Viral-bacterial codetection is common and may increase clinical severity, but evidence in the Peruvian pediatric population is limited. The objective of the present study was to evaluate the clinical, laboratory, and seasonal characteristics associated with viral-bacterial codetection in children hospitalized for lower respiratory tract infections (LRTIs). Cross-sectional secondary database study from a private hospital in Lima, Peru. We included patients < 13 years hospitalized for LRTIs with RT-qPCR results for respiratory viruses and/or bacteria. Viral-bacterial codetection was compared against other detection patterns using Poisson regression for binary outcomes and linear regression for continuous outcomes, with false discovery rate (FDR) correction for multiple comparisons. A total of 548 patients were included (median age 2.0 years; 50.5% female). Viral-bacterial codetection was identified in 21.5% of patients (n = 118), with RSV + Haemophilus influenzae being the most frequent combination. Compared with other detection patterns, viral-bacterial codetection was significantly associated with a higher prevalence of crackles (aPR: 1.30; 95% CI: 1.08-1.57), lower oxygen saturation at admission (β: -0.57; 95% CI: -1.04 to - 0.10), higher platelet counts (β: 30,452; 95% CI: 5,113-55,792), higher hemoglobin levels (β: 0.29 g/dL; 95% CI: 0.03-0.56), and longer hospital stay (β: 0.66 days; 95% CI: 0.02-1.29). However, after FDR correction for multiple comparisons, none of these associations reached statistical significance (q-values: 0.056-0.113). No difference was detected according to seasonality. Viral-bacterial codetection was common and was associated with crackles, lower oxygen saturation, longer hospital stay, and higher platelet counts; however, after FDR correction, none of these associations remained statistically significant, underscoring the exploratory nature of these findings and the need for larger, confirmatory studies.
The expeditious and precise diagnosis of dengue virus (DENV) and chikungunya virus (CHIKV) is paramount for effective patient management and the control of outbreaks. In this study, a duplex reverse transcription multi-enzyme isothermal amplification (RT-MIRA) assay was established for the simultaneous detection of DENV and CHIKV, followed by a nested RT-MIRA assay for DENV serotyping (DENV-1 to -4). Specific primers and probes targeting the DENV 3'-UTR, CHIKV E1 gene, and four DENV serotypes were designed. The duplex RT-MIRA and nested DENV RT-MIRA serotyping reaction systems were optimized at 39 °C with portable fluorescence or lateral flow dipstick readouts. For methodological validation, specificity was evaluated against 35 related pathogens, and the 95% limit of detection (LOD95) was determined via probit regression. For clinical validation, serum samples from 236 suspected patients were tested, benchmarking against RT-qPCR and serology. Statistical analyses included the Wilson score method for calculating 95% confidence intervals (CIs) and Cohen's kappa (κ). For external verification, 12 CHIKV-positive clinical samples and 5 artificially simulated co-infection samples were retrospectively analyzed to validate assay accuracy. The duplex RT-MIRA assay exhibited no cross-reactivity with other pathogens. The LOD95 values were 13.47 copies/μl for DENV and 10.49 copies/μl for CHIKV. Clinical validation demonstrated sensitivities of 96.15% (95% CI: 89.28%-98.67%) for DENV and 88.89% (95% CI: 67.20%-96.90%) for CHIKV. Specificity was 100% (95% CI: 92.87%-100%) for both. Agreement with RT-qPCR was strong for DENV (κ = 0.96) and CHIKV (κ = 0.92). The nested RT-MIRA serotyping assay showed high sensitivity (LOD95: 1.6-18.7 copies/μl) without cross-reactivity, accurately differentiating 75 DENV-positive samples into 71 DENV-1 and 4 DENV-2. In the external verification, the assay accurately detected 10 CHIKV mono-infections and 2 CHIKV/DENV co-infections, and distinguished four DENV serotypes in simulated matrices. A rapid and sensitive integrated method has been developed that combines duplex RT-MIRA for detecting DENV and CHIKV, and nested RT-MIRA for serotyping DENV. The simplicity and speed of the amplification and detection steps demonstrate this platform's potential for use in point-of-care testing and surveillance in areas with limited resources, particularly when used alongside portable extraction methods.
Hepatitis E virus (HEV) infection screening and diagnosis are critical for controlling the HEV epidemic. Serological testing for anti-HEV IgM and IgG is widely used to diagnose acute and past HEV infections, respectively. This study aims to evaluate the analytical performances of the MAGLUMI HEV IgM and IgG assays for detection of HEV antibodies and comparison with the microplate Wantai assay. We assessed the precision, cross-reactivity, and interference of the MAGLUMI HEV IgM and IgG assays, as well as their agreement rates with the Wantai HEV IgM and IgG assays. Method comparison included in total 775 samples from 405 patients with suspected HEV infection and 86 asymptomatic blood donors. Among them, the HEV infection status of 136 individuals was confirmed through HEV RNA results, including 39 cases of acute infection, 3 cases of convalescence, and 94 cases of non-infected individuals. The within-run and between-day imprecision for the MAGLUMI HEV IgM assay ranged from 1.54% to 3.71%, while for the MAGLUMI HEV IgG assay, it ranged from 0.00% to 4.35%. No cross-reactivity was observed in either assay. The positive and negative agreement rates between the MAGLUMI and Wantai IgM assay were 98.61% and 99.31%, respectively. The positive and negative agreement rates between the MAGLUMI and Wantai IgG assay were 94.95% and 97.20%, respectively. Overall, the analytical performance of the MAGLUMI HEV IgM and IgG assays is good, with reactivities comparable to those of the Wantai HEV IgM and IgG assays. However, due to the limited availability of HEV RNA detection results and lack of HEV genotype information, this study could not effectively evaluate the clinical performance of the reagent, and further studies with HEV RNA determination and genotyping are needed for a clinical validation.
The current American Heartworm Society guidelines recommend the concomitant use of an antigen detection test and a microfilariae detection test (MFDT) for diagnosing heartworm infection in dogs. The modified Knott's (MK) test is the preferred MFDT for determining the morphological characteristics of Dirofilaria immitis microfilariae, but it requires extensive microscopy training and can be time-consuming in a clinical setting. The Pluslife Mini Dock is a point-of-care diagnostic platform that uses RNase HII-assisted amplification (RHAM) to eliminate the need for DNA extraction, with results available within 30 min. This study aimed to assess the performance of the Pluslife Mini Dock duplex Dirofilaria immitis/Dirofilaria repens assay in dog blood compared with the MK. Archival, frozen whole-blood samples collected from 250 dogs at shelters in central Texas, USA, were used. Samples were subjected to the MK on the day of collection and stored at 2°C until further processing. The samples were then thawed and subjected to the Pluslife Mini Dock D. immitis/D. repens duplex assay. The results were analyzed using Cohen's kappa coefficient and McNemar's Chi-squared test. Overall, 93.6% of results matched between the two tests; however, the Pluslife assay detected a higher proportion of D. immitis-positive samples (32.4%; 81/250) than the MK (30.0%; 75/250). There was no statistical significance between tests (p = 0.2113). Cohen's kappa statistic indicated almost perfect agreement between the two tests (0.85). Additionally, Acanthocheilonema reconditum was detected in 11 samples in the MK test, without generating false-positive results with the Pluslife assay, indicating its specificity. Our data suggest that the Pluslife Mini Dock D. immitis/D. repens duplex assay provides a novel diagnostic platform and is a suitable option for point-of-care MFDT.
Tick-borne pathogens are increasingly recognized as important threats to veterinary and public health. However, the epidemiological role of cats in the transmission of Anaplasma phagocytophilum-related strains and Hepatozoon spp. remains unclear. In Türkiye, A. phagocytophilum-like 1 (AP-like 1) has been reported in ruminants and ticks; however, molecular evidence in feline hosts has not been documented. This study aimed to investigate the molecular prevalence and genetic diversity of A. phagocytophilum-related strains and Hepatozoon spp. in domestic cats from eastern Turkey. Blood samples from 93 domestic cats from animal shelters in Van and Batman provinces were screened using PCR targeting the 16S rRNA gene of Anaplasma spp. and the 18S rRNA gene of Hepatozoon spp. Positive samples were characterized by restriction fragment length polymorphism and sequencing, followed by phylogenetic analysis using the Maximum Likelihood method. Anaplasma DNA was detected in 78.5% (73/93) of cats, and RFLP analysis indicated that all positive samples were consistent with AP-like 1, representing the first molecular evidence of this variant in domestic cats in Türkiye. Phylogenetic analyses revealed that the feline isolates clustered with ruminant-derived strains, suggesting shared transmission cycles and low host specificity. Hepatozoon spp. DNA was detected in 8.6% (8/93) of the samples. Based on sequencing and phylogenetic analyses of a subset of positive samples, the isolates were identified as Hepatozoon felis genotype I. The absence of other variants in the sequenced subset may suggest regional differences; however, this finding should be interpreted cautiously due to the limited number of sequences analyzed. These findings demonstrate a high prevalence of AP-like 1 and support the circulation of Hepatozoon felis genotype I in domestic cats from eastern Türkiye, highlighting the potential role of feline hosts in regional transmission dynamics. This study expands current knowledge of the host range and epidemiology of A. phagocytophilum-related strains and Hepatozoon spp. and underscores the importance of integrated molecular surveillance and vector control strategies within the One Health framework.
Dirofilariosis, caused by Dirofilaria immitis and Dirofilaria repens, is expanding across Europe because of climate change, raising veterinary and zoonotic concerns. Among over 70 mosquito species considered to be implicated in their transmission, Culex pipiens and Aedes albopictus are recognized as major vectors. In Sardinia, where canine dirofilariosis is endemic, entomological information on Dirofilaria circulation was previously lacking. This study investigated mosquito species composition and assessed the presence of D. immitis and D. repens across five sites on the island. Mosquitoes were collected monthly from August 2022 to October 2023 using BG-Sentinel and CDC light traps. Specimens were morphologically and molecularly identified and screened for Dirofilaria DNA. A total of 1219 mosquitoes were collected, including 945 females belonging to 13 species. The most abundant were Aedes caspius (31.4%), Aedes detritus (28.7%), Culex pipiens (19.5%), and Aedes albopictus (16.6%). Notably, among minor species, the presence of Culex perexiguus in Italy was confirmed for the first time by molecular analysis. Dirofilaria repens and D. immitis DNA were detected in 1.8% and 1.0% of mosquitoes, respectively, with C. pipiens and Ae. albopictus as the main vectors. Other species, including Ae. caspius, Ae. detritus, C. tarsalis, and C. perexiguus, also tested positive, suggesting a potential role in transmission. This study provides the first entomological report of Dirofilaria circulation in Sardinia, revealing a complex vector community and underscoring the need for continuous surveillance of dirofilariosis risk in the Mediterranean region.
Accurate nuchal translucency (NT) measurement for assessing the risk of fetal genetic abnormalities requires precise acquisition of the mid-sagittal plane (MSP). However, achieving an appropriate MSP is technically challenging due to anatomical variability and operator dependence inherent in conventional 2-dimensional (2D) ultrasound. This study aimed to develop and validate a novel deep learning algorithm for automated fetal MSP extraction from 3-dimensional (3D) ultrasound volumes utilizing intracranial structure segmentation to overcome the limitations of conventional methods reliant on facial landmarks. In this prospective study, we developed and evaluated "3D MSP-net," a convolutional neural network (CNN)-based model for automated MSP extraction, involving singleton pregnant women undergoing first-trimester NT screening. Using achieved 3D volume data, 3D MSP-net was validated against the conventional 2D manual method and a commercially available rule-based automated system (5D NT™). Two maternal-fetal medicine (MFM) specialists independently assessed the resulting MPSs to determine the performance for demonstrating the feasibility and high reproducibility of the 3D MSP-net. 3D MSP-net achieved an MSP extraction success rate of 91.6%, comparable to that of the conventional 2D manual method and significantly superior to the rule-based 3D algorithm. NT measurements were comparable between the conventional 2D manual approach and MSPs derived from 3D MSP-net (1.4 ± 0.5 mm versus 1.4 ± 0.4 mm; p = .444). These results were reproducible on external validation. Moreover, the 3D MSP-net maintained robust performance even under challenging conditions, such as increased maternal body mass index and different scan deviation angles. The 3D MSP-net, our artificial intelligence (AI) model that utilizes intracranial landmarks for MSP reconstruction, enables improved efficiency, standardization, and reliability for first-trimester fetal screening addressing a key challenge in prenatal diagnostics.
Bovine tuberculosis (bTB), caused by Mycobacterium bovis, is a zoonosis that threatens public health and causes substantial economic losses in livestock. The suboptimal Escherichia coli-expressed recombinant proteins limit the diagnostic performance of current bTB serological tests. To overcome this limitation, we evaluated Mycolicibacterium smegmatis as an expression host capable of producing recombinant antigens with post-translational modifications comparable to those of M. bovis. Ten antigen candidates were individually expressed in E. coli using the pET-26b( +) vector and in M. smegmatis using the pSOΔBam vector. Their diagnostic performance was evaluated using an enzyme-linked immunosorbent assay (ELISA) with plasma samples from interferon-gamma release assay (IGRA)-negative (n = 30) and -positive (n = 46) cattle in South Korea, followed by receiver operating characteristic (ROC) curve analysis. Among the single antigens, LprA expressed in M. smegmatis demonstrated diagnostic performance comparable to that of the well-established antigen MPB70 (sensitivity: 50.0%, specificity: 96.6%, AUC: 0.791). In addition, several M. smegmatis-derived antigens showed higher concordance with the IGRA results, as assessed by Cohen's kappa and Fisher's exact tests, and a stronger association between age and antigen-specific antibody responses was observed among IGRA-positive cattle. Moreover, a multiple logistic regression model incorporating eight antigens, including those derived from both hosts, achieved high predictive accuracy for IGRA results (sensitivity: 87.0%, specificity: 100%, AUC: 0.991). These findings suggest that M. smegmatis is a promising host for identifying novel antigens and that a multi-host strategy may improve bTB serodiagnosis.
Advancements in high-throughput sequencing have revolutionized transcriptomics, enabling insights into gene expression, splicing, and fusions. However, RNA-seq analysis remains challenging due to complex splice junctions, multi-mapped reads, and chimeric events. We present DeepSAP, which improves RNA-seq alignment by integrating GSNAP's transcriptome-guided genomic alignment with transformer-based splice-junction scoring. This synergy enhances splice-junction detection, indel identification, and resolution of complex splicing patterns. On the Baruzzo human simulated benchmark across complexities, DeepSAP achieves the highest mean F1-score for splice junction detection, outperforming DRAGEN, novoSplice, STAR, HISAT2, and Subjunc. DeepSAP captures intricate sequence patterns surrounding splice donor and acceptor sites, advancing RNA-seq analysis.
Doxorubicin hydrochloride (DOX) remains a cornerstone in the treatment of numerous malignancies. Recent research has focused on improving the efficacy of chemotherapeutic regimens through synergistic combinations. Rifampicin (RFP), in addition to its well-known antimicrobial properties, exhibits chemosensitizing potential, making it a promising adjuvant to DOX therapy. In this study, a robust and eco-friendly HPLC method was developed and optimized for the simultaneous determination of DOX and RFP. Chromatographic separation was achieved on a C18 column (150 × 4.6 mm, 5 μm particle size) at 25 °C, using an isocratic mobile phase composed of acetonitrile and phosphate buffer (0.02 M, pH 5.43) in a 34.85:65.15 (v/v) ratio, with a flow rate of 0.8 mL/min and UV detection at 254 nm. The method was systematically optimized using a Quality-by-Design (QbD) approach with a full factorial design to assess the influence of key variables on chromatographic performance. The optimized chromatographic separation was achieved within 7 min, with retention times (Rts) of 3.26 min of DOX and 6.62 min for RFP. Excellent linearity was obtained over the ranges of 1.0-40.0 µg/mL for DOX and 1.0-30.0 µg/mL for RFP, with high determination coefficients (r² ≥ 0.999) and lower detection limits of 0.44 µg/mL and 0.39 µg/mL and quantitation limits of 1.35 µg/mL and 1.18 µg/mL, respectively. Greenness evaluation using the Analytical Greenness (AGREE) and Analytical Green Star Area (AGSA) metrics confirmed the method's high environmental sustainability. When applied to spiked human plasma samples, recovery values ranged from 92.38% to 107.20% with low %RSD, demonstrating strong accuracy, precision, and bioanalytical applicability. Overall, the developed method represents the first simple, sensitive, and environmentally benign analytical approach for the simultaneous quantification of doxorubicin HCl and rifampicin in biological matrices, making it highly suitable for routine therapeutic drug monitoring (TDM) in clinical and pharmacokinetic studies.
To conduct an epidemiological investigation on a case of severe fever with thrombocytopenia syndrome (SFTS) reported in Longyou County, Zhejiang Province in April 2025, and to perform nucleic acid detection and genetic characterization of Dabie bandavirus (DBV) in the patient's serum sample, aiming to provide scientific evidence and technical support for local SFTS prevention and control. An epidemiological investigation was conducted on the case. Serum samples from the confirmed case and close contacts, as well as related vector host specimens, were collected. DBV-specific nucleic acid was detected using real-time fluorescent quantitative PCR. The viral genome was amplified by RT-PCR and subjected to whole-genome sequencing. Professional computer software (SeqMan Ultra 17.2, Clustal X 2.1, BioEdit V7.2.6.1, and MEGA V7.0.26, etc.) was used for nucleotide and amino acid sequence alignment, phylogenetic tree construction, and calculation of genetic distances and homology percentages. The case had a history of tick bite and no history of travel outside the area. Fluorescence quantitative PCR detection showed that only the case's serum sample was positive for DBV nucleic acid, while serum samples from close contacts and related vector host specimens were all negative for DBV nucleic acid. After amplification and sequencing, the three fragments L, M, and S were successfully obtained. Genetic evolution analysis showed that they belonged to genotypes L (C), M (C), and S (A), respectively. Compared with the closest reference strain HZ2023-16 in the phylogenetic tree, the nucleotide homologies of the L, M, NS, and NP genes were 99.74%, 99.53%, 99.66%, and 99.19%, respectively, and the amino acid homologies were 100%, 99.72%, 99.66%, and 99.59%, respectively. Compared with the type C reference strain AHL/China/2011 (L(C)/M(C)/S(D)), the nucleotide homologies of the L, M, NS, and NP genes were 96.87%, 96%, 93.76%, and 95.66%, respectively, and the amino acid homologies were 99.42%, 98.51%, 98.29%, and 99.18%, respectively. Compared with the type A reference strain JX23XSH (L(A)/M(A)/S(A)), the nucleotide homologies of the L, M, NS, and NP genes were 96.24%, 95.97%, 95.35%, and 97.02%, respectively, and the amino acid homologies were 99.52%, 98.14%, 99.32%, and 99.18%, respectively. The average genetic distances of the L, M, and S genes compared with the HZ2023-16 reference strain were 0.003, 0.005, and 0.005, respectively; compared with the type C reference strain AHL/China/2011 (L(C)/M(C)/S(D)), they were 0.031, 0.04, and 0.053, respectively; compared with the type A reference strain JX23XSH (L(A)/M(A)/S(A)), they were 0.038, 0.04, and 0.038, respectively. Amino acid variation analysis showed that compared with the reference strains, this strain had 0-12 amino acid substitutions in the L protein (e.g., V68I, A140V), 3-20 substitutions in the M protein (e.g., D151E, G863S), 1-5 substitutions in the NS protein, and 1-2 substitutions in the NP protein. Combining the clinical manifestations, epidemiological history, and laboratory test results of the case, this outbreak can be determined as a local case of severe fever with thrombocytopenia syndrome. The strain is genetically closely related to human-derived reference strains and possesses specific genomic characteristics.
Tuberculosis (TB) is one of the deadliest bacterial infectious diseases worldwide, with rising cases of multidrug-resistant (MDR) and extensively drug-resistant (XDR) strains. Bedaquiline (BDQ)-containing regimens have become important for the treatment of MDR/XDR-TB, and resistance to BDQ is increasing. Understanding genetic mutations is crucial for early detection of BDQ-resistant strains and thus maintaining the effectiveness of these drugs. This study aimed to review mutations associated with BDQ-resistant TB globally. This study systematically searched the keywords TB, XDR, MDR, BDQ, and genes in the PubMed, Scopus, Web of Science, and Embase databases for studies reporting BDQ-resistant TB and their associated genes globally from 2014 to 2025. This systematic review included 40 studies and 25,234 patient samples with MDR and XDR-TB from around the world. Results showed significant variation in BDQ resistance across the World Health Organization (WHO) regions, with the highest in the Eastern Mediterranean and the lowest in the Western Pacific. Furthermore, the data collected showed that, among the continents studied, resistance was highest in Africa and lowest in the Americas. The country distribution showed that resistance rates were higher in Iran (n = 24), Moldova (n = 26), and Armenia (n = 35), and lower in Italy (n = 1001) and the Philippines (n = 724) than in other countries in the analysis. Genetically, the most resistance-associated mutations were observed in the Rv0678, atpE, and pepQ genes, respectively. Given the increasing BDQ resistance and regional variability, it is essential to develop early detection systems, genomic surveillance, robust drug policy enforcement, and rapid diagnostics to maintain treatment effectiveness and curb the spread of resistance. Future research should focus on elucidating resistance mechanisms and developing novel therapeutic strategies.
Satellite DNAs (satDNAs) are repetitive sequences that play important roles in chromosomal architecture, genome evolution, and regulation. Here, we present a comprehensive characterization of Tenebrio molitor satellitome, integrating cytogenetic mapping, in silico genome annotation, divergence profiling, screening of extrachromosomal circular DNA (eccDNA), transcription analysis across developmental stages, and phylogenetic and age analyses. SatDNAs exhibited diverse chromosomal organizations, ranging from widespread to chromosome-restricted distributions. Discrepancies between assembly-based and physical mapping highlight limitations of individual approaches and underscore the importance of their integration. Divergence landscape analyses revealed different homogenization efficiencies and turnover rates, reflecting distinct evolutionary trajectories among individual satDNAs. Phylogenetic reconstruction revealed distinct patterns which include clear species-specific clustering of monomers, mixed interspecific clustering, and dispersed topologies. Comparative analyses across insect orders enabled age estimation, identifying both ancient (≥380 MYA) and lineage-specific satDNAs, apparently restricted to T. molitor. We designed and applied an approach that enables the simultaneous detection of multiple satDNAs within the eccDNA fraction which confirmed the presence of six satDNAs in eccDNA. RNA-seq analyses revealed coordinated, stage-specific transcription of all satDNAs, with elevated expression in late male pupae and early male adults. Together, these results reveal a highly dynamic, heterogeneous, and functionally relevant satDNA landscape in T. molitor and demonstrate the importance of integrative approaches for understanding molecular mechanisms and trajectories of satDNA evolution.
Chronic hepatitis B virus (HBV) infection affects nearly 300 million people worldwide and remains a leading cause of the global cancer burden. While its causal relationship with hepatocellular carcinoma (HCC) is well established, accumulating epidemiological and mechanistic evidence suggests that HBV may also contribute to the development of extrahepatic malignancies, including gastric, colorectal, pancreatic, lymphoid, and respiratory cancers. Cohort studies and meta-analyses consistently report an increased risk in individuals infected with HBV, but causality remains controversial due to confounding factors such as coinfection, lifestyle factors, and genetic susceptibility. Possible mechanisms include viral DNA integration into the host genome, HBV X protein (HBx)-mediated activation of oncogenic signaling pathways (PI3K-Akt, Wnt), epigenetic modifications, and alterations in the immune microenvironment that promote tumor immune evasion. Advances in biomarker discovery, imaging technologies, and antiviral and targeted therapies offer opportunities for early detection and improved management of HBV-related cancers. This review summarizes the current knowledge on the epidemiology, pathophysiology, diagnostic strategies, and treatment approaches of HBV-related extrahepatic malignancies, as well as the current controversies, with the aim of guiding future research and clinical practice.
PRAME is a member of the cancer-testis antigen family and exhibits abnormally high expression in various solid tumors. Epigenetic regulation plays a critical role in the development and progression of hepatocellular carcinoma (HCC). Alterations in promoter methylation of specific genes have been widely recognized as valuable biomarkers for predicting cancer risk and assessing prognosis. This study systematically investigated the biological function and clinical relevance of PRAME promoter methylation in HCC. Using data from The Cancer Genome Atlas (TCGA) cohort, we identified methylation-associated prognostic genes and examined the association between PRAME promoter methylation, its mRNA expression levels, overall survival (OS), and clinicopathologic characteristics. We then validated these findings in an independent HCC cohort from Shulan (Hangzhou) Hospital using MethylTarget multiplex gene methylation detection technology. To investigate the potential role of PRAME in vascular invasion and metastasis, we conducted Gene Ontology (GO) enrichment analysis and Gene Set Enrichment Analysis (GSEA) at both histological and cellular levels. Analyses across three HCC cohorts consistently showed that the PRAME promoter is generally hypomethylated in tumor tissues, and this hypomethylation was significantly correlated with elevated PRAME mRNA expression. In both the TCGA and Shulan (Hangzhou) Hospital cohorts, PRAME promoter hypomethylation was associated with more advanced clinicopathologic features and shorter OS. Moreover, PRAME promoter methylation status was strongly linked to vascular invasion and distant metastasis in HCC. Further functional analyses revealed significant upregulation of Wnt signaling and cell adhesion-related pathways in tissues with high PRAME expression, a finding corroborated by single-cell RNA sequencing data. Hypomethylation of the PRAME promoter is closely associated with poor overall survival, adverse clinicopathologic features, vascular invasion, and metastasis in HCC patients. It plays a pivotal role in regulating gene expression and promoting tumor invasiveness. These findings suggest that PRAME promoter methylation may serve as a promising epigenetic biomarker for prognostic evaluation in HCC.
Childhood hypertension is closely associated with cardiovascular disease in adulthood, and the carotid artery is one of the principal target organs affected by elevated blood pressure (BP). The purpose of this study is to assess the association between carotid ultrasound-derived indices and elevated BP in children and to identify early vascular changes using multimodal ultrasound imaging. This case-control observational study included 118 children aged 10-11 years, of whom 55 had elevated BP and 63 had normal BP. Carotid ultrasound measurements included carotid diameter, conventional carotid intima-media thickness (CIMT), ultrahigh frequency CIMT (uhf-CIMT), carotid intima thickness, carotid media thickness (CMT), intima-to-media ratio (I/M), and radiofrequency-derived pulse wave velocity (rf-PWV). Associations between ultrasound parameters and BP were evaluated using multivariate linear regression models adjusted for age, sex, and body mass index. Children with elevated BP had higher values of uhf-CIMT, CMT, rf-PWV and lower values of I/M compared with children with normal BP (all p < 0.05). Systolic BP with the strongest association observed for CMT (r = 0.45). After adjustment for potential confounders, elevated BP remained independently associated with CMT, uhf-CIMT, I/M, and rf-PWV (all p < 0.05). Rf-PWV was significantly associated with carotid structural indices, particularly the I/M ratio (p < 0.05). Elevated BP in children is associated with early carotid structural remodeling and increased arterial stiffness. Ultrahigh frequency ultrasound may facilitate the detection of subtle vascular changes in pediatric with elevated BP.
Avian metapneumovirus (aMPV) is an economically significant respiratory pathogen of poultry that affects performance, egg production, and fertility of breeder flocks. Despite its impact, aMPV continues to be ill-defined in Egypt with no integrated surveillance studies conducted in breeder flocks. Therefore, the current study was designed to provide the first integrated molecular and serological evaluation of aMPV circulation in broiler breeders in Egypt. Between 2024 and 2025, a total of 6,000 serum samples and 800 tracheal swabs were collected from 60 unvaccinated broiler breeder flocks across twelve Egyptian governorates. Sera were obtained at 16 weeks (rearing phase) and 35 weeks (production phase) of age and screened for aMPV subtypes A and B by ELISA. Tracheal swabs collected during the production phase were pooled into 80 composite samples and tested for the viral RNA by reverse transcription quantitative real time PCR (RT-qPCR). Serological analysis revealed widespread aMPV exposure, with governorate-level seroprevalence ranging between 64.4% (Luxor) and 89% (Giza). Antibody titers increased between 16 and 35 weeks of age, reflecting cumulative viral exposure. Molecular testing detected aMPV RNA in 67 (83.75%) of pooled swab samples. Subtype B was the predominant genotype detected solely in 65 (81.25%) sample pools and in co-detection with subtype A in 2 pools (2.5%). Serological and molecular findings were generally aligned, with flocks positive for aMPV RNA often exhibiting higher antibody titers. These findings indicate that aMPV, particularly subtype B, is likely endemic across the Egyptian broiler breeder flocks. The study highlights critical knowledge gaps and emphasizes the need for viral isolation, sequencing, and controlled evaluation of biosecurity and vaccination strategies.
Coarctation of the aorta represents 6-8% of congenital heart defects. Surgical repair is often done in infancy, but late complications such as aneurysm formation may be present decades later. Late aneurysm after childhood coarctation repair remains a recognized complication, but surveillance lapses and complex arch branch involvement can delay detection and complicate repair planning. We present a case of a large descending thoracic aortic aneurysm found incidentally over 60 years after initial coarctation repair, a latency period exceeding many reported cases [5]. A 62-year-old woman with a history of coarctation repairs at 5 months of age and at 10 years of age presented asymptomatically for echocardiogram evaluation of a cardiac murmur. Computed tomography angiography (CTA) revealed a 7.3 cm saccular aneurysm (length 8.2 cm, width 6.5 cm, depth 7.3 cm) of the distal aortic arch and proximal descending aorta, with the left subclavian artery previously ligated and the left carotid artery originating from the aneurysm sac. She underwent a hybrid repair performed in a single setting: right common carotid-to-left common carotid bypass via retro-pharyngeal tunneling followed by thoracic endovascular aortic repair (TEVAR) with overlapping stent grafts. Angiography confirmed complete exclusion of the aneurysm without endoleak. Follow-up revealed no complications, and the patient remained asymptomatic. This case underscores the importance of lifelong surveillance in patients with repaired congenital heart disease, even in the absence of symptoms. Evolution of technology with endovascular repair has provided less invasive options for management of complex aneurysms resulting after repair of aortic coarctation. This case report demonstrates surveillance gaps after repair of thoracic aortic coarctation.
Alzheimer's disease (AD) is one of the most prevalent neurodegenerative disorders worldwide, requiring early identification for timely intervention and to slow disease progression. However, existing diagnostic approaches, while effective at later stages, remain limited in detecting early-stage AD. Handwriting analysis has recently emerged as a non-invasive, cost-effective, and ecologically valid digital behavioral biomarker that reflects neurocognitive impairment. This review examines the role of handwriting as a neurocognitive marker for AD, focusing on integrating deep learning methodologies to enhance early diagnostic accuracy. It also elucidates the neurocognitive mechanisms linking handwriting behavior and AD, addressing current methodological and translational challenges. We performed a PRISMA-informed structured literature search and narrative synthesis of handwriting- and drawing-based studies for detecting AD/mild cognitive impairment (MCI), including offline handwriting images and online pen-stroke kinematics captured by digital devices. Task paradigms, data dimensions, preprocessing pipelines, modeling strategies (traditional machine learning and deep learning), evaluation practices, and translational considerations were summarized, and studies were organized by detection purpose and analytic approach. Our findings show that handwriting-based models generally discriminate AD/MCI from healthy controls with accuracy exceeding 80%, while deep learning models (e.g., convolutional neural network and multimodal Transformer fusion) approach 90% in structured tasks like clock drawing and figure copying. Online kinematic markers (e.g., reduced velocity, prolonged in-air time, increased pausing, and pressure instability) recur across studies, and multimodal integration with speech, gait, or facial signals can further improve sensitivity and ecological validity, although most studies are small and single-center.
Electronic health records (EHR)-based childhood asthma prediction may be a valid, cost-effective, and scalable option for early detection of at-risk children for targeted preventive intervention, but this approach has not been adopted in clinical practice. The purpose of this pilot study was to evaluate the feasibility and acceptability of the Pediatric Asthma Risk Score (PARS) as an EHR-based passive digital marker (PDM) for childhood asthma prognosis in clinical practice. Feasibility was defined by the level of concordance between the PARS and a clinician's classification of a case study's risk of developing school-age asthma. Acceptability was based on clinicians' intentions to use the PARS and its usability as a PDM for childhood asthma prognosis. Blinded to the PARS risk classification, clinicians were asked to predict the case study's risk of school-age asthma based on their professional judgment as either low or high risk and rate their confidence in their prediction. Unblinded to the PARS risk classification, clinicians rated the acceptability of the PARS as a PDM. Logistic regression models were used to summarize and identify correlates of feasibility and acceptability measures. There was 74% (95% CI: 66-81%) concordance between the PARS and clinician prediction of our case study's risk of developing school-age asthma. Risk discordance (26%) was associated with low confidence in a clinician's professional judgment (adjusted odds ratio [aOR]: 4.78; 95% CI: 1.85, 12.34) and PARS (aOR: 4.48; 95% CI: 1.16, 17.33). Unblinded to the PARS risk classification, more than 80% of clinicians ranked their intentions to use the PARS and its usability as a PDM as 4+ on a 0-6 Likert scale. Our findings demonstrate preliminary feasibility and acceptability of the Pediatric Asthma Risk Score (PARS) as a passive digital marker for childhood asthma risk, supporting further evaluation in prospective real-world studies.