Digoxin detection for therapeutic drug monitoring using target-triggered aptamer hairpin switch and nicking enzyme-assisted signal amplification.

Digoxin, a cardiac glycoside drug, is commonly used to treat heart failure and arrhythmias. The therapeutic concentration range of digoxin, with a narrow therapeutic index, is between 0.5 and 2.0 ng mL-1. Hence, it is important for patients to monitor their blood levels after taking medication to achieve effective treatment and reduce the likelihood of experiencing drug side effects. Due to the complex steps and high cost of immunoassays, aptasensors that use aptamers to recognize the targets offer the advantages of low cost and good stability over other analysis methods. Nicking enzyme-assisted signal amplification is a novel isothermal signal amplification technology that relies on nicking enzymes to recognize and cleave restriction sites on one oligonucleotide strand. In this study, we develop a fluorescent aptasensor coupled with target-triggered aptamer hairpin switch and nicking enzyme-assisted signal amplification for digoxin detection in plasma for therapeutic drug monitoring. After optimizing the experimental parameters, we design hairpin probes with ten base pairs of the aptamer sequence and extended sequence complement to react with digoxin in a 10 mM Tris buffer containing 150 mM NaCl and 50 mM MgCl2 (pH 7.4). The signal amplification reactions were performed for 3 hours. The fluorescent aptasensor exhibited high sensitivity with a detection limit of 88 pg mL-1 for detecting digoxin in plasma and a linear range from 0.1 ng mL-1 to 5 ng mL-1. This technology was successfully used for digoxin detection to improve treatment effectiveness and minimize the risk of adverse side effects.

Enhancing Ischemic Stroke Evaluation by a Model-Based Photoacoustic Tomography Algorithm.

Ischemic stroke (IS) is characterized by the sudden interruption of blood supply to the brain, resulting in neurological impairments and even mortality. Photoacoustic computed tomography (PACT) integrates the high contrast of optical imaging and the penetration of ultrasound imaging, enabling non-invasive IS evaluation. However, the image reconstruction quality significantly affects the oxyhemoglobin saturation (sO2) estimation. This study investigates a model-based with total variation minimized by augmented Lagrangian and alternating direction (MB-TVAL3) approach and compared it with the widely used back-projection (BP) and delay-and-sum (DAS) algorithms. Both simulations and in vivo experiments are conducted to validate the performance of the MB-TVAL3 algorithm, showing a higher sO2 estimation accuracy and sensitivity in detecting infarct area compared to BP and DAS. The findings of this study emphasize the impact of acoustic inverse problem on the accuracy of sO2 estimation and the proposed approach offers valuable support for IS evaluation and cerebrovascular diagnosis.

Fabrication of natural enzyme-covered / amino-modified Pd-Pt bimetallic-doped zeolitic imidazolate framework for ultrasensitive detection of metabolites.

The present article introduced an natural enzyme-covered/amino-modified Pd-Pt bimetallic-doped zeolitic imidazolate framework (NAPPZ) for ultrasensitive and specific detection of glucose. The dodecahedral nanomaterial zeolitic imidazolate framework (ZIF-8)-loaded Pd-Pt bimetallic nanoparticles endowed the composite with peroxidase-like activity. The modification with glucose oxidase (GOx) facilitated the rapid access of H2O2 produced through glucose oxidation to the Pd-Pt nanoparticles vicinity reducing diffusion. GOx specifically catalyzes the transformation of glucose into H2O2, which then H2O2 rapidly migrates to the Pd-Pt nanoparticles, catalyzing the oxidation of colorless o-phenylenediamine into the orange-yellow product 2,3-diaminophenazine. Based on the aforementioned cascade reaction, the NAPPZ and NAPPZ based on ChOx were utilized for detecting glucose in human urine samples and cholesterol in milk, respectively. The NAPPZ strategy presented a broad detection range (20-1100 µmol L-1) and a low detection limit (15.9 µmol L-1) for glucose, and the NAPPZ based on ChOx strategy approach offered a broad detection range (10-500 µmol L-1) and low detection limit (6.4 µmol L-1) for cholesterol. Therefore, this novel method holds significant potential in the areas of clinical diagnostics and food safety.

Automated Quantification of HER2 Amplification Levels Using Deep Learning.

HER2 assessment is necessary for patient selection in anti-HER2 targeted treatment. However, manual assessment of HER2 amplification is time-costly, labor-intensive, highly subjective and error-prone. Challenges in HER2 analysis in fluorescence in situ hybridization (FISH) and dual in situ hybridization (DISH) images include unclear and blurry cell boundaries, large variations in cell shapes and signals, overlapping and clustered cells and sparse label issues with manual annotations only on cells with high confidences, producing subjective assessment scores according to the individual choices on cell selection. To address the above-mentioned issues, we have developed a soft-sampling cascade deep learning model and a signal detection model in quantifying CEN17 and HER2 of cells to assist assessment of HER2 amplification status for patient selection of HER2 targeting therapy to breast cancer. In evaluation with two different kinds of clinical datasets, including a FISH data set and a DISH data set, the proposed method achieves high accuracy, recall and F1-score for both datasets in instance segmentation of HER2 related cells that must contain both CEN17 and HER2 signals. Moreover, the proposed method is demonstrated to significantly outperform seven state of the art recently published deep learning methods, including contour proposal network (CPN), soft label-based FCN (SL-FCN), modified fully convolutional network (M-FCN), bilayer convolutional network (BCNet), SOLOv2, Cascade R-CNN and DeepLabv3+ with three different backbones (p ≤ 0.01). Clinically, anti-HER2 therapy can also be applied to gastric cancer patients. We applied the developed model to assist in HER2 DISH amplification assessment for gastric cancer patients, and it also showed promising predictive results (accuracy 97.67 ±1.46%, precision 96.15 ±5.82%, respectively).

Biochar affects organic carbon composition and stability in highly acidic tea plantation soil.

Biochar amendments are effective in stabilizing soil aggregates and improving soil organic carbon (SOC) content. However, the effects of biochar on highly acidic soil and their relation with soil SOC stability remain understudied. The study aimed to investigate the impact of biochar on changes of aggregate distribution and SOC stability in a highly acidic tea plantation soils over an eight-year period. Soil samples were collected from plots with varying biochar application amounts (0, 2.5 t ha-1, 5 t ha-1, 10 t ha-1, 20 t ha-1 and 40 t ha-1). The content of SOC, iron bound organic carbon (OC-Fe), particulate organic carbon (POC), mineral-associated organic carbon (MAOC) and the functional group composition of SOC was analyzed. The results indicated that in the biochar application treatments, the value of soil pH, SOC, POC and MAOC contents were increased from 3.92 to 4.28, 6.68%-187.02%, 8.31%-66.78% and 13.07%-236.47% respectively, compared with CK, while the content of macro-aggregate (particle size>0.25 mm) and soil aggregates mean weight diameter (MWD) significantly increased with higher biochar application amounts. But dissolved organic carbon (DOC) and OC-Fe content exhibited downward trend, decreased from 2.43% to 6.97% and 4.18%-19.91%. Furthermore, aromatic-C levels increased, with increased biochar application amounts. The integration of biochar not only bolstered soil aggregate stability but also amplified the presence of aromatic-C, thereby enhancing the resilience of organic carbon in highly acidic tea garden soil (BC40 > BC20 > BC5>BC2.5 > BC10 > CK), with increases ranging from 6% to 47%. The principal component analysis and structural equation modeling identified soil pH, TN, SOC, POC, MAOC, R > 0.25 and MWD as key factors of soil organic carbon stability. These findings provide crucial insights into the mechanism underlying biochar's efficiency in fortifying organic carbon stability, particularly in the context of highly acidic soil.

Physicians' knowledge, awareness and instructions of home blood pressure monitoring: Asia HBPM survey in Taiwan.

Home blood pressure monitoring (HBPM) has been proven to be reliable for both diagnosis and treatment of hypertension. Experts from 11 Asian countries collaborated to design a questionnaire aimed at healthcare workers, seeking to understand the current status of HBPM. A cross-sectional survey was conducted among healthcare professionals from Taiwan between November 2019 and June 2021. A total of 779 physicians in Taiwan participated in the survey. Nearly all physicians (97.7%) reported recommending HBPM to their patients. There was a 14.6% difference between the HBPM device ownership rate (60.2%) and the proportion of patients who measured their home blood pressures (45.6%). Among physicians who recommended HBPM, only 15.5% cited HBPM diagnostic threshold values consistent with the guidelines. Among all respondents, 85.9% and 48.9% viewed HBPM as highly recognized by physicians and patients, respectively. Lack of guidelines for HBPM and concern to reliability and accuracy of the HBPM devices were identified as key barriers to HBPM recognition. This study indicates that there is still room for improvement among healthcare providers in their understanding and implementation of HBPM in Taiwan.

DOCK1 deficiency drives placental trophoblast cell dysfunction by influencing inflammation and oxidative stress, hallmarks of preeclampsia.

Preeclampsia (PE) is a globally prevalent obstetric disorder, pathologically characterized by abnormal placental development. Dysfunctions of angiogenesis, vasculogenesis and spiral artery remodeling are demonstrated to be involved in PE pathogenesis; however, the underlying mechanisms remain largely unknown. Here, we investigated the role of the dedicator of cytokinesis 1 (DOCK1), crucial molecule in various cellular processes, in PE progression using HTR-8 cells derived from first-trimester placental extravillous trophoblasts. Our analysis revealed an aberrant DOCK1 expression in the placental villi of PE patients and its impact on essential cellular functions for vascular network formation. A deficiency of DOCK1 in HTR-8 cells impaired the vascular network formation, exacerbated the expression of anti-angiogenic factor ENG, and reduced VEGF levels. Moreover, DOCK1 knockout amplified apoptosis, as indicated by an altered BCL2: BAX ratio and enhanced levels of cleaved PARP. DOCK1 depletion also boosted NF-κB activation and pro-inflammatory cytokine production (IL-6 and TNF-α). Furthermore, the mice treated with DOCK1 inhibitor, TBOPP, exhibited PE-like symptoms. These findings highlight the multifaceted roles of DOCK1 in the pathophysiology of PE, demonstrating that its deficiency can lead to placental dysfunction by orchestrating inflammatory responses and oxidative stress. These insights emphasize the pathogenic role of DOCK1 in PE development and suggest potential treatment strategies that require further exploration. In the graphical abstract, a split image of placental villi contrasts the effects of normal and reduced DOCK1 expression on preeclampsia. The left side illustrates adequate DOCK1 levels supporting healthy trophoblast function and effective spiral artery remodeling. The right side highlights the consequences of DOCK1 deficiency, leading to trophoblast dysfunction and impaired spiral artery remodeling, accompanied by angiogenic imbalance, increased inflammation, oxidative stress, and apoptosis, contributing to placental dysfunction and the development of preeclampsia.

Evaluation of an Innovative Postgraduate Medical Education Model Incorporating Social Determinants of Health.

PURPOSE: Incorporating social determinants of health (SDH) into medical education is crucial. However, there are limited data on standard education models and comprehensive SDH curricula in Taiwan are insufficient. This study presents a systematic SDH curriculum instructed primarily by social workers for postgraduate doctors and aims to examine the training outcomes of the innovative curriculum. METHOD: This study assessed training outcomes using Kirkpatrick model levels 1 and 2 regarding trainees' satisfaction and improvement of their knowledge and skills in written and standardized patient (SP) pre- and posttests conducted between 1 August 2021 and 31 July 2022. RESULTS: A total of 28 trainees completed the training. The trainees' overall satisfaction score regarding the curriculum was high (4.6 out of 5). The median pretest scores for the written and SP tests were 66.25 ± 14.38 and 14.50 ± 5.13, respectively, whereas the median posttest scores were 80.00 ± 7.50 and 20.50 ± 6.13, respectively. Both written and SP posttest scores were significantly improved compared to the pretest scores (p < .001). CONCLUSIONS: The presented education model significantly improved postgraduate doctors' SDH knowledge and biopsychosocial assessment skills, and received high satisfaction scores from the trainees. Adopting social workers as primary teachers may enhance interdisciplinary collaboration between social workers and trainee doctors.

Comparative efficacy and choice of lipid-lowering drugs for cardiovascular and kidney outcomes in patients with chronic kidney disease: A systematic review and network meta-analysis.

BACKGROUND: The effect of exact classes of lipid-lowering drugs (LLDs) on preventing major adverse cardiovascular events (MACEs) and poor renal outcomes is not well characterized in the chronic kidney disease (CKD) population. METHODS: We performed a frequentist random-effects network meta-analysis of randomized controlled trials (RCTs) to evaluate the protective effect of the LLDs in non-dialysis CKD patients. The PubMed, Embase, Web of Science, and Cochrane Library databases were systematically searched for relevant trials published before March 31, 2024. The primary outcome was the incidence of MACEs. The secondary outcomes comprised all-cause mortality, end-stage kidney disease, changes in estimated glomerular filtration rate (eGFR) and proteinuria, and safety. RESULTS: Forty-nine eligible RCTs with 77,826 participants with non-dialysis CKD were included. With moderate confidence in the evidence, rosuvastatin and atorvastatin showed statistically significantly more efficacy in reducing the risk of MACE, with a pooled risk ratio of 0.55 (95% CI 0.33-0.91) for rosuvastatin and 0.67 (0.49-0.90) for atorvastatin, respectively, compared with the control group. For the change in the eGFR, atorvastatin (mean difference [MD], 1.40; 95% CI, 0.61 to 2.18), rosuvastatin (MD, 1.73; 95% CI, 0.63 to 2.83), and statin plus ezetimibe (MD, 2.35; 95% CI, 0.44 to 4.26) showed statistically significant increases in the mean eGFR. CONCLUSION: In patients with non-dialysis CKD, there is sufficient evidence to show that rosuvastatin and atorvastatin were statistically significantly more effective and preferable in reducing the risk of MACE and increasing the mean eGFR compared with the control group.

Can routine EPA-based assessments predict OSCE performances of undergraduate medical students?

BACKGROUND: Objective structured clinical examination (OSCE) is used worldwide. This study aims to explore potential alternatives to the OSCE by using entrustable professional activities (EPA)-based assessments in the workplace. METHODS: This study enrolled 265 six-year undergraduate medical students (UGY) from 2021 to 2023. During their rotations, students were assessed using 13 EPAs, with the grading methods modified to facilitate application. Before graduation, they participated in two mock OSCEs and a National OSCE. We used generalized estimating equations to analyze the associations between the EPA assessments and the OSCE scores, adjusting for age and sex, and developed a prediction model. EPA8 and EPA9, which represent advanced abilities that were not significant in the regression models, were removed from the prediction model. RESULTS: Most EPAs were significantly correlated with OSCE scores across the three cohorts. The prediction model for forecasting passing in the three OSCEs demonstrated fair predictive capacity (area under curve = 0.82, 0.66, and 0.71 for students graduated in 2021, 2022, and 2023, respectively all p < 0.05). CONCLUSIONS: The workplace-based assessments (EPA) showed a high correlation with competency-based assessments in simulated settings (OSCE). EPAs may serve as alternative tools to formal OSCE for medical students.

Elucidating the immunomodulatory effects of phytoestrogens and their groundbreaking applications in transplantation medicine.

Phytoestrogens are natural compounds found in plants and plant-based foods. When ingested, they can affect the human body in the same way as estrogen produced by the body. Phytoestrogens affect the regulation, differentiation, and production of immune cells. People who consume polyphenol and flavonoid-rich foods have lower incidences of inflammation, autoimmune diseases, and cancer. In organ transplantation, immune rejection is a lifelong problem for patients. In clinical practice, acute rejection is treated with hormonal shock or immunosuppressive drugs. However, effective reversal measures for chronic rejection, specifically for prevention, are still lacking. Recipients are also prone to post-transplant complications such as new tumors, diabetes, hyperlipidemia, hyperuricemia, and cardiovascular and cerebrovascular diseases, owing to the long-term use of immunosuppressive drugs. Phytoestrogens play a promising role in immune regulation and exert curative effects on cardiovascular diseases and cancer. In this study, we reviewed the use of phytoestrogens in the fields of immune regulation and organ transplantation.

Thyroid Radiofrequency Ablation-Thermal Effects on Recurrent Laryngeal Nerve Using Continuous Intraoperative Neuromonitoring Animal Model.

OBJECTIVE: When performing radiofrequency ablation for thyroid nodules, it is essential to avoid thermal injury to the recurrent laryngeal nerve. This porcine animal model study used continuous intraoperative neuromonitoring to investigate the thermal safety parameters of thyroid radiofrequency ablation. STUDY DESIGN: Porcine animal study. SETTING: University animal laboratory. METHODS: Twelve piglets were tested at different radiofrequency power levels, and the real-time electromyography signal changes were recorded under continuous intraoperative neuromonitoring. The spread heat study (8 piglets) included spontaneous recovery tests and cold water irrigation tests to investigate the safety distance from the recurrent laryngeal nerve to the active tip during 5-second activation with standard stimulation patterns. The residual heat study (4 piglets) investigated the safety cooling durations by touching the recurrent laryngeal nerve with the tip after a 5-second activation. RESULTS: In the spread heat study, substantial signal attenuation events were observed at an spread heat distance of 2, 3, 5, and 5 mm when the power was set as 10, 20, 30, and 50 W, respectively. No signal recovery could be observed in 20 minutes with or without cold water irrigation in the injured recurrent laryngeal nerve area. The residual heat study shows the residual thermal effect of the tip is minimal, and no substantial signal attenuation event was observed at all experiments. CONCLUSIONS: This innovative study established the thermal safety parameters for radiofrequency ablation in a porcine model at various power levels, which can potentially assist operators in delineating a precise ablation field and providing effective thyroid ablation treatment safely.

Maternal total sleep deprivation causes oxidative stress and mitochondrial dysfunction in oocytes associated with fertility decline in mice.

Previous studies have shown sleep deprivation is increasingly reported as one of the causes of female infertility. However, how and by what relevant mechanisms it affects female fertility remains unclear. In this study, female mice underwent 72 hours of total sleep deprivation (TSD) caused by rotating wheel or 2 different controls: a stationary wheel, or forced movement at night. Even though, there was no significant difference in the number of eggs ovulated by the TSD mice compared to the control groups. Overall levels of estrogen and FSH were lower throughout the estrus cycle. A total of 42 genes showed significant differential expression in GV oocytes after TSD by RNA sequencing (RNA-Seq). These included genes were enriched in gene ontology terms of mitochondrial protein complex, oxidoreductase activity, cell division, cell cycle G1/S phase transition, as well as others. The increased concentrations of reactive oxygen species (ROS) in germinal vesicle (GV) and metaphase II (MII) oocytes from TSD mice were observed, which might be induced by impaired mitochondrial function caused by TSD. The GV oocytes displayed increased mitochondrial DNA (mtDNA) copy number and a significant transient increase in inner mitochondrial membrane potential (Δψm) from the TSD mice probably due to compensatory effect. In contrast, MII oocytes in the TSD group showed a decrease in the mtDNA copy number and a lower Δψm compared with the controls. Furthermore, abnormal distribution of mitochondria in the GV and MII oocytes was also observed in TSD mice, suggesting mitochondrial dysfunction. In addition, abnormal spindle and abnormal arrangement of chromosomes in MII oocytes were markedly increased in the TSD mice compared with the control mice. In conclusion, our results suggest that TSD significantly alters the oocyte transcriptome, contributing to oxidative stress and disrupted mitochondrial function, which then resulted in oocyte defects and impaired early embryo development in female mice.

Immune effector cell-associated enterocolitis following chimeric antigen receptor T-cell therapy in multiple myeloma.

We report 14 cases of immune effector cell (IEC)-associated enterocolitis following chimeric antigen receptor T-cell (CAR-T) therapy in multiple myeloma, with a 1.2% incidence overall (0.2% for idecabtagene vicleucel and 2.2% for ciltacabtagene autoleucel). Patients developed acute-onset symptoms (typically non-bloody Grade 3+ diarrhea) with negative infectious workup beginning a median of 92.5 days (range: 22-210 days) after CAR-T therapy and a median of 85 days after cytokine release syndrome resolution. Gut biopsies uniformly demonstrated inflammation, including intra-epithelial lymphocytosis and villous blunting. In one case where CAR-specific immunofluorescence stains were available, CAR T-cell presence was confirmed within the lamina propria. Systemic corticosteroids were initiated in 10 patients (71%) a median of 25.5 days following symptom onset, with symptom improvement in 40%. Subsequent infliximab or vedolizumab led to improvement in 50% and 33% of corticosteroid-refractory patients, respectively. Five patients (36%) have died from bowel perforation or treatment-emergent sepsis. In conclusion, IEC-associated enterocolitis is a distinct but rare complication of CAR-T therapy typically beginning 1-3 months after infusion. Thorough diagnostic workup is essential, including evaluation for potential T-cell malignancies. The early use of infliximab or vedolizumab may potentially hasten symptom resolution and lower reliance on high-dose corticosteroids during the post-CAR-T period.

Zinc finger nuclease-mediated gene editing in hematopoietic stem cells results in reactivation of fetal hemoglobin in sickle cell disease.

BIVV003 is a gene-edited autologous cell therapy in clinical development for the potential treatment of sickle cell disease (SCD). Hematopoietic stem cells (HSC) are genetically modified with mRNA encoding zinc finger nucleases (ZFN) that target and disrupt a specific regulatory GATAA motif in the BCL11A erythroid enhancer to reactivate fetal hemoglobin (HbF). We characterized ZFN-edited HSC from healthy donors and donors with SCD. Results of preclinical studies show that ZFN-mediated editing is highly efficient, with enriched biallelic editing and high frequency of on-target indels, producing HSC capable of long-term multilineage engraftment in vivo, and express HbF in erythroid progeny. Interim results from the Phase 1/2 PRECIZN-1 study demonstrated that BIVV003 was well-tolerated in seven participants with SCD, of whom five of the six with more than 3 months of follow-up displayed increased total hemoglobin and HbF, and no severe vaso-occlusive crises. Our data suggest BIVV003 represents a compelling and novel cell therapy for the potential treatment of SCD.

Home-Based Digital Exercise Training Program to Improve Physical Function of Older Sepsis Survivors: Protocol of the HEAL Sepsis Randomized Clinical Trial.

BACKGROUND: While sepsis, an exaggerated response to infection, can affect people of all age groups, it is more prevalent in middle-aged and older adults. Older adults suffer worse short-term and long-term outcomes than younger patients. Older sepsis survivors are commonly discharged to long-term acute care facilities, where they often die within 1 year. Those who return home from the hospital lose the momentum of physical function improvement after early inpatient rehabilitation, and often face exacerbation of comorbidities and decline in physical function. Additionally, patients who are discharged home often live at distant locations and are not able to commute to rehabilitation centers due to their poor health status. Therefore, remotely delivered exercise interventions tailored to this population hold promise to improve physical function safely and effectively after sepsis. However, this type of intervention has yet to be tested in this population. OBJECTIVE: This study aims to assess the safety, feasibility, and ease of recruitment and retention of participants for a remotely delivered physical activity intervention for improving physical function in middle-aged and older sepsis survivors. METHODS: The proposed intervention will be delivered through a digital health platform that comprises a patient-facing mobile app and a 12-week physical activity program specifically designed for middle-aged and older sepsis survivors with poor health status who may face challenges participating in traditional out-patient or community-based exercise interventions. This study is ongoing and plans to enroll 40 sepsis survivors aged 55 years and older who will be randomized to either a remotely delivered exercise intervention group or a control group (electronic health diary). Both groups will use a tablet containing the Health in Motion app (Blue Marble Health). The intervention group will receive a clinician-designed personalized avatar-guided home exercise program and reminders while the control group will self-report daily activities using the in-app health diary feature. RESULTS: This study is the first to use a home-based, remotely monitored 12-week exercise program to improve physical function in sepsis survivors. This study will evaluate the safety, feasibility, and efficacy, providing the necessary knowledge to design and calculate power for future larger trials. CONCLUSIONS: This study will provide important information for planning a future randomized clinical trial to test the efficacy of a remotely delivered exercise intervention in this high-risk population. TRIAL REGISTRATION: ClinicalTrials.gov NCT05568511; https://clinicaltrials.gov/study/NCT05568511. INTERNATIONAL REGISTERED REPORT IDENTIFIER (IRRID): DERR1-10.2196/60270.

Low magnesium levels and prognosis in newly diagnosed diffuse large B-cell lymphoma.

Magnesium (Mg) is an essential element involved in cellular metabolism. We demonstrated that in patients with diffuse large B-cell lymphoma (DLBCL) undergoing autologous stem cell transplant (SCT), those with a serum Mg < 2.0 mg/dL at the time of transplant had worse outcomes. In this study, we aimed to learn the prognostic value of low serum Mg in patients with untreated DLBCL. We analyzed serum from 408 patients and tested 2 Mg cutpoints-low (<1.7 mg/dL) and low normal (<2.0 mg/dL), a range we found associated with lower survival in the SCT group. We found 3% of patients with low levels and 23% with low normal levels. Low normal serum Mg levels were associated with a higher stage at diagnosis, more extranodal involvement, higher international prognostic index score, lower overall survival (OS), and event-free survival. These data warrant testing Mg replacement to a target of >2.0 mg/dL to learn if survival can be improved.

Mediating role of obstructive sleep apnea in altering slow-wave activity and elevating Alzheimer's disease risk: Pilot study from a northern Taiwan cohort.

OBJECTIVES: Obstructive sleep apnea is associated with alterations in slow-wave activity during sleep, potentially increasing the risk of Alzheimer's disease. This study investigated the associations between obstructive sleep apnea manifestations such as respiratory events, hypoxia, arousal, slow-wave patterns, and neurochemical biomarker levels. METHODS: Individuals with suspected obstructive sleep apnea underwent polysomnography. Sleep disorder indices, oxygen metrics, and slow-wave activity data were obtained from the polysomnography, and blood samples were taken the following morning to determine the plasma levels of total tau (T-Tau) and amyloid beta-peptide 42 (Aß42) by using an ultrasensitive immunomagnetic reduction assay. Subsequently, the participants were categorized into groups with low and high Alzheimer's disease risk on the basis of their computed product Aß42 × T-Tau. Intergroup differences and the associations and mediation effects between sleep-related parameters and neurochemical biomarkers were analyzed. RESULTS: Forty-two participants were enrolled, with 21 assigned to each of the low- and high-risk groups. High-risk individuals had a higher apnea-hypopnea index, oxygen desaturation index (≥3%, ODI-3%), fraction of total sleep time with oxygen desaturation (SpO2-90% TST), and arousal index and greater peak-to-peak amplitude and slope in slow-wave activity, with a correspondingly shorter duration, than did low-risk individuals. Furthermore, indices such as the apnea-hypopnea index, ODI-3% and SpO2-90% TST were found to indirectly affect slow-wave activity, thereby raising the Aß42 × T-Tau level. CONCLUSIONS: Obstructive sleep apnea manifestations, such as respiratory events and hypoxia, may influence slow-wave sleep activity (functioning as intermediaries) and may be linked to elevated neurochemical biomarker levels. However, a longitudinal study is necessary to determine causal relationships among these factors. STATEMENT OF SIGNIFICANCE: This research aims to bridge gaps in understanding how obstructive sleep apnea is associated with an elevated risk of Alzheimer's disease, providing valuable knowledge for sleep and cognitive health.

ALKBH5 modulation of ferroptosis in recurrent miscarriage: implications in cytotrophoblast dysfunction.

Background: As one of the most common and abundant internal modifications of eukaryotic mRNA, N6-methyladenosine (m6A) modifications are closely related to placental development. Ferroptosis is a newly discovered form of programmed cell death. During placental development, placental trophoblasts are susceptible to ferroptosis. However, the interactions of m6A and ferroptosis in trophoblast physiology and injury are unclear. Methods: Recurrent miscarriage (RM) was selected as the main gestational disease in this study. Published data (GSE76862) were used to analyze the gene expression profiles in patients with RM. The extent of m6A modification in total RNA of villous tissues between patients with RM and healthy controls (HC) was compared. ALKBH5 (encoding AlkB homolog 5, RNA demethylase) was selected as the candidate gene for further research. Quantitative real-time reverse transcription PCR, western blotting, and immunohistochemistry (IHC) confirmed the elevated expression of ALKBH5 in the cytotrophoblasts of patients with RM. Then, cell counting kit-8 assays, glutathione disulfide/glutathione quantification, 2',7'-dichlorfluorescein-diacetate staining, and malonaldehyde assays were used to explore the alterations of ferroptosis-related characteristics following RAS-selective lethal (RSL3) stimulation after overexpression of ALKBH5. Thereafter, we re-analyzed the published RNA sequencing data upon knockdown of ALKBH5, combined with published tissue RNA-seq data, and FTL (encoding ferritin light chain) was identified as the ferroptosis-related gene in cytotrophoblasts of patients with RM that is regulated by ALKBH5. Finally, western blotting and IHC confirmed the increased expression of FTL in the cytotrophoblasts from patients with RM. Results: Total m6A levels were decreased in patients with RM. The most significant differentially m6A-related gene was ALKBH5, which was increased in patients with RM. In vitro cell experiments showed that treatment with RSL3 resulted in increased cell death and upregulated ALKBH5 expression. Overexpression of ALKBH5 alleviated RSL3-induced HTR8 cell death and caused decreased levels of intracellular oxidation products. Published transcriptome sequencing revealed that FTL was the major ferroptosis-related gene regulated by ALKBH5 in the villous tissues of patients with RM. Consistent with the expression of ALKBH5, FTL was increased by RSL3-induction and increased in patients with RM. Conclusion: Elevated ALKBH5 alleviated RSL3-induced cytotrophoblast cell death by promoting the expression of FTL in patients with RM. Our results supported the view that ALKBH5 is an important regulator of the ferroptosis-related etiology of RM and suggested that ALKBH5 could be responsible for epigenetic aberrations in RM pathogenesis.