant impact on the precise treatment. This model is used to analyzes the treatment of intramuscular hemangiomas with electroacupuncture electrothermotherapy from two aspects: spatial and temporal domains, providing a theoretical basis for the precise electrothermotherapy for cancer. other: null

Sensing pain: Embodied knowledge in endometriosis.

The article explores how sensations of pain are turned into embodied knowledge in endometriosis, a chronic gynaecological illness characterized by persistent, possibly paralysing pain. While previous studies have shown how people with endometriosis struggle to achieve accurate diagnosis and effective treatment, we examine the ways in which some of these difficulties are rooted in the complexities of embodied experiences of endometriosis pain and the challenges of translating the sensed patterns and shifts in pain into a language acknowledged within a clinical setting. Building on a phenomenologically inspired approach to chronic pain and drawing on interviews with people diagnosed with endometriosis in Finland, we examine how our interlocutors use their embodied sensations of pain to adapt to the evolving biomedical and lived surroundings in which their pain is evaluated and managed. The analysis shows how living with chronic pain involves constantly attuning to the multitude of symptoms as well as developing personal strategies of communicating sensations of pain to gain medical recognition and care. We argue that while the lived complexities of the body with endometriosis may fall outside the scope of medical practices of measuring, such complexities nevertheless require medical acknowledgment and careful attention.

Dental bitewing radiographs segmentation using deep learning-based convolutional neural network algorithms.

Dental radiographs, particularly bitewing radiographs, are widely used in dental diagnosis and treatment Dentalimage segmentation is difficult for various reasons, such as intricate structures, low contrast, noise, roughness, andunclear borders, resulting in poor image quality. Recent developments in deep learning models have improvedperformance in analyzing dental images. In this research, our primary objective is to determine the most effectivesegmentation technique for bitewing radiographs based on different metrics: accuracy, training time, and thenumber of training parameters as a reflection of architectural cost. In this research, we employ several deep learning models, namely Resnet-18, Resnet-50, Xception, InceptionResnet v2, and Mobilenetv2, to segment bitewing radiographs. The process begins by importing the radiographsinto MATLAB®(MathWorks Inc), where the images are first improved, then segmented using the graph cut methodbased on regions to produce a binary mask that distinguishes the background from the original X-ray. The deep learning models were trained on 298 and 99 radiograph training and validation sets and were evaluatedusing 99 images from the testing set. We also compare the segmentation model using several criteria, includingaccuracy, speed, and size, to determine which network is superior. Furthermore, we compare our findings with priorresearch to provide a comprehensive understanding of the advancements made in dental image segmentation. Theaccurate segmentation achieved was 93.67% and 94.42% by the Resnet-18 and Resnet-50 models, respectively. This research advances dental image analysis and facilitates more accurate diagnoses and treatment planning bydetermining the best segmentation technique. The outcomes of this study can guide researchers and practitionersin selecting appropriate segmentation methods for practical dental image analysis.

Foreign buyer taxes and housing affordability

AbstractTo improve housing affordability jurisdictions in different countries has introduced taxes on nonresident home buyers. We use the foreign buyer tax introduced in British Columbia, Canada, in August 2016 to investigate the extent to which such taxes improve housing affordability through their effect on local house prices. Our work uses direct transaction‐level identification of foreign buyers that resulted from policies prior to the announcement and subsequent introduction of the tax. Using a difference in differences methodology, we compare house price changes pre‐ and posttax between high and low foreign buyer concentration neighborhoods. We find that house prices decline by 6% in neighborhoods with above median concentrations of foreign buyers after the tax relative to prices in neighborhoods with below median concentrations of foreign buyers. The quantitative effects are also striking with overall foreign buyer share falling from 13.2% of single‐family transactions in the 6 weeks prior to the announcement of the tax to 1.7% for the 3 months following the tax. The unique contribution of this article is our use of transaction‐level data to explicitly identify the properties purchased by foreign buyers to create more accurate control and treatment groups than found in other analyses.

PP21 Are We Ready For It? Developing Criteria To Include Artificial Intelligence Medical Devices (AI-MDs) For Health Technology Assessment

IntroductionThe increasing pace at which artificial intelligence medical devices (AI-MDs) or digital health technologies (DHTs) have been introduced and integrated in healthcare has not been matched with appropriate selection criteria for health technology assessment (HTA) to inform funding decision-making. To align with international best practice and local regulatory guidance, the Agency for Care Effectiveness (ACE) developed criteria to include AI-MDs as part of its 2022 topic prioritization process for medical technologies. This abstract describes ACE’s approach to develop the inclusion criteria.MethodsTo develop key principles for including AI-MDs in ACE’s topic prioritization process, relevant information from overseas HTA agencies, local regulatory guidelines, and ACE’s existing topic selection criteria were reviewed. A search of international HTA agency websites was conducted in September 2022 to identify relevant information on inclusion of AI-MDs in healthcare for reimbursement recommendations.Additionally, local regulatory guidelines for AI-MDs in healthcare were also identified. The inclusion criteria were then piloted with AI-MDs identified from ACE’s horizon scanning workstream to examine their feasibility for HTA topic selection.ResultsOne overseas framework on DHTs from the National Institute for Health and Care Excellence (NICE) and two local regulatory guidelines were identified. Based on the key finding that the purpose of AI-MD use in guiding clinical management and its associated risks were important considerations, the following criteria were developed: (i) full registration with the regulatory body;(ii) device characteristics should be interventional, have direct impact on patient safety, or support accurate diagnosis or treatment which is critical to avoid death and serious health deterioration; and (iii) the AI algorithm should be fixed as opposed to adaptable as per regulatory requirements. Using this inclusion criteria, eight AI-MDs surfaced from horizon scanning were screened with the above criteria and deemed suitable for HTA topic selection.ConclusionsAs AI technologies are increasingly used to replace or supplement current clinical practice, continuous adaptation of HTA method is needed to ensure appropriate topic selection.

Establishment and validation of a prediction model for myocarditis in Chinese children below 14 years old: a protocol for a retrospective cohort study

IntroductionPaediatric myocarditis, a rare inflammatory disease, often presents without clear early symptoms. Although cardiac troponin I levels can aid in diagnosing myocarditis, they are not definitive indicators. Troponin I levels...

Diagnosing and treating acute myelopathies in dogs and cats

Background: A variety of acute spinal cord disorders (myelopathies) can occur in dogs and cats. Understanding the typical clinical presentation and examination findings of the most common acute myelopathies is the key to formulating an accurate diagnostic and treatment pathway in clinical veterinary practice.Aim of the article: This article describes the characteristic clinical features for the most common acute myelopathies in dogs and cats, as well as outlining a diagnostic approach and important treatment considerations to achieve a successful outcome.

Construction and Clinical Application of Digital Intelligent Diagnosis and Treatment System for Hepatocellular Carcinoma

Abstract: In the past 20 years, with the emergence and update of digital intelligent technology, the diagnosis and treatment of hepatocellular carcinoma (HCC) have undergone profound changes. Three-dimensional visualization technology has revolutionized the traditional two-dimensional diagnosis and treatment model of HCC and realized preoperative visualization of tumors and complex liver anatomy. The emergence of ICG fluorescence imaging has realized intraoperative tumor boundary visualization from the molecular and cellular levels. Augmented reality (AR) and mixed reality (MR) technology can realize the three-dimensional visualization of anatomical structures in surgical navigation. Traditional experiential surgery has been transformed into modern intelligent navigation surgery, and surgery has stepped into a new era of digital intelligent technology. In addition, artificial intelligence, molecular imaging and nanoprobes are also expected to achieve early diagnosis and treatment of HCC and improve the prognosis of patients. This article reviews the latest application of digital intelligent diagnosis and treatment technology related to diagnosing and treating HCC, hoping to help achieve accurate diagnosis and treatment of HCC.

Pseudocystic myxoid liposarcoma of the posterior chest wall – a rare case report

Introduction and importance: Liposarcoma (LPS) is a common soft-tissue sarcoma predominantly diagnosed in adults, arising from malignant adipose cells. Among its various subtypes, myxoid LPS (MLPS) stands out as the second most frequent, accounting for ~30% of all LPS cases. This particular subtype typically manifests in males between the ages of 40 and 50 and is commonly found in the lower extremities. Although rare, MLPS may also occur in the head, neck, and infrequently in the back. Chest wall LPS cases are also sparsely reported. Case presentation: In this report, we present a case of MLPS in a 69-year-old male patient who presented with a complaint of firm swelling on the right posterior chest wall, which was progressively increasing in size over the past 10 years. The tumour was located in the posterior chest wall on the left side, and further diagnostic evaluation using computed tomography (CT) and MRI was conducted to identify its characteristics and extent. Clinical discussion: The use of CT scanning plays a crucial role in differentiating between various lipomatous tumour types, aiding in the identification and classification of MLPS. However, MRI emerges as a more effective technique for detecting microscopic fat compared to CT or ultrasonography, providing valuable insights for accurate diagnosis and treatment planning. Conclusion: Surgery remains the primary therapeutic approach for managing LPSs, including MLPS. Adjuvant preoperative radiation is recommended due to its significant sensitivity and potential for improved outcomes. Given the rarity of this presentation and the varied anatomical locations, a multidisciplinary approach is paramount in effectively managing such cases. Medical practitioners should collaborate closely, considering the unique challenges posed by MLPS to ensure optimal patient care and treatment outcomes.

Copy-Move Forgery Detection in Medical Images Using Handcrafted Features

Abstract: In the realm of medical imaging, the authenticity and integrity of images are paramount for accurate diagnosis and treatment planning. Copy-move forgery, a prevalent form of image tampering, poses a significant threat to the reliability of medical images. This research project focuses on the development and implementation of a robust copy-move forgery detection system tailored specifically for medical images. The proposed methodology leverages handcrafted features, extracting distinctive characteristics from the images to detect instances of forgery. Through a meticulous process of feature engineering and selection, the algorithm aims to enhance sensitivity and specificity in identifying manipulated regions within medical images. The study explores the application of advanced image processing techniques and pattern recognition algorithms to achieve a high level of accuracy in forgery detection.

A Case of Ewing’s Sarcoma Arising from Larynx

Ewing sarcoma is a rare tumor in head and neck area. We demonstrate a rare case of sinonasal Ewing sarcoma initially suspected as olfactory neuroblastoma. After the surgery and immunohistochemical studies, it was accurately diagnosed as Ewing sarcoma. We would like to emphasize the possibility that Ewing sarcoma may originate from the head and neck area, and hence, it is important to use appropriate techniques for accurate diagnosis and treatment.

Calibration of Medical Devices: Method and Impact on Operation Quality

Medical devices play a pivotal role in modern healthcare, aiding in accurate diagnosis, monitoring, and treatment of patients. The reliability and accuracy of these devices are indispensable for ensuring patient safety and well-being. This project highlights the critical importance of calibrating medical devices to maintain their accuracy, precision, and performance, ultimately safeguarding the lives of patients. Calibration of medical devices is a systematic process that verifies and adjusts their measurements to a known standard. This process ensures that medical devices consistently provide accurate data and results. In the absence of proper calibration, there is a significant risk of incorrect readings, misdiagnoses, and inappropriate treatment plans, all of which can lead to patient harm. Regular calibration is essential not only for maintaining patient safety but also for meeting regulatory requirements and quality assurance standards. It helps identify and rectify deviations or drifts in device performance, preventing potential hazards in healthcare settings. Compliance with calibration protocols ensures that medical institutions and professionals can deliver reliable and effective care. Moreover, calibrated medical devices enhance the efficiency and cost-effectiveness of healthcare systems by minimizing the need for repeated tests and procedures due to inaccurate results. Patients can have confidence in the reliability of the devices used in their care, fostering trust in healthcare providers and institutions.

An Ethmoid Sinus Ewing Sarcoma with Exophthalmos in A 21-Year-Old Male

Ewing sarcoma is a rare tumor in head and neck area. We demonstrate a rare case of sinonasal Ewing sarcoma initially suspected as olfactory neuroblastoma. After the surgery and immunohistochemical studies, it was accurately diagnosed as Ewing sarcoma. We would like to emphasize the possibility that Ewing sarcoma may originate from the head and neck area, and hence, it is important to use appropriate techniques for accurate diagnosis and treatment.

Comparison of the diagnostic value of [68Ga]Ga-FAPI-04 PET/MR and [18F]FDG PET/CT in patients with T stage ≤ 2a2 uterine cervical cancer: a prospective study.

To compare the diagnostic value of [68Ga]Ga-FAPI-04 PET/MR and [18F]FDG PET/CT in patients with T stage ≤ 2a2 uterine cervical cancer patients. Patients pathologically diagnosed with cervical cancer and with a T stage ≤ T2a2 were prospectively enrolled. All patients underwent whole-body [68Ga]Ga-FAPI-04 PET/MR and [18F]FDG PET/CT within 2weeks, and surgical treatment was performed within 10days after PET. Twenty-five patients were enrolled. Twenty patients underwent radical hysterectomy, among which all of them underwent pelvic lymphadenectomy, and 10 patients underwent para-aortic lymphadenectomy. Three patients received merely laparoscopic lymphadenectomy without hysterectomy. Two patients with both [18F]FDG and [68Ga]Ga-FAPI-04 lymph node high metabolism were staged as FIGO IIIC1r, and concurrent chemoradiation therapy (CCRT) was performed. [18F]FDG and [68Ga]Ga-FAPI-04 had equivalent detection ability on primary tumors, with a positive detection rate of 96.0%. The accuracy of T staging using [18F]FDG and [68Ga]Ga-FAPI-04 was relatively 50% and 55.0%. Elevated and underrated staging was due to misdiagnosis of either vaginal infiltration or tumor size. In terms of lymph node metastasis detection, the specificity of [68Ga]Ga-FAPI-04 was 100% (95% CI, 84.6% ~ 100.0%), which was significantly higher than [18F]FDG (59.1% (95% CI, 36.4% ~ 79.3%)) (p = 0.004). [68Ga]Ga-FAPI-04 PET/MR and [18F]FDG PET/CT demonstrated an equivalent detection ability on cervical cancer primary tumors. However, [68Ga]Ga-FAPI-04 PET/MR's diagnostic value in lymph node metastasis was significantly higher than [18F]FDG PET/CT. [68Ga]Ga-FAPI-04 PET/MR has the potential for more accurate treatment planning, thus clarifying fertility preservation indications for early-stage young patients.

Multifunctional Nanoplatform for NIR-II Imaging-Guided Synergistic Oncotherapy

Tumors are a major public health issue of concern to humans, seriously threatening the safety of people’s lives and property. With the increasing demand for early and accurate diagnosis and efficient treatment of tumors, noninvasive optical imaging (including fluorescence imaging and photoacoustic imaging) and tumor synergistic therapies (phototherapy synergistic with chemotherapy, phototherapy synergistic with immunotherapy, etc.) have received increasing attention. In particular, light in the near-infrared second region (NIR-II) has triggered great research interest due to its penetration depth, minimal tissue autofluorescence, and reduced tissue absorption and scattering. Nanomaterials with many advantages, such as high brightness, great photostability, tunable photophysical properties, and excellent biosafety offer unlimited possibilities and are being investigated for NIR-II tumor imaging-guided synergistic oncotherapy. In recent years, many researchers have tried various approaches to investigate nanomaterials, including gold nanomaterials, two-dimensional materials, metal sulfide oxides, polymers, carbon nanomaterials, NIR-II dyes, and other nanomaterials for tumor diagnostic and therapeutic integrated nanoplatform construction. In this paper, the application of multifunctional nanomaterials in tumor NIR-II imaging and collaborative therapy in the past three years is briefly reviewed, and the current research status is summarized and prospected, with a view to contributing to future tumor therapy.

PATHOLOGICAL HUMERAL FRACTURE FROM BONE METASTASIS OF A GASTROINTESTINAL STROMAL TUMOR: FIRST REPORTED CASE

Background: Gastrointestinal stromal tumor (GIST) is the most prevalent mesenchymal tumor of the GI tract. Metastases commonly occur in the liver and peritoneum, with very rare reported cases of bone metastases, hence, no therapeutic standard has been established for the management of bone metastases. Modern targeted therapy has improved patient survival, and although there are only a small number of reported GIST bone metastases, the incidence will likely continue to increase. Further understanding of GIST bone metastases is required for the implementation of an accurate and effective treatment plan. Case presentation: We report a 56-year-old patient who has a history of bleeding duodenal GIST resected with a Whipple operation. Post-operatively, Imatinib was started, but she developed liver metastasis 6 years later, requiring segmentectomy. She reported pain over the left humerus 2.5 years later and sustained a trivial injury to the left arm, resulting in a distal humerus shaft spiral fracture associated with a radial nerve palsy (Holstein-Lewis injury). Images showed a lytic lesion with cortical erosion and soft tissue extension at the fracture site. Biopsy confirmed GIST bone metastases. She underwent an open reduction and internal fixation with plating and curettage of the lytic lesion and made a good functional recovery afterward. Conclusion: This is the first reported case of a GIST bone metastases pathological Holstein–Lewis fracture. It highlights the potential for bone metastasis to present a long time after the diagnosis of a GIST. Although bone metastases are rare, they are now increasing due to the prolonged survival of patients and the introduction of targeted therapy. It is important to conduct long-term surveillance in specific patient demographics and future studies should be carried out to facilitate the development of a surveillance protocol in patients with GIST.

Antimicrobial resistance rates of urogenital Mycoplasma hominis and Ureaplasma species before and during the COVID-19 pandemic: results from a Greek survey, 2014 to 2022.

The prevalence of antibiotic-resistant urogenital mycoplasmas and ureaplasmas has been gradually increasing over the years, leading to greater concern for accurate diagnosis and treatment. In this study, the antimicrobial resistance trends in Greece were analyzed using 2992 Ureaplasma spp. and 371 M. hominis isolates collected between 2014 and 2022. Antibiotic sensitivity was determined using eight different antimicrobial agents (josamycin, pristinamycin, clindamycin, ofloxacin, azithromycin, tetracycline, erythromycin, and doxycycline), with the data analyzed using descriptive statistical methods. Resistance rates to clindamycin and erythromycin increased for both M. hominis and Ureaplasma spp., while remaining relatively low for Tetracycline, Doxycycline, and Ofloxacin. For Ureaplasma spp., high susceptibility was observed to pristinamycin, tetracycline, doxycycline, azithromycin, and josamycin, and intermediate susceptibility to erythromycin. However, the resistance rate for clindamycin dramatically increased from 60% in 2014 to a peak of 98.46% in 2021, and the erythromycin resistance rate increased from 9.54% in 2018 to 22.13% in 2021. M. hominis exhibited consistently high resistance rates to Erythromycin, while Azithromycin resistance significantly increased over time, from 52.78% in 2017 to 97.22% in 2022. The alarming escalation in antibiotic-resistant urogenital mycoplasmas and ureaplasmas in the Greek population is a significant concern. Antibiotic overconsumption may have played a crucial role in increasing resistance trends. The implementation of nationwide surveillance systems, proper antibiotic stewardship policies, and appropriate culture-based therapy policies are necessary to effectively control this emerging risk.

Clinical Characteristics and Treatment Outcomes of Idiopathic Multicentric Castleman Disease: Data from a Pediatric Cohort

Background: Idiopathic multicentric Castleman disease (iMCD) is a rare, atypical hematologic disorder characterized by diffuse lymphadenopathy and systemic inflammation causing potentially fatal multi-organ system dysfunction. Though the etiology remains unknown, interleukin-6 (IL-6) has been implicated in disease pathogenesis and siltuximab, a monoclonal antibody that antagonizes IL-6, is the only FDA-approved iMCD treatment. Despite substantial morbidity for children with iMCD, the pediatric experience has not been well characterized. It is unclear whether insights gleaned from adults apply to children or whether siltuximab, which was never prospectively studied in a pediatric population, achieves similar response rates in children. We sought to correct this knowledge gap and better characterize pediatric iMCD. Methods: The ACCELERATE longitudinal Castleman disease (CD) natural history registry was interrogated for patients under 18 years of age at time of diagnosis. For each patient, complete medical history and lymph node biopsy slides were collected and extracted, and a panel of clinicians and pathologists performed independent adjudication of the diagnosis. Demographics - including age at diagnosis, histopathologic subtype, clinical subtype, clinical symptoms, and most severe laboratory parameters present within ± 90 days from date of pathological diagnosis were aggregated. Lastly, treatments administered to the cohort of pediatric patients were inventoried and both clinical response (defined as at least 50% reduction in abnormal clinical and laboratory criteria) and durable response (defined as a positive clinical response lasting ≥ 365 days with no other CD treating drug administered in between) were determined. Results: Nineteen pediatric patients were confirmed to have iMCD, of which 14 (74%) were male and 5 (26%) were female (Table 1A). The mean (standard deviation [SD]) age at diagnosis was 13.6 (4.3), with the youngest patient diagnosed at 2 years of age. Much of the cohort demonstrated hypervascular lymph node histology (89.5%). Clinically, 84% of the patients demonstrated the severe TAFRO (thrombocytopenia, anasarca, fever, renal dysfunction/reticulin fibrosis, organomegaly) subtype while 16% were not otherwise specified. 95% of patients presented with constitutional symptoms and all but one patient presented with fatigue and fevers. All 19 patients presented with organomegaly and fluid retention (anasarca, ascites, pleural effusions, and/or peripheral edema). No patients demonstrated CD-specific skin abnormalities (violaceous lymphocytic papules or cherry hemangiomata) or interstitial lymphocytic pneumonitis. Labs indicated significant systemic inflammation, with mean (SD) C-reactive protein, erythrocyte sedimentation rates, and albumin values of 128.3 (102.7) mg/L, 74.5 (42.5) mm/hr, and 2.7 (0.6) g/dL, respectively. Eighteen (95%) patients presented with anemia, with a mean (SD) hemoglobin value of 9.5 (2.2) g/dL. Sixteen (84%) patients demonstrated platelet abnormalities, with 13 patients presenting with thrombocytopenia and three with thrombocytosis. No patient demonstrated hypergammaglobulinemia. Nine patients received three unique siltuximab regimens: siltuximab ± CS (corticosteroids), siltuximab + sirolimus ± CS, and siltuximab + other (other being defined as all other treatments such as rituximab, eculizumab, and ravulizumab) (Table 1B). Siltuximab ± CS was administered to six patients; 83% (n=5) had a positive response, and 60% of responders achieved a durable response. Lastly, two patients with TAFRO died from their disease. Conclusions: Overall, real-world medical record data from a cohort of pediatric iMCD patients enrolled in ACCELERATE reveals high disease burden within this population. A notable degree of TAFRO, the most severe iMCD subtype, was observed within this pediatric cohort, with clinical manifestations and laboratory abnormalities commonly observed in adult iMCD. This makes this population particularly vulnerable and highlights the need for prompt and accurate treatment. We also observed a high response rate to siltuximab ± CS in this small cohort, supporting its use in pediatric iMCD. Further research examining the effectiveness of siltuximab in a larger cohort of pediatric patients as well as other treatments is needed to identify optimal treatment approaches.

Functional Assessment of VWD Type 2N Variants Using Mammalian Cell Displayed Surface-Tethered Von Willebrand Factor

Background von Willebrand factor (VWF) is a large plasma protein critical in hemostasis and thrombosis. Defects in VWF cause the inherited autosomal bleeding disorder von Willebrand disease (VWD). One critical function of VWF is to bind circulating coagulation factor VIII (FVIII) and protect it from rapid clearance. Type 2N VWD is a recessive form of VWD characterized by low FVIII levels due to impairment in VWF-FVIII binding. Type 2N VWD clinically is similar to mild-to-moderate hemophilia A, an X-linked bleeding disorder caused by deficiency in FVIII. It is critical to distinguish type 2N VWD from hemophilia A because the treatments are different. VWF genotyping can inform accurate diagnoses, treatment, and testing in relatives. With the rapid expansion of DNA sequencing, a growing number of DNA variants are being identified in the VWF gene. However, the interpretation of significance of these variants is often challenged by insufficient evidence to inform confident classification regarding pathogenicity. In vitro functional assays provide an avenue to develop the evidence needed to accurately assign functional significance to DNA variants. We adapted multiplexed surface tethering of extracellular proteins (MultiSTEP), our mammalian cell surface display system for secreted proteins, to display VWF and test the functional impact of VWF variants on VWF-FVIII binding. Methods We constructed a VWF cDNA of common VWF coding sequences by mutating rare variants to the more common nucleotides. The regions encoding the VWF propeptide were deleted (∆Pro-VWF). ∆Pro-VWF was cloned into our MultiSTEP cassette that provides a C-terminal flexible linker, Strep II tag, and CD28 transmembrane domain. Three VWF missense variants implicated in type 2N VWD (Table 1) that had varying levels of evidence to support classification of pathogenicity were chosen for study - VWF c.2561 G>A (p.R854Q), c.2572 T>A (p.C858S), and c.2635 G>A (p.D879N). Variants were subcloned into our ∆Pro-VWF cassette and then recombined into the genomically integrated MultiSTEP landing pad in HEK293 Freestyle cells. Following induction of expression, VWF-displaying cells were incubated with recombinant FVIII. VWF expression was measured using a polyclonal anti-VWF antibody, and VWF-FVIII binding was detected with two different monoclonal antibodies to FVIII. Cells were incubated with fluorescently-conjugated secondary antibodies and cell surface VWF and FVIII abundance determined by flow cytometry. Results We show that ∆Pro-VWF robustly displays in our MultiSTEP system and binds FVIII. All three VWF missense variants express VWF at levels similar to ∆Pro-VWF. Relative to ∆Pro-VWF all three variants show marked impairment in VWF-FVIII binding (Figure 1), providing strong in vitro evidence for pathogenicity in type 2N VWD useful in variant reinterpretation (bold column in Table 1). Conclusions We have developed an in vitro mammalian cell surface display system for the interrogation of VWF variants and adapted it to study VWF-FVIII binding. The Pathogenic variant p.R854Q shows decreased VWF-FVIII binding in this assay, as expected. The Likely Pathogenic variant p.D879N and the Variant of Uncertain Significance p.C858S both exhibit a profound loss of VWF-FVIII binding in this assay. Our results provide evidence for reclassification of p.C858S to Likely Pathogenic for type 2N VWD. We expect our system can be used at scale to similarly interrogate the functional significance of a multiplexed library of VWF variants and improve variant interpretation in VWD.

Comparable b2a2/e13a2 and b3a2/e14a2 Reporting in BCR-ABL Assay When Calibrated By Who Is and By IVT-RNA Copy Number

Introduction: Xpert® BCR-ABL Ultra, automated cartridge-based assay for monitoring BCR-ABL transcript levels, is calibrated by WHO IS to standardize the %reporting for both b2a2/e13a2 BCR-ABL(e13a2) and b3a2/e14a2 BCR-ABL(e14a2) relative to control ABL gene in peripheral blood of patients as a standard management of Chronic Myeloid Leukemia (CML) [1]. Studies have shown differences in patients carrying e13a2 or e14a2 in molecular response to Tyrosine Kinase Inhibitor (TKI) treatment [2-4]. Therefore, it is necessary to understand if there is %reporting differences between two transcripts calibrated by WHO IS[5] and explore the calibration method using breakpoint specific RNA input copy number (CN) for accurate TKI treatment monitoring. Objectives: To establish % reporting method calculated with known CN of e13a2, e14a2 and ABL IVT-RNA within a wide CN range and compare % e13a2 and % e14a2 to WHO IS quantification system. Methods: Three IVT-RNAs, containing e13a2-ABL-BCR, e14a2-ABL-BCR and ABL-BCR, were synthesized as shown in Figure 1. Serial diluted e13a2 RNA, e14a2 RNA and ABL RNA panels were tested on Xpert BCR-ABL Ultra assay to generate individual standard curve of e13a2 Ct, e14a2 Ct and ABL Ct as a function of CN for % reporting calculation. Four levels of IVT-RNA panels with same CN of e13a2 and e14a2 were tested for breakpoint specific % reporting comparison. K562(e14a2), BV173 (e13a2) cell lysates and CML clinical samples carrying e13a2 or e14a2 transcripts were tested to evaluate the %CN reporting compared to WHO IS. Result: Good linearity demonstrated in Ct vs CN input for e13a2, e14a2 and ABL IVT-RNA (Figure2) with comparable efficiency (E) between e13a2 (E=0.992) and e14a2 (E=0.986). %e13a2 reporting was ~1.50-fold (by WHO IS) and ~1.46-fold (by %CN) higher than %e14a2, by testing IVT-RNA panel (Table1). Minor differences in %reporting observed between % CN and WHO IS for e13a2 (84.5%~110.8%) vs e14a2 (82.5%~89.5%) from cell lysates (Table2) and e13a2 (92.6%~105.5%) vs e14a2 (88.1%~92.2%) from clinical samples (Table3). Conclusions: A %CN e13a2/ABL and e14a2/ABL reporting method with known CN of IVT-RNAs has been successfully established in Xpert BCR-ABL Ultra. Both WHO IS and %CN showed very minor differences between e13a2 and e14a2 for % reporting. %CN method demonstrated comparable %reporting to WHO IS in Xpert BCR-ABL Ultra. Recent Publications 1. Dominy KM, Simon IM, Khorashad JS (2021) Evaluation of Xpert® BCR-ABL Ultra for the confirmation of BCR-ABL1 international scale conversion factors for the molecular monitoring of chronic myeloid leukaemia, Int J Lab Hematol, Feb;43(1):e31-e34. 2. Hanfstein B, Lauseker M, Hehlmann R, Saussele S, Erben P, Dietz C (2014) Distinct characteristics of e13a2 versus e14a2 BCR-ABL1 driven chronic myeloid leukemia under first-line therapy with imatinib. Haematologica.99:1441-7. 3. Jain P, Kantarjian H, Patel KP, Gonzalez GN, Luthra R, Shamanna RK (2016) Impact of BCR-ABL transcript type on outcome in patients with chronic-phase CML treated with tyrosine kinase inhibitors. Blood.127:1269-75. 4. Castagnetti F, Gugliotta G, Breccia M, Iurlo A, Levato L, Albano F (2017) The BCR-ABL1 transcript type influences response and outcome in Philadelphia chromosome-positive chronic myeloid leukemia patients treated frontline with imatinib. Am J Hematol. 92:797-805. 5. Matthew S, White HE, Hana Z, Nicholas C.P C (2022) Impact of BCR::ABL1 transcript type on RT-qPCR amplification performance and molecular response to therapy. Leukemia 36(7):1-8