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Diagnostics, Volume 16, Issue 6 (March-2 2026) – 139 articles

Cover Story (view full-size image): Pediatric cholestasis is always pathological and requires rapid and accurate diagnosis to prevent irreversible liver damage. However, its broad range of underlying causes makes interpretation challenging, particularly in liver biopsies from neonates and infants. In this work, we present a practical approach to histological diagnosis, integrating clinical, genetic and pathological features to guide differential diagnosis. Through real clinical cases, we highlight key patterns that help distinguish treatable conditions. This framework supports more confident interpretation and contributes to improved diagnostic accuracy and patient outcomes in pediatric liver disease. View this paper
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11 pages, 397 KB  
Article
Serum Mammalian Target of Rapamycin (mTOR) Levels in Patients with Post-COVID-19 Fibrotic-like Lung Changes: A Cross-Sectional Study
by Mert Gebes, Ozge Oral Tapan, Tuba Edgunlu and Emrah Dogan
Diagnostics 2026, 16(6), 958; https://doi.org/10.3390/diagnostics16060958 - 23 Mar 2026
Viewed by 541
Abstract
Background/Objectives: Post-COVID-19 fibrotic-like lung changes (PC19-FLC), which may represent persistent post-inflammatory abnormalities or early fibrotic remodeling, have emerged as an important long-term pulmonary sequela following SARS-CoV-2 infection. However, the underlying pathogenic mechanisms remain incompletely understood. This study aimed to investigate the potential association [...] Read more.
Background/Objectives: Post-COVID-19 fibrotic-like lung changes (PC19-FLC), which may represent persistent post-inflammatory abnormalities or early fibrotic remodeling, have emerged as an important long-term pulmonary sequela following SARS-CoV-2 infection. However, the underlying pathogenic mechanisms remain incompletely understood. This study aimed to investigate the potential association between mammalian target of rapamycin (mTOR) activity and the presence of PC19-FLC. Methods: This single-center, cross-sectional study included 70 patients who met the predefined inclusion criteria. Participants were categorized according to the presence or absence of PC19-FLC on chest computed tomography. Demographic, laboratory, and radiological data were collected. Serum mTOR levels were measured using enzyme-linked immunosorbent assay (ELISA). Results: Serum mTOR levels and modified Medical Research Council (mMRC) dyspnea scores were significantly higher in patients with PC19-FLC compared with those without fibrotic-like changes. Receiver operating characteristic (ROC) curve analysis identified a serum mTOR cut-off value of 6.15 ng/mL (sensitivity 83%, specificity 94%) for discriminating patients with PC19-FLC in this cohort. Serum mTOR levels were significantly correlated with forced vital capacity (FVC%), mMRC dyspnea score, and peripheral oxygen saturation (SpO2). Conclusions: Increased serum mTOR levels were associated with the presence of fibrotic-like lung changes after COVID-19 and may help distinguish patients with such CT abnormalities in this cohort. Higher mTOR levels were also associated with greater dyspnea severity, lower lung volumes, and reduced peripheral oxygen saturation. These findings suggest a potential role of mTOR signaling in post-COVID-19 pulmonary sequelae and warrant further investigation in larger, multicenter studies. Full article
(This article belongs to the Special Issue Diagnosis and Management of Pulmonary Fibrosis)
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12 pages, 687 KB  
Article
Antinuclear Antibodies Predict Treatment Escalation and Biologic Switching in Rheumatoid Arthritis
by Zeynel Abidin Akar, Dilan Yıldırım, Mehmet Çiftçi, Zeynep Işık Sula, Serap Karaman, Remzi Çevik, Mehmet Karakoç, Serda Em, İbrahim Batmaz, Pelin Oktayoğlu and Mehmet Çağlayan
Diagnostics 2026, 16(6), 957; https://doi.org/10.3390/diagnostics16060957 - 23 Mar 2026
Viewed by 465
Abstract
Background: Antinuclear antibodies (ANAs) are frequently detected in patients with rheumatoid arthritis (RA); however, their prognostic relevance for predicting treatment escalation and biologic therapy initiation remains incompletely understood. Identifying biomarkers associated with earlier transition to advanced therapies may enhance individualized, treat-to-target disease management. [...] Read more.
Background: Antinuclear antibodies (ANAs) are frequently detected in patients with rheumatoid arthritis (RA); however, their prognostic relevance for predicting treatment escalation and biologic therapy initiation remains incompletely understood. Identifying biomarkers associated with earlier transition to advanced therapies may enhance individualized, treat-to-target disease management. Objectives: We aimed to evaluate the association of ANA status and titer levels with clinical characteristics, treatment trajectories, and time to biologic therapy initiation in patients with RA. Methods: In this retrospective cohort study, 223 patients with RA were stratified according to ANA status (112 ANA-positive, 111 ANA-negative). Baseline demographic data, disease activity (DAS28), and serological markers (RF, anti-CCP) were analyzed. Time to biologic therapy initiation, defined from the date of RA diagnosis to first biologic or targeted synthetic DMARD use, was assessed using Kaplan–Meier survival analysis and Cox proportional hazards regression. Multivariate models adjusted for clinically relevant covariates (age, sex, disease duration, RF, anti-CCP). Within the ANA-positive group, exploratory analyses compared low–moderate (1:80–1:320) and high (>1:320) ANA titers, highlighting potential non-linear effects. Results: Baseline demographic and clinical characteristics were comparable between groups (all p > 0.05). ANA-positive patients more frequently initiated biologic therapy (48.2% vs. 24.3%, p < 0.001) and experienced multiple biologic switches (29.5% vs. 16.2%, p = 0.028). In multivariate analysis, ANA positivity independently predicted earlier biologic therapy initiation (adjusted HR 2.14; 95% CI 1.32–3.46; p = 0.002), whereas RF and anti-CCP status were not significant predictors. Exploratory subgroup analysis revealed the “titer paradox,” whereby high ANA titers (>1:320) were associated with a lower hazard of biologic therapy initiation compared with low–moderate titers (HR 0.24; 95% CI 0.06–0.98; p = 0.048). Conclusions: ANA positivity serves as an independent prognostic marker for earlier biologic therapy initiation in RA, providing incremental information beyond traditional serological markers. The observed non-linear association between ANA titers and treatment escalation underscores the need for cautious interpretation and validation in prospective, mechanistic studies, and highlights the potential value of integrating ANA profiling into personalized treatment strategies. Full article
(This article belongs to the Section Clinical Diagnosis and Prognosis)
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12 pages, 740 KB  
Article
Acromegalic Uteropathy: Specific Uterine Ultrasound Findings in Female Patients
by Irina Stanoevich, Aurika Asanova, Svetlana Vorotnikova, Andrey Belov, Ekaterina Grezina, Yulia Fedorova, Ugljesa Stanojevic, Larisa Dzeranova, Ekaterina Pigarova, Galina Melnichenko and Natalya Mokrysheva
Diagnostics 2026, 16(6), 956; https://doi.org/10.3390/diagnostics16060956 - 23 Mar 2026
Viewed by 343
Abstract
Background/Objectives: Acromegaly is a systemic connective tissue disease driven by chronic growth hormone (GH) and insulin-like growth factor-1 (IGF-1) excess; yet, the female reproductive tract—especially the extracellular matrix (ECM)-rich cervix—has been poorly studied. We aimed to compare uterine and cervical morphology in women [...] Read more.
Background/Objectives: Acromegaly is a systemic connective tissue disease driven by chronic growth hormone (GH) and insulin-like growth factor-1 (IGF-1) excess; yet, the female reproductive tract—especially the extracellular matrix (ECM)-rich cervix—has been poorly studied. We aimed to compare uterine and cervical morphology in women with acromegaly versus healthy controls and a gynecologic disease comparator, testing the hypothesis of selective cervical hypertrophy. Methods: We performed a retrospective case–control study of reproductive-age women who underwent pelvic ultrasound: acromegaly (n = 33), healthy controls (n = 45), and adenomyosis without acromegaly (n = 44). Uterine body measurements were obtained by TAUS/TVUS; cervical biometry was performed transvaginally in all cases. Volumes were estimated using the ellipsoid formula, and a uterus-to-cervix (U:C) volume ratio was calculated. Group differences were analyzed with Mann–Whitney tests and Bonferroni correction. Results: A total of 122 women were included. Uterine body length, width, AP size, and volume did not differ between acromegaly and either comparison group (all p-values non-significant). In contrast, cervical length, width, AP thickness, and volume were significantly higher in acromegaly than in healthy controls, with a corresponding reduction in the U:C volume ratio, indicating disproportionate cervical enlargement. Compared with adenomyosis, women with acromegaly again showed larger cervical width, AP thickness, and volume, together with altered U:C indices, whereas cervical length did not differ, suggesting a pattern not explained by nonspecific pelvic pathology. Conclusions: Women with acromegaly demonstrate a distinct uterine phenotype characterized by selective cervical hypertrophy with preserved uterine corpus size—an ECM-centric “acromegalic uteropathy.” This noninvasive morphometric signature may have diagnostic and procedural relevance and warrants confirmation in prospective studies. Full article
(This article belongs to the Special Issue Diagnosis and Prognosis of Gynecological and Obstetric Diseases)
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14 pages, 990 KB  
Article
Endocrine Sequelae of Mild Traumatic Brain Injury in Patients Admitted to the Emergency Department: A 12-Month Study
by Maria Kałas, Mariusz Siemiński and Ewelina Stępniewska
Diagnostics 2026, 16(6), 955; https://doi.org/10.3390/diagnostics16060955 - 23 Mar 2026
Viewed by 473
Abstract
Background/Objectives: Over the last two decades, there has been a substantial change in the understanding of post-traumatic hypopituitarism (PTHP), which is no longer regarded as a marginal phenomenon. Clinical manifestations of pituitary hormone deficiency are frequently nonspecific, with fatigue and cognitive dysfunction predominating. [...] Read more.
Background/Objectives: Over the last two decades, there has been a substantial change in the understanding of post-traumatic hypopituitarism (PTHP), which is no longer regarded as a marginal phenomenon. Clinical manifestations of pituitary hormone deficiency are frequently nonspecific, with fatigue and cognitive dysfunction predominating. Given that head injuries currently constitute a global burden for healthcare systems, the aim of the present study was to determine whether self-reported post-mild traumatic brain injury (mTBI) symptoms that may indicate hypopituitarism reflect true pituitary insufficiency or are attributable to other hormonal aberrations. The study aimed to assess the relationship between self-reported symptoms of PTHP and hormonal test results following mTBI. Setting: Patients were recruited from a tertiary trauma center Emergency Department (ED) in northern Poland from January 2023 to October 2025. Participants: The participants were adult (18 > y.o.) individuals with mTBI who met the inclusion criteria. Design: This was a prospective cohort study. During their post-head injury admission to the ED, patients had a blood sample taken. The procedure was repeated consecutively after 3, 6 and 12 months. After 6 and 12 months, patients were asked to complete a questionnaire. Methods: Pituitary and thyroid hormones were measured using the chemiluminescence immunoassay method and the heterogenous immunochemiluminescence method. The questionnaire used, Questionnaire for the Assessment of Symptoms of Anterior Pituitary Insufficiency in Patients After Mild Traumatic Brain Injury (mTBI) Hospitalized in the Emergency Department, was designed for the purposes of this study. Results: Self-reported symptoms suggestive of anterior pituitary dysfunction following mTBI were not confirmed by laboratory assessment of pituitary hormones. However, after 6 months, a statistically significant correlation was found between the number of reported symptoms and prolactin levels (ρ = 0.730; p = 0.0013), whereas after 12 months a downward trend in free triiodothyronine (fT3) levels was observed compared with the baseline. Conclusions: Persistent symptoms reported by patients following mTBI at 6 and 12 months, particularly fatigue and impaired concentration, showed statistical associations with prolactin levels at 6 months and lower fT3 levels at 12 months. These findings reflect correlations identified in the statistical analysis and do not support inferences regarding causality or the presence of true PTHP. Full article
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12 pages, 1203 KB  
Article
Interpretable Machine Learning for Emergency Department Triage: Clinical Insights from 133,198 Patients Using the Korean Triage and Acuity Scale (KTAS)
by MyoungJe Song, Jongsun Kim, Eun-Chul Jang and SoonChan Kwon
Diagnostics 2026, 16(6), 954; https://doi.org/10.3390/diagnostics16060954 - 23 Mar 2026
Viewed by 641
Abstract
Background/Objectives: Emergency room severity classification (KTAS) is essential for patient safety but has limitations due to its reliance on subjective judgment. Existing machine learning models have not been trusted in clinical settings due to their opaque ‘black box’ nature in decision-making processes. To [...] Read more.
Background/Objectives: Emergency room severity classification (KTAS) is essential for patient safety but has limitations due to its reliance on subjective judgment. Existing machine learning models have not been trusted in clinical settings due to their opaque ‘black box’ nature in decision-making processes. To overcome this, this study aims to develop an explainable machine learning framework that provides a transparent basis for judgment with high accuracy. Method: We retrospectively analyzed 133,198 emergency room visits from 2022 to 2024. We trained Random Forest (RF) and XGBoost models using vital signs and pain scores and applied explainable AI (XAI) techniques to ensure model transparency. Results: Although XGBoost showed the highest predictive performance (94.7% accuracy within a ±1 error margin), we ultimately selected the RF model, which provides a good balance of predictive power (91.6%) and interpretability for clinical use. The results of the XAI analysis confirmed that pain score, age, and systolic blood pressure were the key variables in prediction, strongly aligning with clinical logic. Conclusions: This study demonstrates that explainable AI can provide transparent insights for KTAS prediction beyond the limitations of traditional black-box models. These models may support emergency department triage by improving consistency and assisting clinicians in identifying potentially high-risk patients. However, further external validation is required before routine clinical implementation. Full article
(This article belongs to the Section Machine Learning and Artificial Intelligence in Diagnostics)
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25 pages, 5903 KB  
Case Report
The Efficiency of Allotransplant in a Case of Acute Biphenotypic Myeloid and B-Lymphoid Leukemia (MPAL Myelo/B NOS) That Presented Concurrently with a Mediastinal Granulocytic Sarcoma Co-Expressing Lymphoid Markers Complicated by Cardiac Tamponade
by Alina Camelia Catana, Erzebeth Lazar Benedek, Ioan Zaharie, Liliana Mocanu, Geanina Mera, Cristina Popa and Lidia-Maria Mondoc
Diagnostics 2026, 16(6), 953; https://doi.org/10.3390/diagnostics16060953 - 23 Mar 2026
Viewed by 458
Abstract
Background and Clinical Significance: Mixed-phenotype acute leukemia (MPAL) is a rare hematologic malignancy characterized by the co-expression of myeloid and lymphoid markers and is associated with poor prognosis. Myeloid sarcoma (MS), particularly in the mediastinum, is an uncommon extramedullary manifestation and is [...] Read more.
Background and Clinical Significance: Mixed-phenotype acute leukemia (MPAL) is a rare hematologic malignancy characterized by the co-expression of myeloid and lymphoid markers and is associated with poor prognosis. Myeloid sarcoma (MS), particularly in the mediastinum, is an uncommon extramedullary manifestation and is rarely reported in association with MPAL. Case Presentation: We report a rare case of mediastinal MS with biphenotypic features and pericardial extension occurring concurrently with MPAL, highlighting diagnostic challenges, therapeutic strategies, and long-term outcomes. We describe the clinical course, diagnostic workup, treatment, and follow-up of a 21-year-old woman who presented with cardiac tamponade secondary to a mediastinal mass. Histopathology and immunophenotyping established the diagnosis of mediastinal MS associated with MPAL (B/myeloid, NOS). Management included surgical cytoreduction, intensive induction chemotherapy, and consolidation with allogeneic hematopoietic stem cell transplantation (allo-HSCT) from an unrelated donor. Fertility preservation with oocyte retrieval, in vitro fertilization (IVF), and embryo cryopreservation was performed prior to conditioning. A focused literature review of MPAL cases with extramedullary involvement was conducted. The patient achieved complete remission following induction therapy and underwent allo-HSCT. Despite the historically poor prognosis of mediastinal MS and MPAL, she remains in sustained complete remission 13 years after diagnosis. A literature review identified only eight reported cases of MPAL with extramedullary disease, with mediastinal involvement described in a single case and allo-HSCT performed in only two patients. Conclusions: This case illustrates a rare presentation of MPAL with mediastinal myeloid sarcoma and cardiac tamponade, demonstrating that aggressive multimodal therapy including allo-HSCT may achieve durable remission even in high-risk presentations. Early multidisciplinary management and consideration of fertility preservation are essential in young patients. Full article
(This article belongs to the Special Issue Diagnosis and Management of Hematologic Malignancies)
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13 pages, 747 KB  
Article
Pre-Operative Sonographic Assessment of Ovarian Location and Mobility Predicts Intra-Operative Ovarian Resectability During Vaginal Hysterectomy: A Diagnostic Accuracy Study
by Iakovos Theodoulidis, Nikolaos Roussos, Menelaos Zafrakas, Christos Anthoulakis, Pantelis Trompoukis, Grigorios F. Grimbizis and Themistoklis Mikos
Diagnostics 2026, 16(6), 952; https://doi.org/10.3390/diagnostics16060952 - 23 Mar 2026
Viewed by 333
Abstract
Background/Objectives: This study investigates the predictive role of pre-operative sonographic assessment of ovarian mobility in determining intra-operative ovarian resectability among patients undergoing vaginal hysterectomy for pelvic organ prolapse. Methods: This prospective study was conducted in a tertiary academic urogynecology center. Women [...] Read more.
Background/Objectives: This study investigates the predictive role of pre-operative sonographic assessment of ovarian mobility in determining intra-operative ovarian resectability among patients undergoing vaginal hysterectomy for pelvic organ prolapse. Methods: This prospective study was conducted in a tertiary academic urogynecology center. Women with pelvic organ prolapse scheduled for vaginal hysterectomy were consecutively recruited after providing informed consent. Pre-operatively, all patients had a detailed history, pelvic examination (POP-Q), and pelvic floor ultrasound (including assessment of the mobility of both ovaries and sonographic determination of ovarian descent in relation to the pelvic ischial spines). Patients were planned for vaginal hysterectomy, anterior and posterior colporrhaphy, McCall culdoplasty, and bilateral salpingo-oophorectomy (SO), where feasible. During surgery, the location and mobility of the ovaries, as well as the presence of peri-ovarian adhesions, were recorded. Pair-to-pair comparisons between sonographic and clinical findings were performed. Results: From February 2023 to January 2024, 50 Caucasian adult women underwent reconstructive vaginal surgery for prolapse. Thirty-five patients underwent concomitant bilateral SO via vaginal route, seven underwent unilateral SO, and three under went salpingectomy only. ROC analysis indicated that pre-operative ultrasound assessment of ovarian mobility predicts: (1) intra-operative ovarian mobility (sensitivity 95.6%, specificity 77.8%); (2) the presence of peri-ovarian adhesions (sensitivity 46.1%, specificity 94.2%); and (3) resectability, i.e., the ability to perform SO via the vaginal route (sensitivity 96.4%, specificity 50.0%). The absence of ovarian mobility was not associated with an increased risk of intra-operative and post-operative complications. Conclusions: Pre-operative sonographic assessment of ovarian location and mobility can predict ovarian location and resectability during vaginal surgery with high diagnostic accuracy. Full article
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19 pages, 3121 KB  
Systematic Review
Comparative Diagnostic Performance of TST and IGRAs in the Diagnosis of Latent Tuberculosis Infection: A Systematic Review and Diagnostic Meta-Analysis
by Shyamkumar Sriram, Tareq Abualfaraj, Manal Ali Alsharif, Marwa Zalat, Saad Madani Alawfi, Hammad Ali Fadlalmola and Muayad Albadrani
Diagnostics 2026, 16(6), 951; https://doi.org/10.3390/diagnostics16060951 - 23 Mar 2026
Viewed by 651
Abstract
Background: Patients with latent tuberculosis infection are mainly asymptomatic, but they still carry a notable risk of developing active TB, particularly when the host becomes immunosuppressed. Hence, appropriate diagnosis and management for LTBI are essential. Tuberculin skin test (TST) and interferon-gamma release assays [...] Read more.
Background: Patients with latent tuberculosis infection are mainly asymptomatic, but they still carry a notable risk of developing active TB, particularly when the host becomes immunosuppressed. Hence, appropriate diagnosis and management for LTBI are essential. Tuberculin skin test (TST) and interferon-gamma release assays (IGRAs) are among the most commonly utilized methods for detecting LTBI. Until now, no agreement has been established regarding the most effective diagnostic test, either TST or IGRA, so our study aims to evaluate the diagnostic utility of TST versus IGRA in detecting LTBI. Methods: An extensive literature search was executed in several databases from inception till June 2024. We included all the available studies that compared TST versus IGRA concurrently applied to the same study participants, utilizing one of the following proxy reference standards: previous contact with a tuberculosis patient, tuberculosis history, chest x-ray suggestive of tuberculosis, or a combination of them. The sensitivity (SN) and specificity (SP) were imputed with their 95% confidence interval (CI). A bivariate random-effects model within the OpenMeta-Analyst software was utilized for data analysis. Results: We included 39 studies, and our primary analysis regarding LTBI revealed that TST has an SN of 0.320 (95% CI [0.254–0.393]) and an SP of 0.808 (95% CI [0.752–0.854]). Nevertheless, the IGRA exhibited a higher SN estimated at 0.362 (95% CI [0.295–0.434]) and a lower SP estimated at 0.758 (95% CI [0.700–0.808]). Regarding the adult population, TST consistently showed a lower SN and a higher SP relative to IGRA. However, within the pediatric population, TST showed higher SN and lower SP when compared to IGRA. Furthermore, TST also showed a lower SN and a higher SP within hemodialysis and organ transplant patients than IGRA. Conclusions: Our diagnostic test meta-analysis revealed that TST was associated with a lower SN and a higher SP than IGRA. Clinicians should interpret these findings with caution, considering the substantial heterogeneity observed across the included studies, the reliance on proxy reference standards, the potential influence of BCG vaccination status, and the considerable overlap in confidence intervals between TST and IGRA estimates across most analyses. Full article
(This article belongs to the Section Diagnostic Microbiology and Infectious Disease)
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16 pages, 764 KB  
Article
Integrating Tumor Biology and Host Factors in mCRPC: The Prognostic Value of ‘Time to Castration Resistance’, Systemic Inflammation, and Comorbidity Burden in Patients Treated with Enzalutamide
by Seda Sali, Arife Ulaş, Sibel Oyucu Orhan, Sevgi Topçu, Muharrem Koçar, Mürsel Sali, Birol Ocak, Adem Deligönül, Türkkan Evrensel and Erdem Çubukçu
Diagnostics 2026, 16(6), 950; https://doi.org/10.3390/diagnostics16060950 - 23 Mar 2026
Viewed by 407
Abstract
Background: Outcomes with enzalutamide in metastatic castration-resistant prostate cancer (mCRPC) are influenced by tumor burden, disease kinetics, and host factors. We evaluated the relative prognostic impact of metastatic pattern, laboratory markers, and prostate-specific antigen (PSA) dynamics in a real-world cohort. Methods: We retrospectively [...] Read more.
Background: Outcomes with enzalutamide in metastatic castration-resistant prostate cancer (mCRPC) are influenced by tumor burden, disease kinetics, and host factors. We evaluated the relative prognostic impact of metastatic pattern, laboratory markers, and prostate-specific antigen (PSA) dynamics in a real-world cohort. Methods: We retrospectively analyzed 72 patients with mCRPC treated with enzalutamide. Progression-Free Survival (PFS) and Overall Survival (OS) were estimated using the Kaplan–Meier method. Multivariate Cox proportional hazards models were utilized to identify independent predictors of survival, incorporating clinical variables (visceral metastases, bone tumor burden), kinetic parameters (Time to Castration Resistance [TTCR], Time to PSA Nadir [TTN]), and host factors (Charlson Comorbidity Index [CCI], Eastern Cooperative Oncology Group Performance Status (ECOG PS), Systemic Immune-Inflammation Index [SII], HALP score). Results: Visceral metastasis was a dominant predictor of poor outcomes, increasing the risk of death by 4.0-fold (HR: 4.05; 95% CI: 1.84–8.89; p < 0.001). A high skeletal tumor burden (≥5 bone lesions) was identified as a critical threshold, associated with a 5.5-fold increase in mortality risk (HR: 5.53; p < 0.001). Delays in initiating enzalutamide significantly compromised survival, with each 1-month delay increasing the risk of death by 7.3% (HR: 1.07; p = 0.003). While early PSA decline (≥50% at 3 months) did not independently predict OS, a prolonged TTN (>12 months) was associated with superior survival. Notably, host-related factors, including age, CCI, and ECOG PS, were not found to be significantly associated with survival outcomes in this specific dataset. Conclusions: Our preliminary findings suggest that survival in real-world mCRPC patients treated with enzalutamide may be influenced predominantly by intrinsic tumor biology—specifically anatomical extent and resistance kinetics—rather than host frailty or comorbidity burden. However, given the retrospective and single-center nature of this study, these findings should be considered hypothesis-generating and require validation in larger, multi-center cohorts. Host-related variables (including age and CCI) were evaluated but were not retained as independent predictors in the final multivariable model. Early initiation of therapy and monitoring of kinetic markers like TTN and TTCR offer superior prognostic stratification compared to static baseline characteristics. Full article
(This article belongs to the Special Issue Prostate Cancer: Innovations in Diagnosis and Risk Stratification)
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22 pages, 848 KB  
Article
Digital Specimen Tracking- and ISO 15189-Oriented Risk Management in Anatomic Pathology: A Qualitative Study of Expert Perspectives in Western Austria
by Pius Sommeregger, Natalie Pallua, Bettina Zelger, Riem Kahlil and Johannes Dominikus Pallua
Diagnostics 2026, 16(6), 949; https://doi.org/10.3390/diagnostics16060949 - 23 Mar 2026
Viewed by 474
Abstract
Background: Breakpoints in the pre-examination processes and at organizational interfaces are a significant source of failures in specimen identification and tracking in anatomic pathology. While ISO 15189 emphasizes end-to-end traceability and risk-based quality management, implementing these principles in complex, multi-actor specimen pathways [...] Read more.
Background: Breakpoints in the pre-examination processes and at organizational interfaces are a significant source of failures in specimen identification and tracking in anatomic pathology. While ISO 15189 emphasizes end-to-end traceability and risk-based quality management, implementing these principles in complex, multi-actor specimen pathways remains challenging. This study explores expert perspectives on specimen process chains, tracking mechanisms, and ISO 15189-oriented quality and risk management in pathology. Methods: We conducted 10 semi-structured expert interviews across three settings. Interviews were audio-recorded, transcribed, pseudonymized, and analyzed using structured qualitative content analysis (Mayring) supported by MAXQDA. A deductive category system derived from the theoretical framework and interview guide comprised six main categories and twelve subcategories. Results: Across 512 coded text segments, participants identified several factors as critical for effective implementation, including: (i) interface management along the specimen pathway, with recurrent vulnerabilities at handovers between operating theater/ward/transport and accessioning; (ii) the central role of barcode-based identification and the need for closed-loop traceability; (iii) the importance of measurable quality indicators and incident learning systems to operationalize risk management; (iv) persistent paper–digital handoffs and heterogeneous IT landscapes that undermine data integrity; (v) the need for clearly assigned responsibilities, training, and SOP governance; and (vi) implementation barriers including resources, change management, and vendor integration, alongside practical enablers such as incremental roll-out and cross-professional governance. Conclusions: Experts converge on a pragmatic ISO 15189-aligned roadmap: prioritize interface risks, standardize identifiers and handover rules, define a minimal KPI set for tracking and misidentification events, and reduce paper–digital handoffs by interoperable IT. Future work should quantify baseline error rates and evaluate the impact of digital tracking interventions on patient safety and turnaround times. Full article
(This article belongs to the Section Pathology and Molecular Diagnostics)
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23 pages, 4643 KB  
Article
Assessment of Early Breast Cancer Response to Chemotherapy with Ultrasound Radiomics
by Swapnil Dolui, Basak Dogan, Corinne Wessner, Jessica Porembka, Priscilla Machado, Bersu Ozcan, Nisha Unni, Maysa Abu Khalaf, Flemming Forsberg, Kibo Nam and Kenneth Hoyt
Diagnostics 2026, 16(6), 948; https://doi.org/10.3390/diagnostics16060948 - 23 Mar 2026
Viewed by 549
Abstract
Objective: This prospective study investigated the use of H-scan ultrasound (US) imaging as a novel component of a multiparametric radiomic analysis framework for characterizing human breast cancer response to neoadjuvant chemotherapy (NAC) before and early after treatment initiation. Methods: Thirty breast [...] Read more.
Objective: This prospective study investigated the use of H-scan ultrasound (US) imaging as a novel component of a multiparametric radiomic analysis framework for characterizing human breast cancer response to neoadjuvant chemotherapy (NAC) before and early after treatment initiation. Methods: Thirty breast cancer patients scheduled for NAC were scanned using a clinical US system (Logiq E9, GE HealthCare) equipped with a 9L-D linear array transducer. Radiofrequency (RF) data was obtained at baseline (pre-NAC) and after 10% and 30% of the complete dose of chemotherapy. The RF data was analyzed by a bank of 256 frequency-shifted bandpass filters to form H-scan US frequency images. Grayscale texture features were extracted from both B-scan and H-scan US images. In addition, US attenuation coefficient and speckle statistics based on the Nakagami and Burr distributions were estimated from the RF data. Data classification of tumor and peri-tumoral regions was performed using a novel three-dimensional (3D) score map based on support vector machine (SVM) modeling. Unlike conventional classifiers that report only a single prediction score, a 3D score map provides a visual representation of the classifier decision space, enabling interpretation of class separation and treatment-induced shifts in multiparametric US measurements. Results: The dataset was split into 10 disjoint partitions (90% training, 10% testing) to compute area under the receiver operating characteristic curve (AUC), sensitivity, specificity, and accuracy measures. Actual patient response to NAC was assessed at surgery and categorized as either pathologic complete response (pCR) or non-pCR. Multiparametric US and data classification results at pre-NAC found AUC values of 0.78 after using only tumor information (p < 0.01), which increased to 0.81 with inclusion of peri-tumoral information (p < 0.01). Significant differences in multiparametric US measures from both cancer response types was found after integration of patient data collected at 10% completion of the NAC regimen (i.e., first NAC cycle), yielding an improved AUC of 0.86 (p < 0.001). Conclusions: Multiparametric US imaging with radiomic features from both the tumor and peri-tumoral regions is a promising noninvasive approach for monitoring early breast cancer response to NAC. Full article
(This article belongs to the Section Medical Imaging and Theranostics)
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12 pages, 942 KB  
Article
Cephalometric Assessment of Airway-Related Hyoid Position and Velar Morphology Across Skeletal Malocclusions: A Cross-Sectional Study
by Gizem Yazdan Özen, Ali Kağan Özen and Nebiha Hilal Bilge
Diagnostics 2026, 16(6), 947; https://doi.org/10.3390/diagnostics16060947 - 23 Mar 2026
Viewed by 490
Abstract
Background: The anatomical position of the hyoid bone and the morphological characteristics of the soft palate play a key role in upper airway patency, craniofacial balance, and the coordination of functional structures. These features may vary depending on skeletal pattern and gender. This [...] Read more.
Background: The anatomical position of the hyoid bone and the morphological characteristics of the soft palate play a key role in upper airway patency, craniofacial balance, and the coordination of functional structures. These features may vary depending on skeletal pattern and gender. This study aimed to evaluate the relationship between hyoid bone position, soft palate morphology, skeletal classification, and gender using lateral cephalograms. Methods: A total of 120 individuals (60 females and 60 males) were classified as Skeletal Class I, II, or III based on the ANB (A Point–Nasion–B Point) angle. Measurements reflecting hyoid position and pharyngeal airway width were analyzed, including C3–H (distance from the third cervical vertebra to the hyoid bone), C3–RGn (distance from the third cervical vertebra to retrognathion), H–RGn (distance from the hyoid bone to retrognathion), PNS–UPW (distance from the posterior nasal spine to the upper pharyngeal wall), and U–MPW (distance from the uvula to the middle pharyngeal wall). Soft palate types were classified according to the You classification. Statistical analyses included ANOVA (analysis of variance), the Kruskal–Wallis test, the independent samples t-test, the Mann–Whitney U test, and the chi-square test. Results: Significant differences in C3–RGn, H–RGn, and U–MPW were observed between Skeletal Classes I–III and Classes II–III (p < 0.05). In contrast, C3–H and PNS–UPW did not differ significantly among skeletal classes. Soft palate types showed no significant association with skeletal classification or gender. Gender-based comparisons revealed significant differences in C3–H, C3–RGn, H–RGn, and PNS–UPW (p < 0.05). Conclusions: Mandibular-related hyoid measurements and the U–MPW parameter were associated with skeletal pattern, whereas C3–H and PNS–UPW remained relatively stable. Soft palate morphology was not significantly influenced by skeletal class or gender. These findings suggest that the hyoid–tongue–soft palate complex should be evaluated in conjunction with mandibular position during orthodontic diagnosis and treatment planning. Full article
(This article belongs to the Section Clinical Diagnosis and Prognosis)
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26 pages, 5481 KB  
Review
Intrapartum Ultrasound in Vacuum Operative Delivery: A Comprehensive Review and Proposal of the Novel Ultrasound Flexion Point Method
by Antonio Malvasi, Giorgio Maria Baldini, Tommaso Difonzo, Marco Cerbone, Iris Cara, Marianna Demarco, Ilenia Mappa, Giuseppe Rizzo, Antonella Vimercati, Miriam Dellino, Andrea Tinelli, Edoardo Di Naro and Lorenzo E. Malgieri
Diagnostics 2026, 16(6), 946; https://doi.org/10.3390/diagnostics16060946 - 23 Mar 2026
Viewed by 618
Abstract
Operative vaginal delivery (OVD) via vacuum extraction is a fundamental component of modern obstetric management, yet it carries specific risks of failure and maternal–fetal complications, such as cup detachment, cephalohematoma, and intracranial hemorrhage. The success and safety of the procedure rely heavily on [...] Read more.
Operative vaginal delivery (OVD) via vacuum extraction is a fundamental component of modern obstetric management, yet it carries specific risks of failure and maternal–fetal complications, such as cup detachment, cephalohematoma, and intracranial hemorrhage. The success and safety of the procedure rely heavily on the correct application of the vacuum cup over the “flexion point” of the fetal head. Traditional identification of this landmark via digital examination is often hindered by caput succedaneum and cranial molding, leading to high rates of diagnostic error, particularly in dystocic labor, due to fetal head malpositions and malpresentation. Intrapartum ultrasound (ITU) has demonstrated superior accuracy compared to clinical examination in assessing fetal head position and station and internal rotation. This expert commentary and technical proposal analyzes the current literature regarding vacuum extraction application and failures, focusing on the predictive value of ITU parameters (e.g., Angle of Progression, Midline Angle, Head-Symphysis Distance) and the impact of ITU on cup placement and delivery outcomes. Furthermore, we propose a novel technique: the “Ultrasound Flexion Point” (UFP). This method utilizes translabial ultrasound to identify the specific intersection of the fetal midline and the biparietal diameter as an objective sonographic proxy for the classical flexion point. By providing spatial orientation guidance immediately before the procedure, this technique aims to guide the operator in aligning the cup’s notch with the sonographically identified target zone, using the midline angle as orientation reference, thereby potentially minimizing paramedian or deflexing applications and reducing the incidence of vacuum detachment and associated neonatal trauma. This expert commentary and technical proposal synthesizes current evidence and proposes a protocol requiring prospective validation through randomized controlled trials. Full article
(This article belongs to the Special Issue Advances in Ultrasound Diagnosis in Maternal Fetal Medicine Practice)
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17 pages, 3545 KB  
Article
A Radiomics–Clinical Nomogram for Pre-Treatment Prediction of Neoadjuvant Chemotherapy Response in Locally Advanced Gastric Cancer
by Qianzheng Zhou, Jun Xu, Qiong Li, Fengyuan Li and Hao Xu
Diagnostics 2026, 16(6), 945; https://doi.org/10.3390/diagnostics16060945 - 23 Mar 2026
Cited by 1 | Viewed by 440
Abstract
Objective: To develop and evaluate a nomogram integrating radiomic features from contrast-enhanced CT with clinical variables for pre-treatment predictions of the response to neoadjuvant therapy (NAT) in locally advanced gastric cancer (LAGC). Methods: In this retrospective multicenter study, 183 LAGC patients from the [...] Read more.
Objective: To develop and evaluate a nomogram integrating radiomic features from contrast-enhanced CT with clinical variables for pre-treatment predictions of the response to neoadjuvant therapy (NAT) in locally advanced gastric cancer (LAGC). Methods: In this retrospective multicenter study, 183 LAGC patients from the First Affiliated Hospital of Nanjing Medical University (2014–2023) were included. Radiomic features were extracted from manually delineated pre-treatment CT regions of interest. A machine learning-based predictive model combining radiomic scores and clinical data was constructed. Model performance was assessed using receiver operating characteristic (ROC) analysis, calibration curves, and decision curve analysis (DCA). Results: Multivariate analysis identified the radiomic score, preoperative N stage, and neoadjuvant regimen as independent predictors of NAT responses (all p < 0.05). The integrated nomogram achieved an area under the ROC curve of 0.807 and showed a moderate net benefit in DCA compared with the radiomics-only model. Conclusions: The radiomics–clinical nomogram demonstrates moderate predictive performance for pre-treatment stratification of NAT responses in LAGC. These findings are exploratory and hypothesis-generating, and further validation in independent cohorts is required before clinical application. Full article
(This article belongs to the Special Issue Imaging Research on Gastrointestinal Disorders: Second Edition)
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15 pages, 1686 KB  
Article
Establishment and Temporal Validation of Next-Generation Reference Intervals for Routine Hematological Parameters Using Large-Scale Data
by Chaochao Ma, Lihua Guan, Qian Chen, Rongrong Cheng, Wei Wu and Ling Qiu
Diagnostics 2026, 16(6), 944; https://doi.org/10.3390/diagnostics16060944 - 23 Mar 2026
Viewed by 370
Abstract
Background: Conventional reference intervals (RIs) are typically expressed as fixed limits and may not adequately reflect continuous biological variation across age and sex. Next-generation reference intervals (NGRIs) allow dynamic modeling of laboratory parameters across the lifespan. This study aimed to establish age- [...] Read more.
Background: Conventional reference intervals (RIs) are typically expressed as fixed limits and may not adequately reflect continuous biological variation across age and sex. Next-generation reference intervals (NGRIs) allow dynamic modeling of laboratory parameters across the lifespan. This study aimed to establish age- and sex-specific NGRIs for routine hematological parameters using large-scale health examination data and to evaluate their temporal stability. Methods: Health examination records were linked with laboratory data, and a relatively healthy reference population was defined based on age (18–80 years), normal body mass index, normal blood pressure, and absence of documented disease history. NGRIs were constructed using generalized additive models for location, scale, and shape (GAMLSS) with the Box–Cox Cole and Green distribution. Age-dependent percentile curves (2.5th–97.5th) were generated using bootstrap resampling (100 iterations). Temporal external validation was conducted in five independent annual cohorts (2019–2023). Results: Age- and sex-dependent distributional patterns were observed across multiple hematological parameters, particularly RBC, HGB, and HCT. Continuous percentile curves demonstrated gradual age-related trends, with more pronounced changes in females after midlife. In temporal validation cohorts, the proportion of individuals classified outside the reference intervals remained consistently below 10% across years and sexes, indicating stable performance. Conclusions: Using large-scale real-world health examination data and a flexible distributional modeling framework, we established stable age-continuous next-generation reference intervals for routine hematological parameters. The proposed approach provides a reproducible strategy for modernizing laboratory reference interval construction and supports broader implementation in population-based laboratory medicine. Full article
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21 pages, 3822 KB  
Article
Uncertainty-Aware Framework for CT Radiation Dose Optimization in the Active Surveillance of Small Renal Masses: Clinical and Radiological Considerations
by M. A. Elsabagh, Amira Samy Talaat, Dalia Elwi, Shaimaa M. Hassan, Sameer Alqassimi and Esraa Hassan
Diagnostics 2026, 16(6), 943; https://doi.org/10.3390/diagnostics16060943 - 23 Mar 2026
Viewed by 413
Abstract
Background: Active surveillance of small renal masses is challenged by cumulative radiation exposure from repeated CT imaging, raising long-term health concerns. Low-dose CT protocols offer a strategy to mitigate this risk but are limited by uncertainty regarding measurement accuracy and potential effects on [...] Read more.
Background: Active surveillance of small renal masses is challenged by cumulative radiation exposure from repeated CT imaging, raising long-term health concerns. Low-dose CT protocols offer a strategy to mitigate this risk but are limited by uncertainty regarding measurement accuracy and potential effects on clinical decision-making. Methods: We propose an uncertainty-aware analytical framework using a multi-observer dataset of 40 paired CT cases (low-dose vs. standard-dose). The methodology combines statistical agreement assessment (concordance correlation coefficient, intraclass correlation coefficient), multi-algorithm machine learning prediction (linear regression, random forest, gradient boosting, and SVR), and integrated uncertainty quantification to evaluate equivalence across imaging protocols. Results: Comparative analysis demonstrates near-perfect concordance between protocols (concordance correlation coefficient = 0.9930). Linear regression achieved the highest predictive performance (R2 = 0.9933, MAE = 0.4239 mm, MAPE = 2.07%), outperforming more complex ensemble models, highlighting that interpretable models can achieve superior accuracy without compromising reliability. Conclusions: Clinically, the framework supports the safe adoption of low-dose CT for longitudinal tumor assessment, preserving measurement fidelity and diagnostic confidence essential for timely intervention or continued surveillance. Radiologically, it ensures robust lesion characterization across protocols while minimizing cumulative radiation exposure, particularly in younger patients. By integrating uncertainty quantification, this approach enhances transparency, informs clinical decision-making, and facilitates personalized, evidence-based surveillance strategies, promoting safer, dose-optimized imaging in the management of small renal masses. Full article
(This article belongs to the Section Medical Imaging and Theranostics)
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15 pages, 339 KB  
Article
Short-Term Heart Rate Variability Dynamics and Mortality Risk After Acute Coronary Syndrome
by Nikola Marković, Maša Petrović, Silvana Babić, Milovan Bojić and Branislav Milovanović
Diagnostics 2026, 16(6), 942; https://doi.org/10.3390/diagnostics16060942 - 23 Mar 2026
Viewed by 415
Abstract
Background/Objectives: Heart rate variability (HRV) is a non-invasive marker of autonomic nervous system function with established prognostic value after acute coronary syndrome (ACS). The clinical relevance of temporal changes in short-term HRV remains insufficiently defined. This study evaluated short-term HRV dynamics and their [...] Read more.
Background/Objectives: Heart rate variability (HRV) is a non-invasive marker of autonomic nervous system function with established prognostic value after acute coronary syndrome (ACS). The clinical relevance of temporal changes in short-term HRV remains insufficiently defined. This study evaluated short-term HRV dynamics and their association with mortality after ACS. Methods: This retrospective–prospective study included 230 patients with acute myocardial infarction. Five-minute resting ECG recordings were obtained on day 1 and day 21. Time- and frequency-domain HRV parameters were analyzed, and delta values were calculated. The primary endpoint was overall mortality. Survival was assessed using Kaplan–Meier analysis and Cox regression. Results: Patients who died during follow-up had lower HRV values on day 21 and more pronounced declines in selected parameters. In multivariable analysis, decreased ΔLF and shorter RR intervals independently predicted overall mortality. Conclusions: Short-term HRV provides a practical bedside assessment of autonomic function after ACS. Unfavorable temporal changes likely reflect persistent autonomic imbalance and may offer additional prognostic insight. Larger contemporary studies are needed to confirm these findings. Full article
(This article belongs to the Section Medical Imaging and Theranostics)
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6 pages, 674 KB  
Interesting Images
Brachial Artery Pseudoaneurysm as a Complication of Osteochondral Exostosis of the Humerus in Computed Tomography Angiography Images
by Paweł Gać, Michał Wesołowski, Kamil Biedka and Rafał Poręba
Diagnostics 2026, 16(6), 941; https://doi.org/10.3390/diagnostics16060941 - 22 Mar 2026
Viewed by 403
Abstract
We present computed tomography angiography images of a rare pseudoaneurysm of the left brachial artery, a complication of idiopathic injury to the artery caused by an osteochondral exostosis of the left humerus. A 22-year-old Caucasian man with no significant medical history was admitted [...] Read more.
We present computed tomography angiography images of a rare pseudoaneurysm of the left brachial artery, a complication of idiopathic injury to the artery caused by an osteochondral exostosis of the left humerus. A 22-year-old Caucasian man with no significant medical history was admitted to the emergency department due to sudden, intense pain in his left arm, numbness, and pallor of his left forearm and hand. The patient’s consulting vascular surgeon referred him to the computed tomography (CT) laboratory for a computed tomography angiography (CTA) of the arteries of his left upper limb. In the CTA examination, at the level of the proximal segment of the left brachial artery, an excess of contrast was visualized, measuring up to approximately 1.5 × 1.2 cm in cross-sections and up to approximately 0.7 cm in the craniocaudal dimension. The CTA image was suggestive of a pseudoaneurysm of the left brachial artery. Laterally, the pseudoaneurysm was adjacent to the apex of the imaged osteochondral exostosis on the medial surface of the proximal shaft of the left humerus. A surgical procedure was performed to repair the pseudoaneurysm of the left brachial artery, including removal of the bony exostosis of the left humerus. In summary, relatively common, benign bone lesions can occasionally result in serious vascular complications. CTA is the gold standard for diagnosing these complications. Full article
(This article belongs to the Section Medical Imaging and Theranostics)
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6 pages, 2697 KB  
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The Diagnostic Challenge of Rapunzel Syndrome: Multimodal Imaging Findings in a Child
by Chih-Hao Wang and Shu-Chao Weng
Diagnostics 2026, 16(6), 940; https://doi.org/10.3390/diagnostics16060940 - 22 Mar 2026
Viewed by 361
Abstract
We report the case of a school-aged patient with attention-deficit/hyperactivity disorder who presented with a palpable epigastric mass. The initial abdominal ultrasonography indicated the presence of a heterogeneous space-occupying lesion in the upper abdomen. Subsequent computed tomography revealed a large intragastric mass with [...] Read more.
We report the case of a school-aged patient with attention-deficit/hyperactivity disorder who presented with a palpable epigastric mass. The initial abdominal ultrasonography indicated the presence of a heterogeneous space-occupying lesion in the upper abdomen. Subsequent computed tomography revealed a large intragastric mass with a mottled air-containing density, an imaging feature characteristic of a bezoar. Esophagogastroduodenoscopy confirmed a massive trichobezoar extending beyond the pylorus into the duodenum, consistent with Rapunzel syndrome. Although the endoscopic removal proved unsuccessful, a subsequent surgical extraction via laparotomy yielded a 22 cm trichobezoar. Further history revealed prior hair-picking behavior approximately one year earlier, with a localized bald patch noted by her parents. This case highlights the characteristic multimodal imaging findings of trichobezoars and serves to emphasize the diagnostic challenge posed by a limited clinical history in children with psychiatric comorbidities presenting with abdominal masses. Full article
(This article belongs to the Section Medical Imaging and Theranostics)
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17 pages, 900 KB  
Article
Association of Serum Glucose/Potassium Ratio with Injury Severity and Transfusion Requirements in Traumatic Pelvic Fractures: A Retrospective Cohort Study
by Abdullah Alper Sahin, Yunus Emre Özbilen and Çağrı Akalın
Diagnostics 2026, 16(6), 939; https://doi.org/10.3390/diagnostics16060939 - 22 Mar 2026
Viewed by 358
Abstract
Background: We evaluated the association between admission serum glucose-to-potassium ratio (GPR) and injury severity as well as early transfusion requirements in patients with traumatic pelvic fractures. Methods: This single-center, retrospective cohort study included 84 adult patients with isolated or predominantly pelvic fractures admitted [...] Read more.
Background: We evaluated the association between admission serum glucose-to-potassium ratio (GPR) and injury severity as well as early transfusion requirements in patients with traumatic pelvic fractures. Methods: This single-center, retrospective cohort study included 84 adult patients with isolated or predominantly pelvic fractures admitted between January 2020 and December 2024. Patients with concomitant non-pelvic skeletal fractures were excluded to isolate the metabolic response attributable to pelvic injury. GPR was calculated from admission serum glucose and potassium levels. Higher transfusion requirement (HT) was defined as ≥4 units of packed red blood cells within 24 h. Receiver operating characteristic (ROC) analysis identified the optimal GPR cut-off using the Youden index. Internal validation was performed using bootstrap resampling (1000 iterations), and model calibration was assessed with the Hosmer–Lemeshow test. The incremental discriminatory value of GPR beyond the Injury Severity Score (ISS) was evaluated by comparing AUC values using the DeLong test, and reclassification metrics including the category-free net reclassification improvement (NRI) and integrated discrimination improvement (IDI) were calculated. Sensitivity analyses were conducted using alternative transfusion thresholds (≥6 and ≥10 units). Results: The optimal GPR cut-off was 34 (area under the curve (AUC) = 0.730; 95% CI: 0.593–0.853; sensitivity 78.8%; specificity 59.0%). Patients with GPR ≥ 34 (n = 43) had significantly higher ISS values (median 25 [IQR: 16–34] vs. 9 [5–17]; p < 0.001), greater transfusion volumes (median 3 [0–6] vs. 0 [0–1] units; p < 0.001), and longer intensive care unit (ICU) stays (3 (0–6) vs. 0 (0–1) days; p < 0.001). In univariable logistic regression, GPR was significantly associated with HT (OR = 1.059 per unit increase; 95% CI: 1.015–1.104; p = 0.008); however, significance was not retained in the multivariable model after adjustment for ISS (p = 0.194). ISS remained the sole independent predictor (OR = 1.128; p < 0.001). The combined ISS + GPR model yielded an AUC of 0.857, representing a modest increment over ISS alone (AUC = 0.849; ΔAUC = 0.009; DeLong p = 0.566). Bootstrap-corrected AUCs confirmed minimal optimism (GPR alone: 0.726; ISS + GPR: 0.847). The Hosmer–Lemeshow test indicated adequate calibration for all models (p > 0.05). The category-free NRI was 0.627 (p = 0.009), whereas the IDI did not reach significance (0.017; p = 0.290). Sensitivity analysis at the ≥6-unit threshold yielded consistent results (GPR AUC = 0.709). Conclusions: Admission GPR is significantly associated with injury severity, hemorrhagic burden, and transfusion requirements in patients with traumatic pelvic fractures. Although GPR does not independently predict transfusion needs beyond ISS, it yields significant reclassification improvement and may serve as a practical, rapidly obtainable adjunct for early risk stratification in the acute trauma setting. Level of Evidence: III (retrospective prognostic study). Full article
(This article belongs to the Section Clinical Diagnosis and Prognosis)
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17 pages, 1021 KB  
Article
Pre-Treatment Breast MRI Features and ADC Values as Predictors of Pathologic Complete Response in Breast Cancer: A Molecular Subtype-Based Analysis
by Ela Kaplan, Hüseyin Alakus and Selcuk Kaplan
Diagnostics 2026, 16(6), 938; https://doi.org/10.3390/diagnostics16060938 - 22 Mar 2026
Viewed by 473
Abstract
Background/Objectives: The role of pre-treatment breast magnetic resonance imaging (MRI) findings and apparent diffusion coefficient (ADC) values in predicting pathologic complete response (pCR) in breast cancer patients receiving neoadjuvant chemotherapy (NAC) has not yet been sufficiently clarified, especially in the context of [...] Read more.
Background/Objectives: The role of pre-treatment breast magnetic resonance imaging (MRI) findings and apparent diffusion coefficient (ADC) values in predicting pathologic complete response (pCR) in breast cancer patients receiving neoadjuvant chemotherapy (NAC) has not yet been sufficiently clarified, especially in the context of molecular subtype differences. In this study, we questioned whether these imaging parameters were independent predictors of pCR. Methods: This study retrospectively explored MRI characteristics of 188 patients who underwent NAC from 2015 to 2023. The patients were divided into the pCR-positive and pCR-negative groups—the latter comprising patients with partial response (n = 61) and stable disease (n = 90)—and were classified into four molecular subtypes: Luminal A/B, HER2-enriched, and triple-negative breast cancer (TNBC). The MRI parameters included pre-chemotherapy T2-weighted signal characteristics, shape features, contrast kinetics, peritumoral edema, and ADC MIN/ADC MAX. Post-treatment ADC and ΔADC were the post-chemotherapy MRI parameters. Independent predictors were evaluated by logistic regression and discriminant performance by ROC analysis. Results: The overall pCR rate was 19.7%. In multivariate analysis, T2-weighted isointense signal (OR = 4.50), uniform tumor shape (OR = 12.83), HER2-enriched subtype (OR = 6.03), TNBC (OR = 5.15), ADC MIN (OR = 1.41), tumor size (OR = 1.28), and kinetic Type 3 pattern (OR = 3.21) were identified as independent predictors. Pre-treatment ADC MIN yielded an AUC of 0.724, while post-treatment ADC achieved 100% sensitivity and 96.7% specificity (AUC = 0.967). Conclusions: MRI morphology and ADC values may make a meaningful contribution to the prediction of pCR when evaluated in the context of molecular subtype. Post-treatment ADC demonstrated particularly strong discriminatory performance; however, external validation in multicenter cohorts is required before clinical implementation. Full article
(This article belongs to the Section Medical Imaging and Theranostics)
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19 pages, 1284 KB  
Systematic Review
Human In Vivo Cardiac Magnetic Resonance Imaging at 7 T: Feasibility, Applications, and Current Limitations—A Systematic Review
by Arosh S. Perera Molligoda Arachchige, Gabriel Amorim Moreira Alves, Ayça Zal, Giulia D’Acunto, Maciej Węglarz, Oana-Georgiana Voicu, Erica Maffei and Filippo Cademartiri
Diagnostics 2026, 16(6), 937; https://doi.org/10.3390/diagnostics16060937 - 22 Mar 2026
Viewed by 561
Abstract
Background/Objectives: Cardiac magnetic resonance (CMR) imaging at 7 Tesla provides a substantially higher intrinsic signal-to-noise ratio compared with conventional 1.5 T and 3 T systems, potentially enabling higher spatial resolution, improved tissue contrast, and advanced metabolic imaging. However, clinical translation remains limited by [...] Read more.
Background/Objectives: Cardiac magnetic resonance (CMR) imaging at 7 Tesla provides a substantially higher intrinsic signal-to-noise ratio compared with conventional 1.5 T and 3 T systems, potentially enabling higher spatial resolution, improved tissue contrast, and advanced metabolic imaging. However, clinical translation remains limited by technical challenges associated with ultra-high-field operation. This systematic review aimed to synthesize current human in vivo evidence on the feasibility, applications, and methodological limitations of 7-T cardiovascular MRI. Methods: A PRISMA-guided systematic search of PubMed, Cochrane Library, Web of Science, and Scopus was conducted from database inception through January 2025. Studies reporting human in vivo cardiovascular MRI at 7 Tesla were included. Data regarding study design, sample characteristics, imaging applications, feasibility, quantitative findings, and reported limitations were extracted and qualitatively synthesized. Results: Sixty-five studies met inclusion criteria, predominantly small prospective cohorts (mean sample size = 13), largely involving healthy volunteers. Across diverse applications—including coronary MR angiography, cine imaging, valvular assessment, vascular imaging, flow quantification, myocardial tissue characterization, and multinuclear (31P, 23Na, 39K) imaging—7-T CMR was consistently feasible and capable of producing high-quality images. Quantitative ventricular and vascular measurements were generally concordant with lower field strengths. Incremental benefits were most apparent in high-resolution structural imaging and metabolic applications, whereas routine functional and flow assessments showed limited additional advantages. No serious adverse events were reported. Conclusions: Human cardiovascular MRI at 7 Tesla represents a technically feasible research and early translational platform with selective advantages over established field strengths. Further advances in radiofrequency technology, protocol harmonization, and larger disease-focused studies are required to clarify its potential clinical role. Full article
(This article belongs to the Special Issue Cardiovascular Imaging, 2nd Edition)
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11 pages, 683 KB  
Article
Optical Coherence Tomography Angiography Findings in Primary Progressive Multiple Sclerosis Patients Receiving Ocrelizumab Treatment
by Burçin Çakır, Seren Kaplan Güngördü, Nilgün Özkan Aksoy and Dilcan Kotan
Diagnostics 2026, 16(6), 936; https://doi.org/10.3390/diagnostics16060936 - 22 Mar 2026
Viewed by 378
Abstract
Objectives: The aim of this study was to evaluate macular vessel area densities (superficial and deep) and foveal avascular zone (FAZ) measurements using OCT-A in the eyes of primary progressive multiple sclerosis (PPMS) patients receiving Ocrelizumab treatment with or without optic nerve [...] Read more.
Objectives: The aim of this study was to evaluate macular vessel area densities (superficial and deep) and foveal avascular zone (FAZ) measurements using OCT-A in the eyes of primary progressive multiple sclerosis (PPMS) patients receiving Ocrelizumab treatment with or without optic nerve involvement. Methods: The medical records of PPMS patients who received Ocrelizumab treatment at least once were reviewed. Retinal nerve fiber layer (RNFL) thickness measurements and OCT-A analysis were conducted on the PPMS patients and on age-matched healthy individuals. The patient group was divided into two subgroups: eyes with optic neuritis (PPMS+ON) and eyes without ON (PPMS-ON). Central and mean superficial vessel area (SVA) and deep vessel area (DVA) densities, as well as foveal avascular zone (FAZ) measurements, were analyzed. All parameters were statistically compared between groups and subgroups. Results: A total of 38 PPMS patients receiving Ocrelizumab treatment and 31 healthy individuals were included in this study. Statistically significant differences were observed between the groups in terms of best corrected visual acuity (BCVA), RNFL thickness, and the superficial vessel area densities for all parts except for the central part. In terms of deep vessel area densities, differences were found in the central and inferior parts. The mean FAZ area also showed a statistically significant difference between groups. Mean RNFL thickness differed significantly between the subgroups. Mean nasal, temporal, inferior part, and total superficial vessel area densities were statistically different between the subgroups. The central and inferior parts of the deep vessel area densities showed statistically significant differences. The mean FAZ area was also statistically different between the subgroups. Conclusions: The findings suggest that macular superficial and deep vascular densities are affected in PPMS patients receiving the same therapy modality and that previous optic neuritis may influence the results. Full article
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43 pages, 6336 KB  
Systematic Review
A Systematic Literature Review of You Only Look Once Architectures (v1–v12) in Healthcare Systems
by Ozgur Koray Sahingoz, Gozde Karatas Baydogmus and Emin Kugu
Diagnostics 2026, 16(6), 935; https://doi.org/10.3390/diagnostics16060935 - 22 Mar 2026
Cited by 1 | Viewed by 878
Abstract
Background/Objectives: The integration of deep learning and computer vision into healthcare has improved medical diagnosis and image analysis. Among object detection algorithms, the YOLO family has attracted substantial attention due to its ability to analyze images in real time with reported improvements [...] Read more.
Background/Objectives: The integration of deep learning and computer vision into healthcare has improved medical diagnosis and image analysis. Among object detection algorithms, the YOLO family has attracted substantial attention due to its ability to analyze images in real time with reported improvements in detection performance across multiple studies. This systematic review examines the evolution of YOLO algorithms for diagnostic applications in healthcare from YOLOv1 to YOLOv12. Methods: Peer-reviewed scientific articles published up to 1 January 2026 were retrieved from major scientific databases in accordance with PRISMA 2020 guidelines. The included studies applied YOLO models to medical imaging tasks, including disease and lesion detection and support for clinical procedures. Performance was synthesized using reported metrics such as average precision, accuracy, inference time, and computational efficiency. Results: The reviewed literature suggests progressive architectural refinements associated with reported improvements in diagnostic performance. YOLOv5 and YOLOv8 are the most frequently used architectures in diagnostic settings, reflecting a favorable trade-off between accuracy and computational complexity. YOLO-based methods have demonstrated strong performance across radiological, pathological, ophthalmological, and endoscopic applications. Conclusions: YOLO models have matured into robust and optimized solutions for medical image analysis; however, challenges remain in interpretability, cross-institution generalization, and deployment on edge devices. Future work on explainable YOLO-based diagnostics and energy-efficient model design will be particularly valuable. Full article
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16 pages, 1810 KB  
Article
Local Versus Global Binarization Techniques After Frangi Filtering for Optical Coherence Tomography Angiography Based Retinal Vessel Density Assessment in Diabetic Retinopathy
by Andrada-Elena Mirescu, Ioana Teodora Tofolean, Sanda Jurja, Florian Balta, Alina Popa-Cherecheanu, Ruxandra Angela Pirvulescu, Gerhard Garhofer, George Balta, Irina-Elena Cristescu and Dan George Deleanu
Diagnostics 2026, 16(6), 934; https://doi.org/10.3390/diagnostics16060934 - 21 Mar 2026
Viewed by 476
Abstract
Background/Objectives: Optical coherence tomography angiography (OCTA) enables noninvasive quantitative assessment of the retinal microvasculature and is widely used in diabetic retinopathy (DR). However, OCTA-derived metrics are highly dependent on post-processing techniques, particularly vessel binarization. This study aimed to compare local and global binarization [...] Read more.
Background/Objectives: Optical coherence tomography angiography (OCTA) enables noninvasive quantitative assessment of the retinal microvasculature and is widely used in diabetic retinopathy (DR). However, OCTA-derived metrics are highly dependent on post-processing techniques, particularly vessel binarization. This study aimed to compare local and global binarization methods applied after Frangi filtering for vessel enhancement in parafoveal vessel density analysis. Methods: This cross-sectional study included 69 participants: 17 healthy controls and 52 diabetic patients, classified as the following: no DR (n = 14), non-proliferative DR (NPDR, n = 18), or proliferative DR (PDR, n = 20). All subjects underwent comprehensive ophthalmological examination and OCTA imaging of the superficial capillary plexus using a Topcon OCTA system. Images were processed using a custom MATLAB protocol. Following Frangi filtering, five binarization methods were applied: three local (Phansalkar, local Otsu, adaptive mean) and two global (global mean and global Otsu). Parafoveal vessel density was quantified within the four inner quadrants of the ETDRS grid. Results: Statistically significant differences in vessel density were consistently observed between PDR group and both the control and no DR groups across all local binarization methods. Among global methods, only global Otsu thresholding detected a significant difference between PDR and control. The most robust differences were predominantly identified in the nasal and inferior quadrants. Conclusions: Local adaptive binarization methods demonstrated superior sensitivity and structural preservation for parafoveal vessel density analysis in DR. Global methods showed limited discriminative capability. These findings support the preferential use of local adaptive techniques for reliable OCTA-based vascular assessment in diabetic retinopathy. Full article
(This article belongs to the Special Issue Diagnosing, Treating, and Preventing Eye Diseases)
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16 pages, 1835 KB  
Article
Potential Diagnostic and Prognostic Value of Neutrophil Counts, Neutrophil–Lymphocyte Ratio, Pan-Immunoinflammatory Score, Systemic Immunoinflammatory Index, and Systemic Inflammatory Response Index in Brain Tumors
by Murat Ozcan Yay, Adem Keskin, Mahmut Ali Osman Eryilmaz and Cagatay Kaya
Diagnostics 2026, 16(6), 933; https://doi.org/10.3390/diagnostics16060933 - 21 Mar 2026
Viewed by 459
Abstract
Background/Objectives: The aim of this study is to investigate the possible association between systemic inflammation parameters and tumor presence in brain tumor patients with high morbidity and mortality rates, to examine the discriminative performance of these markers and, furthermore, to investigate the [...] Read more.
Background/Objectives: The aim of this study is to investigate the possible association between systemic inflammation parameters and tumor presence in brain tumor patients with high morbidity and mortality rates, to examine the discriminative performance of these markers and, furthermore, to investigate the relationship of these markers with the clinical findings of patients at different time points. Methods: This study included 99 patients with brain tumors as the case group and 99 healthy individuals as the control group. Neutrophil, lymphocyte, monocyte, and platelet levels, as well as indices and ratios derived from these parameters, were compared among the participants. Binary logistic regression and ROC analyses were applied to variables showing significant differences, and the relationship between these variables and demographic and clinical findings was also evaluated. Results: The neutrophil count, neutrophil–lymphocyte ratio (NLR), pan-immunoinflammatory value (PIV), systemic immunoinflammatory index (SII), and systemic inflammatory response index (SIRI) values of the case group were higher compared to the control group. While these parameters were associated with the presence of brain tumors, the highest odds ratio, area under the curve, and specificity were found in the neutrophil count, and the highest sensitivity was found in the SIRI parameter. Some or all of these parameters differed according to tumor type, localization, motor weakness, 3-month adjuvant treatment, 6-month recurrence, postoperative day 1, 3-month and 6-month neurological deficits and 3-month and 6-month Karnofsky Performance Status. Conclusions: These parameters can be considered useful biomarkers that show moderate discrimination in patients with brain tumors and can support clinical evaluation. Full article
(This article belongs to the Section Clinical Diagnosis and Prognosis)
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18 pages, 2594 KB  
Article
APSified Peripapillary Vessel Density in Glaucoma Suspects and Open-Angle Glaucoma
by Michael Moritz, Julia Schottenhamml, Marius Muenk, Meike Müller, Christian Mardin and Bettina Hohberger
Diagnostics 2026, 16(6), 932; https://doi.org/10.3390/diagnostics16060932 - 21 Mar 2026
Viewed by 423
Abstract
Background/Objectives: Optical coherence tomography-angiography (OCT-A) is a non-invasive method of visualizing the capillary system. As vascular dysregulation impacts glaucoma pathogenesis, the aim of this study was to evaluate APSified-BMO-based-peripapillary vessel density (VD) in patients with ocular hypertension (OHT), pre-perimetric-open-angle glaucoma, as well [...] Read more.
Background/Objectives: Optical coherence tomography-angiography (OCT-A) is a non-invasive method of visualizing the capillary system. As vascular dysregulation impacts glaucoma pathogenesis, the aim of this study was to evaluate APSified-BMO-based-peripapillary vessel density (VD) in patients with ocular hypertension (OHT), pre-perimetric-open-angle glaucoma, as well as primary (POAG) and secondary (SOAG) open-angle glaucoma in comparison to healthy controls using OCT-A. Methods: The present study included 180 eyes from 115 patients of the Erlangen Glaucoma Registry, divided into 35 eyes with OHT, 16 pre-perimetric-OAG eyes, 64 OAG eyes—which were subdivided into 37 POAG and 27 SOAG eyes—and 65 healthy controls. All subjects underwent measurements of the retinal nerve fiber layer (RNFL), inner nuclear layer (INL), retinal ganglion cell (RGC) layer, and Bruch membrane opening–minimum rim width (BMO-MRW). APSified-BMO-based-peripapillary vessel density (VD) was visualized by using OCT-A and quantified using the Erlangen Angio Tool. Results: Mean APSified-BMO-based peripapillary VD showed a significant correlation with age (p < 0.0001). Considering the age effect, mean APSified-BMO-based peripapillary VD of OAG was significantly lower compared to healthy eyes (p < 0.0001) and OHT (p = 0.016). Subgroup analysis yielded a significant difference in mean APSified-BMO-based peripapillary VD between controls and POAG (p = 0.001) and SOAG (p = 0.018), respectively. In addition, a significant difference was observed between OHT and POAG patients (p = 0.036). No significant differences were observed between the OHT, pre-perimetric-OAG, and healthy eyes, respectively. Conclusions: As peripapillary VD was significantly decreased in glaucoma patients compared to controls, the data might suggest that peripapillary VD might be useful for monitoring glaucoma progress. Full article
(This article belongs to the Section Biomedical Optics)
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20 pages, 1203 KB  
Article
Proteomics-Based Study of Potential Emphysema Biomarkers Reveals Systemic Redox System and Extracellular Matrix Component Dysregulation
by Grgur Salai, Ruđer Novak, Stela Hrkač, Václav Pustka, David Potěšil, Zbyněk Zdráhal, Divo Ljubicic and Lovorka Grgurević
Diagnostics 2026, 16(6), 931; https://doi.org/10.3390/diagnostics16060931 - 21 Mar 2026
Viewed by 604
Abstract
Objective: Emphysema is an important chronic obstructive pulmonary disease (COPD) phenotype characterized by the destruction of air spaces distal to the terminal bronchiole. Aiming to detect potential emphysema biomarkers and to assess the systemic effects of emphysema in blood plasma, we conducted a [...] Read more.
Objective: Emphysema is an important chronic obstructive pulmonary disease (COPD) phenotype characterized by the destruction of air spaces distal to the terminal bronchiole. Aiming to detect potential emphysema biomarkers and to assess the systemic effects of emphysema in blood plasma, we conducted a small cross-sectional shotgun proteomics study. Methods: This study included N = 40 participants divided into four subgroups (N = 10 per group): patients with emphysema and COPD (CE), patients with COPD but without emphysema (CN), healthy smokers (HS) and healthy never-smokers (HN). The participants were sampled non-probabilistically to be similar in terms of age, sex and comorbidities. Participants’ blood plasma was analyzed using liquid chromatography–mass spectrometry. Bioinformatic analysis included detection of differentially expressed proteins (DEPs) and overrepresentation analysis (ORA). Results: Across all groups, a total of 994 proteins were identified, with NADP-dependent malic enzyme (NADP-ME; encoded by ME1) being the only DEP in the CE vs. CN contrast. Proteins such as BMP1, ADAMTSL-2, -4 and IGFBP4, -5, 6 were identified to be upregulated in CE vs. HN. Fibulin-1, -3 and several immunoglobulin components were identified to be downregulated in the CE vs. HN contrast. ORA revealed several enriched processes, including serine-type endopeptidase activity, insulin-like growth factor I and II binding, and signaling receptor binding. Conclusion: We propose NADP-ME, an important enzyme of intermediary metabolism and redox homeostasis, as a potential biomarker candidate of emphysema. Notably, NADP-ME is also implicated in anoikis resistance. Additionally, changes in the expression levels of BMP1, ADAMTSL-2 and -4, and fibulin suggest potential major systemic effects of extracellular matrix perturbation. As all data was derived from LC-MS analysis, these findings need to be further evaluated with complementary methods. Full article
(This article belongs to the Special Issue Diagnosis and Management of Lung Diseases)
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24 pages, 323 KB  
Review
Molecular Point-of-Care Testing for Respiratory Infections: A Comprehensive Literature Review (2006–2026)
by Ahmed J. Alzahrani
Diagnostics 2026, 16(6), 930; https://doi.org/10.3390/diagnostics16060930 - 20 Mar 2026
Viewed by 908
Abstract
Molecular point-of-care testing (POCT) for respiratory infections has undergone remarkable advancement over the past two decades, driven by technological innovation and urgent clinical needs highlighted by the COVID-19 pandemic. This comprehensive systematic review was conducted following PRISMA 2020 guidelines, synthesizing evidence from 254 [...] Read more.
Molecular point-of-care testing (POCT) for respiratory infections has undergone remarkable advancement over the past two decades, driven by technological innovation and urgent clinical needs highlighted by the COVID-19 pandemic. This comprehensive systematic review was conducted following PRISMA 2020 guidelines, synthesizing evidence from 254 peer-reviewed studies published between 2006 and 2026, with detailed analysis of the 30 most relevant papers selected through a rigorous four-stage screening process. The review examines the evolution of molecular POCT technologies, including reverse transcription polymerase chain reaction (RT-PCR), loop-mediated isothermal amplification (LAMP), recombinase polymerase amplification (RPA), and CRISPR-based detection systems. Key findings demonstrate that modern molecular POCT platforms achieve diagnostic performance comparable to laboratory-based testing, with sensitivities ranging from 88% to 100% and specificities from 98% to 100%, while delivering results in 15 to 80 min. These technologies enable rapid, accurate detection of major respiratory pathogens, including SARS-CoV-2, influenza A/B, respiratory syncytial virus (RSV), and atypical bacteria. The integration of microfluidic systems, portable devices, and smartphone-based analysis has expanded access to testing in resource-limited settings, emergency departments, and wearable platforms. This review provides critical insights for clinicians, researchers, and policymakers regarding the current state, clinical applications, and future directions of molecular POCT for respiratory infections. Full article
(This article belongs to the Special Issue Advances in Infectious Disease Diagnosis Technologies)
28 pages, 5094 KB  
Review
Mixed Lymphocyte Reaction: Functional Immune Profiling in Transplantation and Beyond
by Nurtilek Galimov, Aruzhan Asanova, Sholpan Altynova and Aidos Bolatov
Diagnostics 2026, 16(6), 929; https://doi.org/10.3390/diagnostics16060929 - 20 Mar 2026
Viewed by 983
Abstract
The mixed lymphocyte reaction (MLR) is a classic functional assay that models in vitro interactions between responder T cells and allogeneic antigen-presenting cells (APCs). It quantifies the magnitude and quality of alloreactivity, integrating signals from allorecognition, co-stimulation, inflammatory context, and minor histocompatibility antigens [...] Read more.
The mixed lymphocyte reaction (MLR) is a classic functional assay that models in vitro interactions between responder T cells and allogeneic antigen-presenting cells (APCs). It quantifies the magnitude and quality of alloreactivity, integrating signals from allorecognition, co-stimulation, inflammatory context, and minor histocompatibility antigens that may not be captured by molecular matching alone. This review is narrative in nature and is intended as a practical, non-systematic synthesis of the field. To provide a modern, practice-oriented synthesis of MLR designs, readouts, and translational uses, highlighting how new technologies have expanded MLR from bulk proliferation into multidimensional immune profiling.We summarize why MLR remains valuable as a functional compatibility probe beyond HLA typing, including the high baseline frequency of alloreactive T cells that produces robust signals without priming. We then review key design options (one-way vs. two-way formats; stimulator inactivation; responder definition; APC source and maturation) and how these choices affect interpretation for rejection and graft-versus-host disease risk modeling, tolerance-focused studies, and immunomodulatory screening. Next, we outline major readouts—radiometric and flow cytometric proliferation (dye dilution, Ki-67), cytokine/chemokine profiling, cytotoxicity adaptations, and next-generation add-ons (e.g., scRNA-seq, TCR sequencing)—emphasizing complementary strengths and common pitfalls. Finally, we consolidate practical quality and reproducibility controls (donor variability, dynamic range, timing, batch effects, and acceptance criteria) to improve cross-study comparability and translational readiness. Modern MLR platforms combine controllable allogeneic stimulation with scalable, high-resolution readouts for mechanistic discovery, immune monitoring and translational immune profiling. Standardized modular design and rigorous quality control can improve reproducibility and support broader adoption across transplantation, immunotherapy, and immune-modulation research. Full article
(This article belongs to the Section Pathology and Molecular Diagnostics)
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