Sample Cohort Research Articles

Performance of a blood-based screening test for the early detection of colorectal cancer.

232 Background: While the clinical benefit of highly sensitive screening tools for colorectal cancer (CRC) has been established, adherence rates remain low. The convenience and familiarity of blood-based testing may reduce disease burden through greater access and compliance to regular and recommended asymptomatic screening. We report on preliminary findings from a minimally invasive blood-based early cancer detection (ECD) screening test for CRC. Methods: Plasma samples from healthy individuals and CRC patients sourced from biobanks were included. A targeted panel composed of differentially methylated genomic regions was developed. Using an independent cohort of plasma samples, a machine-learning model was trained to classify patient samples into cancer-free and CRC based on observed differential methylation patterns in circulating cell-free DNA (cfDNA). The assay performance was calculated based on concordance with clinical diagnosis (CRC, cancer-free). Results: The test set included 440 individuals: 127 with a confirmed CRC diagnosis (47% stage I/II, mean age 65±12 years, male 46%) and 313 without cancer (mean age 56±10 years, male 41%). Overall sensitivity was 95% (95% CI: 88-100%), and specificity was 91% (95% CI: 84-98%). Among patients with stage I disease, sensitivity was 92% overall, and 88% in screen-detected individuals. Sensitivity was 100% (n=14/14) in microsatellite-high tumors and 95% (n=107/113) in microsatellite-stable tumors. Multivariate analysis demonstrated associations between methylation signal and tumor size, cancer stage, and sex. Compared with a tumor-informed, variant-based circulating tumor DNA (ctDNA) test (Signatera), we observed a PPA (positive percent agreement) of 100%. Moreover, the differentially methylated allele fraction (DMAF) across target regions strongly correlated with variant allele frequencies (VAFs) from the tumor-informed test (Spearman’s correlation coefficient, 0.85). Conclusions: Using epigenetic markers specific for CRC, we developed a sensitive and specific blood-based test to detect cfDNA from early-stage and asymptomatic disease. The methylation signal strongly correlated with ctDNA VAFs from a clinically validated tumor-informed test and increased with cancer stage and disease burden. This test could serve as a means to increase patient adherence rates for CRC screening, thereby improving patient outcomes.

Trajectory Analysis in FBG and the Incidence of Chronic Kidney Disease: A Nationwide Population-Based Study

Objectives: This study aimed to classify fasting blood glucose (FBG) trajectories by sex and examine their associations with the risk of chronic kidney disease (CKD). Methods: Using data from the National Health Insurance Service-National Sample Cohort in Korea, participants aged 40 years and above, without CKD or diabetes mellitus (DM), were followed from 2002 to 2009. Based on their FBG trajectories, participants were categorized into two classes and stratified by sex. CKD incidence rates were analyzed according to these FBG trajectories, and the impact of additional risk factors on CKD incidence was assessed. Results: A total of 91,131 participants were analyzed. Among individuals classified in Class 1, FBG levels gradually increased from 90.7 (men) and 88.7 (women) in 2002 to 96.6 (men) and 93.2 (women) in 2009. In contrast, participants classified as Class 2 exhibited a rapid increase in FBG levels, rising from 106 (men) and 106 (women) in 2002 to 144 (men) and 132 (women) in 2009. The incidence of CKD increased over time in both men and women classified as Class 2 compared to Class 1, with respective hazard ratios (HR) of 1.35 for men and 1.53 for women. Additionally, increased age, hypertension, and body mass index (BMI) were independently associated with an elevated risk of CKD. Conclusions: The Class 2 group demonstrated a significantly higher incidence of CKD compared to the Class 1 group. This finding indicates the need for the proactive management of individuals with relatively high FBG levels featuring rapid FBG increases in order to mitigate the risk of CKD development.

Multi-site blinded validation of a deep learning approach for clinical-grade MSI/dMMR detection in colorectal cancer from H&E-stained pathology images.

44 Background: Testing for microsatellite instability (MSI) or mismatch repair deficiency (dMMR) is part of the diagnosis and clinical management of patients with colorectal cancer (CRC). Healthcare services recommend MSI or dMMR testing for all CRC patients to guide therapeutic choices and assist in identifying Lynch Syndrome. However, in clinical practice, high costs and the demand for timely test results, combined with the rising prevalence of CRC and a shrinking pathology workforce, present a barrier to universal adoption. This highlights the need for rapid and affordable alternatives. PANProfiler CRC (PPC) is a deep learning-based solution for detecting MSI/dMMR in CRC tumors that only requires whole slide images (WSIs) of haematoxylin and eosin (H&E)-stained tissue to provide test results. Using only WSIs, PPC offers an efficient alternative to standard testing. This study evaluates PPC's performance in a multi-site blinded setting. Methods: Blinded validation was performed using 3246 WSIs of H&E-stained CRC specimens. PPC provided outputs as "Stable", "Unstable", or "Indeterminate", with "Unstable" indicating dMMR or MSI-High, and "Stable" indicating proficient mismatch repair or non-MSI-High. "Indeterminate" was returned when PPC did not have a definitive result. PPC was evaluated by comparison to standard MSI/dMMR tests. Validation data spanned three cohorts from two sites (Table). St James’s University Hospital (SJUH), Leeds, UK, supplied Cohorts 1 and 2; Cohort 3 was sourced from Wales Cancer Biobank (WCB), UK. Blinded analysis was performed at SJUH. Results: Results are given (Table). PPC demonstrated an overall percent agreement of 93.91%, a positive percent agreement of 92.17%, and a negative percent agreement of 94.15%, returning a definitive result for 88.05% of WSIs. Conclusions: This real-world, multi-site, blinded validation study demonstrates PPC’s remarkable performance, comparable to standard tests for detecting MSI/dMMR in CRC, with high test replacement rates. In the clinical setting, PPC could significantly accelerate testing and enable timely delivery of stratified treatment plans. This accurate and cost-effective diagnostic solution promises to revolutionize MSI/dMMR testing in CRC. Blinded validation results of PPC with confidence intervals (CI) at 95%. Site Cohort Sample Size (Unstable; Stable) Overall Percent Agreement % (CI) Positive Percent Agreement % (CI) Negative Percent Agreement %(CI) Test Replacement Rate % SJUH 1 488 (78; 410) 92.79 (89.86-95.08) 90.16 (79.81-96.30) 93.24(90.11-95.62) 85.25 SJUH 2 2704 (318; 2386) 94.31(93.31-95.21) 92.31(88.48-95.18) 94.57(93.52-95.50) 88.42 WCB 3 54 (11; 43) 84.31 (71.41-92.98) 100.00(71.51-100.00) 80.00(64.35-90.95) 94.44 All 3246 (407; 2839) 93.91 (92.97-94.76) 92.17(88.82-94.78) 94.15(93.16-95.04) 88.05

P-2142. Cross-Reactivity of Fungal Enzyme Immunoassays (EIA): Implications for Clinical Diagnosis in Patients with Autoimmune Disease

Abstract Background The immune system exhibits remarkable discriminatory power, distinguishing between self and non-self to mount protective responses against infections. In clinical diagnostics targeting dimorphic fungi, this immunological discrimination is leveraged to detect specific antibodies indicative of disease states. However, reliance on immune modulation for diagnostic purposes presents inherent challenges due to the intricate nature of antigen-antibody interactions. This complexity is particularly pronounced in patients with a history of autoimmune disorders, where the risk of false positive results in immunoassays is heightened due to the dysregulated immune response characteristic of autoimmune conditions. Methods We assessed the potential for cross-reactivity of serum samples from patients with autoimmune disease in dimorphic fungal Enzyme Immunoassays (EIAs) by analyzing a cohort of clinical serum samples sourced from the Florida region, which is non-endemic for dimorphic fungal diseases. Samples were commercially sourced as remnant-banked serum samples (Medix Biochemica, Florida, USA). 25 serum samples obtained from distinct patients were subjected to testing using Coccidioides, Blastomyces, and Histoplasma EIA (MiraVista Diagnostics, Indiana, USA). This ncluded 5 individuals devoid of autoimmune disorders, cohorts of 5 patients each diagnosed with Rheumatoid Arthritis and Multiple Sclerosis, and 10 patients with Systemic Lupus Erythematosus. Samples were tested in triplicate. Results Preliminary findings from our study unveiled nuanced levels of cross-reactivity across the dimorphic fungal EIAs utilized for the three autoimmune disorders examined. Depending on the specific dimorphic fungal EIA assay and autoimmune disorder, approximately 10–30% of serum samples from patients tested intermediate or weakly positive in the respective fungal EIAs. All control patient samples tested negative. Conclusion These preliminary findings underscore the complexity of diagnosing dimorphic fungal infections, highlighting the significance of the patient's immune status as a confounding factor. Clinical practitioners should consider the possibility of autoimmunity when encountering weakly positive or intermediate dimorphic fungal EIA results. Disclosures Arryn Craney, PhD D(ABMM), MiraVista Diagnostics: Employee; Salary Kendra Carlson, n/a, MiraVista Diagnostics: Employee; Salary Kyla Seyfried, n/a, MiraVista Diagnostics: Employee; Salary John Witt, n/a, MiraVista Diagnostics: Employee; Salary Andrew Hanzlicek, DVM, MiraVista Diagnostics: Employee; Salary Nicolas Barros, MD, MiraVista Diagnostics: Advisor/Consultant Lawrence J. Wheat, MD, MiraVista Diagnostics: Employee; Salary|MiraVista Diagnostics: Ownership Interest

Metabolic adaptations to acute glucose uptake inhibition converge upon mitochondrial respiration for leukemia cell survival

One hallmark of cancer is the upregulation and dependency on glucose metabolism to fuel macromolecule biosynthesis and rapid proliferation. Despite significant pre-clinical effort to exploit this pathway, additional mechanistic insights are necessary to prioritize the diversity of metabolic adaptations upon acute loss of glucose metabolism. Here, we investigated a potent small molecule inhibitor to Class I glucose transporters, KL-11743, using glycolytic leukemia cell lines and patient-based model systems. Our results reveal that while several metabolic adaptations occur in response to acute glucose uptake inhibition, the most critical is increased mitochondrial oxidative phosphorylation. KL-11743 treatment efficiently blocks the majority of glucose uptake and glycolysis, yet markedly increases mitochondrial respiration via enhanced Complex I function. Compared to partial glucose uptake inhibition, dependency on mitochondrial respiration is less apparent suggesting robust blockage of glucose uptake is essential to create a metabolic vulnerability. When wild-type and oncogenic RAS patient-derived induced pluripotent stem cell acute myeloid leukemia (AML) models were examined, KL-11743 mediated induction of mitochondrial respiration and dependency for survival associated with oncogenic RAS. Furthermore, we examined the therapeutic potential of these observations by treating a cohort of primary AML patient samples with KL-11743 and witnessed similar dependency on mitochondrial respiration for sustained cellular survival. Together, these data highlight conserved adaptations to acute glucose uptake inhibition in diverse leukemic models and AML patient samples, and position mitochondrial respiration as a key determinant of treatment success.

A Fast, High-Sensitivity 96-Well Plate-Based MICROFASP Method for Processing Low Microgram Proteomics Sample within 1.5 h.

A rapid, sensitive, and high-throughput sample preparation method is of paramount significance for proteomics analysis. Here, we report a fast, high-sensitivity MICROFASP method that is capable of completing sample preparation within 1.5 h, enhancing the throughput by over 13 times compared to the previous reports. Protein digestion time was significantly cut from 17 h to 20 min in a limited volume. Simultaneous reduction and alkylation occurred within 30 min. The label-free quantitation intensities of proteins from the fast and conventional MICROFASP methods were highly correlated (r = 0.91), validating the reliability of the fast-MICROFASP method. When starting with 1 μg of K562 cell lysate, the fast-MICROFASP method identified over 6 times more protein groups and 19 times more peptides than did the iST method. A 96-well plate-based version was developed to process 8 brain tissue samples from APP/PS1 transgenic mice in parallel. Averagely, with only 1 μg of protein lysate, 2826 protein groups (n = 8, RSD = 0.7%) and 12,972 peptides (n = 8, RSD = 1.5%) were identified from each sample. Amyloid-beta protein was successfully identified as a highly expressed protein, which shows its potential for detecting diagnostic markers and proteome profiling with low-microgram samples. We anticipate the high-sensitivity 96-well plate-based fast-MICROFASP method will have wide application in high-throughput and rapid preparation of large cohorts of low-microgram samples (e.g., clinical biopsy) for comprehensive proteome profiling. Data are available via ProteomeXchange with the identifier PXD053720.

Aberrant promoter methylation of CTHRC1 gene and its clinicopathological characteristics in head and neck cancer.

Head and neck squamous cell carcinoma (HNSCC) is genetically complex and difficult to treat. Detection in the early stage is challenging, leading to diagnosis at advanced stages with limited treatment options. This study examined the collagen triple helix repeat containing 1 gene (CTHRC1) as a potential biomarker and therapeutic target in HNSCC. Despite documented CTHRC1 upregulation in various cancers, the underlying causes remain unclear. The objective was to investigate potential epigenetic regulation of CTHRC1 expression through the analysis of promoter methylation. CTHRC1 DNA methylation, mRNA, and its protein expression were analysed using The Cancer Genome Atlas (TCGA) HNSCC cohort and oral squamous cell carcinoma (OSCC) patient samples. Functional analysis included scrutinizing the protein-protein interaction network and associations with DisGeNET (disease gene network). Various statistical methods were employed for analysis. HNSCC tumours exhibited significant hypomethylation of CTHRC1 DNA, correlating with advanced disease features. Elevated mRNA and protein expression of CTHRC1 further support its role in disease progression. High CTHRC1 gene expression was associated with a poorer prognosis. The protein interaction network implicated crucial pathways in cancer development and links to oral submucous fibrosis. Despite the limitations of this study, including the use of retrospective data and need for functional experiments, CTHRC1 shows potential as a prognostic predictor and target for therapeutic applications in HNSCC, paving the way for further research and improved patient management.

Prevalence of the oral pathogen Filifactor alocis and its FtxA toxin related to clinical parameters and presence of Aggregatibacter actinomycetemcomitans

The Gram-positive organism Filifactor alocis is implicated in multiple oral diseases including periodontitis, and approximately 50% of known strains encode and produce a recently identified repeat-in-toxin (RTX) protein, FtxA, partly homologous to the Aggregatibacter actinomycetemcomitans leukotoxin. By assessing a longitudinal Ghanaian study population of adolescents, we recently identified a possible correlation between F. alocis levels, ftxA gene carriage, and progression of clinical attachment loss (CAL). To extend knowledge on the possible significance of F. alocis and its FtxA in periodontal disease, we have in the present work analyzed saliva samples in an independent cohort of periodontitis (n=156), collected at two private periodontal specialist practices in Perth, Western Australia. The present results corroborate that high loads of F. alocis and the presence of its ftxA gene together are associated with parameters of periodontal tissue destruction and severity. Moreover, among the individuals carrying A. actinomycetemcomitans, a majority also exhibited an ftxA-positive F. alocis, supporting the notion of the synergistic behavior of these two species. This emphasizes that F. alocis and its ftxA are involved in the pathogenesis of periodontitis and may have ecological roles, with diagnostic and prognostic implications for the disease.

Integrated multi-omics analyses of oral squamous cell carcinoma reveal precision patient stratification and personalized treatment strategies.

Oral cavity squamous cell carcinoma (OSCC), a leading subtype of head and neck cancer, exhibits high global incidence and mortality rates. Despite advancements in surgery and radiochemotherapy, approximately one-third of patients experience relapse. To improve current targeted and immunotherapy strategies for recurrent OSCC, we conducted multi-omics analyses on pretreatment OSCC samples (cohorts 1 and 2, n=137) and identified A3A and EGFR, both at the RNA and protein levels, as inversely expressed markers for patient stratification and response prediction. Survival analysis demonstrated that elevated A3A or PD-L1 expression levels correlated to improved responses to anti-PD-1 therapy in patients (cohort 3a, n=50, IHC). In contrast, high RRAS expression (cohort 4, n=252, qRT-PCR) was significantly associated with OSCC recurrence. Cell-based experiments revealed that RRAS was involved in radiotherapy and cisplatin resistance through the EGFR/RRAS/AKT/ERK signaling pathway. In OSCC patient-derived xenograft (PDX) mouse models, treatments with cisplatin and cetuximab (anti-EGFR) effectively reduced tumor size in EGFR-high-derived (#34) but not A3A-high-derived (#22) PDX tumors. Our study demonstrated that A3A-high tumors were immune-hot and responsive to anti-PD-1 therapy, whereas EGFR-high tumors exhibited chr.7p11.2 gains and DNA repair alterations. Additionally, RRAS-high tumors were associated with OSCC recurrence via AKT and ERK phosphorylation and demonstrate improved clinical outcomes with cetuximab therapy (cohort 3b, n=49, IHC). This study emphasizes the significance of A3A and EGFR expression levels in OSCC patient stratification and precision therapy, suggesting the use of anti-PD-1 or anti-EGFR treatments, respectively based on these biomarkers. Furthermore, RRAS emerges as a novel prognostic marker for local recurrence.

Risk of Osteoporotic Fractures among Patients with Thyroid Cancer: A Nationwide Population-Based Cohort Study.

The associations between thyroid cancer and skeletal outcomes have not been thoroughly investigated. We aimed to investigate the risk of osteoporotic fractures in patients with thyroid cancer compared to that in a matched control group. This retrospective cohort study included 2,514 patients with thyroid cancer and 75,420 matched controls from the Korean National Health Insurance Service-National Sample Cohort (NHIS-NSC, 2006-2019). The rates of osteoporotic fractures were analyzed, and associations with the levothyroxine dose were evaluated. Patients with thyroid cancer had a significantly lower risk of fracture than did the control group (hazard ratio [HR], 0.81; 95% confidence interval [CI], 0.69 to 0.94; P=0.006). Patients diagnosed with thyroid cancer after the age of 50 years (older cancer group) had a significantly lower risk of fracture than did those in the control group (HR, 0.72; 95% CI, 0.6 to 0.85; P<0.001), especially those diagnosed with spinal fractures (HR, 0.66; 95% CI, 0.51 to 0.85; P=0.001). Patients in the older cancer group started osteoporosis treatment earlier than did those in the control group (65.5±7.5 years vs. 67.3±7.6 years, P<0.001). Additionally, a lower dose of levothyroxine was associated with a reduced risk of fractures. In the clinical setting, the risk of fracture in women diagnosed with thyroid cancer after the age of 50 years was lower than that in the control group, which was caused by more proactive osteoporosis treatment in postmenopausal women with thyroid cancer.

Dynamic chronological changes of upper reference limits for liver enzymes in the nationwide population: the Korean National Health Insurance Service-National Sample Cohort 2002–2013

Dynamic chronological changes of upper reference limits for liver enzymes in the nationwide population: the Korean National Health Insurance Service-National Sample Cohort 2002–2013

Detection of GFAP and D‐Dimer in a Point‐of‐Care Test for Large Vessel Occlusion Ischemic Stroke

BACKGROUND The blood biomarkers GFAP (glial fibrillary acidic protein) and D‐dimer have been shown to accurately diagnose large vessel occlusion ischemic stroke. The Upfront DX LVOne is a novel lateral flow‐based point‐of‐care test used for the detection of GFAP and D‐dimer. This study sought to (1) assess the performance of LVOne against commercially available point‐of‐care test platforms, which independently detect GFAP and D‐dimer (Abbott iSTAT TBI Plasma and LumiraDx D‐dimer), and to (2) assess the diagnostic performance of LVOne in large vessel occlusion detection. METHODS Manufacturer‐reported detection thresholds of the LVOne test for GFAP and D‐dimer (213 pg/mL and 600 ng/mL, respectively) were compared against commercial hospital‐based point‐of‐care test platforms using 20 randomly selected plasma samples for each biomarker. D‐dimer correlation between LVOne and LumiraDx was assessed using Spearman's Rank correlation coefficient; GFAP correlation between LVOne and the Abbott iSTAT TBI results were assessed using point biserial correlation. Diagnostic performance of the LVOne point‐of‐care test in conjunction with clinical assessment for detection of large vessel occlusion in plasma samples from 210 patients with suspected stroke was validated. RESULTS There was a strong positive correlation (Rho = 0.86, P &lt;0.001) between the qualitative scoring of the lateral flow test and serum GFAP concentrations. Similarly, there was a significant positive correlation between the D‐dimer quantification and LVOne semiquantitative measurements (Rho = 0.90; P &lt;0.0001). In our cohort of 210 patients with suspected stroke, the LVOne test in conjunction with clinical assessment had a sensitivity of 75% (95% CI, 51%–91%), a specificity of 92% (95% CI, 87%–95%), a positive predictive value of 48% (95% CI, 30%–67%), and a negative predictive value of 97% (95% CI, 94%–99%) for detection of large vessel occlusion. CONCLUSION Our results validate detection thresholds for the LVOne test in a representative cohort of plasma samples of patients with suspected stroke. Further studies are required to demonstrate the efficacy, safety, and diagnostic performance of LVOne in capillary blood during emergency care/clinical triage.

EZH2 modulates mRNA splicing and exerts part of its oncogenic function through repression of splicing factors in CML.

The polycomb protein EZH2 is up-regulated in Chronic Myeloid Leukaemia (CML) and associated with transcriptional reprogramming. Here we tested whether EZH2 might also act as a modulator of the mRNA splicing landscape to elicit its oncogenic function in CML. We treated CML cell lines with EZH2 inhibitors and detected differential splicing of several hundreds of events, potentially caused by the transcriptional regulation of splicing factors. Amongst those genes, CELF2 was identified as a candidate to mediate part of the EZH2 inhibitor induced phenotype. Upon over-expression, we observed (1) reduced cell growth, viability, and colony formation of CML cell lines, (2) a change in the splicing landscape, partially overlapping with EZH2 mediated changes, (3) the down-regulation of MYC signalling. Importantly, these findings were successfully validated in a cohort of CML patient samples, confirming the role of CELF2 as EZH2-regulated tumour-suppressor, contributing to the severe splicing de-regulation present in CML. Based on this we propose that EZH2 exerts part of its oncogenic function in CML through the transcriptional repression of splicing factors. Finally, analysis of publicly available datasets suggests that splicing modulation by EZH2 might not be restricted to CML.

Children's emerging concepts of resilience: insights from using body mapping in an East London cohort sample of 7-10-year-old children.

Understanding resilience factors in children is essential for developing early mental health interventions. Middle childhood is an understudied developmental stage, with many quantitative measures lacking validation for this age group and not capturing diverse experiences. This study aimed to use body mapping, an arts-based method, as a novel approach to understand 7-10-year-old children's concepts of resilience (including definitions and factors that contribute to resilience) in East London. An advisory group of six children commented on the findings. Body mapping was included in the Development of Emotional Resilience (DEER) Study. Participants drew a resilience symbol, wrote recent worries and colored on an A4-sized body map to signal where they embody stress. Demographic data were collected via self- and parent-report surveys and school records. Manifest content analysis identified four thematic categories related to worries, somatic stress and resilience. 196 children (48.47% boys, 46.43% girls; 35.20% White, 30.10% Asian, 11.22% Black) across school years 3-5 completed body mapping. Concepts of resilience included perseverance and metaphorical representations of personal strength. We also identified socioecological factors that contributed to resilience, mainly at the individual and interpersonal levels. Boys more often depicted Sports whilst more girls depicted Engagement in the arts and Social networks. 11 worry categories emerged, including education, relationships and physical health. Of the body categories colored (n = 51), the most common were the head, hands and abdomen/stomach. Children expressed dominant and abstract symbols of resilience and identified factors that contributed to resilience. Hobbies and strong relationships may be particularly important in middle childhood, corroborated by the advisory group's experiences. Body mapping revealed diverse worries (e.g., education, change and uncertainty and global and societal concerns) and somatic experiences of stress (e.g., the head, chest and torso). Through prioritising children's perspectives, body mapping holds promise in clinical and educational settings.

Role of Gut and Urinary Microbiome in Children with Urinary Tract Infections: A Systematic Review.

Background: The complex interaction between the gut and urinary microbiota underscores the importance of understanding microbial dysbiosis in pediatric urinary tract infection (UTI). However, the literature on the gut-urinary axis in pediatric UTIs is limited. This systematic review aims to summarize the current literature on the roles of gut and urinary dysbiosis in pediatric UTIs. Methods: This systematic review was conducted in accordance with the Preferred Reporting Items for Systematic Reviews and Meta-Analyses guidelines. A comprehensive literature search was performed across four databases, including PubMed, Web of Science, Scopus, and EMBASE. All studies published between January 2003 and December 2023 utilizing 16S rRNA sequencing to profile the gut or urinary microbiome in children with UTIs were included. Heat map visualization was used to compare microbial profiles between UTI and control cohorts. The methodological quality assessment was performed using the Newcastle-Ottawa scale (NOS). Results: Eight studies were included in this review. While five studies compared the microbiota signatures between patients and controls, three studies focused solely on the UTI cohort. Also, the gut and urinary microbiome profiles were investigated by four studies each. The consistent loss of microbiome alpha-diversity with an enrichment of specific putative pathobiont microbes was observed among the included studies. Escherichia coli consistently emerged as the predominant uropathogen in pediatric UTIs. In addition to this, Escherichia fergusonii, Klebsiella pneumoniae, and Shigella flexneri were isolated in the urine of children with UTIs, and enrichment of Escherichia, Enterococcus, Enterobacter, and Bacillus was demonstrated in the gut microbiota of UTI patients. On the contrary, certain genera, such as Achromobacter, Alistipes, Ezakiella, Finegoldia, Haemophilus, Lactobacillus, Massilia, Prevotella, Bacteroides, and Ureaplasma, were isolated from the controls, predominantly in the fecal samples. The methodological quality of the included studies was variable, with total scores (NOS) ranging from 5 to 8. Conclusions: The enrichment of specific pathobionts, such as Escherichia coli, in the fecal or urinary samples of the UTI cohort, along with the presence of core microbiome-associated genera in the non-UTI population, underscores the critical role of the gut-urinary axis in pediatric UTI pathogenesis. These findings highlight the potential for microbiome-based strategies in pediatric UTIs. Further studies with larger cohorts, standardized healthy controls, and longitudinal profiling are essential to validate these observations and translate them into clinical practice.

Micro-RNA Signature in CSF Before and After Autologous Hematopoietic Stem Cell Transplantation for Multiple Sclerosis.

MicroRNAs (miRNAs) are regulators of gene expression and have been reported to be dysregulated in people with multiple sclerosis (pwMS). Autologous hematopoietic stem cell transplantation (aHSCT) is an immune-ablative treatment intervention for pwMS. Currently, it is unknown if aHSCT affects expression levels of miRNAs in CSF. We explored the ability of circulating miRNA to discriminate between pwMS and healthy controls (HCs) and investigated whether these miRNAs were affected by treatment with aHSCT. Using quantitative reverse transcription PCR, 87 miRNAs were analyzed in CSF samples of a discovery cohort (baseline: 4 & HC: 4). The top 22 miRNAs discriminating between pwMS and HCs were then analyzed in 187 CSF samples of a validation cohort (pwMS: 50, HC: 32). Samples, failing quality control or being follow-ups to baseline samples with quality control issues, were excluded from further analyses. The remaining 133 samples (HC: 29, MS: baseline: 33, 1 year: 30, 2 years: 26, 3-5 years: 15) were analyzed for expression of the top 22 miRNAs. Twelve miRNAs were dysregulated in pwMS compared with HC (q < 0.05). Associations with clinical and analytical parameters were observed in relation to all 12 miRNAs; however, a cluster of 4 miRNAs (miR-16-5p, miR-21-5p, miR-150-5p, and miR-146a-5p) with strong correlations (r > 0.60, p < 0.001) with multiple parameters was identified. Of the 12 miRNAs, 8 were differentially expressed in pwMS with gadolinium-enhancing lesions at baseline and 4 by prior disease-modifying treatment class (p < 0.05). These 4 miRNAs correlated strongly with each other, decreased after aHSCT, and remained low throughout the follow-up period (p < 0.05). Target and pathway analysis of these revealed association with biological processes affecting cytokine production, inflammatory response, and regulation of myelin maintenance. miRNAs are dysregulated in CSF from pwMS and particularly in patients with less effective treatments and/or higher inflammatory disease activity. A 4-miRNA signature with elevated expression of miR-16-5p, miR-21-5p, miR-150-5p, and miR-146a-5p was recurring in multiple analyses. After intervention with aHSCT, the expression levels approached the levels of the HCs, suggesting a potent treatment effect.

Equilibrium mechanical properties of the human uterus in tension and compression.

A successful pregnancy relies on the proper cellular, biochemical, and mechanical functions of the uterus. A comprehensive understanding of nonpregnant and pregnant uterine mechanical properties is key to understanding different obstetric and gynecological disorders such as preterm birth, placenta accreta, uterine rupture, leiomyoma, adenomyosis, and endometriosis. This study sought to characterize the macro-scale equilibrium material behaviors of the human uterus in nonpregnancy and late pregnancy under both compressive and tensile loading. Forty four human uterine specimens from 16 patients (8 nonpregnant [NP] and 8 pregnant [PG]) were tested using spherical indentation and uniaxial tension coupled with digital image correlation (DIC). A three-strain level incremental load-hold protocol was applied to both tests. A microstructurally-inspired material model considering fiber architecture was applied to this dataset. Inverse finite element analysis (IFEA) was then performed to generate a single set of mechanical parameters to describe compressive and tensile behaviors. The freeze-thaw effect on uterine mechanical properties was also evaluated. For this cohort of tissue samples, the fiber network of the PG uterus was more extensible than in the NP tissue. The initial fiber stiffness and ground substance compressibility were similar between NP and PG uterine tissue. Lastly, a single freeze-thaw cycle did not systematically alter the mechanical behavior of the human uterus under indentation.

Screening and identification of gene expression in large cohorts of clinical tissue samples unveils the major involvement of EZH2 and SOX2 in lung cancer

Lung adenocarcinoma (LUAD), the primary subtype of Non-Small Cell Lung Cancer (NSCLC), accounts for 80% to 85% of cases. Due to suboptimal screening method, LUAD is often detected in late stage, leading to aggressive progression and poor outcomes. Therefore, early disease prognosis for the LUAD is high priority. In order to identify early detection biomarkers, we conducted a meta-analysis of mRNA expression TCGA and GTEx datasets from LUAD patients. A total of 795 differentially expressed genes (DEGs) were identified by exploring the Network-Analyst tool and utilizing combined effect size methods. DEGs refer to genes whose expression levels are significantly different (either higher or lower) compared to their normal baseline expression levels. KEGG pathway enrichment analysis highlighted the TNF signaling pathway as being prominently associated with these DEGs. Subsequently, using the MCODE and CytoHubba plugins in Cytoscape software, we filtered out the top 10 genes. Among these, SOX2 was the only gene exhibiting higher expression, while the others were downregulated. Consequently, our subsequent research focused on SOX2. Further transcription factor-gene network analysis revealed that enhancer of zeste homolog 2 (EZH2) is a significant partner of SOX2, potentially playing a crucial role in euchromatin-heterochromatin dynamics. Structure of SOX2 protein suggest that it is a non-druggable transcription factor, literature survey suggests the same. SOX2 is considered challenging to target directly, or "non-druggable," because of several intrinsic properties that make it difficult to design effective therapeutic agents against it. The primary function of SOX2 is to bind DNA and regulates gene expression. Unlike enzymes or receptors with defined active sites or binding pockets, transcription factors typically have relatively flat or diffuse surfaces that do not offer obvious "pockets" for small molecules to bind effectively. Hence, we drove our focus to investigate on potential drug(s) targeting EZH2. Molecular docking analyses predicted most probable inhibitors of EZH2. We employed several predictive analysis tools and identified GSK343, as a promising inhibitor of EZH2.

A Quantitative Method for Assessing Treatment-Related Changes within the Airway Mucosa in Patients with Chronic Bronchitis

No standardized method has yet been established for evaluating airway mucosal aberrancies associated with chronic obstructive pulmonary disease (COPD) or chronic bronchitis (CB). While goblet cell hyperplasia (GCH) is an established pathognomonic hallmark of the CB disease process, no standardized method exists for acquiring mucosal biopsies and assessing morphologic airway mucosa alterations. Additionally, the impacts from interventions targeting the airway mucosa are not well defined. In this context, a reliable and robust measure for assessing airway mucosa at baseline and subsequent to an intervention is critical for characterizing treatment-related changes. Can standardizing airway biopsy tissue collection and histopathological assessment methods generate a robust and repeatable measure to assess airway mucosa tissue characteristics in the setting of COPD/CB? Initial tissue collection and histological assessment methods were designed by integrating various aspects from previously published evaluations, applied to an initial tissue sample cohort, and then iteratively refined by independent pathologists. A standardized metric for histologic airway mucosa assessments was developed that specified tissue collection methods, including re-sampling airways at multiple time points to enable evaluation of treatment-related effects by incorporating scores for GCH, eosinophilia, and chronic inflammation, and the degree of GCH heterogeneity present within each sample. This multi-center study generated a robust, reproducible approach for assessing airway mucosa aberrancies in the setting of COPD/CB. The iterative approach established consistent tissue specimen recovery and a granular scoring matrix that enabled quantitative scoring of the various tissue findings with substantial histopathologic interrater reliability. ClinicalTrials.gov; NCT03107494, NCT04677465; URL: www. gov. Australian New Zealand Clinical Trials Registry (ANZCTR); ACTRN12617000330347; URL: anzctr.org.au.

Attributable Costs of Clostridioides difficile Infections in Korea.

Clostridioides difficile infection (CDI) is one of the most common hospital-acquired infections, with its incidence and disease burden increasing markedly worldwide over the past decade. To assess the attributable costs of CDI in Korea, the expenses related to hospital management of CDI cases were computed. This analysis used data from the National Health Insurance Service-National Sample Cohort spanning a decade (2010-2019). The annual national burden of CDI was determined by combining the attributable cost per CDI case with the number of patients with CDI obtained from the Health Insurance Review and Assessment Service data. The attributable costs of CDI were determined based on variations in the length of hospital stay and medical costs between patients with CDI and control patients. The mean length of hospital stay was significantly longer for patients with CDI than that for control patients: 43.06 vs. 14.76 days (a difference of 28.30 days, P < 0.001). The adjusted medical costs (2019 = 100) for cases of CDI and controls were 11,162 USD and 3,318 USD, respectively, with a significant difference of 7,843 USD (P < 0.001). The cost of CDI per case exhibited a noticeable annual increase from 2010 to 2019, despite an annual decreasing trend in length of hospital stay. The estimated national cost attributed to CDI was $28.9 million in 2010; however, it increased gradually each year, reaching $205.6 million in 2019 (a 600% increase over 10 years). CDI is associated with substantial healthcare costs in Korea. The economic burden of CDI has gradually increased in South Korea.