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16 pages, 611 KB  
Article
Dissociation of Clinical Outcomes and CSF Proteinopathy Biomarkers in Parkinson’s Disease: Cognitive–Affective Dissociation with Specificity for Tau
by João Paulo Mota Telles, Lucas Camargo, Anna Carolyna Gianlorenço and Felipe Fregni
Biomedicines 2025, 13(10), 2478; https://doi.org/10.3390/biomedicines13102478 (registering DOI) - 11 Oct 2025
Viewed by 20
Abstract
Background: Parkinson’s disease (PD) is characterized not only by motor dysfunction but also by widespread degeneration across cortico-striatal, limbic, and cortical circuits. Mounting evidence suggests that tau and α-synuclein pathology underlie these processes, though how these proteinopathies translate into affective and cognitive outcomes [...] Read more.
Background: Parkinson’s disease (PD) is characterized not only by motor dysfunction but also by widespread degeneration across cortico-striatal, limbic, and cortical circuits. Mounting evidence suggests that tau and α-synuclein pathology underlie these processes, though how these proteinopathies translate into affective and cognitive outcomes remains uncertain. Depression and anxiety are highly prevalent in PD, yet the biological correlates of these affective disturbances are poorly defined. Methods: This is a retrospective analysis of existing data from the Parkinson’s Progression Markers Initiative (PPMI). Montreal Cognitive Assessment (MoCA), geriatric depression scale (GDS), and State-Trait Anxiety Inventory (STAI) were used to assess cognition, depression, and anxiety in PD, respectively. The CSF biomarkers evaluated were Aβ42, t-tau, and p-tau181, using Elecsys electro-chemiluminescence immunoassays on the cobas e601 platform (Roche Diagnostics). Results: From the 4380 patients who had GDS information, the MoCA test was collected from 438 patients, and 445 from the GDS test for depression, and the STAI screening for anxiety. There were no significant differences in biomarker levels between patients with depression (GDS ≥ 5) and those without (GDS < 5), nor between patients with anxiety (STAI > 40) and those with lower anxiety scores (STAI ≤ 40). In contrast, cognitive outcomes showed clear associations. Patients with cognitive impairment (MoCA < 26) demonstrated higher levels of pTau (p = 0.02) and tTau (p = 0.01), as well as elevated pTau/Aβ42 (p = 0.003) and tTau/Aβ42 (p = 0.002) ratios compared to those with MoCA ≥ 26. In multivariate analysis, both pTau/Aβ42 > 0.022 (OR 4.64, 95% CI 1.67–13.8) and tTau/Aβ42 > 0.26 (OR 4.18, 95% CI 1.6–11.5) remained significantly associated with cognitive decline. In a longitudinal analysis in the first 3 years of follow-up, cognition in PD remained lower than in controls, while CSF p-tau and Aβ42 remained higher in controls. Conclusions: In our cohort, no associations were found between CSF biomarkers and depression or anxiety, underscoring that mood disturbances in PD are likely mediated by alternative mechanisms such as monoaminergic dysregulation, neuroinflammation, and psychosocial factors. By contrast, cognitive performance (MoCA) was clearly linked to tau-related pathology, rather than α-synuclein or Aβ42 alone. While Aβ42 and α-synuclein remain useful for staging and assessing global disease risk, our findings highlight the specificity of tau-related pathology for cognitive outcomes in PD. Full article
(This article belongs to the Section Neurobiology and Clinical Neuroscience)
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10 pages, 1038 KB  
Article
Cytokine Profiling of Exudates from Periapical Lesions and the Efficacy of CXCL10 as a Healing Marker
by Kazuhisa Ouhara, Yuri Taniguchi, Ruoqi Zhai, Katsuhiro Takeda, Ryousuke Fujimori, Naoya Kuwahara, Shoya Ueda, Yitong Hou, Nomi Honoka, Masaru Shimizu, Shoko Kono, Tomoyuki Iwata, Shinji Matsuda and Noriyoshi Mizuno
Pathogens 2025, 14(10), 1013; https://doi.org/10.3390/pathogens14101013 - 7 Oct 2025
Viewed by 225
Abstract
This study aimed to evaluate cytokine profiling in a periapical lesion to provide a rationale for future treatment strategies for periapical lesions. Thirteen samples of exudative fluid were collected from such a lesion directly through the root canal. Cytokine profiling was performed using [...] Read more.
This study aimed to evaluate cytokine profiling in a periapical lesion to provide a rationale for future treatment strategies for periapical lesions. Thirteen samples of exudative fluid were collected from such a lesion directly through the root canal. Cytokine profiling was performed using the Bio-Plex system. CXCL10 (C-X-C motif chemokine ligand 10, IP10) was found to be elevated in apical exudates of patients exhibiting favorable healing. To evaluate the role of CXCL10 in cell migration, a Transwell assay was conducted using bone marrow-derived mononuclear cells (BMMCs). Different types of cytokines were detected from the samples of periapical lesion at the initial visit. However, cytokine production varied across patient samples. Release of interleukin (IL)-1β, IL-2, IL-4, IL-5, IL-6, IL-8, IL-10, IL-12, IL-13, IL-17, granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage colony-stimulating factor (GM-CSF), interferon gamma (IFN-γ), monocyte chemoattractant protein (MCP)-1, macrophage inflammatory protein (MIP)-1α, MIP-1β, and tumor necrosis factor (TNF)-α showed differential expression. Comparison of cytokine profiles indicated that cytokine production was variable before and after root canal treatment. In vitro, CXCL10 significantly improved BMMC migration in a dose-dependent manner, supporting clinical findings that elevated CXCL10 levels are associated with favorable healing in apical lesions. Although this study was limited by the small sample size and exploratory design, the cytokine profile of periapical lesions may be useful for assessing the condition of periapical lesions and modulating the immune response to bacterial infection. Full article
(This article belongs to the Section Bacterial Pathogens)
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17 pages, 1876 KB  
Article
Shunt Dysfunction Assessment in Shunted Patients via Multiparametric Non-Invasive ICP Waveform Monitoring
by Raphael Bertani, Gabriel Semione, Milene Zanella Capitanio, Sérgio Brasil, Sávio Batista, Gabriel André da Silva Mendes, Caio Perret, Christian Ferreira, Wellingson Silva Paiva and Fernando Campos Gomes Pinto
Biomedicines 2025, 13(10), 2436; https://doi.org/10.3390/biomedicines13102436 - 7 Oct 2025
Viewed by 259
Abstract
Background/Objectives: Ventriculoperitoneal shunts are the standard treatment for adults diagnosed with several CSF disorders, but often face dysfunction, leading interest in non-invasive methods for diagnosing shunt issues. This study evaluates the potential of non-invasive intracranial pressure waveform monitoring (nICPw) with the brain4care [...] Read more.
Background/Objectives: Ventriculoperitoneal shunts are the standard treatment for adults diagnosed with several CSF disorders, but often face dysfunction, leading interest in non-invasive methods for diagnosing shunt issues. This study evaluates the potential of non-invasive intracranial pressure waveform monitoring (nICPw) with the brain4care (B4C) system to distinguish overdrainage, underdrainage, and normal shunt function in patients with CSF disorders. Methods: In this single-center, observational study at Hospital das Clínicas, Brazil, adult patients with CSF shunts were enrolled. Patients were categorized as Overdrainage or Underdrainage, based on clinical parameters, with an Asymptomatic group. The B4C system provided nICPw monitoring, and six parameters (including various P2/P1 ratios) were analyzed via MANOVA and ANOVA. Results: Among 30 patients (6 overdrainage, 6 underdrainage, 18 asymptomatic), five asymptomatic patients were excluded from the main analysis due to incomplete data collection, leaving 25 patients. Overdrainage patients had significantly higher ΔP2/P1 values (0.618 ± 0.210) than asymptomatic ones (0.227 ± 0.171). After excluding outliers, differences were more pronounced (H = 10.89; p < 0.01). Underdrainage patients had intermediate ΔP2/P1 values (0.387 ± 0.179) and consistently higher P2/P1 averages (>1.3). ROC analysis indicated that ΔP2/P1 > 0.3 suggested shunt dysfunction (AUC = 0.731), while the highest P2/P1 offered stronger discrimination (AUC = 0.782). A global average P2/P1 > 1.3 was linked to underdrainage, with the lowest P2/P1 values differentiating overdrainage (0.948 ± 0.321) from underdrainage (1.143 ± 0.156). Conclusions: nICPw monitoring with the B4C system demonstrated potential for detecting shunt dysfunction. Combining parameters, especially ΔP2/P1 and highest P2/P1, improves diagnostic accuracy, offering a non-invasive method that may aid in distinguishing normal from abnormal shunt function. Full article
(This article belongs to the Section Molecular and Translational Medicine)
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19 pages, 2543 KB  
Article
Impact of Nutritional Status on Clinical Outcomes of Patients Undergoing PRGF Treatment for Knee Osteoarthritis—A Prospective Observational Study
by Paola De Luca, Giulio Grieco, Simona Landoni, Eugenio Caradonna, Valerio Pascale, Enrico Ragni and Laura de Girolamo
Nutrients 2025, 17(19), 3134; https://doi.org/10.3390/nu17193134 - 30 Sep 2025
Viewed by 182
Abstract
Background: Osteoarthritis (OA) is a major global health issue, increasing with aging and obesity. Current therapies mainly address symptoms without modifying disease progression. Platelet-rich growth factor (PRGF) therapy has potential regenerative effects through high cytokines and growth factors, but the outcomes of these [...] Read more.
Background: Osteoarthritis (OA) is a major global health issue, increasing with aging and obesity. Current therapies mainly address symptoms without modifying disease progression. Platelet-rich growth factor (PRGF) therapy has potential regenerative effects through high cytokines and growth factors, but the outcomes of these therapies remain heterogeneous. This study explores the relationship between patient nutritional status, PRGF characteristics, and clinical outcomes in knee OA treatment. Methods: Baseline anthropometric, metabolic, and nutritional assessments of 41 patients with knee OA who underwent PRGF treatment were conducted. Blood samples were analyzed for metabolic and inflammatory markers. PRGF composition was assessed by protein content and extracellular vesicle (EV) markers. KOOS and VAS pain scores were collected at 2, 6, and 12 months. Responders improved KOOS by ≥10 points. An elastic-net regularized logistic model allowed the identification of the predictors of treatment response. Results: KOOS and VAS scores improved significantly at all follow-ups. At 2 months, the PRGF of responder patients showed higher PRGF G-CSF levels; at 12 months, increased CD49e and HLA-ABC expression. Higher BMI correlated with increased IL-6, IL-1ra, and resistin in PRGF samples. Hypercholesterolemic patients displayed altered EV profiles, with elevated levels of CD8 but reduced CD49e, HLA-ABC, CD42a, and CD31. Multivariate analysis identified BMI, biceps fold, fat percentage, red blood cell, platelet, and neutrophil counts as predictors of early response. Conclusions: Metabolic and immunological factors influence PRGF composition and clinical efficacy in knee OA. Baseline body composition and hematological parameters as key predictors of response, highlighting the potential of personalized PRGF therapy. Full article
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17 pages, 722 KB  
Article
Association of Plasma Placental Growth Factor with White Matter Hyperintensities in Alzheimer’s Disease
by Kazuya Igarashi, Tamao Tsukie, Kazuo Washiyama, Kiyoshi Onda, Yuki Miyagi, Shoya Inagawa, Soichiro Shimizu, Akinori Miyashita, Osamu Onodera, Takeshi Ikeuchi and Kensaku Kasuga
Biomolecules 2025, 15(10), 1367; https://doi.org/10.3390/biom15101367 - 26 Sep 2025
Viewed by 328
Abstract
Autopsy studies have shown that Alzheimer’s disease (AD) often coexists with cerebrovascular injury, affecting cognitive outcomes and the effectiveness of anti-amyloid-beta (Aβ) drugs. No fluid biomarkers of cerebrovascular injury have been identified yet. We investigated the association between white matter hyperintensities (WMH) severity [...] Read more.
Autopsy studies have shown that Alzheimer’s disease (AD) often coexists with cerebrovascular injury, affecting cognitive outcomes and the effectiveness of anti-amyloid-beta (Aβ) drugs. No fluid biomarkers of cerebrovascular injury have been identified yet. We investigated the association between white matter hyperintensities (WMH) severity and fluid biomarkers, including cerebrospinal fluid (CSF) neurofilament light chain and plasma placental growth factor (PlGF) levels. This study included 242 patients from memory clinics. Magnetic resonance imaging (MRI), CSF, and plasma samples were collected. Patients were classified as AD+ or non-AD based on the CSF Aβ42/Aβ40 ratio. In the discovery cohort (79 AD+ and 20 non-AD patients with 3D-T1 images), we analyzed the association between WMH volume and plasma PlGF. In the validation cohort (54 AD+ patients without 3D-T1 images), we analyzed the association between WMH grading and plasma PlGF. Among AD+ patients in the discovery cohort, plasma PlGF levels remained significantly associated with WMH volume and grading after adjusting for age, sex, and global cognition. Among the AD+ patients in the validation cohort, the high-PlGF (above median) group had significantly greater WMH volumes and a higher number of patients with a high WMH grading than the low-PlGF (below median) group. Plasma PlGF is a promising marker of cerebrovascular injury in AD. Full article
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14 pages, 1505 KB  
Article
Bcl-2 and FAS as Apoptosis-Related Markers in Patients with Convulsive Status Epilepticus
by Lejla Ćorić, Slavica Sović, Brankica Šimac, Iva Mihaljević, Ines Vukasović, Zrinka Čolak Romić, Ivana Šušak Sporiš and Željka Petelin Gadže
J. Clin. Med. 2025, 14(19), 6734; https://doi.org/10.3390/jcm14196734 - 24 Sep 2025
Viewed by 338
Abstract
Background: Status epilepticus (SE) is a neurological emergency associated with neuronal injury and activation of apoptotic pathways. While these mechanisms are well described in experimental models, evidence in humans is limited. This study evaluated Bcl-2 and FAS—key apoptosis-related proteins—in the serum and [...] Read more.
Background: Status epilepticus (SE) is a neurological emergency associated with neuronal injury and activation of apoptotic pathways. While these mechanisms are well described in experimental models, evidence in humans is limited. This study evaluated Bcl-2 and FAS—key apoptosis-related proteins—in the serum and cerebrospinal fluid (CSF) of patients with convulsive SE. Methods: Between February 2024 and January 2025, CSF and serum samples were collected from 18 adults with convulsive SE within 48 h of onset, and from 15 control subjects. Patients with acute brain injury, stroke, tumors, or central nervous system infections were excluded. Bcl-2 and FAS concentrations were quantified using ELISA. Serum samples were obtained at diagnosis (S1), 24 h (S2), and 7 days (S3). Results: CSF Bcl-2 levels were significantly higher in SE patients compared with controls (z = 4.1, p < 0.001). CSF FAS levels did not differ significantly (z = 0.07, p = 0.94). No differences in serum Bcl-2 were observed. In contrast, serum FAS concentrations were significantly elevated at all three time points in SE patients compared with controls (S1–S3; all p < 0.001). Conclusions: Convulsive SE is associated with distinct apoptotic responses in the central nervous system and periphery. Elevated CSF Bcl-2 may reflect acute neuroprotective or stress-related responses, whereas persistently increased serum FAS suggests systemic apoptotic activation. These findings highlight the potential prognostic and therapeutic relevance of apoptosis-related biomarkers in SE. Full article
(This article belongs to the Section Clinical Laboratory Medicine)
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11 pages, 605 KB  
Article
A Ten-Year Retrospective Review of Medical Records of Patients Admitted with Meningitis or Encephalitis at Five Hospitals in the United States Highlights the Potential for Under-Ascertainment of Invasive Meningococcal Disease
by Julio Ramirez, Stephen Furmanek, Thomas Chandler, Josue Prado, Lisa R. Harper, Steven Shen, Raffaella Iantomasi, Jessica V. Presa, Mohammad Ali, Jamie Findlow, Jennifer C. Moïsi and Frederick J. Angulo
Pathogens 2025, 14(10), 962; https://doi.org/10.3390/pathogens14100962 - 24 Sep 2025
Viewed by 388
Abstract
Laboratory confirmation of invasive meningococcal disease (IMD) relies on detection of Neisseria meningitidis in a biological specimen. Clinical management guidelines for patients presenting with signs and/or symptoms of meningitis and encephalitis emphasize the need for appropriate specimen collection for laboratory testing. To explore [...] Read more.
Laboratory confirmation of invasive meningococcal disease (IMD) relies on detection of Neisseria meningitidis in a biological specimen. Clinical management guidelines for patients presenting with signs and/or symptoms of meningitis and encephalitis emphasize the need for appropriate specimen collection for laboratory testing. To explore the potential for IMD under-diagnosis, we reviewed medical records of patients admitted with signs and/or symptoms of meningitis or encephalitis at five hospitals in Louisville, Kentucky, in 2014 to 2023. Among 675 patients admitted with meningitis and/or encephalitis with cerebrospinal fluid (CSF) cultures who received antibiotics, 300 (44.4%) received antibiotics before CSF collection. Among 431 with blood cultures who received antibiotics, 133 (30.9%) received antibiotics before blood collection. Among 751 patients with CSF collected, 651 (86.7%) CSF specimens were tested using polymerase chain reaction (PCR) for N. meningitidis detection. No blood specimens were PCR-tested. These findings indicated that current standard-of-care practices may lead to IMD under-diagnosis. Since public health surveillance relies on IMD laboratory diagnosis, these findings highlight the potential for under-ascertained IMD by surveillance. Full article
(This article belongs to the Special Issue Cutting-Edge Research on Pathogenic Neisseria)
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16 pages, 56922 KB  
Article
Translational Evaluation of an Intraparenchymal Collagen Matrix Tamponade: Initial Preclinical and Clinical Experiments to Prevent CSF Reflux Following Endoscopic Brain Surgery
by Yasuo Aihara, Kentaro Chiba, Yuichi Oda, Kevin Browne, Dmitriy Petrov, Takakazu Kawamata and John C. O’Donnell
Int. J. Mol. Sci. 2025, 26(18), 9081; https://doi.org/10.3390/ijms26189081 - 18 Sep 2025
Viewed by 385
Abstract
Transparent polymer sheaths are often utilized in neuroendoscopic procedures to minimize intraventricular bleeding and parenchymal injuries. However, cerebrospinal fluid (CSF) leakage remains a common complication following neuroendoscopic surgery for intraventricular and deep-seated lesions. We investigated an innovative technique to prevent postoperative CSF leakage [...] Read more.
Transparent polymer sheaths are often utilized in neuroendoscopic procedures to minimize intraventricular bleeding and parenchymal injuries. However, cerebrospinal fluid (CSF) leakage remains a common complication following neuroendoscopic surgery for intraventricular and deep-seated lesions. We investigated an innovative technique to prevent postoperative CSF leakage through the tract using a collagen matrix dural graft. A rolled collagen matrix (DuraGen®) was used as a parenchymal tract tamponade to seal the tract created by an angiocatheter (preclinical pilot) or neuroendoscopic sheath (clinical case studies). A small pilot study using a juvenile pig model was first conducted to test the implantation technique and to evaluate the inflammatory response to, and absorption of intraparenchymal DuraGen. The efficacy of this approach was then assessed in two clinical cases using MRI at postoperative days 1, 7, 40, and 60. The outer segment of the graft was unfurled to cover the dural defect for clinical application. In the pig model, histological analysis showed healing with minimal inflammation in DuraGen®-implanted hemispheres, while untreated control tracts exhibited parenchymal scarring and chronic inflammation. In both patients, postoperative MRI demonstrated resolution of subdural fluid collections and progressive absorption of DuraGen® with no complications. This technique ameliorated CSF leakage and enhanced parenchymal healing after neuroendoscopic surgery. DuraGen® may modulate the local environment for tissue repair beyond its use in dural grafting. Full article
(This article belongs to the Special Issue Regenerative Medicine: Biomaterials and Stem Cell Research)
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9 pages, 1367 KB  
Case Report
Case Report of Salmonella and HHV-6 Meningitis in an Infant
by Sara Abed, Tahani Asiri, Razan Alzahrani and Wujud Hunjur
Pediatr. Rep. 2025, 17(5), 94; https://doi.org/10.3390/pediatric17050094 - 15 Sep 2025
Viewed by 530
Abstract
Bacterial meningitis is one of the most serious infections. Salmonella meningitis is associated with a high prevalence of long-term adverse outcomes, often linked to acute complications and a broad range of potential neurological sequelae following the infection. Acute complications such as brain abscesses [...] Read more.
Bacterial meningitis is one of the most serious infections. Salmonella meningitis is associated with a high prevalence of long-term adverse outcomes, often linked to acute complications and a broad range of potential neurological sequelae following the infection. Acute complications such as brain abscesses and chronic complications such as hearing loss and developmental delay. In this report, we present a case of a 2-month-old male patient with seizures, hypoactivity and respiratory symptoms, who was found to have Salmonella bacteremia complicated by Salmonella and Human Herpes Virus-6 (HHV-6) meningitis, as well as rhinovirus bronchiolitis, along with follow-up findings. The patient’s data, including demographics, presenting symptoms, physical examination findings, and whole exome sequence results, as well as investigations such as complete blood count (CBC), cerebrospinal fluid (CSF) analysis, liver enzyme levels, and imaging findings, were collected from the electronic medical record system using a case report form. In addition, immunological workups were performed, as serious Salmonella infections were more common in immunocompromised patients. In the literature, there was no clear correlation between Salmonella and HHV-6 meningitis, rhinovirus bronchiolitis, and the complications that developed in this infant. This case report provides valuable insights into the clinical spectrum and long-term outcomes of patients with Salmonella meningitis. Full article
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13 pages, 1451 KB  
Article
Glutamine and Albumin Levels in Cerebrospinal Fluid Are Correlated with Neurological Severity in an Experimental Model of Acute Hepatic Encephalopathy
by Pedro Arend Guazzelli, Felipe dos Santos Fachim, Anderson Santos Travassos, Caroline Casagrande Schaukoski, Pâmela Cristina Lukasewicz Ferreira, Fernanda Uruth Fontella, Marco Antônio de Bastiani, Adriano Martimbianco de Assis and Diogo Onofre Souza
Metabolites 2025, 15(9), 598; https://doi.org/10.3390/metabo15090598 - 8 Sep 2025
Viewed by 411
Abstract
Background/Objectives: Hepatic encephalopathy (HE) is a severe neurological complication of acute liver failure (ALF) characterized by the accumulation of neurotoxic metabolites and impaired cerebral function. We aimed to examine the correlation between HE severity and cerebrospinal fluid (CSF) biomarker levels in a rat [...] Read more.
Background/Objectives: Hepatic encephalopathy (HE) is a severe neurological complication of acute liver failure (ALF) characterized by the accumulation of neurotoxic metabolites and impaired cerebral function. We aimed to examine the correlation between HE severity and cerebrospinal fluid (CSF) biomarker levels in a rat model of ALF induced by subtotal hepatectomy. Methods: Male Wistar rats underwent 92% hepatectomy and were monitored for neurological impairment via a standardized HE score. At twenty-four hours post surgery, CSF and blood were collected for biochemical analysis. Results: We found a significant positive correlation between neurological severity and CSF levels of glutamine (r = 0.929, p < 0.001) and albumin (r = 0.869, p < 0.001), both with HE grade I scores, highlighting their prominent role as HE biomarkers. Other amino acids, including aspartate (r = 0.790, p < 0.001), glutamate (r = 0.853, p < 0.001), isoleucine (r = 0.834, p < 0.001), leucine (r = 0.813, p < 0.001), lysine (r = 0.861, p < 0.001), methionine (r = 0.889, p < 0.001), phenylalanine (r = 0.916, p < 0.001), ornithine (r = 0.775, p < 0.001), tryptophan (r = 0.814, p < 0.001), and valine (r = 0.721, p < 0.001), also showed significant correlations with HE severity but not with HE grade I scores. Conclusions: These findings underscore the potential of glutamine and albumin in CSF as key biomarkers for assessing neurological severity in ALF patients. Full article
(This article belongs to the Special Issue Metabolic Profiling in Neurometabolisms)
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20 pages, 1819 KB  
Article
Cerebrospinal Fluid MicroRNAs as Early Biomarker Candidates for Predicting Vasospasm Following Aneurysmal Subarachnoid Hemorrhage
by Emre Ozkara, Ozlem Aykac, Ebru Erzurumluoglu Gokalp, Nazli Durmaz Celik, Sara Khadem Ansari, Zehra Uysal Kocabas, Ertugrul Colak, Sinem Kocagil, Zuhtu Ozbek, Oguz Cilingir, Ali Arslantas, Atilla Ozcan Ozdemir and Sevilhan Artan
Genes 2025, 16(9), 1025; https://doi.org/10.3390/genes16091025 - 29 Aug 2025
Viewed by 707
Abstract
Background/Objectives: Aneurysmal subarachnoid hemorrhage (aSAH) is frequently complicated by cerebral vasospasm, a major contributor to delayed cerebral ischemia and poor neurological outcomes. Early prediction remains challenging, and there is a critical need for reliable biomarkers. MicroRNAs (miRNAs) in cerebrospinal fluid (CSF) have [...] Read more.
Background/Objectives: Aneurysmal subarachnoid hemorrhage (aSAH) is frequently complicated by cerebral vasospasm, a major contributor to delayed cerebral ischemia and poor neurological outcomes. Early prediction remains challenging, and there is a critical need for reliable biomarkers. MicroRNAs (miRNAs) in cerebrospinal fluid (CSF) have emerged as promising indicators of acute neuropathological changes. This study aimed to evaluate CSF miRNA expression profiles in patients with aSAH to identify early predictors of vasospasm and improve clinical risk stratification. Methods: We conducted a prospective observational study involving 48 patients (40 patients with aSAH (20 who developed vasospasm, 20 who did not) and 8 healthy controls). A panel of 20 candidate miRNAs was analyzed in CSF samples collected on days 1 and 5 post−hemorrhage using quantitative real−time PCR. Expression differences between groups were assessed, and receiver operating characteristic (ROC) curves were used to evaluate diagnostic performance. Results: Several miRNAs were differentially expressed in patients who developed vasospasm. On day 1, miR−221−3p and miR−183−5p were significantly upregulated (p = 0.014, p = 0.009), while miR−126, miR−29a, and miR−27b−3p were downregulated (p = 0.006, 0.021, 0.028) compared with controls. MiR−126 remained suppressed on day 5 (p = 0.002). These early changes showed high predictive accuracy (e.g., day 1 AUC for miR−221−3p ≈ 0.98, 95% CI 0.83–1.00). Compared with non−vasospasm patients, miR−221−3p was lower (0.12−fold), while miR−9−3p and miR−183−5p were higher (13.4−fold and 2.7−fold, respectively; all p < 0.01). MiR−24 and miR−21−5p correlated with more severe grades and poorer outcomes (p < 0.05). Conclusions: Specific CSF miRNAs—particularly miR−221−3p, miR−9−3p, and miR−183−5p—may serve as early biomarkers for vasospasm, warranting further validation in larger cohorts. Full article
(This article belongs to the Section Molecular Genetics and Genomics)
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25 pages, 433 KB  
Review
The Quest for Non-Invasive Diagnosis: A Review of Liquid Biopsy in Glioblastoma
by Maria George Elias, Harry Hadjiyiannis, Fatemeh Vafaee, Kieran F. Scott, Paul de Souza, Therese M. Becker and Shadma Fatima
Cancers 2025, 17(16), 2700; https://doi.org/10.3390/cancers17162700 - 19 Aug 2025
Viewed by 1557
Abstract
Background: Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumour, associated with poor survival outcomes and significant clinical challenges. Conventional diagnostic methods, including MRI, CT, and histopathological analysis of tissue biopsies, are limited by their inability to reliably distinguish [...] Read more.
Background: Glioblastoma multiforme (GBM) is the most common and aggressive primary brain tumour, associated with poor survival outcomes and significant clinical challenges. Conventional diagnostic methods, including MRI, CT, and histopathological analysis of tissue biopsies, are limited by their inability to reliably distinguish treatment effects from true tumour progression, often resulting in misdiagnosis and delayed intervention. Repeated tissue biopsies are also invasive and unsuitable for longitudinal monitoring. Liquid biopsy, a minimally invasive approach analysing tumour-derived material in biofluids such as blood and cerebrospinal fluid (CSF), offers a promising alternative. This review aims to evaluate current evidence on circulating biomarkers including circulating tumour cells (CTCs), circulating tumour DNA (ctDNA), microRNAs (miRNAs), extracellular vesicles (EVs), and proteins in GBM diagnosis and monitoring, and to assess the potential role of artificial intelligence (AI) in enhancing their clinical application. Methods: A narrative synthesis of the literature was undertaken, focusing on studies that have investigated blood- and CSF-derived biomarkers in GBM patients. Key aspects evaluated included biomarker biology, detection techniques, diagnostic and prognostic value, current technical challenges, and progress towards clinical translation. Studies exploring AI and machine learning (ML) approaches for biomarker integration and analysis were also reviewed. Results: Liquid biopsy enables repeated and minimally invasive sampling of tumour-derived material, reflecting the genetic, epigenetic, proteomic, and metabolomic landscape of GBM. Although promising, its translation into routine clinical practice is hindered by the low abundance of circulating biomarkers and lack of standardised collection and analysis protocols. Evidence suggests that combining multiple biomarkers improves sensitivity and specificity compared with single-marker approaches. Emerging AI and ML tools show significant potential for improving biomarker discovery, integrating multi-omic datasets, and enhancing diagnostic and prognostic accuracy. Conclusions: Liquid biopsy represents a transformative tool for GBM management, with the capacity to overcome limitations of conventional diagnostics and provide real-time insights into tumour biology. By integrating multiple circulating biomarkers and leveraging AI-driven approaches, liquid biopsy could enhance diagnostic precision, enable dynamic disease monitoring, and improve clinical decision-making. However, large-scale validation and standardisation are required before routine clinical adoption can be achieved. Full article
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14 pages, 1026 KB  
Article
Targeted Whole Genome Sequencing of African Swine Fever Virus and Classical Swine Fever Virus on the MinION Portable Sequencing Platform
by Chester D. McDowell, Taeyong Kwon, Patricia Assato, Emily Mantlo, Jessie D. Trujillo, Natasha N. Gaudreault, Leonardo C. Caserta, Igor Morozov, Jayme A. Souza-Neto, Roman M. Pogranichniy, Diego G. Diel and Juergen A. Richt
Pathogens 2025, 14(8), 804; https://doi.org/10.3390/pathogens14080804 - 13 Aug 2025
Viewed by 972
Abstract
African swine fever virus (ASFV) and classical swine fever virus (CSFV) are important transboundary animal diseases (TADs) affecting swine. ASFV is a large DNA virus with a genome size of 170–190+ kilobases (kB) belonging to the family Asfarviridae, genus Asfivirus. CSFV is [...] Read more.
African swine fever virus (ASFV) and classical swine fever virus (CSFV) are important transboundary animal diseases (TADs) affecting swine. ASFV is a large DNA virus with a genome size of 170–190+ kilobases (kB) belonging to the family Asfarviridae, genus Asfivirus. CSFV is a single-stranded RNA virus with a genome size of approximately 12 kB, belonging to the family Flaviviridae, genus Pestivirus. Outbreaks involving either one of these viruses result in similar disease syndromes and significant economic impacts from: (i) high morbidity and mortality events; (ii) control measures which include culling and quarantine; and (iii) export restrictions of swine and pork products. Current detection methods during an outbreak provide minimal genetic information on the circulating virus strains/genotypes that are important for tracing and vaccine considerations. The increasing availability and reduced cost of next-generation sequencing (NGS) allow for the establishment of NGS protocols for the rapid identification and complete genetic characterization of outbreak strains during an investigation. NGS data provides a better understanding of viral spread and evolution, facilitating the development of novel and effective control measures. In this study, panels of primers spanning the genomes of ASFV and CSFV were independently developed to generate approximately 10 kB and 6 kB amplicons, respectively. The primer panels consisted of 19 primer pairs for ASFV and 2 primer pairs for CSFV, providing whole genome amplification of each pathogen. These primer pools were further optimized for batch pooling and thermocycling conditions, resulting in a total of 5 primer pools/reactions used for ASFV and 2 primer pairs/reactions for CSFV. The ASFV primer panel was tested on viral DNA extracted from blood collected from pigs experimentally infected with ASFV genotype I and genotype II viruses. The CSFV primer panel was tested on 11 different strains of CSFV representing the three known CSFV genotypes, and 21 clinical samples collected from pigs experimentally infected with two different genotype 1 CSF viruses. ASFV and CSFV amplicons from optimized PCR were subsequently sequenced on the Oxford Nanopore MinION platform. The targeted protocols for these viruses resulted in an average coverage greater than 1,000X for ASFV, with 99% of the genome covered, and 10,000X–20,000X for CSFV, with 97% to 99% of the genomes covered. The ASFV targeted whole genome sequencing protocol has been optimized for genotype II ASF viruses that have been responsible for the more recent outbreaks outside of Africa. The CSFV targeted whole genome sequencing protocol has universal applications for the detection of all CSFV genotypes. Protocols developed and evaluated here will be essential complementary tools for early pathogen detection and differentiation, as well as genetic characterization of these high-consequence swine viruses, globally and within the United States, should an outbreak occur. Full article
(This article belongs to the Section Viral Pathogens)
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14 pages, 3149 KB  
Article
CSF-Exosomal miRNAs and Delayed Cerebral Ischemia: Insights into Pathophysiology but No Definitive Biomarkers
by Chathathayil M. Shafeeque, Devin W. McBride, Yuanqing Yan, Hussein A. Zeineddine, John P. Hagen, H. Alex Choi, Jude P. Savarraj, Ari Dienel, Spiros L. Blackburn and Peeyush Kumar Thankamani
Biomolecules 2025, 15(8), 1161; https://doi.org/10.3390/biom15081161 - 13 Aug 2025
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Abstract
Background: Aneurysmal subarachnoid hemorrhage (aSAH) is notoriously known for its high mortality and morbidity. Approximately one-third of the patients who survive aneurysm rupture are reported to develop delayed cerebral ischemia (DCI), which contributes to a poor clinical outcome. Currently, there are no biomarkers [...] Read more.
Background: Aneurysmal subarachnoid hemorrhage (aSAH) is notoriously known for its high mortality and morbidity. Approximately one-third of the patients who survive aneurysm rupture are reported to develop delayed cerebral ischemia (DCI), which contributes to a poor clinical outcome. Currently, there are no biomarkers for identifying which aSAH patients are at risk of developing DCI. We aimed to determine the feasibility of cerebrospinal fluid (CSF) exosomal microRNAs (miRNAs) for predicting DCI post-aSAH. Methods: aSAH patients were prospectively enrolled, and CSF samples were collected at two time points (<24 h and 72 h post-aSAH) from individuals undergoing external ventricular drainage. Exosomal miRNAs were isolated from the CSF for analysis. In the initial group of patients (discovery cohort), an exploratory analysis was conducted using a CSF panel containing 84 miRNAs, assessed by quantitative real-time PCR (RT-qPCR). Based on this analysis, 27 miRNAs were selected for further evaluation in a second group of patients (validation cohort). Among these, 10 miRNAs had previously been reported in SAH-related CSF studies, supporting their relevance for continued investigation. Results: In this study, RT-qPCR analysis of 84 miRNAs in CSF samples from aSAH patients (n = 10 DCI, n = 16 no DCI) and non-aSAH controls (n = 5) identified 9 upregulated and 13 downregulated miRNAs in the DCI group, and 7 upregulated and 18 downregulated miRNAs in the no-DCI group, compared to the controls. When comparing DCI to no-DCI patients, 13 miRNAs were found to be upregulated in the DCI group. Additionally, seven miRNAs showed temporal upregulation in DCI patients between early (<24 h/T1) and later (72 h/T3) time points across both discovery and validation cohorts. However, no miRNAs were uniquely expressed in either DCI or no-DCI groups, limiting their potential as specific biomarkers for DCI. Conclusions: Despite analyses in both the discovery and validation phases, no miRNAs emerged as consistent and reliable biomarkers for distinguishing DCI from no-DCI patients. However, the identified miRNAs are involved in the key KEGG pathways that regulate vascular integrity, neuronal survival, and inflammatory processes central to DCI pathophysiology. These findings highlight the complexity of miRNA regulation following aSAH, as reflected by the variability in differentially expressed miRNAs between cohorts. This variability may be influenced by factors such as limited sample size, patient heterogeneity, individual biological differences, and experimental variability. Comprehensive profiling using larger, well-characterized cohorts, along with rigorous validation, is essential to determine the predictive value and mechanistic significance of candidate miRNAs in DCI. Full article
(This article belongs to the Special Issue Barrier Formation and Homeostasis in the Vertebrate Brain)
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16 pages, 3925 KB  
Communication
Identifying Angiogenic Factors in Pediatric Choroid Plexus Papillomas
by Nurfarhanah Bte Syed Sulaiman, Sofiah M. Y. Sng, Khurshid Z. Merchant, Lee Ping Ng, David C. Y. Low, Wan Tew Seow and Sharon Y. Y. Low
NeuroSci 2025, 6(3), 76; https://doi.org/10.3390/neurosci6030076 - 11 Aug 2025
Viewed by 745
Abstract
(1) Background: Choroid plexus papillomas (CPPs) are rare brain tumors that tend to occur in very young children. Mechanisms of CPP development remain unelucidated. Separately, the process of angiogenesis has been implicated in other primary brain tumors. We hypothesize that angiogenesis is a [...] Read more.
(1) Background: Choroid plexus papillomas (CPPs) are rare brain tumors that tend to occur in very young children. Mechanisms of CPP development remain unelucidated. Separately, the process of angiogenesis has been implicated in other primary brain tumors. We hypothesize that angiogenesis is a hallmark of CPP biology. This study aims to identify and validate angiogenic factors in CPPs. (2) Methods: Cerebrospinal fluid (CSF) and CPP tumor samples are collected. A multiplex immunoassay panel is used to identify differentially expressed cytokines in the CSF samples. Concurrently, patient-derived primary cell cultures and their supernatants are derived from CPP samples. Targeted proteome blot arrays and human umbilical vein endothelial cell (HUVEC) angiogenesis assays are used for validation studies. (3) Results: CSF profiling showed higher expressions of VEGF-A, MCP-1, MMP-1, TNF-α, and CD40L in CPP patient samples versus non-tumor controls. Next, assessment via online protein–protein network platforms reports that these cytokines are associated with endothelial cell regulation. Using an angiogenesis-focused approach, CPP-derived cell lines and supernatants showed similarly higher expressions of VEGF, MCP-1, and MMP-1. Next, sprouting of nodes and tubule formation were observed in HUVEC angiogenesis assay cultures when conditioned CPP cell culture media was added. (4) Conclusions: This proof-of-concept study demonstrates potential to explore angiogenesis in CPP. Full article
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