Diagnosis Research Articles

Considerations for reporting variants in novel candidate genes identified during clinical genomic testing.

Since the first novel gene discovery for a Mendelian condition was made via exome sequencing (ES), the rapid increase in the number of genes known to underlie Mendelian conditions coupled with the adoption of exome (and more recently, genome) sequencing by diagnostic testing labs has changed the landscape of genomic testing for rare disease. Specifically, many individuals suspected to have a Mendelian condition are now routinely offered clinical ES. This commonly results in a precise genetic diagnosis but frequently overlooks the identification of novel candidate genes. Such candidates are also less likely to be identified in the absence of large-scale gene discovery research programs. Accordingly, clinical laboratories have both the opportunity, and some might argue a responsibility, to contribute to novel gene discovery which should in turn increase the diagnostic yield for many conditions. However, clinical diagnostic laboratories must necessarily balance priorities for throughput, turnaround time, cost efficiency, clinician preferences, and regulatory constraints, and often do not have the infrastructure or resources to effectively participate in either clinical translational or basic genome science research efforts. For these and other reasons, many laboratories have historically refrained from broadly sharing potentially pathogenic variants in novel genes via networks like Matchmaker Exchange, much less reporting such results to ordering providers. Efforts to report such results are further complicated by a lack of guidelines for clinical reporting and interpretation of variants in novel candidate genes. Nevertheless, there are myriad benefits for many stakeholders, including patients/families, clinicians, researchers, if clinical laboratories systematically and routinely identify, share, and report novel candidate genes. To facilitate this change in practice, we developed criteria for triaging, sharing, and reporting novel candidate genes that are most likely to be promptly validated as underlying a Mendelian condition and translated to use in clinical settings.

Critical care specialists, the missing link in organ procurement for transplantation

The procurement process for organ donation begins with the identification of potential organ donors in emergency or critical care units (CCU), followed by their clinical evaluation, diagnostic procedures, and therapeutic interventions, mostly conducted in CCUs. It concludes with the request for organ donation and, if accepted, the retrieval of organs. Despite most interventions occurring in detection units, there has been a neglect of the strategic role played by critical care specialists (CCS) in managing and caring for brain-dead or near-brain-death patients. Questions arise: Are they willing to undertake this responsibility? Do they fully comprehend the nature of organ procurement? Are they aware of the specific interventions required to maintain possible organ donors in optimal physiological condition? Our objective is to examine the role of CCS in organ procurement and propose ways to enhance it, ultimately aiming to increase and enhance organ donation rates.

Improving Breast Cancer Detection with Convolutional Neural Networks and Modified ResNet Architecture.

The pathogenesis of breast cancer is characterized by dysregulated cell proliferation, leading to the formation of a neoplastic mass. Conventional methodologies for analyzing carcinomatous distal areas within whole-slide images (WSIs) tissue regions may lack comprehensive insights. This study aims to introduce an innovative methodology based on convolutional neural networks (CNN), specifically employing a CNN Modified ResNet architecture for breast cancer detection. The research seeks to address the limitations of existing approaches and provide a robust solution for the comprehensive analysis of tissue regions. The dataset utilized in this study comprises approximately 275,000 RGB image patches, each standardized at 50x50 pixels. The CNN Modified ResNet architecture is implemented, and a comparative evaluation against diverse architectures is conducted. Rigorous validation tests employing established performance metrics are carried out to assess the proposed methodology. The proposed architecture achieves a notable 89% accuracy in breast cancer detection, surpassing alternative methods by 2%. The results signify the efficacy and superiority of the CNN Modified ResNet model in analyzing carcinomatous distal areas within WSIs tissue regions. In conclusion, this study demonstrates the potential of the CNN Modified ResNet architecture as an effective tool for breast cancer detection. The enhanced accuracy and comprehensive analysis capabilities make it a promising approach for advancing the understanding of neoplastic masses in WSIs tissue regions. Further research and validation could solidify its role in clinical applications and diagnostic procedures.

Effects of fixation and demineralization on histomorphology and DNA amplification of canine bone marrow.

Fixation and demineralization protocols for bone marrow (BM) across diagnostic laboratories are not standardized. How different protocols affect histomorphology and DNA amplification is incompletely understood. In this study, 2 fixatives and 3 demineralization methods were tested on canine BM samples. Twenty replicate sternal samples obtained within 24 hours of death were fixed overnight in either acetic acid-zinc-formalin (AZF) or 10% neutral-buffered formalin (NBF) and demineralized with formic acid for 12 hours. Another 53 samples were fixed in AZF and demineralized with hydrochloric acid for 1-hour, formic acid for 12 hours, or ethylenediamine tetraacetic acid (EDTA) for 24 hours. Histologic sections were scored by 4 raters as of insufficient, marginal, good, or excellent quality. In addition, DNA samples extracted from sections treated with the different fixation and demineralization methods were amplified with 3 sets of primers to conserved regions of T cell receptor gamma and immunoglobulin heavy chain genes. Amplification efficiency was graded based on review of capillary electrophoretograms. There was no significant difference in the histomorphology scores of sections fixed in AZF or NBF. However, EDTA-based demineralization yielded higher histomorphology scores than demineralization with hydrochloric or formic acid, whereas formic acid resulted in higher scores than hydrochloric acid. Demineralization with EDTA yielded DNA amplification in 29 of 36 (81%) samples, whereas demineralization with either acid yielded amplification in only 2 of 72 (3%) samples. Although slightly more time-consuming and labor-intensive, tissue demineralization with EDTA results in superior morphology and is critical for polymerase chain reaction (PCR) amplification with the DNA extraction method described in this article.

Isolation, identification, antibiotic resistance profile and molecular analysis of Ornithobacterium rhinotracheal isolates from turkeys.

Ornithobacterium rhinotracheal (ORT) infects numerous birds, particularly chickens and turkeys. ORT is an emerging bacterial pathogen of global concern in the poultry industry. As ORT is rapidly spreading throughout commercial poultry, it requires intensive studies of its epidemiology, diagnostic procedures, molecular typing, virulence genes and antimicrobial resistance. The present study was conducted in isolation and identification of ORT from slaughtered turkeys. Cleft palate swabs of 200 were collected from slaughtered turkeys and cultured on blood agar. ORT was characterized using biochemical tests and PCR targeting the ORT 16S rRNA gene. Virulence genes of isolates were determined targeting adenylate kinase (adk), copA and virulence-associated protein D (vapD) genes. Additionally, diversity of ORT isolates was performed by enterobacterial repetitive intergenic consensus (ERIC) and RAPD PCR. Disk diffusion was used to determine the antibiotic sensitivity of the isolates. ORT was identified in 23 (11.5%) samples using both the biochemical tests and PCR. The result of detecting virulence genes showed that all the isolates (23: 100%) had the adk gene, whereas two (8.7%) isolates had the copA gene, and seven (30.43%) isolates had the vapD gene. Molecular typing of isolates revealed 21 different patterns by RAPD PCR assay using M13 primer and 20 distinct patterns by ERIC PCR test. Both ERIC and RAPD PCR were distinctive methods for investigating the genetic diversity of ORT isolates. The antibiotic resistance test showed that 18 (78.26%) isolates were resistant to gentamicin, amikacin, cefazolin, streptomycin and penicillin. All isolates (100%) were resistant to cloxacillin and fosfomycin. This study showed the prevalence of ORT in turkey and high resistance of this bacterium to many common veterinary antibiotics. Moreover, both ERIC and RAPD PCR are distinctive methods for investigating the genetic diversity of ORT isolates. These data may help monitor antibiotic resistance and typing of ORT in epidemiological studies and serve as the foundation for designing region-specific vaccines for future use.

Anesthesiologist-directed care for elective gastrointestinal endoscopy: results of an Italian multicentric prospective observational study.

Sedation, ranging from minimal, moderate and deep sedation to general anesthesia, improves patient comfort and procedure quality in gastrointestinal endoscopy (GIE). There are currently no comprehensive recommendations on sedation practice in diagnostic and therapeutic GIE. We aimed to investigate real-life sedation practice in elective GIE. We performed a multicentric observational study across 14 Endoscopy Units in Italy. We recorded consecutive data on all diagnostic procedures performed with Anesthesiologist-directed care (ADC) and all therapeutic procedures performed with ADC or non-Anesthesiologist sedation (NAS) over a three-month period. Dedicated ADC is available five days/week in 28.6% (4/14), four days/week in 21.5% (3/14), three days/week in 35.7% (5/14), two days/week in 7.1% (1/14) and one day/week in 7.1% (1/14) of participating Centers. ADC use for elective diagnostic GIE varied from 18.2% to 75.1% of the total number of procedures performed with ADC among different Centers. ADC use for elective therapeutic GIE varied from 10.8% to 98.9% of the total number of elective therapeutic procedures performed among different Centers. Our study highlights the lack of standardization and consequent great variability in sedation practice for elective GIE, with ADC being potentially overused for diagnostic procedures and underused for complex therapeutic procedures. A collaborative effort involving Endoscopists, Anesthesiologist and Institutions is needed to optimize sedation practice in GIE.

Functional state of the kidneys in patients with acute coronary syndrome against the background of newly identified disorders of carbohydrate metabolism: a multidisciplinary problem

ABSTRACT Background Determining the relationship between cardiovascular risks, carbohydrate metabolism disorders, and renal dysfunction can help in creating new tools for their management and for better interaction of specialists in a multidisciplinary team. The purpose of this study was to determine the functional state of the kidneys and carbohydrate metabolism in patients with acute coronary syndrome without a history of such disorders. Methods 200 patients of the cardiology department of the City Clinical Hospital No. 7 in Almaty were examined using laboratory diagnostics and subsequent statistical data processing. Results Acute coronary syndrome develops in 63% of cases against the background of previous disorders of carbohydrate metabolism – prediabetes (45.5%) and type 2 diabetes mellitus (17.5%). In this group of patients, in the presence of disorders of carbohydrate metabolism, in all cases, it is accompanied by acute renal damage. It was noted that diabetes mellitus in newly diagnosed patients actually had a certain duration and occurred much earlier than cardiovascular complications – this was evidenced by an increased level of glycated hemoglobin. Conclusions Such results indicate the need for early diagnosis of cardio-reno-metabolic syndrome in patients with cardiovascular complications, as well as timely administration of drugs that simultaneously have antidiabetic, cardio- and nephroprotective effects.

Changing Dermatologic Procedural Patterns in the Medicare Population: A National Trend Analysis.

Dermatologists are trained in diagnostic and therapeutic procedures for cutaneous lesions, yet comparative trends for basic dermatologic procedures across dermatology providers are lacking at the national level. To trend common dermatologic procedures among general dermatologists, Mohs surgeons, primary care providers or primary care physicians (PCPs), and nonphysician clinicians (NPCs). Longitudinal analysis of 2016 to 2021 Medicare Public Use Files. Aggregate dermatologic procedural volume decreased 3.0%. Procedural volume declined among general dermatologists (-11.7%), Mohs surgeons (-16.7%), and PCPs (-41.7%) but increased among NPCs (+57.5%). The proportion of procedures by general dermatologists decreased substantially for premalignant destructions (-6.2%), skin biopsies and shave removals (-4.7%), and malignant excisions (-4.1%) and more notably in counties that were nonmetro (-7.1%), low in income (-6.1%), and with <4.0 dermatologists per 100,000 population (-7.0%). Aggregate procedural volume decreased across the study period with general dermatologists, Mohs surgeons, and PCPs performing a progressively smaller proportion. The increase in procedures by NPCs may help to address demand but underscores the value of formalized procedural training. The procedural decline by general dermatologists in rural and low-income counties and those with baseline dermatologist shortages may exacerbate existing unmet need. Primary limitation included lack of commercial data.

The importance of multimodal CT examination in stroke mimics diagnosis: design of prospective observational multicentre study.

Non-contrast computed tomography (CT) and CT angiography are the gold standard in neuroimaging diagnostics in the case of suspected stroke. CT perfusion (CTP) may play an important role in the diagnosis of stroke mimics (SM), but currently, it is not a standard part of the stroke diagnostic procedure. The project is a multicentre prospective observational clinical research focused on refining the diagnostics of stroke and stroke mimics (SM) in hospital care. This study aimed to evaluate the degree of specificity and sensitivity of multimodal CT (NCCT, CTA, and CTP) in the diagnosis of SM versus stroke. In this study, we will include 3,000 patients consecutively admitted to the comprehensive stroke centres with a diagnosis of suspected stroke. On the basis of clinical parameters and the results of multimodal CT and magnetic resonance imaging (MRI), the diagnosis of stroke and SM will be established. To clarify the significance of the use of the multimodal CT scan, the analysis will include a comparison of the blinded results for each imaging scan performed by radiologists and AI technology and a comparison of the initial and final diagnosis of the enrolled patients. Based on our results, we will compare the economic indicators and costs that would be saved by not providing inadequate treatment to patients with SM. The expected outcome is to present an optimised diagnostic procedure that results in a faster and more accurate diagnosis, thereby eliminating the risk of inadequate treatment in patients with SM. clinicaltrials.gov, NCT06045455.

Characteristics of Drug Hypersensitivity Reactions in Children: A Retrospective Analysis in an Allergy Outpatient Clinic

Objective: Confirmation of drug hypersensitivity reactions (DHRs) is crucial—as various nonallergic reactions, such as viral infections, in children can mimic such reactions. This study aimed to evaluate the characteristics of children with suspected DHRs applying to an allergy outpatient clinic. Material and Methods: This study involved children who visited our hospital’s pediatric allergy outpatient clinic between April 1 and December 31, 2023, with suspected DHRs . The data of patients analyzed retrospectively. Patient demographics, reaction characteristics, culprit drugs, diagnostic procedures (including skin and/or provocation tests), and final diagnoses were recorded. Results: The study included 163 reactions of 140 patients with 176 suspected drugs. The median age was 7.7 years (interquartile range [IQR]; 5.1–12 years), with an equal gender distribution. Notably, 27.1% of the patients presented with concurrent atopic diseases. The median age at the onset of reaction was 72 months (IQR; 34.5–108 months), with 16% of reactions occurring within hospital settings and the remainder at home. Oral administration accounted for 84.7% of the reactions, with antibiotics being the most common culprit drug group (75.5%). Immediate reactions constituted 41.1% (n = 67) of reactions, while delayed reactions accounted for 58.9% (n = 96). Skin symptoms were predominant (97.5%). DHRs were excluded in 75.5% (n = 123) of reactions but confirmed by diagnostic drug allergy tests in 4.9% (n = 8). Conclusion: A through evaluation of suspected DHRs in children is essential. Despite high suspicion rates, confirmation via diagnostic tests was low, emphasizing the need for referral to specialized clinics and appropriate diagnostics for accurate management.

Phenotypic Screening and Antibiogram of Infectious Ulcerative Keratitis Cases in Tripoli Eye Hospital, Libya

Aims: This study to investigate the most prevalent pathogens that cause corneal ulcer in Tripoli-Libya, and to recognize the antibiogram of the isolated pathogens Study Design: This study was a prospective study performed in eye hospital (Zawyet-Aldahmany eye hospital). Place and Duration of Study: It was conducted at the microbiology laboratory of the hospital over a year from December 2018 to January 2020, The overall size of the sample enrolled in this study was 50 inpatients in the admission department, which was taken according to the incidence of the corneal ulceration cases admitted to the hospital. Methodology: The corneal scrapings and swabs were collected and subjected to microbiological examination. Demographic data and risk factors were collected for each patient. The antibiotic susceptibility of the bacterial microorganisms was determined by the Kirby-Bauer disk diffusion method, to identify the most appropriate antibiotic for each bacterial isolate. Results: 60% of the patients were males, the mean age of the patients was 46.5 years, unilateral right eye keratitis was involved in (54%) of cases, trauma was the major risk factor (24%), followed by systemic disease (14%). Concerning the seasonal distribution of the cases between months, our study revealed that summer and winter seasons in Libya (January, July, August, and September) involved most of the cases, and interestingly, fungal keratitis (Aspergillus) was higher in (July, August), which is the summer season in Libya. The culture positive rate was (80%), of which (36%) were bacterial, and (22%) were fungal pathogens, and the other (22%) were mixed fungal and bacterial cultures. The most common bacterial isolate was (26%) P.aeruginosa and (16%) staphylococcus aureus, while Aspergillus (40%) was the most common fungal isolate, followed by Penicillium and fusarium species. Our results showed that P.aeruginosa was the responsible microorganism for rapid keratitis progression in three cases who were using cosmetic lenses promptly. Concerning antibiotic sensitivity, all gram-negative bacteria and positive isolates were susceptible to old-generation fluoroquinolones such as ciprofloxacin and levofloxacin, while ceftazidime showed resistance to penicillins. Conclusion: In conclusion, with early suspicion and microbiological diagnostic procedures, appropriate therapy can only reduce the incidence of this threatening ocular case.

Pitfalls in gynecological cytology - review of the common and less frequent entities in Pap test.

Pitfalls in Pap test could be defined as false positive, false negative or underdiagnosed results which can lead to unnecessary diagnostic procedures or delayed and inadequate treatment. It can be a consequence of misinterpretation of certain morphological entities which are described in this paper. the paper presents the overview of the morphological features and look-alikes of the common sources of pitfalls such as atrophy, repair, IUD change, tubal metaplasia, hyperchromatic crowded groups, and radiation changes. Rare causes of pitfalls such as Arias-Stella changes, pemphigus, tumor diathesis per se, rare types of cervical cancer, including verrucous and papillary squamous cell cancer, gastric type and endometrioid adenocarcinoma are also described. The awareness of pitfalls in cervical cytology is important for cytopathologists and clinicians to avoid future errors. Review of Pap tests with erroneous diagnosis is important for quality control in cytology laboratory, and it must be considered as educational and experience building procedure. Cytopathologist should not pull back in significant diagnoses, especially in HPV negative cases.

Reference Material Production and Milk Protein Concentration as Elements to Improve Bluetongue Serological Diagnosis in Bulk Tank Milk.

The serological surveillance of bluetongue in bulk tank milk is an efficient and cost-effective method for the early detection of bluetongue virus incursions in unvaccinated free areas of the disease. In addition, the availability of standardized and reliable reagents and refined diagnostic procedures with high sensitivity and specificity are essential for surveillance purposes. However, no available reference materials for bluetongue virus serological surveillance in bulk tank milk exist. This study shows the production and characterization of reference material for the implementation of a commercially available bluetongue milk ELISA test in official laboratories, as well as the evaluation of a procedure to increase the sensitivity in samples with low levels of antibodies. This procedure, based on milk protein concentration, allowed us to notably increase the ELISA test's analytical sensitivity, which is useful for milk samples from farms with low within-herd prevalence or pools of bulk tank milk samples. The standardized milk reference material produced here, together with the evaluated procedure to improve analytical sensitivity, could be applied as tools to ensure an accurate diagnosis by official laboratories in bluetongue unvaccinated free areas.

EXPERTISE OF A SPECIALISED FIXTURE WITH A VARIABLE VERTICAL PUSHING EFFECT FOR DIAGNOSTICS OF MEASURING DEVICES

Sensor diagnostics is an increasingly hot topic nowadays. It helps to improve their operational accuracy, analyze and evaluate their lifetime and monitor wear rates. A diagnostic laboratory has been constructed for force-measuring sensors with the ambition of future experiments. A current topic is the monitoring of the influence of deformation of progressive materials in the measurement and diagnostic process. The deformation and elasticity of these materials is evaluated by measuring the forces in the diagnostics laboratory. To perform the experiment, a specialized jig was made in which measurement segments of different materials were implemented. The main objective of the experiment is the expertise of the influence of elastic material in diagnostics. The expected logical negative factor of using flexible segments is the reduction of the measurable range due to the deformation of the flexible element. An expected important conclusion of the experiment is the increase of the sensitivity and especially the stability of the measurement. The aim of the expertise is to quantify the percentage values of the measurement deviations.

Changes in echocardiographic indices and left ventricular strain values by two-dimensional speckle-tracking echocardiography following pre-anesthetic oral pimobendan administration compared with intravenous pimobendan in dogs.

The effects of pre-anesthetic single-dose oral pimobendan during inhalational anesthesia, including the comparison with the effects of single intravenous pimobendan under anesthesia, remain unexplored. Therefore, this study aimed to determine changes in hemodynamic and echocardiographic parameters induced by pre-anesthetic administration of oral pimobendan under isoflurane general anesthesia and to compare them with those induced by intravenous pimobendan. Thirteen clinically normal dogs (4 laboratory and 9 client-owned dogs) with no clinical signs and not on any medical treatment were included. Anesthesia was performed three times: no pimobendan (Control), oral pimobendan (PIMO PO, 0.3 mg/kg), and intravenous pimobendan (PIMO IV, 0.15 mg/kg). Echocardiographic and hemodynamic parameters were monitored at 30-min intervals in all groups. Compared to the Control group, end-systolic volume index (ESVI) and normalized left ventricular internal diameter at end-systole (LVIDSN) were significantly lower, and fractional shortening (FS) and ejection fraction (EF) were significantly higher in the PIMO PO and IV groups (p < 0.001). Global radial strain (GRS) was significantly higher in the PIMO PO and IV groups (p = 0.015). Under general anesthesia, oral pimobendan preserved LV systolic and myocardial function in a manner comparable to intravenous pimobendan. Pre-anesthetic administration of oral pimobendan can be used to compensate for cardiac systolic function in dogs who require therapeutic and diagnostic procedures under general anesthesia with potential risk of circulatory failure.

Process improvement in thyroid fine needle aspiration: Standardizing number of smears for enhanced adequacy and diagnosis.

Fine-needle aspiration (FNA) is a safe, cost-effective diagnostic procedure used in the evaluation of thyroid nodules. The number of thyroid FNAs has dramatically increased over the past few years. In the absence of standardized procedures regarding the number of needle passes needed for diagnosis and the lack of clarity on the use of conventional smears (CS) versus liquid-based preparations (LBP), the demand of thyroid FNAs has led to increased workload on cytology laboratories, which can negatively affect patient safety. We implemented a standardized two needle passes for CS and collection of all needle rinses and additional pass material in CytoRich Red for ThinPrep LBP and compared the non-diagnostic and diagnostic rates before and after this intervention. There were 290 pre-intervention cases and 348 post-intervention cases; of which, there were 17 (5.9%) non-diagnostic cases of the pre-intervention group and 27 (7.8) non-diagnostic cases of the post-intervention group. There was no statistically significant difference in non-diagnostic and diagnostic rates before and after the change (p = 0.347 by two-tailed Z test).

Utilizing deep learning for automated detection of oral lesions: A multicenter study

ObjectivesWe aim to develop a YOLOX-based convolutional neural network model for the precise detection of multiple oral lesions, including OLP, OLK, and OSCC, in patient photos. Materials and methodsWe collected 1419 photos for model development and evaluation, conducting both a comparative analysis to gauge the model’s capabilities and a multicenter evaluation to assess its diagnostic aid, where 24 participants from 14 centers across the nation were invited. We further integrated this model into a mobile application for rapid and accurate diagnostics. ResultsIn the comparative analysis, our model overperformed the senior group (comprising three most experienced experts with more than 10 years of experience) in macro-average recall (85 % vs 77.5 %), precision (87.02 % vs 80.29 %), and specificity (95 % vs 92.5 %). In the multicenter model-assisted diagnosis evaluation, the dental, general, and community hospital groups showed significant improvement when aided by the model, reaching a level comparable to the senior group, with all macro-average metrics closely aligning or even surpassing with those of the latter (recall of 78.67 %, 74.72 %, 83.54 % vs 77.5 %, precision of 80.56 %, 76.42 %, 85.15 % vs 80.29 %, specificity of 92.89 %, 91.57 %, 94.51 % vs 92.5 %). ConclusionOur model exhibited a high proficiency in detection of oral lesions, surpassing the performance of highly experienced specialists. The model can also help specialists and general dentists from dental and community hospitals in diagnosing oral lesions, reaching the level of highly experienced specialists. Moreover, our model’s integration into a mobile application facilitated swift and precise diagnostic procedures.

Immune subtyping of ER-positive, PR-negative and HER2-negative breast cancers by immunohistochemistry: Molecular basis and clinical relevance.

e12553 Background: The diversity and plasticity behind ER+/PR−/HER2− breast cancer (BC) remain underexplored. In our previous work, an immune subtype classification for ER+/PR-/HER2- BC with RNA-seq data was established to distinguish between immune-hot and -cold tumors to provide guidance for immune checkpoint inhibitor (ICI) therapy. However, considering the economic costs, the clinical application of sequencing methods is limited. This study aimed to identify biomarkers of the immune subtype classification and develop a diagnostic procedure for ER+/PR-/HER2- BC immune subtyping based on the immunohistochemistry (IHC) technique, thereby identifying candidates of ICI therapy and refining treatment strategies for ER+/PR-/HER2- BC. Methods: RNA-seq data for 411 patients diagnosed with ER+/PR-/HER2- BC were obtained from the METABRIC database (METABRIC cohort). The immune subtype classification was established by consensus clustering with 1480 immune-related genes. Biomarkers for each immune subtype were identified from differentially expressed genes and validated using logistic regression and ROC curve analysis. Validation of the immune classification was conducted using RNA-seq data from the GEO and TCGA databases. Transcriptomic gene chip sequencing was performed on surgical specimens from 15 ER+/PR-/HER2- BC patients in our center (Chip cohort) to classify the immune subtypes. Biomarkers identified were validated and a diagnostic procedure was designed using IHC techniques. Results: In the METABRIC cohort, immune subtypes including Clusters 1-5 were established. Five biomarkers including DCN, SPP1, CXCL9, STC2, and SRGAP1 were identified, with the AUC values of ROC curves being 0.90, 0.82, 0.99, 0.70, and 0.89, respectively. Notably, Cluster 3 exhibited higher infiltration of CD8+ T cells, B cells, and M1 macrophages, with a greater association with immune activation pathways, thus being defined as the immune-hot subtype. Cluster 1, also showing a moderate immune cell infiltration, was characterized by a more pronounced stromal infiltration. In the Chip cohort, IHC revealed that Cluster 3 exhibited the highest levels of CD8+ T cell and B cell infiltration as well as PD-L1 expression, suggesting a potential benefit from ICI therapy. In contrast, Cluster 1 demonstrated the most extensive infiltration of endothelial and fibroblast cells. The other subtypes were characterized by a suppressed immune microenvironment, especially in Cluster 2, where an infiltration of SPP1+ macrophages was observed. A diagnostic procedure was designed with the biomarkers using IHC techniques. Conclusions: This study validated the immune subtype classification for ER+/PR-/HER2- BC. An easy-to-use IHC-based diagnostic procedure was developed, facilitating the identification of potential ICI therapy candidates in clinical practice at a lower cost.

Diagnostic Strategy Using Color Doppler Ultrasound of Temporal Arteries in Patients With High Clinical Suspicion of Giant Cell Arteritis : A Prospective Cohort Study.

Giant cell arteritis (GCA) is the most prevalent systemic vasculitis in people older than 50 years. Any delay in diagnosis impairs patients' quality of life and can lead to permanent damage, particularly vision loss. To evaluate a diagnostic strategy for GCA using color Doppler ultrasound of the temporal artery as a first-line diagnostic test, temporal artery biopsy (TAB) as a secondary test, and physician expertise as the reference method. Prospective multicenter study with a 2-year follow-up. (ClinicalTrials.gov: NCT02703922). Patients were referred by their general practitioner or ophthalmologist to a physician with extensive experience in GCA diagnosis and management in one of the participating centers: 4 general and 2 university hospitals. 165 patients with high clinical suspicion of GCA, aged 79 years (IQR, 73 to 85 years). The diagnostic procedure was ultrasound, performed less than 7 days after initiation of corticosteroid therapy. Only ultrasound-negative patients underwent TAB. Bilateral temporal halo signs seen on ultrasound were considered positive. Ultrasound and TAB results were compared with physician-diagnosed GCA based on clinical findings and other imaging. Diagnosis of GCA was confirmed in 44%, 17%, and 21% of patients by ultrasound, TAB, and clinical expertise and/or other imaging tests, respectively. Their diagnosis remained unchanged at 1 month, and 2 years for those with available follow-up data. An alternative diagnosis was made in 18% of patients. The proportion of ultrasound-positive patients among patients with a clinical GCA diagnosis was 54% (95% CI, 45% to 62%). Small sample size, no blinding of ultrasound and TAB results, lack of an objective gold-standard comparator, and single diagnostic strategy. By using ultrasound of the temporal arteries as a first-line diagnostic tool in patients with high clinical suspicion of GCA, further diagnostic tests for patients with positive ultrasound were avoided. Tender "Recherche CH-CHU Poitou-Charentes 2014."

Workforce evaluation in the field of radiation oncology: National data from a limited-resource setting.

e13576 Background: The workforce shortage in the field of radiation oncology is a critical challenge in developing countries. This study's main objective was to evaluate the workload of radiation oncologists (RO) in Armenia. Methods: In this cross-sectional study, the adapted questionnaire from Gunasekera et al. (2021) survey was used. The study setting was the only two RO centers operating in Armenia (public and private). For data collection purposes, census sampling was done and self-administered questionnaires were distributed. The data collection time frame was 15-25 July 2023. Descriptive statistics was done to analyze and present data through portions. Results: In total, data on 16 ROs and residents was collected. The ratio of ROs per million population was 6 in 2023, in Armenia. Primary consultations in median lasted 25 minutes (range: 10-40 mins). Similarly, consults of the patients on active treatment lasted in median of 12.5 minutes (min-60 mins). The median duration of follow-up oncology care consults was 7.5 minutes per day (range: 10- 40 mins). In terms of clinic hours, physicians in public settings reported spending a median of 50 hours per week. According to the respondents, on average radiation therapy specialists allocated 50% of their working time to providing direct patient care, 35% to fulfilling administrative responsibilities, 5% to educational and teaching endeavors, and 10% to conducting scientific research. The median radiation therapy delay was 1 week. Namely, treatment delay in the non-metastatic setting was 1․19, and in the metastatic setting 1 week. By the ROs, the most common barriers to delivering cancer care were treatment high cost (68%, n=11), lack of access to imaging (18.7%, n=3) and pathological (18.7%, n=3) diagnostic procedures, and lack of physicians' time to dedicate to patients (31%, n=5). Conclusions: Our study offers some insight into the radiation oncology workload in Armenia. From this research, we conclude that Armenian healthcare is understaffed in terms of radiation oncologists and medical physicists compared to other upper-middle-income and high-income income countries. Moreover, the median daily workload of the ROs exceeds the upper limit of the amount of consults suggested by the international societies. Hence, further research is required for a detailed evaluation of the RO workforce and specific intervention development that will be able to address this issue in Armenia and other resource-limited settings.