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Search Results (3,035)

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14 pages, 310 KiB  
Article
Clinical Outcomes of Patients with Achalasia Following Pneumatic Dilation Treatment: A Single Center Experience
by Viktorija Sabljić, Dorotea Božić, Damir Aličić, Žarko Ardalić, Ivna Olić, Damir Bonacin and Ivan Žaja
J. Clin. Med. 2025, 14(15), 5448; https://doi.org/10.3390/jcm14155448 (registering DOI) - 2 Aug 2025
Abstract
Background/Objectives: Pneumatic dilation (PD) is a widely used treatment modality in the management of achalasia. It is particularly relevant in regions where many centers lack access to advanced therapeutic modalities. Therefore, we aimed to assess the effectiveness and safety of PD in [...] Read more.
Background/Objectives: Pneumatic dilation (PD) is a widely used treatment modality in the management of achalasia. It is particularly relevant in regions where many centers lack access to advanced therapeutic modalities. Therefore, we aimed to assess the effectiveness and safety of PD in our local region. Methods: This study retrospectively analyzed patients with achalasia that underwent PD from 1/2013 to 12/2019. The diagnosis of achalasia was established on the grounds of clinical symptoms, radiological and endoscopic findings, and esophageal manometry. Data on patient’s clinical characteristics, dilation technique and postprocedural follow-up were collected and statistically analyzed. Procedure effectiveness was defined as the postprocedural Eckardt score ≤ 3. Results: PD significantly reduced frequency of dysphagia, regurgitation, and retrosternal pain (p < 0.001). Body-weight increased significantly one month and one year after the procedure (p < 0.001). The procedural success rate was 100%. No severe complications were reported. Conclusions: PD is an effective and safe treatment modality in the management of achalasia. The study limitations include a single center design with the small number of participants, not all of whom underwent manometry, gender disproportion, absence of non-responders, and a short follow-up. Full article
(This article belongs to the Special Issue Clinical Advances in Gastrointestinal Endoscopy)
13 pages, 769 KiB  
Article
A Novel You Only Listen Once (YOLO) Deep Learning Model for Automatic Prominent Bowel Sounds Detection: Feasibility Study in Healthy Subjects
by Rohan Kalahasty, Gayathri Yerrapragada, Jieun Lee, Keerthy Gopalakrishnan, Avneet Kaur, Pratyusha Muddaloor, Divyanshi Sood, Charmy Parikh, Jay Gohri, Gianeshwaree Alias Rachna Panjwani, Naghmeh Asadimanesh, Rabiah Aslam Ansari, Swetha Rapolu, Poonguzhali Elangovan, Shiva Sankari Karuppiah, Vijaya M. Dasari, Scott A. Helgeson, Venkata S. Akshintala and Shivaram P. Arunachalam
Sensors 2025, 25(15), 4735; https://doi.org/10.3390/s25154735 (registering DOI) - 31 Jul 2025
Abstract
Accurate diagnosis of gastrointestinal (GI) diseases typically requires invasive procedures or imaging studies that pose the risk of various post-procedural complications or involve radiation exposure. Bowel sounds (BSs), though typically described during a GI-focused physical exam, are highly inaccurate and variable, with low [...] Read more.
Accurate diagnosis of gastrointestinal (GI) diseases typically requires invasive procedures or imaging studies that pose the risk of various post-procedural complications or involve radiation exposure. Bowel sounds (BSs), though typically described during a GI-focused physical exam, are highly inaccurate and variable, with low clinical value in diagnosis. Interpretation of the acoustic characteristics of BSs, i.e., using a phonoenterogram (PEG), may aid in diagnosing various GI conditions non-invasively. Use of artificial intelligence (AI) and improvements in computational analysis can enhance the use of PEGs in different GI diseases and lead to a non-invasive, cost-effective diagnostic modality that has not been explored before. The purpose of this work was to develop an automated AI model, You Only Listen Once (YOLO), to detect prominent bowel sounds that can enable real-time analysis for future GI disease detection and diagnosis. A total of 110 2-minute PEGs sampled at 44.1 kHz were recorded using the Eko DUO® stethoscope from eight healthy volunteers at two locations, namely, left upper quadrant (LUQ) and right lower quadrant (RLQ) after IRB approval. The datasets were annotated by trained physicians, categorizing BSs as prominent or obscure using version 1.7 of Label Studio Software®. Each BS recording was split up into 375 ms segments with 200 ms overlap for real-time BS detection. Each segment was binned based on whether it contained a prominent BS, resulting in a dataset of 36,149 non-prominent segments and 6435 prominent segments. Our dataset was divided into training, validation, and test sets (60/20/20% split). A 1D-CNN augmented transformer was trained to classify these segments via the input of Mel-frequency cepstral coefficients. The developed AI model achieved area under the receiver operating curve (ROC) of 0.92, accuracy of 86.6%, precision of 86.85%, and recall of 86.08%. This shows that the 1D-CNN augmented transformer with Mel-frequency cepstral coefficients achieved creditable performance metrics, signifying the YOLO model’s capability to classify prominent bowel sounds that can be further analyzed for various GI diseases. This proof-of-concept study in healthy volunteers demonstrates that automated BS detection can pave the way for developing more intuitive and efficient AI-PEG devices that can be trained and utilized to diagnose various GI conditions. To ensure the robustness and generalizability of these findings, further investigations encompassing a broader cohort, inclusive of both healthy and disease states are needed. Full article
(This article belongs to the Special Issue Biomedical Signals, Images and Healthcare Data Analysis: 2nd Edition)
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21 pages, 799 KiB  
Review
The Molecular Diagnosis of Invasive Fungal Diseases with a Focus on PCR
by Lottie Brown, Mario Cruciani, Charles Oliver Morton, Alexandre Alanio, Rosemary A. Barnes, J. Peter Donnelly, Ferry Hagen, Rebecca Gorton, Michaela Lackner, Juergen Loeffler, Laurence Millon, Riina Rautemaa-Richardson and P. Lewis White
Diagnostics 2025, 15(15), 1909; https://doi.org/10.3390/diagnostics15151909 - 30 Jul 2025
Viewed by 310
Abstract
Background: Polymerase chain reaction (PCR) is highly sensitive and specific for the rapid diagnosis of invasive fungal disease (IFD) but is not yet widely implemented due to concerns regarding limited standardisation between assays, the lack of commercial options and the absence of [...] Read more.
Background: Polymerase chain reaction (PCR) is highly sensitive and specific for the rapid diagnosis of invasive fungal disease (IFD) but is not yet widely implemented due to concerns regarding limited standardisation between assays, the lack of commercial options and the absence of clear guidance on interpreting results. Objectives and Methods: This review provides an update on technical and clinical aspects of PCR for the diagnosis of the most pertinent fungal pathogens, including Aspergillus, Candida, Pneumocystis jirovecii, Mucorales spp., and endemic mycoses. Summary: Recent meta-analyses have demonstrated that quantitative PCR (qPCR) offers high sensitivity for diagnosing IFD, surpassing conventional microscopy, culture and most serological tests. The reported specificity of qPCR is likely underestimated due to comparison with imperfect reference standards with variable sensitivity. Although the very low limit of detection of qPCR can generate false positive results due to procedural contamination or patient colonisation (particularly in pulmonary specimens), the rates are comparable to those observed for biomarker testing. When interpreting qPCR results, it is essential to consider the pre-test probability, determined by the patient population, host factors, clinical presentation and risk factors. For patients with low to moderate pre-test probability, the use of sensitive molecular tests, often in conjunction with serological testing or biomarkers, can effectively exclude IFD when all tests return negative results, reducing the need for empirical antifungal therapy. Conversely, for patients with high pre-test probability and clinical features of IFD, qPCR testing on invasive specimens from the site of infection (such as tissue or bronchoalveolar lavage fluid) can confidently rule in the disease. The development of next-generation sequencing methods to detect fungal infection has the potential to enhance the diagnosis of IFD, but standardisation and optimisation are essential, with improved accessibility underpinning clinical utility. Full article
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10 pages, 1153 KiB  
Article
Clinical Trends and Hospital Mortality of Transjugular Intrahepatic Portosystemic Shunt (TIPS) in Germany: A Descriptive Analysis Between 2019 and 2023
by Sven H. Loosen, Christian Weigel, Anselm Kunstein, Peter Minko, Gerald Antoch, Johannes G. Bode, Tom Luedde, Christoph Roderburg and Karel Kostev
Diagnostics 2025, 15(15), 1902; https://doi.org/10.3390/diagnostics15151902 - 29 Jul 2025
Viewed by 158
Abstract
Background/Objectives: The transjugular intrahepatic portosystemic shunt (TIPS) is an established treatment for complications of portal hypertension in patients with liver cirrhosis. While its use has increased and indications have broadened in recent years, recent comprehensive data on patient characteristics, trends, and in-hospital mortality [...] Read more.
Background/Objectives: The transjugular intrahepatic portosystemic shunt (TIPS) is an established treatment for complications of portal hypertension in patients with liver cirrhosis. While its use has increased and indications have broadened in recent years, recent comprehensive data on patient characteristics, trends, and in-hospital mortality in Germany are lacking. This study aimed to evaluate current clinical patterns and mortality outcomes associated with TIPS. Methods: This nationwide cross-sectional study used anonymized hospital data from the German InEK database between 2019 and 2023. TIPS procedures were identified using relevant OPS codes. Patient demographics, liver cirrhosis stage (Child–Pugh), hepatic encephalopathy grade, comorbid conditions, and in-hospital mortality were analyzed descriptively. Analyses were conducted using SAS 9.4. Results: A total of 12,905 TIPS procedures were documented. Annual case numbers rose from 2180 in 2019 to 2954 in 2023. Most patients were male (66.3%) and aged 60–74 years. Ascites (68.6%) was the most frequent associated diagnosis, followed by variceal bleeding (16.4%) and hepatorenal syndrome (14.9%). The average hospital stay decreased from 19.6 to 16.8 days. Overall in-hospital mortality was 8.5%, increasing with age (13.0% in ≥75 years), Child–Pugh C cirrhosis (14.9%), PCCL grade 4 (17.6%), hepatorenal syndrome (16.7%), and grade 4 hepatic encephalopathy (56.1%). Conclusions: TIPS usage in Germany has increased over the past five years, with a shift toward earlier disease stages. Higher in-hospital mortality in clinically complex patients underscores the importance of careful patient selection and tailored management strategies in high-risk groups. Full article
(This article belongs to the Special Issue Diagnosis and Management of Liver Diseases, Third Edition)
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18 pages, 1965 KiB  
Article
Barriers to Seeking Medical Care for Hemorrhoidal Symptoms: A Cross-Sectional Observational Study
by Adrian Cote, Roxana Loriana Negrut, Bogdan Feder, Ioan Andrei Antal, Maur Sebastian Horgos, Emilia Tomescu and Adrian Marius Maghiar
J. Clin. Med. 2025, 14(15), 5361; https://doi.org/10.3390/jcm14155361 (registering DOI) - 29 Jul 2025
Viewed by 135
Abstract
Background: Despite their high prevalence and potential for significant morbidity, hemorrhoidal symptoms remain underreported and undertreated. Misconceptions and stigma may delay care-seeking behaviors and negatively influence patient outcomes. Methods: We conducted a cross-sectional, questionnaire-based study in Romania to assess public awareness, attitudes, and [...] Read more.
Background: Despite their high prevalence and potential for significant morbidity, hemorrhoidal symptoms remain underreported and undertreated. Misconceptions and stigma may delay care-seeking behaviors and negatively influence patient outcomes. Methods: We conducted a cross-sectional, questionnaire-based study in Romania to assess public awareness, attitudes, and barriers related to hemorrhoidal disease. The survey included 185 participants and evaluated variables such as symptom severity, understanding of the condition, perceived stigma, and willingness to consult a physician. Results: Only 30.8% of participants had sought medical advice for hemorrhoidal symptoms. Younger age (p < 0.001), male sex (p = 0.013), and lower levels of perceived severity were significantly associated with reluctance to seek medical care. The most frequently reported barriers were embarrassment and fear of invasive diagnostic procedures. Colonoscopy and digital rectal examination were identified as major deterrents by 39.5% and 38.9% of respondents, respectively. Educational level influenced both the perceived understanding of the disease (p = 0.001) and comfort in discussing anal symptoms (p = 0.002). Gender preference for physicians was significantly associated with respondent sex (p = 0.007) but not with education or age. Conclusions: Hemorrhoidal disease remains a stigmatized and underestimated condition. Public health efforts should prioritize educational interventions, destigmatization campaigns, and improved physician–patient communication to facilitate earlier diagnosis and better disease management. Full article
(This article belongs to the Section Gastroenterology & Hepatopancreatobiliary Medicine)
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16 pages, 1795 KiB  
Article
Hospital Coordination and Protocols Using Serum and Peripheral Blood Cells from Patients and Healthy Donors in a Longitudinal Study of Guillain–Barré Syndrome
by Raquel Díaz, Javier Blanco-García, Javier Rodríguez-Gómez, Eduardo Vargas-Baquero, Carmen Fernández-Alarcón, José Rafael Terán-Tinedo, Lorenzo Romero-Ramírez, Jörg Mey, José de la Fuente, Margarita Villar, Angela Beneitez, María del Carmen Muñoz-Turrillas, María Zurdo-López, Miriam Sagredo del Río, María del Carmen Lorenzo-Lozano, Carlos Marsal-Alonso, Maria Isabel Morales-Casado, Javier Parra-Serrano and Ernesto Doncel-Pérez
Diagnostics 2025, 15(15), 1900; https://doi.org/10.3390/diagnostics15151900 - 29 Jul 2025
Viewed by 180
Abstract
Background/Objectives: Guillain–Barré syndrome (GBS) is a rare autoimmune peripheral neuropathy that affects both the myelin sheaths and axons of the peripheral nervous system. It is the leading cause of acute neuromuscular paralysis worldwide, with an annual incidence of less than two cases per [...] Read more.
Background/Objectives: Guillain–Barré syndrome (GBS) is a rare autoimmune peripheral neuropathy that affects both the myelin sheaths and axons of the peripheral nervous system. It is the leading cause of acute neuromuscular paralysis worldwide, with an annual incidence of less than two cases per 100,000 people. Although most patients recover, a small proportion do not regain mobility and even remain dependent on mechanical ventilation. In this study, we refer to the analysis of samples collected from GBS patients at different defined time points during hospital recovery and performed by a medical or research group. Methods: The conditions for whole blood collection, peripheral blood mononuclear cell isolation, and serum collection from GBS patients and volunteer donors are explained. Aliquots of these human samples have been used for red blood cell phenotyping, transcriptomic and proteomic analyses, and serum biochemical parameter studies. Results: The initial sporadic preservation of human samples from GBS patients and control volunteers enabled the creation of a biobank collection for current and future studies related to the diagnosis and treatment of GBS. Conclusions: In this article, we describe the laboratory procedures and the integration of a GBS biobank collection, local medical services, and academic institutions collaborating in its respective field. The report establishes the intra-disciplinary and inter-institutional network to conduct long-term longitudinal studies on GBS. Full article
(This article belongs to the Section Clinical Laboratory Medicine)
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15 pages, 1262 KiB  
Article
Bycatch in Cetaceans from the North-Western Mediterranean Sea: Retrospective Study of Lesions and Utility of Bycatch Criteria
by Laura Martino, Mariona Leiva Forns, Marina Cid Cañete, Lola Pérez, Cèlia Pradas and Mariano Domingo
Vet. Sci. 2025, 12(8), 711; https://doi.org/10.3390/vetsci12080711 - 29 Jul 2025
Viewed by 382
Abstract
Bycatch is the most common cause of death of small delphinids worldwide, including the Mediterranean Sea. The diagnosis of bycatch as cause of death in stranded cetaceans depends on the cumulative presence of multiple findings, termed bycatch criteria. In this study, we retrospectively [...] Read more.
Bycatch is the most common cause of death of small delphinids worldwide, including the Mediterranean Sea. The diagnosis of bycatch as cause of death in stranded cetaceans depends on the cumulative presence of multiple findings, termed bycatch criteria. In this study, we retrospectively evaluated the presence of bycatch criteria in 138 necropsied cetaceans, 136 stranded and 2 confirmed bycaught, in the Catalan Mediterranean Sea across a 13-year period. With the aim of identifying the most specific and reliable bycatch criteria, the animals’ cause of death was classified as either bycaught or other causes. Animals were necropsied according to standard procedures with complete histopathological examination and ancillary diagnostic techniques. We reviewed the necropsy reports and photographs of 138 cetaceans of seven species. Bycatch had been determined as the cause of death/stranding in 40 (29%) necropsied cetaceans. Both sexes were equally represented in the bycatch group. Bycatch was diagnosed in the Mediterranean common bottlenose dolphin (10/14; 71.4%), striped dolphin (29/108; 26.9%), and Risso’s dolphin (1/11; 9.1%). Sixty-seven out of 98 (68.3%) cetaceans that had been classified as non-bycatch had one or two bycatch criteria. Cetaceans with two and three major criteria had an overlap of causes of death, as some animals were diagnosed with bycatch and others with other causes of mortality. Animals with four criteria were invariably diagnosed as being bycaught. Recent feeding, absence of disease, good nutritional status, marks of fishing gear, multiorgan intravascular gas bubbles, hyphema and amputations or sharp incisions presumably inflicted by humans were significantly more likely to result in a diagnosis of bycatch, while loss of teeth and cranial fractures were not. None of the dolphins diagnosed as bycatch had ingested fishing gear. Our results highlight the relevance of bycatch as the cause of death of dolphins in the Mediterranean and suggest that some criteria traditionally linked to bycatch are not specific for bycatch in our region. Full article
(This article belongs to the Special Issue Pathology of Marine Large Vertebrates)
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11 pages, 556 KiB  
Article
Added Value of SPECT/CT in Radio-Guided Occult Localization (ROLL) of Non-Palpable Pulmonary Nodules Treated with Uniportal Video-Assisted Thoracoscopy
by Demetrio Aricò, Lucia Motta, Giulia Giacoppo, Michelangelo Bambaci, Paolo Macrì, Stefania Maria, Francesco Barbagallo, Nicola Ricottone, Lorenza Marino, Gianmarco Motta, Giorgia Leone, Carlo Carnaghi, Vittorio Gebbia, Domenica Caponnetto and Laura Evangelista
J. Clin. Med. 2025, 14(15), 5337; https://doi.org/10.3390/jcm14155337 - 29 Jul 2025
Viewed by 188
Abstract
Background/Objectives: The extensive use of computed tomography (CT) has led to a significant increase in the detection of small and non-palpable pulmonary nodules, necessitating the use of invasive methods for definitive diagnosis. Video-assisted thoracoscopic surgery (VATS) has become the preferred procedure for nodule [...] Read more.
Background/Objectives: The extensive use of computed tomography (CT) has led to a significant increase in the detection of small and non-palpable pulmonary nodules, necessitating the use of invasive methods for definitive diagnosis. Video-assisted thoracoscopic surgery (VATS) has become the preferred procedure for nodule resections; however, intraoperative localization remains challenging, especially for deep or subsolid lesions. This study explores whether SPECT/CT improves the technical and clinical outcomes of radio-guided occult lesion localization (ROLL) before uniportal video-assisted thoracoscopic surgery (u-VATS). Methods: This is a retrospective study involving consecutive patients referred for the resection of pulmonary nodules who underwent CT-guided ROLL followed by u-VATS between September 2017 and December 2024. From January 2023, SPECT/CT was systematically added after planar imaging. The cohort was divided into a planar group and a planar + SPECT/CT group. The inclusion criteria involved nodules sized ≤ 2 cm, with ground glass or solid characteristics, located at a depth of <6 cm from the pleural surface. 99mTc-MAA injected activity, timing, the classification of planar and SPECT/CT image findings (focal uptake, multisite with focal uptake, multisite without focal uptake), spillage, and post-procedure complications were evaluated. Statistical analysis was performed, with continuous data expressed as the median and categorical data as the number. Comparisons were made using chi-square tests for categorical variables and the Mann–Whitney U test for procedural duration. Cohen’s kappa coefficient was calculated to assess agreement between imaging modalities. Results: In total, 125 patients were selected for CT-guided radiotracer injection followed by uniportal-VATS. The planar group and planar + SPECT/CT group comprised 60 and 65 patients, respectively. Focal uptake was detected in 68 (54%), multisite with focal uptake in 46 (36.8%), and multisite without focal uptake in 11 patients (8.8%). In comparative analyses between planar and SPECT/CT imaging in 65 patients, 91% exhibited focal uptake, revealing significant differences in classification for 40% of the patients. SPECT/CT corrected the classification of 23 patients initially categorized as multisite with focal uptake to focal uptake, improving localization accuracy. The mean procedure duration was 39 min with SPECT/CT. Pneumothorax was more frequently detected with SPECT/CT (43% vs. 1.6%). The intraoperative localization success rate was 96%. Conclusions: SPECT/CT imaging in the ROLL procedure for detecting pulmonary nodules before u-VATs demonstrates a significant advantage in reclassifying radiotracer positioning compared to planar imaging. Considering its limited impact on surgical success rates and additional procedural time, SPECT/CT should be reserved for technically challenging cases. Larger sample sizes, multicentric and prospective randomized studies, and formal cost–utility analyses are warranted. Full article
(This article belongs to the Section Nuclear Medicine & Radiology)
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16 pages, 5818 KiB  
Case Report
Novel Sonoguided Digital Palpation and Ultrasound-Guided Hydrodissection of the Long Thoracic Nerve for Managing Serratus Anterior Muscle Pain Syndrome: A Case Report with Technical Details
by Nunung Nugroho, King Hei Stanley Lam, Theodore Tandiono, Teinny Suryadi, Anwar Suhaimi, Wahida Ratnawati, Daniel Chiung-Jui Su, Yonghyun Yoon and Kenneth Dean Reeves
Diagnostics 2025, 15(15), 1891; https://doi.org/10.3390/diagnostics15151891 - 28 Jul 2025
Viewed by 741
Abstract
Background and Clinical Significance: Serratus Anterior Muscle Pain Syndrome (SAMPS) is an underdiagnosed cause of anterior chest wall pain, often attributed to myofascial trigger points of the serratus anterior muscle (SAM) or dysfunction of the Long Thoracic Nerve (LTN), leading to significant disability [...] Read more.
Background and Clinical Significance: Serratus Anterior Muscle Pain Syndrome (SAMPS) is an underdiagnosed cause of anterior chest wall pain, often attributed to myofascial trigger points of the serratus anterior muscle (SAM) or dysfunction of the Long Thoracic Nerve (LTN), leading to significant disability and affecting ipsilateral upper limb movement and quality of life. Current diagnosis relies on exclusion and physical examination, with limited treatment options beyond conservative approaches. This case report presents a novel approach to chronic SAMPS, successfully diagnosed using Sonoguided Digital Palpation (SDP) and treated with ultrasound-guided hydrodissection of the LTN using 5% dextrose in water (D5W) without local anesthetic (LA), in a patient where conventional treatments had failed. Case Presentation: A 72-year-old male presented with a three-year history of persistent left chest pain radiating to the upper back, exacerbated by activity and mimicking cardiac pain. His medical history included two percutaneous coronary interventions. Physical examination revealed tenderness along the anterior axillary line and a positive hyperirritable spot at the mid axillary line at the 5th rib level. SDP was used to visualize the serratus anterior fascia (SAF) and LTN, and to reproduce the patient’s concordant pain by palpating the LTN. Ultrasound-guided hydrodissection of the LTN was then performed using 20–30cc of D5W without LA to separate the nerve from the surrounding tissues, employing a “fascial unzipping” technique. The patient reported immediate pain relief post-procedure, with the pain reducing from 9/10 to 1/10 on the Numeric Rating Scale (NRS), and sustained relief and functional improvement at the 12-month follow-up. Conclusions: Sonoguided Digital Palpation (SDP) of the LTN can serve as a valuable diagnostic adjunct for visualizing and diagnosing SAMPS. Ultrasound-guided hydrodissection of the LTN with D5W without LA may provide a promising and safe treatment option for patients with chronic SAMPS refractory to conservative management, resulting in rapid and sustained pain relief. Further research, including controlled trials, is warranted to evaluate the long-term efficacy and generalizability of these findings and to compare D5W to other injectates. Full article
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13 pages, 1650 KiB  
Article
A Fast TaqMan® Real-Time PCR Assay for the Detection of Mitochondrial DNA Haplotypes in a Wolf Population
by Rita Lorenzini, Lorenzo Attili, Martina De Crescenzo and Antonella Pizzarelli
Genes 2025, 16(8), 897; https://doi.org/10.3390/genes16080897 - 28 Jul 2025
Viewed by 168
Abstract
Background/Objectives: The gene pool of the Apennine wolf is affected by admixture with domestic variants due to anthropogenic hybridisation with dogs. Genetic monitoring at the population level involves assessing the extent of admixture in single individuals, ranging from pure wolves to recent [...] Read more.
Background/Objectives: The gene pool of the Apennine wolf is affected by admixture with domestic variants due to anthropogenic hybridisation with dogs. Genetic monitoring at the population level involves assessing the extent of admixture in single individuals, ranging from pure wolves to recent hybrids or wolf backcrosses, through the analysis of nuclear and mitochondrial DNA (mtDNA) markers. Although individually non-diagnostic, mtDNA is nevertheless essential for completing the final diagnosis of genetic admixture. Typically, the identification of wolf mtDNA haplotypes is carried out via sequencing of coding genes and non-coding DNA stretches. Our objective was to develop a fast real-time PCR assay to detect the mtDNA haplotypes that occur exclusively in the Apennine wolf population, as a valuable alternative to the demanding sequence-based typing. Methods: We validated a qualitative duplex real-time PCR that exploits the combined presence of diagnostic point mutations in two mtDNA segments, the NDH-4 gene and the control region, and is performed in a single-tube step through TaqMan-MGB chemistry. The aim was to detect mtDNA multi-fragment haplotypes that are exclusive to the Apennine wolf, bypassing sequencing. Results: Basic validation of 149 field samples, consisting of pure Apennine wolves, dogs, wolf × dog hybrids, and Dinaric wolves, showed that the assay is highly specific and sensitive, with genomic DNA amounts as low as 10−5 ng still producing positive results. It also proved high repeatability and reproducibility, thereby enabling reliable high-throughput testing. Conclusions: The results indicate that the assay presented here provides a valuable alternative method to the time- and cost-consuming sequencing procedure to reliably diagnose the maternal lineage of the still-threatened Apennine wolf, and it covers a wide range of applications, from scientific research to conservation, diagnostics, and forensics. Full article
(This article belongs to the Section Animal Genetics and Genomics)
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25 pages, 2913 KiB  
Review
The Art of Interpreting Antinuclear Antibodies (ANAs) in Everyday Practice
by Marcelina Kądziela, Aleksandra Fijałkowska, Marzena Kraska-Gacka and Anna Woźniacka
J. Clin. Med. 2025, 14(15), 5322; https://doi.org/10.3390/jcm14155322 - 28 Jul 2025
Viewed by 230
Abstract
Background: Antinuclear antibodies (ANAs) serve as crucial biomarkers for diagnosing systemic autoimmune diseases; however, their interpretation can be complex and may not always correlate with clinical symptoms. Methods: A comprehensive narrative review was conducted to evaluate the peer-reviewed literature published between 1961 and [...] Read more.
Background: Antinuclear antibodies (ANAs) serve as crucial biomarkers for diagnosing systemic autoimmune diseases; however, their interpretation can be complex and may not always correlate with clinical symptoms. Methods: A comprehensive narrative review was conducted to evaluate the peer-reviewed literature published between 1961 and 2025. Databases, including PubMed and Scopus, were searched using combinations of controlled vocabulary and free-text terms relating to antinuclear antibodies and their clinical significance. The objective was to gather and synthesize information regarding the diagnostic utility and interpretation of ANA testing in routine medical practice. Discussion: The indirect immunofluorescence assay (IIF) on HEp-2 cells is established as the gold standard for detecting ANAs, facilitating the classification of various fluorescent patterns. While a positive ANA test can suggest autoimmune disorders, the presence and titre must be interpreted alongside clinical findings, as low titres often lack diagnostic significance. Findings indicate that titres higher than 1:160 may provide greater specificity in differentiating true positives from false positives in healthy individuals. The study also emphasizes the relevance of fluorescence patterns, with specific patterns linked to particular diseases, although many do not have strong clinical correlations. Moreover, certain autoantibodies demonstrate high specificity for diseases like systemic lupus erythematosus (SLE) and mixed connective tissue disease (MCTD). Ultimately, while ANA testing is invaluable for diagnosing connective tissue diseases, healthcare providers must consider its limitations to avoid misdiagnosis and unnecessary treatment. Conclusions: ANA testing is a valuable tool in the diagnosis of connective tissue diseases, but its interpretation must be approached with caution. Clinical context remains crucial when evaluating ANA results to avoid misdiagnosis and overtreatment. This review is about the diagnostic aspects and clinical consequences of ANA testing, as well as highlighting both the diagnostic benefits and the potential limitations of this procedure in everyday clinical practice. The review fills a gap in the literature by integrating the diagnostic and clinical aspects of ANA testing, with a focus on real-world interpretation challenges. Full article
(This article belongs to the Section Immunology)
8 pages, 855 KiB  
Case Report
Severe Malaria Due to Plasmodium falciparum in an Immunocompetent Young Adult: Rapid Progression to Multiorgan Failure
by Valeria Sanclemente-Cardoza, Harold Andrés Payán-Salcedo and Jose Luis Estela-Zape
Life 2025, 15(8), 1201; https://doi.org/10.3390/life15081201 - 28 Jul 2025
Viewed by 200
Abstract
Plasmodium falciparum malaria remains a major cause of morbidity and mortality, particularly in endemic regions. We report the case of a 21-year-old male with recent travel to an endemic area (Guapi, Colombia), who presented with febrile symptoms, severe respiratory distress, and oxygen saturation [...] Read more.
Plasmodium falciparum malaria remains a major cause of morbidity and mortality, particularly in endemic regions. We report the case of a 21-year-old male with recent travel to an endemic area (Guapi, Colombia), who presented with febrile symptoms, severe respiratory distress, and oxygen saturation below 75%, necessitating orotracheal intubation. During the procedure, he developed pulseless electrical activity cardiac arrest, achieving return of spontaneous circulation after advanced resuscitation. Diagnosis was confirmed by thick blood smear, demonstrating P. falciparum infection. The patient progressed to multiorgan failure, including acute respiratory distress syndrome with capillary leak pulmonary edema, refractory distributive shock, acute kidney injury with severe hyperkalemia, and consumptive thrombocytopenia. Management included invasive mechanical ventilation, vasopressor support, sedation-analgesia, neuromuscular blockade, methylene blue, unsuccessful hemodialysis due to hemorrhagic complications, and platelet transfusions. Despite these interventions, the patient experienced a second cardiac arrest and died. This case highlights the severity and rapid progression of severe malaria with multisystem involvement, underscoring the critical importance of early diagnosis and intensive multidisciplinary management. It also emphasizes the need for preventive strategies for travelers to endemic areas and the development of clinical protocols to improve outcomes in complicated malaria. Full article
(This article belongs to the Section Medical Research)
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21 pages, 14138 KiB  
Case Report
Multi-Level Oncological Management of a Rare, Combined Mediastinal Tumor: A Case Report
by Vasileios Theocharidis, Thomas Rallis, Apostolos Gogakos, Dimitrios Paliouras, Achilleas Lazopoulos, Meropi Koutourini, Myrto Tzinevi, Aikaterini Vildiridi, Prokopios Dimopoulos, Dimitrios Kasarakis, Panagiotis Kousidis, Anastasia Nikolaidou, Paraskevas Vrochidis, Maria Mironidou-Tzouveleki and Nikolaos Barbetakis
Curr. Oncol. 2025, 32(8), 423; https://doi.org/10.3390/curroncol32080423 - 28 Jul 2025
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Abstract
Malignant mediastinal tumors are a group representing some of the most demanding oncological challenges for early, multi-level, and successful management. The timely identification of any suspicious clinical symptomatology is urgent in achieving an accurate, staged histological diagnosis, in order to follow up with [...] Read more.
Malignant mediastinal tumors are a group representing some of the most demanding oncological challenges for early, multi-level, and successful management. The timely identification of any suspicious clinical symptomatology is urgent in achieving an accurate, staged histological diagnosis, in order to follow up with an equally detailed medical therapeutic plan (interventional or not) and determine the principal goals regarding efficient overall treatment in these patients. We report a case of a 24-year-old male patient with an incident-free prior medical history. An initial chest X-ray was performed after the patient reported short-term, consistent moderate chest pain symptomatology, early work fatigue, and shortness of breath. The following imaging procedures (chest CT, PET-CT) indicated the presence of an anterior mediastinal mass (meas. ~11 cm × 10 cm × 13 cm, SUV: 8.7), applying additional pressure upon both right heart chambers. The Alpha-Fetoprotein (aFP) blood levels had exceeded at least 50 times their normal range. Two consecutive diagnostic attempts with non-specific histological results, a negative-for-malignancy fine-needle aspiration biopsy (FNA-biopsy), and an additional tumor biopsy, performed via mini anterior (R) thoracotomy with “suspicious” cellular gatherings, were performed elsewhere. After admission to our department, an (R) Video-Assisted Thoracic Surgery (VATS) was performed, along with multiple tumor biopsies and moderate pleural effusion drainage. The tumor’s measurements had increased to DMax: 16 cm × 9 cm × 13 cm, with a severe degree of atelectasis of the Right Lower Lobe parenchyma (RLL) and a pressure-displacement effect upon the Superior Vena Cava (SVC) and the (R) heart sinus, based on data from the preoperative chest MRA. The histological report indicated elements of a combined, non-seminomatous germ-cell mediastinal tumor, posthuberal-type teratoma, and embryonal carcinoma. The imminent chemotherapeutic plan included a “BEP” (Bleomycin®/Cisplatin®/Etoposide®) scheme, which needed to be modified to a “VIP” (Cisplatin®/Etoposide®/Ifosfamide®) scheme, due to an acute pulmonary embolism incident. While the aFP blood levels declined, even reaching normal measurements, the tumor’s size continued to increase significantly (DMax: 28 cm × 25 cm × 13 cm), with severe localized pressure effects, rapid weight loss, and a progressively worsening clinical status. Thus, an emergency surgical intervention took place via median sternotomy, extended with a complementary “T-Shaped” mini anterior (R) thoracotomy. A large, approx. 4 Kg mediastinal tumor was extracted, with additional RML and RUL “en-bloc” segmentectomy and partial mediastinal pleura decortication. The following histological results, apart from verifying the already-known posthuberal-type teratoma, indicated additional scattered small lesions of combined high-grade rabdomyosarcoma, chondrosarcoma, and osteosarcoma, as well as numerous high-grade glioblastoma cellular gatherings. No visible findings of the previously discovered non-seminomatous germ-cell and embryonal carcinoma elements were found. The patient’s postoperative status progressively improved, allowing therapeutic management to continue with six “TIP” (Cisplatin®/Paclitaxel®/Ifosfamide®) sessions, currently under his regular “follow-up” from the oncological team. This report underlines the importance of early, accurate histological identification, combined with any necessary surgical intervention, diagnostic or therapeutic, as well as the appliance of any subsequent multimodality management plan. The diversity of mediastinal tumors, especially for young patients, leaves no place for complacency. Such rare examples may manifest, with equivalent, unpredictable evolution, obliging clinical physicians to stay constantly alert and not take anything for granted. Full article
(This article belongs to the Section Thoracic Oncology)
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11 pages, 924 KiB  
Article
Radial-Probe Endobronchial Ultrasound as Part of Different Navigational Bronchoscopy Modalities in Combination with Cryobiopsy Could Be More than a Confirmation Tool: A Case Series
by Nevenka Piskac Zivkovic, Maja Karaman Ilic, Suncana Divosevic, Hrvoje Feljan, Igor Nikolic, Zrinka Juros, Ana-Marija Sola, Sven Seiwerth, Dragan Schwarz and Ivica Mazuranic
Diagnostics 2025, 15(15), 1884; https://doi.org/10.3390/diagnostics15151884 - 27 Jul 2025
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Abstract
Background: As part of different navigational bronchoscopy (NVB) modalities, radial-probe endobronchial ultrasound (rEBUS) is used to confirm the peribronchial localization of peripheral pulmonary nodules (PPNs) immediately before collecting samples for histopathological analysis. Methods: This retrospective case series study presents the results [...] Read more.
Background: As part of different navigational bronchoscopy (NVB) modalities, radial-probe endobronchial ultrasound (rEBUS) is used to confirm the peribronchial localization of peripheral pulmonary nodules (PPNs) immediately before collecting samples for histopathological analysis. Methods: This retrospective case series study presents the results of en bloc cryobiopsy of PPNs using a flexible 1.1-mm cryoprobe with different NVB modalities. For PPNs classified as adjacent or eccentric lesions by rEBUS (ES-rEBUS), the cryoprobe’s position was adjusted by 90–180° in relation to the ultrasound image of the lesion during the first and second biopsies. Results: All patients with a final histopathologically confirmed diagnosis of PPNs had positive rEBUS findings, regardless of the navigation modality, eccentric (18/42 patients, 43%) and concentric (24/42 patients, 57%) rEBUS view. In 5 out of 6 patients without a histopathological diagnosis, PPNs were not visualized by radial ultrasound. In the (ES-rEBUS) group of patients, 4 out of 18 had fewer than three biopsy samples collected per procedure, which means only an adjusted probe position has been applied, although diagnostic outcomes were achieved. Common Terminology Criteria for Adverse Events (CTCAE) grade 2 complications were reported in 10.4% of the patients, and grade 3 complications in 2% of the patients. Conclusions: Confirming the localization of nodules by rEBUS and properly adjusting the cryoprobe immediately before cryobiopsy of PPNs resulted in a diagnostic yield meeting the literature standards. Full article
(This article belongs to the Special Issue New Advances in Diagnostic Bronchoscopy)
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14 pages, 3811 KiB  
Article
Ultrasonography Elastography to Predict the Diagnosis, Severity, and Treatment Indication of Esophageal Varices in Patients with Chronic Liver Diseases
by Azusa Wada, Yasunobu Yamashita, Mikitaka Iguchi, Yoshiyuki Ida, Takao Maekita, Reiko Ashida and Masayuki Kitano
Diagnostics 2025, 15(15), 1867; https://doi.org/10.3390/diagnostics15151867 - 25 Jul 2025
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Abstract
Background/Objectives: Esophageal varices (EVs) are a serious complication of liver cirrhosis. Guidelines for cirrhosis/chronic liver diseases (CLDs) do not specify a follow-up period or the need for esophagogastroduodenoscopy (EGD). EGD is a useful but uncomfortable procedure for the assessment of varices. Follow-up [...] Read more.
Background/Objectives: Esophageal varices (EVs) are a serious complication of liver cirrhosis. Guidelines for cirrhosis/chronic liver diseases (CLDs) do not specify a follow-up period or the need for esophagogastroduodenoscopy (EGD). EGD is a useful but uncomfortable procedure for the assessment of varices. Follow-up with abdominal ultrasonography (AUS) is recommended in patients with CLDs. If EVs are assessed by AUS, more patients eligible for endoscopic screening of EVs can be selected. We aimed to investigate whether AUS elastography [shear wave (Vs) and F-index] can predict the diagnosis, severity, and treatment indication of EVs. Methods: Between April 2018 and October 2022, we retrospectively collected data of 194 patients who underwent elastography and EGD for CLDs. The correlations between Vs/F-index values and presence/severity of EVs were evaluated. Each cut-off value for diagnosis and treatment indication of EVs was investigated. Results: 85 patients without exclusion criteria were enrolled. Vs and F-index values were significantly higher in patients with EVs than in patients without EVs (p = 0.0005 and 0.0021, respectively) and positively correlated with severity of EVs. The cut-off Vs and F-index values for the presence of EVs were 1.63 m/s and 1.88, respectively, with 88.1%/83.3% sensitivity, 48.8%/51.2% specificity, and 0.71/0.70 area under the curve (AUC). The cut-off Vs and F-index values for treatment indication were 1.71 m/s and 2.08, respectively, with 100%/88.2% sensitivity, 45.6%/52.9% specificity, and 0.69/0.70 AUC. There were no significant differences between the two modalities. Conclusions: Elastography may provide objective assessment and thus be a non-invasive screening tool for diagnosis and treatment indication of EVs. Full article
(This article belongs to the Special Issue New Insights into Endoscopy-Guided Diagnosis)
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