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9 pages, 192 KiB  
Review
Underdiagnosed and Misunderstood: Clinical Challenges and Educational Needs of Healthcare Professionals in Identifying Autism Spectrum Disorder in Women
by Beata Gellert, Janusz Ostrowski, Jarosław Pinkas and Urszula Religioni
Behav. Sci. 2025, 15(8), 1073; https://doi.org/10.3390/bs15081073 - 7 Aug 2025
Abstract
Autism Spectrum Disorder (ASD) remains significantly underdiagnosed in women, resulting in a persistent gender gap with important clinical, functional, and psychosocial implications. This narrative review explores the multifactorial barriers contributing to diagnostic disparities, including the male-oriented structure of current diagnostic criteria, the prevalence [...] Read more.
Autism Spectrum Disorder (ASD) remains significantly underdiagnosed in women, resulting in a persistent gender gap with important clinical, functional, and psychosocial implications. This narrative review explores the multifactorial barriers contributing to diagnostic disparities, including the male-oriented structure of current diagnostic criteria, the prevalence of co-occurring psychiatric conditions, and the phenomenon of social camouflaging shaped by culturally reinforced gender norms. These factors frequently lead to delayed identification, clinical misinterpretation, and suboptimal care. The review synthesizes evidence from clinical, psychological, and sociocultural research to demonstrate how the under-recognition of ASD in women impacts mental health outcomes, access to education, occupational stability, and overall quality of life. Special emphasis is placed on the consequences of missed or late diagnoses for healthcare delivery and the educational needs of clinicians involved in ASD assessment and care. This article concludes with actionable, evidence-based recommendations for enhancing diagnostic sensitivity, developing gender-responsive screening strategies, and integrating training on female autism presentation into medical and allied health education. Addressing these challenges is essential to reducing diagnostic inequities and ensuring timely, accurate, and person-centered care for autistic women throughout their lifespan. Full article
12 pages, 1394 KiB  
Article
Integrating Cartilage Explant Culture with Simulated Digestion and Hepatic Biotransformation Refines In Vitro Screening of Joint Care Nutraceuticals
by Michelina Crosbie, Kailey Vanderboom, Jamie Souccar-Young and Wendy Pearson
Methods Protoc. 2025, 8(4), 91; https://doi.org/10.3390/mps8040091 - 6 Aug 2025
Abstract
In vitro cartilage explant culture has been used to assess nutraceuticals on cartilage responses to inflammatory stimuli. However, applying extracts of nutraceuticals directly to cartilage explants does not account for effects of digestion and hepatic biotransformation, or selective exclusion of product metabolites from [...] Read more.
In vitro cartilage explant culture has been used to assess nutraceuticals on cartilage responses to inflammatory stimuli. However, applying extracts of nutraceuticals directly to cartilage explants does not account for effects of digestion and hepatic biotransformation, or selective exclusion of product metabolites from joint fluid by the synovial membrane. The current study produced a simulated biological extract of a common nutraceutical (glucosamine; Gsim) by exposing it to a simulated upper gastrointestinal tract digestion, hepatic biotransformation by liver microsomes, and purification to a molecular weight cut-off of 50 kDa. This extract was then used to condition cartilage explants cultured for 120 h in the presence or absence of an inflammatory stimulus (lipopolysaccharide). Media samples were analyzed for prostaglandin E2 (PGE2), glycosaminoglycan (GAG), and nitric oxide (NO). Tissue was digested and analyzed for GAG content and stained for viability. Conditioning of explants with Gsim significantly reduced media GAG in stimulated and unstimulated explants and reduced nitric oxide production in unstimulated explants. These data provide evidence for the value of glucosamine in protecting cartilage from deterioration following an inflammatory challenge, and the model improves applicability of these in vitro data to the in vivo setting. Full article
(This article belongs to the Section Biomedical Sciences and Physiology)
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18 pages, 4635 KiB  
Article
Nylon Affinity Networks Capture and Sequester Two Model Bacteria Spiked in Human Plasma
by Fatema Hashemi, Silvia Cachaco, Rocio Prisby, Weidong Zhou, Gregory Petruncio, Elsa Ronzier, Remi Veneziano, Barbara Birkaya, Alessandra Luchini and Luisa Gregori
Pathogens 2025, 14(8), 778; https://doi.org/10.3390/pathogens14080778 - 6 Aug 2025
Abstract
Ensuring bacterial safety of blood transfusions remains a critical focus in medicine. We investigated a novel pathogen reduction technology utilizing nylon functionalized with synthetic dyes (nylon affinity networks) to capture and remove bacteria from plasma. In the initial screening process, we spiked phosphate [...] Read more.
Ensuring bacterial safety of blood transfusions remains a critical focus in medicine. We investigated a novel pathogen reduction technology utilizing nylon functionalized with synthetic dyes (nylon affinity networks) to capture and remove bacteria from plasma. In the initial screening process, we spiked phosphate buffer solution (PBS) and human plasma (1 mL each) with 10 or 100 colony forming units (cfu) of either Escherichia coli or Staphylococcus epidermidis, exposed the suspensions to affinity networks and assessed the extent of bacterial reduction using agar plate cultures as the assay output. Nineteen synthetic dyes were tested. Among these, Alcian Blue exhibited the best performance with both bacterial strains in both PBS and plasma. Next, bacterial suspensions of approximately 1 and 2 cfu/mL in 10 and 50 mL, respectively, were treated with Alcian Blue affinity networks in three sequential capture steps. This procedure resulted in complete bacterial depletion, as demonstrated by the lack of bacterial growth in the remaining fraction. The viability of the captured bacteria was confirmed by plating the post-treatment affinity networks on agar. Alcian Blue affinity networks captured and sequestered a few plasma proteins identified by liquid chromatography tandem mass spectrometry. These findings support the potential applicability of nylon affinity networks to enhance transfusion safety, although additional investigations are needed. Full article
(This article belongs to the Section Bacterial Pathogens)
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23 pages, 787 KiB  
Systematic Review
Beyond Construction Waste Management: A Systematic Review of Strategies for the Avoidance and Minimisation of Construction and Demolition Waste in Australia
by Emma Heffernan and Leela Kempton
Sustainability 2025, 17(15), 7095; https://doi.org/10.3390/su17157095 - 5 Aug 2025
Abstract
The construction sector is responsible for over 40% of waste generated in Australia. Construction materials are responsible for around 11% of global carbon dioxide emissions, and a third of these materials can end up wasted on a construction site. Attention in research and [...] Read more.
The construction sector is responsible for over 40% of waste generated in Australia. Construction materials are responsible for around 11% of global carbon dioxide emissions, and a third of these materials can end up wasted on a construction site. Attention in research and industry has been directed towards waste management and recycling, resulting in 78% of construction and demolition waste being diverted from landfill. However, the waste hierarchy emphasises avoiding the generation of waste in the first place. In this paper, the PRISMA approach is used to conduct a systematic review with the objective of identifying waste reduction strategies employed across all stages of projects in the Australian construction industry. Scopus and Web of Science databases were used. The search returned 523 publications which were screened and reviewed; this resulted in 24 relevant publications from 1998 to 2025. Qualitative analysis identifies strategies categorised into five groupings: pre-demolition, design, culture, materials and procurement, and on-site activities. The review finds a distinct focus on strategies within the materials and procurement category. The reviewed literature includes fewer strategies for the avoidance of waste than for any of the other levels of the waste hierarchy, evidencing the need for further focus in this area. Full article
(This article belongs to the Special Issue Waste Management for Sustainability: Emerging Issues and Technologies)
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18 pages, 3891 KiB  
Review
Navigating Brain Organoid Maturation: From Benchmarking Frameworks to Multimodal Bioengineering Strategies
by Jingxiu Huang, Yingli Zhu, Jiong Tang, Yang Liu, Ming Lu, Rongxin Zhang and Alfred Xuyang Sun
Biomolecules 2025, 15(8), 1118; https://doi.org/10.3390/biom15081118 - 4 Aug 2025
Viewed by 266
Abstract
Brain organoid technology has revolutionized in vitro modeling of human neurodevelopment and disease, providing unprecedented insights into cortical patterning, neural circuit assembly, and pathogenic mechanisms of neurological disorders. Critically, human brain organoids uniquely recapitulate human-specific developmental processes—such as the expansion of outer radial [...] Read more.
Brain organoid technology has revolutionized in vitro modeling of human neurodevelopment and disease, providing unprecedented insights into cortical patterning, neural circuit assembly, and pathogenic mechanisms of neurological disorders. Critically, human brain organoids uniquely recapitulate human-specific developmental processes—such as the expansion of outer radial glia and neuromelanin—that are absent in rodent models, making them indispensable for studying human brain evolution and dysfunction. However, a major bottleneck persists: Extended culture periods (≥6 months) are empirically required to achieve late-stage maturation markers like synaptic refinement, functional network plasticity, and gliogenesis. Yet prolonged conventional 3D culture exacerbates metabolic stress, hypoxia-induced necrosis, and microenvironmental instability, leading to asynchronous tissue maturation—electrophysiologically active superficial layers juxtaposed with degenerating cores. This immaturity/heterogeneity severely limits their utility in modeling adult-onset disorders (e.g., Alzheimer’s disease) and high-fidelity drug screening, as organoids fail to recapitulate postnatal transcriptional signatures or neurovascular interactions without bioengineering interventions. We summarize emerging strategies to decouple maturation milestones from rigid temporal frameworks, emphasizing the synergistic integration of chronological optimization (e.g., vascularized co-cultures) and active bioengineering accelerators (e.g., electrical stimulation and microfluidics). By bridging biological timelines with scalable engineering, this review charts a roadmap to generate translationally relevant, functionally mature brain organoids. Full article
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20 pages, 1376 KiB  
Article
Comienzo Saludable Puerto Rico: A Community-Based Network of Care to Improve Maternal, Newborn, and Child Health Outcomes
by Edna Acosta-Pérez, Cristina Díaz, Atisha Gómez-Reyes, Samaris Vega, Carlamarie Noboa Ramos, Rosario Justinianes-Pérez, Glamarie Ferran, Jessica Carnivali-García, Fabiola J. Grau, Lili M. Sardiñas, Maribel Campos and Marizaida Sánchez Cesareo
Int. J. Environ. Res. Public Health 2025, 22(8), 1204; https://doi.org/10.3390/ijerph22081204 - 31 Jul 2025
Viewed by 192
Abstract
Background: Maternal and newborn health disparities remain a challenge in Puerto Rico, especially in underserved communities. Comienzo Saludable Puerto Rico, sponsored by the U.S. Department of Health and Human Services’ Healthy Start Initiative (HRSA), addresses these gaps through an integrated Networks of Care [...] Read more.
Background: Maternal and newborn health disparities remain a challenge in Puerto Rico, especially in underserved communities. Comienzo Saludable Puerto Rico, sponsored by the U.S. Department of Health and Human Services’ Healthy Start Initiative (HRSA), addresses these gaps through an integrated Networks of Care model known as Cuidado Compartido. Comienzo Saludable Puerto Rico is a maternal, paternal, and child health program aimed at improving the health and well-being of pregnant women, mothers, fathers, newborns, and children in Puerto Rico, particularly those from disadvantaged communities. Methods: This paper presents the Comienzo Saludable Puerto Rico program’s Cuidado Compartido model to integrate a network of healthcare providers and services across hospitals, community organizations, and families. This model aims to improve maternal and newborn/child health outcomes by focusing on the importance of integrated, hospital-community-based care networks. Results: Participants experienced significant improvements in key birth outcomes: low birth weight prevalence declined by 27.2% compared to the community baseline, premature birth rates decreased by 30.9%, and infant mortality dropped by 75%, reaching 0% by 2021 and remaining there through 2023. These results were complemented by increases in maternal mental health screening, paternal involvement, and breastfeeding practices. Conclusions: The Cuidado Compartido model demonstrates a scalable, culturally responsive strategy to improve maternal, newborn, and child health outcomes. It offers critical insights for implementation in other high-need contexts. Full article
(This article belongs to the Special Issue Community Interventions in Health Disparities)
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11 pages, 415 KiB  
Article
A Nosocomial Outbreak of Burkholderia cepacia complex Linked to Contaminated Intravenous Medications in a Tertiary Care Hospital
by Hanife Nur Karakoc Parlayan, Firdevs Aksoy, Masite Nur Ozdemir, Esra Ozkaya and Gurdal Yilmaz
Antibiotics 2025, 14(8), 774; https://doi.org/10.3390/antibiotics14080774 - 31 Jul 2025
Viewed by 260
Abstract
Objectives: Burkholderia cepacia complex (Bcc), a Gram-negative organism, is a well-recognized cause of hospital outbreaks, often linked to a contaminated shared source, such as multidose medications. In this study, we report an outbreak of Bcc infections in a tertiary care hospital, associated with [...] Read more.
Objectives: Burkholderia cepacia complex (Bcc), a Gram-negative organism, is a well-recognized cause of hospital outbreaks, often linked to a contaminated shared source, such as multidose medications. In this study, we report an outbreak of Bcc infections in a tertiary care hospital, associated with the intrinsic contamination of a prepared solution used in interventional radiology (IR) procedures. Additionally, we provide a detailed explanation of the interventions implemented to control and interrupt the outbreak. Methods: Records from the infection control committee from 1 January 2023 to 31 October 2024 were screened to identify cases with Bcc growth in cultured blood, urine, or respiratory samples. Clinical and laboratory data were collected in March 2025. Bacterial identification was performed using conventional methods and MALDI-TOF (Bruker Daltonics, Bremen, Germany). Controls were matched to cases by ward, date of initial growth, and duration of hospitalization. Demographic and clinical data of these patients were systematically collected and analyzed. Microbiological cultures were obtained from environmental objects of concern and certain medications. Results: A total of 82 Burkholderia species were identified. We enrolled 77 cases and 77 matched controls. The source of contamination was identified in ready-to-use intravenous medications (remifentanil and magnesium preparations) in the IR department. These preparations were compounded in advance by the team and were used repeatedly. Although the outbreak originated from contaminated IV medications used in IR, secondary transmission likely affected 28 non-IR patients via fomites, shared environments, and possible lapses in isolation precautions. The mortality rate among the cases was 16.9%. Infection with Bcc was associated with prolonged intensive care unit stays (p = 0.018) and an extended overall hospitalization duration (p < 0.001); however, it was not associated with increased mortality. The enforcement of contact precautions and comprehensive environmental decontamination successfully reduced the incidence of the Bcc outbreak. No pathogens were detected in cultures obtained after the disinfection. Conclusions: The hospital transmission of Bcc is likely driven by cross-contamination, invasive medical procedures, and the administration of contaminated medications. Implementing stringent infection control measures such as staff retraining, updated policies on medication use, enhanced environmental decontamination, and strict adherence to isolation precautions has proven effective in curbing the spread of virulent and transmissible Bcc. Full article
(This article belongs to the Section Antibiotics Use and Antimicrobial Stewardship)
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11 pages, 551 KiB  
Article
Artificial Neural Network for the Fast Screening of Samples from Suspected Urinary Tract Infections
by Cristiano Ialongo, Marco Ciotti, Alfredo Giovannelli, Flaminia Tomassetti, Martina Pelagalli, Stefano Di Carlo, Sergio Bernardini, Massimo Pieri and Eleonora Nicolai
Antibiotics 2025, 14(8), 768; https://doi.org/10.3390/antibiotics14080768 - 30 Jul 2025
Viewed by 264
Abstract
Background: Urine microbial analysis is a frequently requested test that is often associated with contamination during specimen collection or storage, which leads to false-positive diagnoses and delayed reporting. In the era of digitalization, machine learning (ML) can serve as a valuable tool to [...] Read more.
Background: Urine microbial analysis is a frequently requested test that is often associated with contamination during specimen collection or storage, which leads to false-positive diagnoses and delayed reporting. In the era of digitalization, machine learning (ML) can serve as a valuable tool to support clinical decision-making. Methods: This study investigates the application of a simple artificial neural network (ANN) to pre-identify negative and contaminated (false-positive) specimens. An ML model was developed using 8181 urine samples, including cytology, dipstick tests, and culture results. The dataset was randomly split 2:1 for training and testing a multilayer perceptron (MLP). Input variables with a normalized importance below 0.2 were excluded. Results: The final model used only microbial and either urine color or urobilinogen pigment analysis as inputs; other physical, chemical, and cellular parameters were omitted. The frequency of positive and negative specimens for bacteria was 6.9% and 89.6%, respectively. Contaminated specimens represented 3.5% of cases and were predominantly misclassified as negative by the MLP. Thus, the negative predictive value (NPV) was 96.5% and the positive predictive value (PPV) was 87.2%, leading to 0.82% of the cultures being unnecessary microbial cultures (UMC). Conclusions: These results suggest that the MLP is reliable for screening out negative specimens but less effective at identifying positive ones. In conclusion, ANN models can effectively support the screening of negative urine samples, detect clinically significant bacteriuria, and potentially reduce unnecessary cultures. Incorporating morphological information data could further improve the accuracy of our model and minimize false negatives. Full article
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16 pages, 1591 KiB  
Article
Molecular and Drug Resistance Characteristics of Haemophilus influenzae Carried by Pediatric Patients with Adenoid Hypertrophy
by Nan Xiao, Jia-Hao Qin, Xiu-Ying Zhao and Lin Liu
Microorganisms 2025, 13(8), 1764; https://doi.org/10.3390/microorganisms13081764 - 29 Jul 2025
Viewed by 235
Abstract
Purpose: The adenoid microbiota plays a key role in adenoid hypertrophy (AH). This study explored the molecular epidemiology and antimicrobial resistance of Haemophilus. Influenzae (H. influenzae) strains in pediatric AH patients. Methods: Retrospective analysis of pediatric AH patients undergoing endoscopic adenoidectomy. [...] Read more.
Purpose: The adenoid microbiota plays a key role in adenoid hypertrophy (AH). This study explored the molecular epidemiology and antimicrobial resistance of Haemophilus. Influenzae (H. influenzae) strains in pediatric AH patients. Methods: Retrospective analysis of pediatric AH patients undergoing endoscopic adenoidectomy. Adenoid tissue samples were cultured to screen for pathogens. H. influenzae strains were identified by 16S rRNA sequencing and serotyped via q-PCR. Multilocus sequence typing (MLST) and ftsI gene analysis were conducted using PubMLST. β-lactamase genes (blaTEM-1, blaROB-1) were detected by PCR, and antibiotic susceptibility testing (AST) was performed using the Etest method. For imipenem-resistant strains, the acrRAB efflux pump gene cluster and ompP2 porin gene were sequenced and compared with those of the wild-type strain Rd KW20. Results: Over 8 months, 56 non-duplicate H. influenzae strains were isolated from 386 patients. The detection rate was highest in children under 5 years (30.5%) compared to those aged 5–10 years (13.4%) and 10–15 years (8.7%). Of 49 sub-cultured strains, all were non-typeable H. influenzae (NTHi). MLST identified 22 sequence types (STs) and 13 clonal complexes (CCs), with CC11 (26.5%), CC3 (14.3%), and CC107 (14.3%) being predominant. Common STs included ST103 (22.4%), ST57 (10.2%), and ST107 (10.2%). Most strains belonged to the ftsI group III-like+ (57.1%). β-lactamase positivity was 98.0% (48/49), with blaTEM-1 (95.9%) and blaROB-1 (18.4%) detected. AST showed low susceptibility to ampicillin (10.2%), amoxicillin–clavulanate (34.7%), azithromycin (12.2%), and trimethoprim–sulfamethoxazole (14.3%). Among the β-lactamase-positive strains, 44/48 were β-lactamase-positive ampicillin-resistant (BLPAR); none were β-lactamase-negative ampicillin-resistant (BLNAR). Imipenem susceptibility was 91.8% (45/49). No carbapenemases were found in the imipenem-resistant strains, but mutations in acrRAB (88.12–94.94% identity) and ompP2 (77.10–82.94% identity) were observed. Conclusions: BLPAR NTHi strains of CC11 are major epidemic strains in pediatric AH. Imipenem resistance in H. influenzae likely results from porin mutations rather than carbapenemase activity. Enhanced surveillance of H. influenzae’s role in AH and its resistance patterns is warranted. Full article
(This article belongs to the Section Medical Microbiology)
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18 pages, 1278 KiB  
Review
Metabolic Maturation in hiPSC-Derived Cardiomyocytes: Emerging Strategies for Inducing the Adult Cardiac Phenotype
by Daniela Malan, Maria Pia Gallo, Federica Geddo, Renzo Levi and Giulia Querio
Pharmaceuticals 2025, 18(8), 1133; https://doi.org/10.3390/ph18081133 - 29 Jul 2025
Viewed by 327
Abstract
Human induced pluripotent stem cells (hiPSCs) are widely used in basic research because of their versatility and ability to differentiate into multiple cell types. In particular, differentiating hiPSCs into cardiac cells (hiPSC-CMs) has been an important milestone in cardiac pathophysiology studies. Although hiPSC-CMs [...] Read more.
Human induced pluripotent stem cells (hiPSCs) are widely used in basic research because of their versatility and ability to differentiate into multiple cell types. In particular, differentiating hiPSCs into cardiac cells (hiPSC-CMs) has been an important milestone in cardiac pathophysiology studies. Although hiPSC-CMs offer a model for human cardiomyocytes, they still exhibit characteristics linked to the fetal cardiac cell phenotype. One important feature that prevents hiPSC-CMs from being identified as adult cells relates to their metabolism, which is a key factor in defining a mature phenotype capable of sustaining the workload requirements characteristic of fully differentiated cardiomyocytes. This review aims to present the most relevant strategies in terms of culture medium composition, culture times, and 3D culture methods that have been developed to promote the metabolic maturation of hiPSC-CMs, which are now widely used. Defining a standardized and universally accepted protocol would enable the creation of a cellular model for studies of cardiac pathophysiology from a patient-specific perspective and for drug screening. Full article
(This article belongs to the Special Issue Cell Therapy for Cardiac Disease)
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16 pages, 646 KiB  
Article
Psychometric Properties of the Diabetes Eating Problem Survey—Revised in Arab Adolescents with Type 1 Diabetes: A Cross-Cultural Validation Study
by Abdullah M. Alguwaihes, Shuliweeh Alenezi, Renad Almutawa, Rema Almutawa, Elaf Almusahel, Metib S. Alotaibi, Mohammed E. Al-Sofiani and Abdulmajeed AlSubaihin
Behav. Sci. 2025, 15(8), 1026; https://doi.org/10.3390/bs15081026 - 29 Jul 2025
Viewed by 324
Abstract
Objectives: The objective of this manuscript is to translate, adapt, and validate an Arabic version of the Diabetes Eating Problem Survey—Revised (DEPS-R) questionnaire to assess disordered eating behaviors (DEBs) in adolescents with T1D in Saudi Arabia. Additionally, the study sought to estimate the [...] Read more.
Objectives: The objective of this manuscript is to translate, adapt, and validate an Arabic version of the Diabetes Eating Problem Survey—Revised (DEPS-R) questionnaire to assess disordered eating behaviors (DEBs) in adolescents with T1D in Saudi Arabia. Additionally, the study sought to estimate the prevalence of DEBs and analyze its associations with glycemic control and diabetes-related complications. Methods: A cross-cultural validation study was conducted following the COSMIN guidelines. The DEPS-R questionnaire was translated into Arabic through forward and backward translation involving expert panels, including psychiatrists, diabetologists, and linguists. A sample of 409 people with type 1 diabetes (PwT1D) (58.4% females) aged 12–20 years was recruited from outpatient diabetes clinics in the five main regions of Saudi Arabia. Participants completed the Arabic DEPS-R and the validated Arabic version of the SCOFF questionnaire. Sociodemographic, anthropometric, and biochemical data were collected, and statistical analyses, including confirmatory factor analysis (CFA) and internal consistency tests, were conducted. Results: The Arabic DEPS-R exhibits strong internal consistency (Cronbach’s alpha = 0.829) and high test–retest reliability (ICC = 0.861), with a CFA supporting a three-factor structure, namely body weight perception, disordered eating behaviors (DEBs), and bulimic tendencies. Notably, higher DEPS-R scores are significantly linked to elevated HbA1c levels, increased BMI, and more frequent insulin use. Alarmingly, 52.8% of participants show high-risk DEB, which is directly associated with poor glycemic control (HbA1c ≥ 8.1%) and a heightened risk of diabetic ketoacidosis (DKA). Conclusions: The Arabic DEPS-R is a valid and reliable tool for screening DEBs among Saudi adolescents with T1D. Findings underscore the necessity for early identification and intervention to mitigate the impact of EDs on diabetes management and overall health outcomes. Full article
(This article belongs to the Section Child and Adolescent Psychiatry)
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28 pages, 2854 KiB  
Article
Real-Time Functional Stratification of Tumor Cell Lines Using a Non-Cytotoxic Phospholipoproteomic Platform: A Label-Free Ex Vivo Model
by Ramón Gutiérrez-Sandoval, Francisco Gutiérrez-Castro, Natalia Muñoz-Godoy, Ider Rivadeneira, Adolay Sobarzo, Jordan Iturra, Ignacio Muñoz, Cristián Peña-Vargas, Matías Vidal and Francisco Krakowiak
Biology 2025, 14(8), 953; https://doi.org/10.3390/biology14080953 - 28 Jul 2025
Viewed by 280
Abstract
The development of scalable, non-invasive tools to assess tumor responsiveness to structurally active immunoformulations remains a critical unmet need in solid tumor immunotherapy. Here, we introduce a real-time, ex vivo functional system to classify tumor cell lines exposed to a phospholipoproteomic platform, without [...] Read more.
The development of scalable, non-invasive tools to assess tumor responsiveness to structurally active immunoformulations remains a critical unmet need in solid tumor immunotherapy. Here, we introduce a real-time, ex vivo functional system to classify tumor cell lines exposed to a phospholipoproteomic platform, without relying on cytotoxicity, co-culture systems, or molecular profiling. Tumor cells were monitored using IncuCyte® S3 (Sartorius) real-time imaging under ex vivo neutral conditions. No dendritic cell components or immune co-cultures were used in this mode. All results are derived from direct tumor cell responses to structurally active formulations. Using eight human tumor lines, we captured proliferative behavior, cell death rates, and secretomic profiles to assign each case into stimulatory, inhibitory, or neutral categories. A structured decision-tree logic supported the classification, and a Functional Stratification Index (FSI) was computed to quantify the response magnitude. Inhibitory lines showed early divergence and high IFN-γ/IL-10 ratios; stimulatory ones exhibited a proliferative gain under balanced immune signaling. The results were reproducible across independent batches. This system enables quantitative phenotypic screening under standardized, marker-free conditions and offers an adaptable platform for functional evaluation in immuno-oncology pipelines where traditional cytotoxic endpoints are insufficient. This approach has been codified into the STIP (Structured Traceability and Immunophenotypic Platform), supporting reproducible documentation across tumor models. This platform contributes to upstream validation logic in immuno-oncology workflows and supports early-stage regulatory documentation. Full article
(This article belongs to the Section Cancer Biology)
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23 pages, 4112 KiB  
Article
Metabolic Culture Medium Enhances Maturation of Human iPSC-Derived Cardiomyocytes via Cardiac Troponin I Isoform Induction
by Daria V. Goliusova, Agnessa P. Bogomolova, Alina V. Davidenko, Kristina A. Lavrenteva, Margarita Y. Sharikova, Elena A. Zerkalenkova, Ekaterina M. Vassina, Alexandra N. Bogomazova, Maria A. Lagarkova, Ivan A. Katrukha and Olga S. Lebedeva
Int. J. Mol. Sci. 2025, 26(15), 7248; https://doi.org/10.3390/ijms26157248 - 26 Jul 2025
Viewed by 483
Abstract
Human induced pluripotent stem cell-derived cardiomyocytes (iCMs) provide a powerful platform for investigating cardiac biology. However, structural, metabolic, and electrophysiological immaturity of iCMs limits their capacity to model adult cardiomyocytes. Currently, no universally accepted criteria or protocols for effective iCMs maturation exist. This [...] Read more.
Human induced pluripotent stem cell-derived cardiomyocytes (iCMs) provide a powerful platform for investigating cardiac biology. However, structural, metabolic, and electrophysiological immaturity of iCMs limits their capacity to model adult cardiomyocytes. Currently, no universally accepted criteria or protocols for effective iCMs maturation exist. This study aimed to identify practical culture conditions that promote iCMs maturation, thereby generating more physiologically relevant in vitro cardiac models. We evaluated the effects of short- and long-term culture in media supplemented with various stimulatory compounds under 2D conditions, focusing on intracellular content and localization of slow skeletal troponin I (ssTnI) and cardiac troponin I (cTnI) isoforms. Our findings demonstrate that the multicomponent metabolic maturation medium (MM-1) effectively enhances the transition toward a more mature iCM phenotype, as evidenced by increased cTnI expression and formation of cross-striated myofibrils. iCMs cultured in MM-1 more closely resemble adult cardiomyocytes and are compatible with high-resolution single-cell techniques such as electron microscopy and patch-clamp electrophysiology. This work provides a practical and scalable approach for advancing the maturation of iPSC-derived cardiac models, with applications in disease modeling and drug screening. Full article
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33 pages, 1777 KiB  
Review
Immunomodulatory Natural Products in Cancer Organoid-Immune Co-Cultures: Bridging the Research Gap for Precision Immunotherapy
by Chang-Eui Hong and Su-Yun Lyu
Int. J. Mol. Sci. 2025, 26(15), 7247; https://doi.org/10.3390/ijms26157247 - 26 Jul 2025
Viewed by 619
Abstract
Natural products demonstrate potent immunomodulatory properties through checkpoint modulation, macrophage polarization, and T cell/natural killer (NK) cell activation. While cancer organoid-immune co-culture platforms enable physiologically relevant modeling of tumor–immune interactions, systematic investigation of natural product immunomodulation in these systems remains entirely unexplored. We [...] Read more.
Natural products demonstrate potent immunomodulatory properties through checkpoint modulation, macrophage polarization, and T cell/natural killer (NK) cell activation. While cancer organoid-immune co-culture platforms enable physiologically relevant modeling of tumor–immune interactions, systematic investigation of natural product immunomodulation in these systems remains entirely unexplored. We conducted a comprehensive literature analysis examining natural products tested in cancer organoids, immunomodulatory mechanisms from traditional models, technical advances in organoid-immune co-cultures, and standardization requirements for clinical translation. Our analysis reveals a critical research gap: no published studies have investigated natural product-mediated immunomodulation using organoid-immune co-culture systems. Even though compounds like curcumin, resveratrol, and medicinal mushroom polysaccharides show extensive immunomodulatory effects in two-dimensional (2D) cultures, and organoid technology achieves high clinical correlation for drug response prediction, all existing organoid studies focus exclusively on direct cytotoxicity. Technical challenges include compound stability, limited matrix penetration requiring substantially higher concentrations than 2D cultures, and maintaining functional immune populations in three-dimensional (3D) systems. The convergence of validated organoid-immune co-culture platforms, Food and Drug Administration (FDA) regulatory support through the Modernization Act 2.0, and extensive natural product knowledge creates unprecedented opportunities. Priority research directions include systematic screening of immunomodulatory natural products in organoid-immune co-cultures, development of 3D-optimized delivery systems, and clinical validation trials. Success requires moving beyond cytotoxicity-focused studies to investigate immunomodulatory mechanisms in physiologically relevant 3D systems, potentially unlocking new precision cancer immunotherapy approaches. Full article
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14 pages, 3991 KiB  
Article
Detection of Pestalotiopsis abbreviata sp. nov., the Causal Agent of Pestalotiopsis Leaf Blight on Camellia japonica Based on Metagenomic Analysis
by Sung-Eun Cho, Ki Hyeong Park, Keumchul Shin and Dong-Hyeon Lee
J. Fungi 2025, 11(8), 553; https://doi.org/10.3390/jof11080553 - 25 Jul 2025
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Abstract
Tree diseases affecting Camellia japonica have emerged as a significant threat to the health and longevity of this ornamental tree, particularly in countries where this tree species is widely distributed and cultivated. Among these, Pestalotiopsis spp. have been frequently reported and are considered [...] Read more.
Tree diseases affecting Camellia japonica have emerged as a significant threat to the health and longevity of this ornamental tree, particularly in countries where this tree species is widely distributed and cultivated. Among these, Pestalotiopsis spp. have been frequently reported and are considered one of the most impactful fungal pathogens, causing leaf blight or leaf spot, in multiple countries. Understanding the etiology and distribution of these diseases is essential for effective management and conservation of C. japonica populations. The traditional methods based on pathogen isolation and pure culture cultivation for diagnosis of tree diseases are labor intensive and time-consuming. In addition, the frequent coexistence of the major pathogens with other endophytes within a single C. japonica tree, coupled with inconsistent symptom expression and the occurrence of pathogens in asymptomatic hosts, further complicates disease diagnosis. These challenges highlight the urgent need to develop more rapid, accurate, and efficient diagnostic or monitoring tools to improve disease monitoring and management on trees, including C. japonica. To address these challenges, we applied a metagenomic approach to screen fungal communities within C. japonica trees. This method enabled comprehensive detection and characterization of fungal taxa present in symptomatic and asymptomatic tissues. By analyzing the correlation between fungal dominance and symptom expression, we identified key pathogenic taxa associated with disease manifestation. To validate the metagenomic approach, we employed a combined strategy integrating metagenomic screening and traditional fungal isolation to monitor foliar diseases in C. japonica. The correlation between dominant taxa and symptom expression was confirmed. Simultaneously, traditional isolation enabled the identification of a novel species, Pestalotiopsis, as the causal agent of leaf spot disease on C. japonica. In addition to confirming previously known pathogens, our study led to the discovery and preliminary characterization of a novel fungal taxon with pathogenic potential. Our findings provide critical insights into the fungal community of C. japonica and lay the groundwork for developing improved, rapid diagnostic tools for effective disease monitoring and management of tree diseases. Full article
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