Serum Of Patients Research Articles

Analysis of a mouse germ cell tumor model establishes pluripotency-associated miRNAs as conserved serum biomarkers for germ cell cancer detection

Malignant testicular germ cells tumors (TGCTs) are the most common solid cancers in young men. Current TGCT diagnostics include conventional serum protein markers, but these lack the sensitivity and specificity to serve as accurate markers across all TGCT subtypes. MicroRNAs (miRNAs) are small non-coding regulatory RNAs and informative biomarkers for several diseases. In humans, miRNAs of the miR-371-373 cluster are detectable in the serum of patients with malignant TGCTs and outperform existing serum protein markers for both initial diagnosis and subsequent disease monitoring. We previously developed a genetically engineered mouse model featuring malignant mixed TGCTs consisting of pluripotent embryonal carcinoma (EC) and differentiated teratoma that, like the corresponding human malignancies, originate in utero and are highly chemosensitive. Here, we report that miRNAs in the mouse miR-290-295 cluster, homologs of the human miR-371-373 cluster, were detectable in serum from mice with malignant TGCTs but not from tumor-free control mice or mice with benign teratomas. miR-291-293 were expressed and secreted specifically by pluripotent EC cells, and expression was lost following differentiation induced by the drug thioridazine. Notably, miR-291-293 levels were significantly higher in the serum of pregnant dams carrying tumor-bearing fetuses compared to that of control dams. These findings reveal that expression of the miR-290-295 and miR-371-373 clusters in mice and humans, respectively, is a conserved feature of malignant TGCTs, further validating the mouse model as representative of the human disease. These data also highlight the potential of serum miR-371-373 assays to improve patient outcomes through early TGCT detection, possibly even prenatally.

Microbial players in autoimmunity: multicentric analysis of the association between Mycoplasma hominis serostatus and rheumatoid arthritis.

Resident mucosal pathogens may induce immune tolerance breach, specific autoimmune response, and the development of rheumatoid arthritis (RA) in susceptible individuals. Despite a number of studies linking infections by Mollicutes bacteria to autoimmune disorders' onset and progression, the role of Mycoplasma hominis, a common urogenital mucosa colonizing bacterium, in inducing a specific humoral response in RA has been seldom addressed. This study reports M. hominis seroprevalence in RA patients compared to healthy controls (HC) by testing two separate cohorts sampled in two distinct geographical settings (Italy and Vietnam). The amount of circulating anti-lipid-associated membrane proteins (LAMPs) antibodies was significantly higher in RA patients than HC in both cohorts. Also, a significantly higher seroprevalence of anti-M. hominis antibodies in RA patients compared to HC in both cohorts was observed. Notably, neither ELISA OD values nor positivity of anti-LAMPs were significantly associated with RA-specific variables. Further studies are essential to elucidate the underlying mechanisms by which Mycoplasma species may contribute to the pathogenesis of RA, thereby advancing our understanding of the potential causal links between Mollicutes and autoimmune disorders.IMPORTANCEMycoplasmas may cause persistent asymptomatic mucosal infections and elicit chronic host immune responses. In this study, we evaluated the prevalence of serological response to the sexually transmitted bacterium Mycoplasma hominis in patients with rheumatoid arthritis. We show that sera of patients with rheumatoid arthritis are enriched with antibodies specifically recognizing microbial surface antigens compared with the general population. This suggests that M. hominis genital infection, with its peculiar host immunity subversion mechanisms, might play a role in predisposing to the development and progression of chronic arthritis in susceptible individuals. Thus, the range of microbes with a role as triggers of autoimmune disease (P. gingivalis, A. actinomycetemcomitans, Streptococcus spp., and F. nucleatum, among others) might have a new member in M. hominis. The potential role of the interactions taking place at the host-pathogen interface during persistent M. hominis infections in inducing autoimmunity should be further explored and characterized.

Serum N-Glycomics with Nano-LC-QToF LC-MS/MS Reveals N-Glycan Biomarkers for Glioblastoma, Meningioma, and High-Grade Meningioma.

Alteration of glycosylation in cancer cells leads to the expression of tumor-associated glycans, which can be used as biomarkers for diagnosis and prognostic prediction of diseases. In this study, we used nano-LC-QToF to identify serum N-glycan biomarkers for the detection of brain tumors. We observed an increase in sialylated N-glycans and a decrease in fucosylated N-glycans in the serum of patients with glioblastoma (GBM) and meningioma (MG) compared to healthy individuals. In GBM, a combination of increased serum sialylated N-glycan (6_4_0_2 compound) and decreased fucosylated N-glycan (4_4_1_0 compound) was identified as the most appropriate panel, with an area under the curve (AUC) of 0.8660, 78.95% sensitivity, 84.21% specificity, and 82.89% accuracy. For MG, a combination of decreased 6_6_2_0 and 5_5_2_0 compounds and increased 4_4_1_1 compound achieved an AUC of 0.9260, 82.35% sensitivity, 78.57% specificity, and 80.26% accuracy for diagnosis of MG. Additionally, an increase in 5_5_1_0 and 4_3_0_0 compounds combined with a decrease in 7_7_4_3 was associated with high-grade MG (WHO grades II-III). In conclusion, we identified serum N-glycan profiles associated with brain tumors, highlighting their potential as biomarkers for the diagnosis and prognosis of these diseases.

Antinuclear Antibodies Staining Patterns and Titers in Juvenile Idiopathic Arthritis.

The importance of antinuclear antibodies (ANA) patterns and titers in juvenile idiopathic arthritis (JIA) patients is not clearly determined in the literature. In our study, we evaluated the ANA staining patterns and titers in JIA patients. JIA patients were retrospectively assessed. ANA was identified by using indirect immunofluorescence assay on HEp-2 cells, with a positivity threshold set at a titer of 1/100 or higher. Eight hundred-five patients with JIA were evaluated (oligoarticular JIA [OJIA, n=396], enthesitis-related arthritis [ERA, n=195], polyarticular JIA [PJIA, n=132], systemic JIA [SJIA, n=53], psoriatic arthritis [PsA, n=20], unclassified JIA [n=9]). Their median (min-max) age at diagnosis was 7.9 (4.6-12.1) years (F/M=1.2). The most frequently observed ANA nuclear staining patterns were AC-4/5 (fine or large speckled) in 29.7% of patients and AC-1 (homogeneous) in 25.9%. The most common ANA cytoplasmic staining pattern was AC-19 (dense fine speckled) (17.6%). Most SJIA and unclassified JIA patients were ANA negative. The most frequently detected ANA titer in OJIA and ERA patients was 1/160 (p=0.026 and p=0.018, respectively), while in PsA patients, it was 1/320 (p<0.001). In addition, uveitis and inflammatory bowel disease, which were comorbidities associated with JIA, were most frequently associated with the nuclear AC-4/5 pattern and the cytoplasmic AC-19 pattern, as well as with an ANA titer of 1/160 (all p<0.001). Our study showed that many JIA subtypes and JIA-related comorbidities were associated with the AC-4/5, AC-1, and AC-19 patterns of ANA. However, multicenter studies in larger cohorts are needed to generalize these results.

Exosomal mir-126-3p derived from endothelial cells induces ion channel dysfunction by targeting RGS3 signaling in cardiomyocytes: a novel mechanism in Takotsubo cardiomyopathy

BackgroundTakotsubo cardiomyopathy (TTC) is marked by an acute, transient, and reversible left ventricular systolic dysfunction triggered by stress, with endothelial dysfunction being one of its pathophysiological mechanisms. However, the precise molecular mechanism underlying the interaction between endothelial cells and cardiomyocytes during TTC remains unclear. This study reveals that exosomal miRNAs derived from endothelial cells exposed to catecholamine contribute to ion channel dysfunction in the setting of TTC.MethodsHuman-induced pluripotent stem cell-derived cardiomyocytes (hiPSC-CMs) were treated with epinephrine (Epi) or exosomes (Exo) from Epi-treated human cardiac microvascular endothelial cells (HCMECs) or Exo derived from HCMECs transfected with miR-126-3p. The immunofluorescence staining, flow cytometry, qPCR, single-cell contraction, intracellular calcium transients, patch-clamp, dual luciferase reporter assay and western blot were performed for the study.ResultsModeling TTC with high doses of epinephrine (Epi) treatment in hiPSC-CMs shows suppression of depolarization velocity (Vmax), prolongation of action potential duration (APD), and induction of arrhythmic events. Exo derived from HCMECs treated with Epi (Epi-exo) mimicked or enhanced the effects of Epi. Epi exposure led to elevated levels of miR-126-3p in both HCMECs and their exosomes. Exo enriched with miR-126-3p demonstrated similar effects as Epi-exo, establishing the crucial role of miR-126-3p in the mechanism of Epi-exo. Dual luciferase reporter assay coupled with gene mutation techniques identified that miR-126-3p was found to target the regulator of G-protein signaling 3 (RGS3) gene. Western blot and qPCR analyses confirmed that miR-126-3p-mimic reduced RGS3 expression in both HCMECs and hiPSC-CMs, indicating miR-126-3p inhibits RGS3 signaling. Additionally, miR-126-3p levels were significantly higher in the serum of TTC patients compared to healthy controls and patients who had recovered from TTC.ConclusionsOur study is the first to reveal that exosomal miR-126-3p, originating from endothelial cells, contributes to ion channel dysfunction by regulating RGS3 signaling in cardiomyocytes. These findings provide new perspectives on the pathogenesis of TTC and suggest potential therapeutic targets for treatment.Graphical

BRD9 promotes the malignant phenotype of thyroid cancer by activating the MAPK/ERK pathway

Thyroid cancer is one of the most common endocrine gland malignancies in China. During gene transcription, the bromodomain and extraterminal domain (BET) proteins perform epigenome interpretation tasks. Bromodomain-containing protein 9 (BRD9) is one of the BET family members. Increasing evidence has implicated that BRD9 plays significant roles in multiple malignancies. However, its role in thyroid cancer is still not fully understood. In this research, our results demonstrated that high expression of BRD9 can facilitate the malignant phenotype of thyroid cancer cell lines, while low expression of BRD9 can impede the malignant phenotype of thyroid cancer cell lines. Pharmacologically, I-BRD9 treatment inhibits the proliferation and promotes the rate of apoptosis in thyroid cancer cell lines. Moreover, our results also revealed that BRD9 promoted xenograft tumor growth. In addition, our study showed that the expression of mitogen-activated protein kinase (MAPK)/extracellular signal-regulated protein kinase (ERK) pathway-related proteins was decreased in BRD9 knockdown thyroid cancer cells, such as Raf, ERK, p-ERK, c-Fos, and c-Myc, which could be significantly reversed by overexpressing the BRD9 in different thyroid cancer cells. After the specific inhibitor of ERK (SCH772984) was applied to thyroid cancer cells (BCPAP cells) overexpressing the BRD9 gene, the results suggested that SCH772984 reverses the high expression of MAPK/ERK pathway-associated protein in BCPAP cells (over-expression BRD9 cells). In conclusion, this study demonstrated that BRD9 was highly expressed in serum and malignant tumor tissues of thyroid cancer patients and further promoted the development of the malignant phenotype of thyroid cancer by activating the MAPK/ERK signaling pathway.

Integration of transcriptomics and metabolomics data revealed role of insulin resistant SNW1 gene in the pathophysiology of gestational diabetes.

Gestational Diabetes Mellitus (GDM) is an emerging maternal health problem with increasing incidences. The lack of complete understanding of its pathophysiological mechanisms and novel regulatory biomarkers makes early diagnosis difficult. High-throughput RNA sequencing and computational bioinformatics analyses were conducted to identify novel hub genes, and their regulatory mechanisms were validated through qRT-PCR, western blot, and siRNA-mediated knockdown studies. Intermediate metabolites and circulatory levels of amino acids in the serum of GDM patients and healthy controls were measured. Transcriptomic studies identified SNW1 as the most sensitive and specific biomarker, significantly up-regulated in GDM (fold change = 1.09; p < 0.001). Metabolomic studies indicated significantly elevated gluconeogenesis in GDM, evidenced by decreased levels of alanine and increased levels of pyruvate and glucose compared to controls. siRNA-mediated knockdown of SNW1 in PANC1 cells resulted in significant down-regulation of alanine aminotransferase (ALT/GPT) and insulin receptor substrate (IRS1), while glucose transporters (GLUT2/GLUT4) and insulin (INS) were significantly up-regulated at both mRNA and protein levels. This study identified SNW1 as a novel insulin-resistant gene that induces hyperglycemia by elevating gluconeogenesis and decreasing glucose uptake. SNW1 may be considered a potential therapeutic target with clinical utility for the management of GDM.

Identification of novel autoantibodies in Sjögren’s disease

IntroductionThe diagnosis of Sjögren’s disease (SjD) in patients without autoantibodies against Ro/SSA is a major challenge. We aimed to identify novel autoantibodies in SjD that may facilitate the diagnostic procedure for Ro/SSA negative SjD.MethodsIgG and IgA autoantibody reactivity of 94 potential candidate autoantigens for SjD, selected from a discovery screen of 1,629 human antigens coupled to Luminex beads and prior knowledge about potential biological relevance, were examined in serum of SjD patients (n=347) using Luminex and ELISA technology. Healthy (HC, n=118) and non-Sjögren’s sicca syndrome (NSS, n=44) individuals served as controls. To assess disease specificity, the novel autoantibodies were also measured in serum of patients with Rheumatoid Arthritis (RA, n=50), Systemic Lupus Erythematosus (SLE, n=49), and Systemic Sclerosis (SSc, n=37). Results45 novel autoantibodies were significantly (p ≤ 0.05) more prevalent in SjD than in HC and were detected in up to 19% of the SjD cohort. The most common autoantibodies were against CCL4, M5, TMPO and OAS3. Some of the novel autoantibodies were associated with extraglandular disease manifestations, such as anti-TONSL or anti-IL6 with pulmonary involvement. We have developed a three and five marker panel for the detection of Ro/SSA negative patients, consisting of anti-FNBP4, anti-SNRPC, anti-CCL4, anti-M3 and anti-KDM6B, which had a sensitivity of up to 46% with a specificity of 95% (SjD vs. HC). Both panels discriminate these patients from HC, whereas the three-marker more effectively differentiates between Ro/SSA negative patients and NSS.DiscussionNovel autoantibodies will facilitate the diagnosis of Ro/SSA negative patients with SjD, in particular our predictive panel will be useful in the diagnosis and differentiation of these patients from healthy and NSS individuals in a clinical context. In addition, the autoantibodies may also be useful for risk stratification of extraglandular manifestations.

The Assessment of Serum Cytokeratin 18 and Liver Function Tests in Chronic hepatitis B patients.

Background Chronic hepatitis is a clinical and pathological disorder, some biochemical testes like serum Alanine Amino-transferase (ALT), Aspartate Amino-transferase (AST) and total serum bilirubin (TSB) used to monitoring hepatitis diseases. Albumin levels in serum fall with liver disease progression, representing decreased synthesis. Prothrombin time evaluate the level of converted prothrombin to thrombin and this represent the synthetic function of the liver. In acute on chronic liver Disease non apoptotic pattern of cell death is predominant .It is confirmed by cleaved K18. Cytokeratin 18 was measured by ELISA Objectives: This study aimed to measure the ALT ,AST ,TSB, Albumin ,PT and K18 levels inthe serum of Iraqi patients who were having Liver disease And study the correlation between the variables mentioned above Materials and Methods: The study included approximately One hundred subjects, ages 20 to 60 year, who were split into two groups , Fifty patient with chronic hepatitisB virus,The second group comprises of 50 subjects who look to be in good health and serves as the control group. Results Serum levels of ALT, AST, TSB, PT and albumin displayed a significant increase in chronic hepatitis B virus group compare with control group . Also, the current study revealed non-significant association of liver function tests with different age groups, except total serum Bilirubin values decline significantly with increasing age, and there was a significant gender difference in AST concentration. While, there was non-significant gender difference in further liver parameters. Conclusion Among chronic hepatitis B patientin Babylon province, abnormality of liver function tests in different age and gender groups; indicat the significant linke with the development of chronic hepatitis B virus.

Exosomes Derived from Dialysis Patients Serum enhance Endothelial-Mesenchymal Transition and Calcification in Endothelial Cells.

Vascular calcification (VC) is prevalent among patients with end stage kidney disease (ESKD). Exosomes, small extracellular vesicles actively secreted by cells, contain proteins, nucleic acids, lipids and other bioactive substances and are considered major mediators of cell-cell interactions. Endothelial-Mesenchymal Transition (EndMT) has been observed in a variety of pathological conditions, such as abnormal shear stress, vascular damage, and chronic inflammation. The aim of this research was to assess the effects of serum-derived exosomes from ESKD patients with VC on the induction of EndMT in endothelial cells and their potential role in accelerating VC. Twenty hemodialysis patients with VC and 10 healthy volunteers were recruited. Cardiac and brain VC were assessed among patients with ESKD treated with dialysis. Serum samples were taken at dialysis initiation for exosome isolation. Human umbilical vein endothelial cells (HUVEC) were treated with 100 µg/mL exosomes for 24-96 hours. At the end of incubation, cells were collected for mRNA and protein analysis. Exosomes isolated from dialysis patients with VC induced EndMT in HUVECs. After 24h, endothelial markers CD31 and VE-cadherin were decreased (31% and 51%, respectively; P<0.001) and the mesenchymal proteins Vimentin and N-cadherin were increased (283% and 156%, respectively; P<0.001), compared to healthy exosomes. After 96h of incubation, expression of genes essential for osteoblast differentiation, including the bone morphogenetic genes (BMP2, BMPR2, BMP4 and BMP9), and the transcription factor RUNX2 were significantly elevated. Exosomes derived from the serum of dialysis patients with VC, induced EndMT and contributed to calcification. The vicious cycle highlighted the intricate interplay between exosomes, EC and VC, emphasizing the critical necessity for therapeutic strategies to disrupt this pathway and mitigate calcification advancement.

Atezolizumab neutralizing assay development: a novel way of evaluating assay cutpoint after sample pretreatment

Atezolizumab, a biotherapeutic monoclonal antibody directed against PD-L1, has been shown to be efficacious in multiple oncology indications. In clinical trials, a subset of patients developed anti-atezolizumab antibodies, necessitating the development of a neutralizing antibody (NAb) assay. A bead-based sample pretreatment method was developed to reduce high levels of therapeutic in patient serum samples. Untreated sample variability is reduced by this sample pretreatment, based on this, a novel approach was taken for determining an appropriate assay decision threshold (cutpoint). The therapeutic was added to the samples to mimic typical patient samples. The resulting assay was successfully validated and applied to sample analysis from multiple clinical trials.

DNA-Assembled Size-Permeable Microcapsule Bioreactor Enables Direct and Rapid Sensing of Trace Circulating MicroRNA Biomarkers in Undiluted Cancer Patient Serum.

Sensitive and direct detection of circulating microRNAs (miRNAs) in undiluted human serums can significantly facilitate the diagnosis of different diseases in the early stages; however, the achievement of such a goal is highly challenged by the extremely complex sample matrix nature of serums. Here, we show that sensitive and rapid detection of the circulating let-7a miRNA biomarker in undiluted cancer patient serums can be realized by a new size-permeable microcapsule bioreactor. The Cas13a/crRNA-dsDNA@polyethylene glycol core/shell biomicrocapsules are synthesized by simple encapsulation and etching steps. Such biomicrocapsules exhibit high stability, enhanced protein antifouling capability, and size-selective permeability that only allows short miRNA sequences to enter the core cavities, thereby enabling highly sensitive and direct detection of let-7a miRNA in undiluted serums down to 0.21 pM within 20 min via signal amplification by Cas13a/crRNA and near-infrared (NIR) fluorescence signals. The biomicrocapsule also allows unambiguous discrimination of the clear cell renal cell carcinoma patients from the healthy groups via direct detection of let-7a miRNA in the serum samples, thereby highlighting its robust clinical potential for early disease diagnoses.

Progranulin Protects Against Osteoporosis by Regulating Osteoclast and Osteoblast Balance via TNFR Pathway.

Osteoporosis is a disease of bone metabolism caused by an imbalance between osteoclast-mediated bone destruction and osteoblast-mediated bone formation. Tumour necrosis factor α (TNFα) has been reported to promote osteoclast generation and inhibit osteoblast generation. Progranulin (PGRN), which has a strong anti-inflammatory effect, interacts with tumour necrosis factor receptor (TNFR). Serum and bone tissues from patients with or without osteoporosis were collected to analyse the relationship between PGRN content and bone metabolic markers. The role of TNFα and PGRN in osteoclast differentiation was explored by using RAW 264.7 cells and BMMs. MC3T3-E1 cells and BMSCs were used to observe the role of TNFα and PGRN in osteoblast differentiation. The PGRN content in the serum and bone tissues of osteoporosis patients was lower than that in the serum and bone tissues of nonosteoporosis patients. TNFα promoted osteoclast differentiation, while PGRN inhibited this effect by interacting with TNFR1. PGRN inhibited TNFα-mediated attenuation of osteoblast differentiation by interacting with TNFR1. Moreover, PGRN alone promoted osteoblast differentiation by interacting with TNFR2. Our findings reveal that PGRN can effectively inhibit TNFα-induced osteoporosis and has a certain osteogenic effect. This discovery might provide a potential target for osteoporosis treatment.

Mediation of the liver protective effect of the anti-cancer drug candidate namodenoson via adiponectin.

604 Background: Namodenoson (CF102) is a small, orally available, anti-cancer drug candidate, currently in pivotal phase 3 clinical study for the treatment of hepatocellular carcinoma. Adiponectin is a positive cytokine released by adipocytes and endothelial cells, known to induce liver-, cardio-, and neuro-protective effects. We evaluated the potential role of adiponectin as the mediator of the observed namodenoson-induced protective effects. Methods: Proteins extracted fromN1S1 tumor-bearing mice and from murine adipocyte culture treated with namodenoson or vehicle were analyzed using Western blot. Apoptosis was determined in the N1S1 tumor-bearing mice using the TUNEL assay. The effect of namodenoson on hepatic ischemia/reperfusion (IR) was evaluated in Wistar rats by temporarily clamping the hepatic vasculature; the effect on hepatic inflammation was evaluated in Con. A-induced hepatitis in mice. The STAM murine in-vivo model was used to study the effect of namodenoson on metabolic dysfunction-associated steatohepatitis (MASH). Adiponectin levels were determined in the serum of patients in the phase 2 trial evaluating namodenoson vs placebo in patients with metabolic dysfunction-associated steatotic liver disease (MASLD)/MASH. Results: The anti-cancer effect of namodenoson was attributed to its capability to bind to the A3 adenosine receptor, inducing inhibition of PI3K and subsequent de-regulation of the Wnt/β-catenin and NF-kB signal transduction pathways, resulting in apoptosis of liver tumor cells. At the same time, namodenoson demonstrated a liver protective effect manifested by prevention of hepatic IR injury, suppression of liver inflammation, and induction of anti-fibrotic and anti-steatotic effects. In adipocyte cell culture, namodenoson stimulated adiponectin production. An increase in adiponectin levels was also shown in the murine MASH model (STAM). These in-vitro and in-vivo results were consistent with translational data from the phase 2 study of namodenoson in MASLD/MASH where upregulation of adiponectin was observed with namodenoson (mean change from baseline of 539 ng/mL for namodenoson 12.5 mg BID vs -78 ng/mL for placebo, P = 0.032). Conclusions: The anti-cancer effect of namodenoson combined with its protective effects, position it as a unique drug inducing a dual effect in patients with advanced liver cancer.

Proteomic changes in serum of patients with Erythema migrans, Anaplasma phagocytophilum infection and co-infection

Proteomic changes in serum of patients with Erythema migrans, Anaplasma phagocytophilum infection and co-infection

Extracellular vesicles released by ALL patients contain HNE-adducted proteins: implications of collateral damage

Off-target neuronal injury is a serious side-effect observed in cancer survivors. It has previously been shown that pediatric acute lymphoblastic leukemia (ALL) survivors have a decline in neurocognition compared to healthy age-matched counterparts. Elevated oxidative stress has been documented to be a mediator in off-target tissue damage in cancer survivors. Early detection of oxidative stress markers may provide an opportunity to prevent off-target tissue damage. Extracellular vesicles (EVs) have surfaced as a potential diagnostic tool due to molecular cargo they contain. We investigated the potential for EVs to be a sensitive indicator of oxidative stress and off-target tissue damage by isolating EVs from pediatric ALL patients throughout their first 2 months of treatment. EVs were measured throughout the collection points for: 1) number of EV particles generated using nanoparticle tracking analysis (NTA); 2) markers of neurons (NeuN), astrocyte activation (GFAP), neuronal stability (BDNF), 3) markers of pre-B cell ALL (CD19 and CD22); and) 4-hydroxy-2-nonenal (HNE) adducted proteins. HNE protein adductions were measured in the patient sera and CSF. Pro-inflammatory cytokine levels were also measured in patient sera because of their contribution to oxidative stress and neuronal injury. Our results: 1) demonstrate EVs are a sensitive indicator of oxidative damage; 2) suggest EVs as a marker of a decline in neuronal stability; and 3) show the presence of leukemia has a greater contribution to pro-inflammatory cytokine production in the patient's serum than the cancer treatment. Specifically, we observed a significant decrease in cytokine levels (e.g., TNF-α, IL-1β, IL-6, and IL-8) following the initiation of treatment, highlighting the influence of leukemia burden on systemic inflammation. The results support the utilization of EVs as a sensitive marker of oxidative stress and off-target tissue damage.

Ultrasensitive and label-free electrochemical immunosensor using gold nanoparticles deposited on a carbon electrode for the quantification of osteopontin: A serum-based oncomarker.

Early detection of cancer biomarkers is crucial for effective diagnosis and treatment, prompting the development of an ultrasensitive label-free electrochemical immunosensor. In this study, we fabricated an ultrasensitive label-free electrochemical immunosensor using a glassy carbon electrode/gold nanoparticles (GCE/AuNPs) modification for quantification of osteopontin (OPN), an oncomarker. The surface features of the modified electrodes were confirmed using scanning electron microscopy (SEM) and X-ray photoelectron spectroscopy (XPS) methods. The electrochemical behavior of the bare and modified electrode was characterized using cyclic voltammetry (CV) and electrochemical impedance spectroscopy (EIS). The quantification of the OPN antigen was achieved through the differential pulse voltammetry (DPV) method. The fabricated immunosensor demonstrated excellent detection capabilities in both commercial serum samples and phosphate-buffered saline (PBS). It showed sensitive quantification of OPN in the range of 0.001 to 1000ng/mL with a limit of detection (LOD) of 0.005ng/mL in PBS. Furthermore, the immunosensor retained approximately 89.3% of its initial signal after storage for up to 8weeks. The results were validated by detecting OPN-spiked commercial serum samples with a satisfactory recovery rate. The potential of this immunosensor makes it suitable for assaying OPN in real cancer patient's serum samples with minimal interference from complex sample matrices.

Surveillance Donor-Specific Antibody and Pathologic Antibody Mediated Rejection Testing in Heart Transplant Patients in the Contemporary Era

Surveillance Donor-Specific Antibody and Pathologic Antibody Mediated Rejection Testing in Heart Transplant Patients in the Contemporary Era

Underappreciated Effects of Delayed Graft Function in Pediatric Kidney Transplantation.

In kidney transplantation (KT), delayed graft function (DGF) is associated with worse outcomes. However, it is unclear what effect DGF plays in long-term survival compared to the impact of the various transplant, donor, and recipient risk factors associated with DGF. This study aims to determine the effect of DGF alone on long-term survival in pediatric deceased donor kidney transplant recipients (DDKTRs). Using a national database, 1728 pediatric patients who received a kidney transplant between 1994 and 2009 (n = 978) and 2010 and 2022 (n = 750) were identified. Cases and controls were matched by DGF status for transplant date, cold ischemia time, distance from the transplant center, HLA mismatch level, PRA, KDPI, recipient and donor age, BMI, sex assigned at birth, and ethnicity. Kaplan-Meier curves were analyzed with a log-rank test. Pearson's chi-squared tests were used to compare survival proportions between groups at time benchmarks. Pediatric DDKTRs with DGF had worse graft (p < 0.001) and patient (p < 0.001) survival compared to non-DGF patients in both eras. Between 1994 and 2009, graft survival rates were lower in the DGF cohort at 1 (p < 0.001), 3 (p < 0.001), 5 (p < 0.001), and 10 years (p = 0.010). Patient survival rates were lower in the DGF cohort at 1 (p = 0.017) and 5 years (p = 0.019) post-DDKT. There was no difference in 3 (p = 0.071) and 10-year patient survival rates (p = 0.141). Between 2010 and 2022, graft and patient survival rates were lower in the DGF cohort at 1, 3, 5, and 10 years (all p < 0.001). There was no difference in rates of retransplantation (p = 0.307 and p = 0.771) between groups for either era. DGF alone is associated with worse outcomes in pediatric DDKTRs. DGF should be taken seriously, especially in cases where pediatric DDKTRs do not possess known, attributable risk factors for DGF.

MiR-92a aggravates metabolic syndrome via KLF2/miR-483 axis.

To exam the role of miR-92a/KLF2/miR-483 in the pathogenesis of metabolic syndrome. In this study, the serum of healthy controls and patients with metabolic syndrome were collected to detect the circulating level of miR-92a and miR-483. In vitro cultured HUVECs, overexpression or suppression of miR-92a, miR-483 or KLF2 to determine the relationship among miR-92a, KLF2 and miR-483. Ang II, ox-LDL, or high glucose treatment were used to mimic the metabolic syndrome. HUVECs or HepG2 cells were treated with Telmisartan, Atorvastatin, or metformin, the miR-483 and its target gene expression was detected. In animal experiment, ob/ob mice were chose to confirm the changes of miR-92a, KLF2, and miR-483. Compared with the healthy controls, the level of miR-92a was significantly increased, while miR-483 level was remarkably decreased in the patients with metabolic syndrome. In vitro cultured HUVECS, overexpression of miR-92a significantly reduced the expression of miR-483, but overexpression of miR-483 had no effect on miR-92a. Overexpression of KLF2 could downregulate miR-483 level, while inhibition of KLF2 had the opposite effect. When HUVECs and HepG2 were stimulated with Ang II, ox-LDL and high glucose, the expression of miR-483 was significantly decreased and its target genes was increased. Anti-miR-92a could reverse the effect. Furthermore, Telmisartan, Atorvastatin, and Metformin significantly increased miR-483 expression and decreased its target gene expression, which could be reversed by miR-92a mimic. The level of miR-92a was significantly increased in HepG2 cells, which were treated with exosomes derived from endothelial cells with miR-92a overexpression. ob/ob mice showed the similar effects. Endothelial dysfunction and fatty liver are critically involved in the pathogenesis of metabolic syndrome. MicroRNAs can mediate the cellular communication between vascular endothelial cells (ECs) and distal cell. Serum miR-92a level was higher in metabolic syndrome patients than controls. KLF2 is the target gene of miR-92a, which can increase the production of miR-483, miR-483 acts on its target genes CTGF, ET-1, and β-catenin to protect cell function. EC miR-92a is secreted out of cells into the blood, circulates through the blood to the liver, and continues to exert its biological effects after being absorbed by hepatocytes. LNA-miR-92a administration reversed endothelial cell damage and fatty liver caused by metabolic syndrome by affecting the KLF2/miR-483 pathway.