YTHDF3 Modulates EGFR/ATK/ERK/p21 Signaling Axis to Promote Cancer Progression and Osimertinib Resistance of Glioblastoma Cells.

BACKGROUND/AIM: Despite recent advances in EGFR-tyrosine kinase inhibitor (TKI) drugs for glioblastoma multiforme (GBM), intrinsic EGFR alterations in GBM have resulted in drug resistance and unsatisfactory clinical development of EGFR-TKIs. Determining the unknown mechanisms underlying EGFR-TKI drug resistance is an urgent, but unmet, medical need for GBM. Although several m6A RNA methylation regulators, such as reader YTHDF1/2, were recently predicted to be related to GBM recurrence, none was associated with resistance to the 3rd generation EGFR-TKI osimertinib. MATERIALS AND METHODS: Osimertinib-resistant GBM cells (U87OSR) were established to ascertain the correlation between m6A expression and osimertinib resistance, prior to systemic analyses on m6A writers, erasers, and readers. YTHDF3-silencing was employed to reveal changes in IC50, cellular migration, cancer stemness, and p21-guided senescence in U87OSR cells. Signaling pathways and an in vivo xenograft model of U87OSR cells were investigated to delineate the influence of osimertinib-resistance and elevated YTHDF3 expression. RESULTS: YTHDF3 played a crucial role in inducing cellular proliferation, migration, and stemness in U87OSR GBM cells. Importantly, silencing of YTHDF3 markedly reduced the activation of certain signaling pathways, including EGFR- or ITGA7- AKT, and ERK in U87OSR cells. Our study also revealed the oncogenic function of YTHDF3 in inducing senescence escape via p21 down-regulation. In contrast, silencing of YTHDF3 resulted in increased p21 expression, senescence, and suppressed tumor growth in our osimertinib-resistant preclinical model. CONCLUSION: Overall, our research underscores the novel potential of YTHDF3 as a new pharmacological target in GBM treatment, specifically for patients with osimertinib-resistant or refractory tumors.

Lymph Node Follicle-Targeting STING Agonist Nanoshells Enable Single-Shot M2e Vaccination for Broad and Durable Influenza Protection.

The highly conserved matrix protein 2 ectodomain (M2e) of influenza viruses presents a compelling vaccine antigen candidate for stemming the pandemic threat of the mutation-prone pathogen, yet the low immunogenicity of the diminutive M2e peptide renders vaccine development challenging. A highly potent M2e nanoshell vaccine that confers broad and durable influenza protectivity under a single vaccination is shown. Prepared via asymmetric ionic stabilization for nanoscopic curvature formation, polymeric nanoshells co-encapsulating high densities of M2e peptides and stimulator of interferon genes (STING) agonists are prepared. Robust and long-lasting protectivity against heterotypic influenza viruses is achieved with a single administration of the M2e nanoshells in mice. Mechanistically, molecular adjuvancy by the STING agonist and nanoshell-mediated prolongation of M2e antigen exposure in the lymph node follicles synergistically contribute to the heightened anti-M2e humoral responses. STING agonist-triggered T cell helper functions and extended residence of M2e peptides in the follicular dendritic cell network provide a favorable microenvironment that induces Th1-biased antibody production against the diminutive antigen. These findings highlight a versatile nanoparticulate design that leverages innate immune pathways for enhancing the immunogenicity of weak immunogens. The single-shot nanovaccine further provides a translationally viable platform for pandemic preparedness.

Mitochondrial AAA protease gene associated with immune infiltration is a prognostic biomarker in human ovarian cancer.

Early diagnosis and identification of prognostic markers for ovarian cancer can significantly improve survival and reduce mortality. The role of the YME1L1 signaling axis in genetic alterations and immune infiltration of the tumor microenvironment remains unclear. Bioinformatics web resources, including GEPIA2, cBioPortal, Oncomine, Kaplan-Meier Plotter, and TIMER, were used to analyze the expression profile, prognostic value, and immune infiltration of YME1L1. We further performed a tissue microarray analysis of paraffin-embedded tissues from 60 patients with ovarian cancer, recorded at the FIGO/TNM cancer staging. Here, we used multi-omics analysis of multiple histological datasets to map the role of epigenetic and genetic alterations of YME1L1 in tumor immune infiltration and the prognosis of cancer patients. We explored YME1L1 gene expression profiles by systematically analyzing the association between YME1L1 expression and the prognosis of patients with ovarian cancer confirmed in multiple databases. High expression levels of YME1L1 were associated with poor overall and disease-free survival. Together, our studies suggest that YME1L1 may modulate tumor survival and immune features and contribute to tumor immune invasion, poor prognosis, and immunotherapy failure. Our findings may have clinical implications for the design of treatment strategies, prognosis assessment, and follow-up management of patients receiving immunotherapy for various cancers.

Detecting Genetic Variation of Colonizing Streptococcus agalactiae Genomes in Humans: A Precision Protocol.

Deciphering the genotypic diversity of within-individual pathogens and verifying the evolutionary model can help elucidate resistant genotypes, virulent subpopulations, and the mechanism of opportunistic pathogenicity. However, observed polymorphic mutations (PMs) are rare and difficult to be detected in the "dominant-lineage" model of bacterial infection due to the low frequency. The four pooled group B Streptococcus (GBS) samples were collected from the genital tracts of healthy pregnant women, and the pooled samples and the isogenic controls were genomically sequenced. Using the PMcalling program, we detected the PMs in samples and compared the results between two technical duplicates, GBS-M001T and GBS-M001C. Tested with simulated datasets, the PMcalling program showed high sensitivity especially in low-frequency PMs and reasonable specificity. The genomic sequence data from pooled samples of GBS colonizing carrier pregnant women were analyzed, and few high-frequency PMs and some low-frequency PMs were discovered, indicating a dominant-lineage evolution model. The PMs mainly were nonsynonymous and enriched in quorum sensing, glycolysis/gluconeogenesis, ATP-binding cassette (ABC) transporters, etc., suggesting antimicrobial or environmental selective pressure. The re-analysis of the published Burkholderia dolosa data showed a diverse-community model, and only a few low-frequency PMs were shared between different individuals. Genes of general control non-repressible 5-related N-acetyltransferases family, major facilitator superfamily (MFS) transporter, and ABC transporter were positive selection candidates. Our findings indicate an unreported nature of the dominant-lineage model of GBS colonization in healthy women, and a formerly not observed mutation pool in a colonized microbial community, possibly maintained by selection pressure.

Higher risk of type 2 diabetes in young women with polycystic ovary syndrome: A 10-year retrospective cohort study.

BACKGROUND: Polycystic ovary syndrome (PCOS) is a common disorder in women of reproductive age. Over the last few decades, research studies have revealed that PCOS is strongly associated with metabolic disorders, including metabolic syndrome, obesity, insulin resistance and prediabetes. Clinical observation has shown that women with PCOS are expected to have an increased risk of developing type 2 diabetes (T2DM) in the future. AIM: To assess the hazard ratio (HR) of T2DM between women with/without PCOS. METHODS: This population-based, retrospective cohort study evaluated data retrieved from the National Health Insurance Research Database. The subjects were women with PCOS (n = 2545) identified on the basis of diagnosis, testing, or treatment codes, and women without PCOS as controls (n = 2545). The HR of T2DM between women with or without PCOS was the main outcome measure analyzed. RESULTS: Our study found that, during a 10-year follow-up period, the overall incidence of T2DM was 6.25 per 1000 person-years in the PCOS group compared with 1.49 in the control group. After adjustment for potential confounding variables, the overall incidence of T2DM was higher in the PCOS group vs the control group (HR = 5.13, 95%CI: 3.51-7.48, P < 0.0001). The risk of developing T2DM subsequent to PCOS decreased with increasing diagnosis age: the adjusted HR was 10.4 in the 18-24-year age group, 5.28 in the 25-29-year age group, and 4.06 in the 29-34-year age group. However, no such significant association was noted in women older than 35 years. CONCLUSION: These findings highlight the importance of prompting a more aggressive treatment to prevent diabetes in women diagnosed with PCOS at a young age, and, in contrast, the lessened importance of this type of intervention in women diagnosed with PCOS at a late reproductive age.

Genomic Analysis of Group B Streptococcus from Neonatal Sepsis Reveals Clonal CC17 Expansion and Virulence- and Resistance-Associated Traits After Intrapartum Antibiotic Prophylaxis.

BACKGROUND: Group B Streptococcus (GBS) is a leading cause of invasive neonatal infections. This study aimed to investigate the trend of GBS serotype and genotype change and their correlation with antimicrobial resistance before and after implementation of intrapartum antibiotic prophylaxis (IAP). METHODS: We performed serotyping, whole-genome sequencing, antimicrobial susceptibility testing, and single-nucleotide polymorphism (SNP)-based phylogenetic analysis on 238 invasive GBS isolates collected from October 1998 to February 2020 in Taiwan. RESULTS: There were 7 serotypes and 6 clonal complexes (CCs) among the 238 GBS isolates, and more than half of the isolates carried multiple antimicrobial resistance genes. The expansion of CC17 strains and the increase in late-onset disease occurred synchronously after the implementation of IAP. Analysis of the carriage isolates from pregnant women showed diverse serotype distribution in the IAP era. The antimicrobial susceptibility testing showed that all 238 strains were susceptible to ampicillin and penicillin, while the number of various resistance genes in GBS genomes was found increased with the expansion of CC17. Compared with reference genomes, 697 nonsynonymous SNPs in 443 protein-coding genes were CC17 specific. CONCLUSIONS: The study revealed the expansion of GBS CC17 and the increase of late-onset disease that occurred simultaneously with the implementation of IAP. Although the susceptibility of CC17 to antimicrobial agents is not different from that of other sequence types at present, GBS with phenotypic resistance to antimicrobials may emerge in the future, given the environmental selection pressure and the continued accumulation of SNP mutations.

Urinary Biomarkers for Detection of Clinical Endometriosis or Adenomyosis.

Endometriosis or adenomyosis can be clinically diagnosed by ultrasound, symptoms, physical examination, and serum CA125. The urinary markers need to be investigated. The aim of our study was to investigate the urinary markers of clinical endometriosis/adenomyosis, and the correlation of serum CA125 was also studied. From the literature, alpha-1 antitrypsin (A1AT), enolase-1, vitamin D binding protein (VDBP), and CA125 in urine and serum were used in our study and measured by enzyme-linked immunosorbent assays (ELISA). Further clinical correlation and detection performance were evaluated. We enrolled 19 normal controls and 33 patients clinically diagnosed with endometriosis/adenomyosis. There were significant differences between studied patients and normal controls, as follows: serum CA125 (130.91 vs. 19.75 U/mL, p = 0.004); urinary CA125-creatinine ratio (5.591 vs. 0.254 ng/mg, p = 0.028); and urinary VDBP-creatinine ratio (28.028 vs. 7.301 ng/mg, p = 0.018). For diagnostic performances, serum CA125 provided the best results, with an area under curve (AUC) of 0.888 (p = 0.001) and accuracy of 86.5%. Other excellent results were also found using urinary VDBP (AUC 0.841, p = 0.001) and A1AT (AUC 0.722, p = 0.011) creatinine ratio. Using three combined biomarkers, serum CA125, urinary VDBP, and A1AT creatinine ratio, provided good detection power (AUC 0.913, p = 0.001, sensitivity 90.9%, specificity 76.5%). Double urine markers used in combination with VDBP and A1AT creatinine ratio also provided good diagnostic performance (AUC 0.809, p = 0.001, sensitivity 81.8%, specificity 76.5%, accuracy 80%). Further development of non-invasive point-of-care tests using these biomarkers could be a fruitful future endeavor.

Emotion Regulation in Close Relationships: The Role of Individual Differences and Situational Context.

A substantial amount of research has examined the role of individual differences in the regulation of emotion and the impact of emotion regulation on mental health; however, few studies have covered the role of situational context in the selection of emotion regulation strategies. In this paper, we investigate the extent to which an individual's choice of emotion regulation strategy is affected by factors such as emotional intelligence, the person with whom one is in conflict, situational sense of control, and the individual's aim in dealing with the conflict. A total of 300 participants (46.67% female) between the ages of 21 and 35 were recruited from the community (female's mean age = 28.14, SD = 4.49; male's mean age = 28.12, SD = 4.32). Participants filled out a set of questionnaires related to their emotion intelligence and emotion regulations they used in two interpersonal incidents with parents and partner. Structural equation modeling was used for data analyses. Results showed that positive correlation between emotional intelligence and cognitive reappraisal, in contrast to previous studies, a positive correlation between emotional intelligence and repression was found. Moreover, the person one is interacting with influences the degree to which one's sense of control impacts the choice of emotion regulation strategy. For example, in the event of conflict with one's parents, the degree of situational control has little impact on emotion regulation; however, in conflicts with spouses or partners, women have more situational control and are more likely to use cognitive reappraisal or suppression. Regarding the relationship between the goal of emotion regulation and the strategies used, this study found that they are moderated by gender and the persons involved; for example, when maintaining the relationship is the primary goal of emotion regulation, cognitive reappraisal is more likely the strategy of choice for men involved in a conflict with their partner and for women involved in a conflict with their parents. Overall, the results confirm that emotion regulation is affected by both individual and situational factors, indicating the importance of adopting a dynamic approach when investigating emotion regulation.

MicroRNA­10b modulates cisplatin tolerance by targeting p53 directly in lung cancer cells.

MicroRNA (miRNA or miR)­10b is an oncogenic miRNA associated with metastasis that is present in various types of tumor, including lung cancer. However, whether miR­10b is involved in different malignant characteristics, such as drug resistance or stemness, remains unclear. Therefore, the present study investigated whether miR­10b is an upstream regulator of p53. Ectopic expression of miR­10b­agomir decreased the expression of p53 and its downstream effectors, such as Bax and p53 upregulated modulator of apoptosis. Two non­canonical sites, including 1,580­1,587 and 2,029­2,035, located in p53 3'­untranslated region (UTR) were affected by the presence of miR­10b. In functional assays, upregulation of the p53 signaling pathway following cisplatin treatment was associated with decreased levels of miR­10b and upregulation of the luciferase activity of wild­type, but not 1,584, 2,032­dual­mutant, p53 3'­UTR. The ectopic expression of miR­10b­agomir attenuated the stability of p53 3'­UTR and the expression of p53 and its downstream effectors induced by cisplatin. By contrast, the knockdown of miR­10b induced the stability of p53 3'­UTR and increased levels of p53 and the sensitivity of A549 cells to cisplatin treatment. Similar results were also observed for Beas 2B cells. In the clinical investigation, p53 exhibited two distinct associations (cocurrent and countercurrent) with miR­10b in patients with lung cancer. Patients with lung cancer with low p53 and high miR­10b levels exhibited the poorest prognosis, while those with high p53 and low miR­10b exhibited the most favorable prognosis. These findings indicate a novel pathway in which cisplatin induces the levels of p53 by increasing mRNA stability via miR­10b, indicating a novel oncogenic role of miR­10b in promoting the malignant characteristics of non­small cell lung carcinoma.

Design and characterization of a low-vibration laboratory with cylindrical inertia block geometry.

Many modern nanofabrication and imaging techniques require an ultra-quiet environment to reach optimal resolution. Isolation from ambient vibrations is often achieved by placing the sensitive instrument atop a massive block that floats on air springs and is surrounded by acoustic barriers. Because typical building noise drops off above 120 Hz, it is advantageous to raise the flexural resonance frequencies of the inertia block and instrument far above 120 Hz. However, it can be challenging to obtain a high fundamental frequency of the floating block using a simple rectangular design. Here, we design, construct, and characterize a vibration isolation system with a cylindrical inertia block, whose lowest resonance frequency of 249 Hz shows good agreement between finite element analysis simulation and directly measured modes. Our simulations show that a cylindrical design can achieve a higher fundamental resonance frequency than a rectangular design of the same mass.

Detection of Microorganisms in Body Fluids via MTT-PMS Assay.

Early detection of microorganisms is essential for the management of infectious diseases. However, this is challenging, as traditional culture methods are labor-intensive and time-consuming. The 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide-phenazine methosulfate (MTT-PMS) assay has been used to evaluate the metabolic activity in live cells and can thus be used for detecting living microorganisms. With the addition of NaOH and Tris-EDTA, the same approach can be accelerated (within 15 min) and used for the quick detection of common bacterial pathogens. The assay results can be evaluated colorimetrically or semi-quantitatively. Here, the quick detection by MTT-PMS assay was further investigated. The assay had a detection limit of approximately 104 CFU/mL. In clinical evaluations, we used the MTT-PMS assay to detect clinical samples and bacteriuria (>105 CFU/mL). The negative predictive value of the MTT-PMS assay for determining bacteriuria was 79.59% but was 100% when the interference of abnormal blood was excluded. Thus, the MTT-PMS assay might be a potential "rule-out" tool for bacterial detection in clinical samples, at a cost of approximately USD 1 per test. Owing to its low cost, rapid results, and easy-to-use characteristics, the MTT-PMS assay may be a potential tool for microorganism detection.

Home Sample Self-Collection for COVID-19 Patients.

Real-time reverse transcription-polymerase chain reaction (qRT-PCR) using specimens collected from nasopharyngeal and/or oropharyngeal swabs is the standard screening approach for coronavirus disease 2019 (COVID-19). While PCR is rapid and highly accurate, it requires costly laboratory equipment and healthcare professionals that limit its use for large-scale screening of mild or asymptomatic patients. Self-collection kits for use in the home could remedy this and have consequently received great attention. In April, 2020, a self-collection kit from LapCorp was the first such kit to be approved by the FDA. In the following month, May 2020, another kit developed by Everlywell received FDA approval, and more kits are evidently on their way to the market in the United Kingdom and elsewhere. Because these home-based, self-collection kits are easy to use and may be more acceptable for patients, they provide a superior screening option for mild or asymptomatic patients under self-quarantine. These kits conserve personal protective equipment and healthcare manpower already in short supply. The primary issues affecting the efficacy of this approach are the potential for inappropriate sampling and insufficient clinical examination. A detailed review of the commercially available kits currently available is provided and their prospective impact is noted during the current pandemic.

Differential Markers of Bacterial and Viral Infections in Children for Point-of-Care Testing.

Children suffering from infectious diseases, both bacterial and viral, are often treated with empirical antibiotics. Keeping in mind both the menace of microorganisms and antibiotic toxicity, it is imperative to develop point-of-care testing (POCT) to discriminate bacterial from viral infections, and to define indications for antibiotic treatment. This article reviews potential protein biomarkers and host-derived gene expression signatures for differentiating between bacterial and viral infections in children, and focuses on emerging multiplex POCT devices for the simultaneous detection of sets of protein biomarkers or streamlined gene expression signatures that may provide rapid and cost-effective pathogen-discriminating tools.

Rapid detection of biofilm with modified alcian blue staining: In-vitro protocol improvement and validation with clinical cases.

For chronic wounds, biofilm infection is a critical issue because it can tip the scales toward an unhealing state. Biofilm-based wound therapy has been extensively advocated. However, point-of-care biofilm diagnosis still largely relies on clinical judgment. In this study, we aimed to develop a rapid tool for diagnosing wound biofilm presence by alcian blue staining. First, we sought to optimize alcian blue staining using a colorimetric-based approach to detect the biofilm, specifically targeting polysaccharides in the extracellular polymeric substances. Among examined transfer membranes and cationic detergents at various concentrations, we selected a positively charged nylon transfer membrane for sample loading, and 1% cetyl trimethyl ammonium chloride (CTAC) as the blocking solution. After sample loading and blocking, the membrane was immersed in alcian blue solution for staining, followed by immersion in 1% CTAC to decrease background noise. Each step required only 30 seconds, and the whole procedure was completed within a few minutes. In the second part of this study, we enrolled 31 patients with chronic wounds to investigate the predictive validity of biofilm detection for unhealed wounds at a 1-month follow-up visit. Among the 18 cases with positive wound biofilm staining, 15 wounds (83.3%) were not healed at the 1-month follow-up visit. Only three unhealed wounds (30%) produced in negative staining cases. This finding indicates that biofilm infection is associated with poor healing outcome for chronic wounds. Moreover, our staining results correlated well with the clinical microbiological culture assessment (83.9% consistency; 95.2% sensitivity, and 60% specificity). In conclusion, the modified alcian blue staining protocol used here represents a rapid and sensitive procedure for detecting biofilm in chronic wounds. This technique provides a practical point-of-care approach for detection of wound biofilm, the implementation of which may improve clinical outcomes for chronic wound patients. Additional studies are required to validate this method.

Clinicopathological and prognostic significance and molecular mechanisms governing uveal melanoma.

Uveal melanoma (UM) is the most common primary intraocular malignancy in adults. Although UM and cutaneous melanoma are derived from melanocytes, UM differs clinically and biologically from its more common skin counterparts. More than half of primary UMs metastasize. However, there is currently no effective treatment for metastatic UM. Therefore, studying mutations related to the metastasis, growth, proliferation, and survival of UM can help researchers understand its pathogenesis and metastatic mechanism, thereby leading to a more effective treatment. In addition, we provide an overview of the recent basic and clinical studies to provide a strong foundation for developing novel anti-carcinogenesis targets for future interventions.

Activation of cannabinoid type 1 receptors decreases the synchronization of local field potential oscillations in the hippocampus and entorhinal cortex and prolongs the interresponse time during a differential-reinforcement-of-low-rate task.

Marijuana intoxication impairs neurocognitive functions. Common side effects of consuming cannabis include time distortion and memory loss. However, the underlying neurophysiological mechanisms involved in these effects remain unclear. We hypothesized that communication between the hippocampal CA1 region and medial entorhinal cortex (MEC) is essential for the transmission of temporal-associated information. We used a differential-reinforcement-of-low-rate (DRL) task, which requires subjects to press a lever at an optimal time point, to correlate the distributions of interresponse time (IRT) with local field potentials (LFPs) recorded in the CA1 and MEC under the effects of a cannabinoid type 1 (CB1) receptor agonist. We used a DRL 10-s schedule and trained the rats to withhold for 10 s before pressing a lever. Our data showed that the percentage of 12.4- to 14-s IRT events rose after activation of CB1 receptors in the MEC. In addition, gamma amplitude synchronization and CA1 theta phase-MEC gamma amplitude coupling decreased during the 6- to 14-s IRT events. These results suggest that activation of CB1 receptors in the MEC disrupt the functional connectivity between the CA1 and the MEC. This inefficient communication may result in increased IRT during a DRL schedule. Overall, we postulate that marijuana intoxication impairs the communication between the CA1 and MEC and influences behavioral performances that require precise timing ability.

An Envenoming Syndrome from Massive Vespa Stings Induces Multiple Organ Failure.

Envenoming syndrome is a systemic reaction induced by inoculation of large volumes of Hymenoptera venom. The clinical manifestations range from skin allergic reactions to multiple organ failure. Vespid venom-induced toxic reactions and anaphylaxis are the most common lethal mechanism of death, involving acute respiratory failure, acute liver failure, rhabdomyolysis, acute kidney injury, and severe coagulopathy. Multiple organ failure as a consequence of severe venom toxicity is a rare but dangerous complication in victims. Delay of intervention to correct vespid venom-induced toxic reactions may cause catastrophic complications. Here, we describe a case presenting a rare vespid venom-induced multiple organ failure with systemic coagulopathy after massive Vespa attack.

Spontaneous Splenic Rupture as a Rare Initial Presentation in an Acute Lymphoblastic Leukemia Patient.

A spontaneous rupture of the spleen is a rare but critical diagnosis of an acute abdomen, which may accompany unspecific symptoms mimicking acute pancreatitis, rupture of aortic aneurism, or acute coronary syndrome, delaying diagnosis and treatment. In patients that have experienced a severe spleen rupture, hypovolemic shock may cause catastrophic clinical outcomes. Therefore, early diagnosis is very important in order for physicians to declare the etiology for prevention and timely correction of the shock status. Several causes of spontaneous splenic rupture have been reported, including infection, vasculitis, pancreatitis, or hematological malignancies. Acute lymphoblastic leukemia (ALL) remains a rare but important cause of non-traumatic splenic rupture that physicians are required to assess for. Here, we describe a case presenting an acute abdomen due to spontaneous spleen rupture as the first manifestation. The purpose of this case report was to highlight the importance of considering spontaneous ruptures of the spleen as a rare but critical differential diagnosis of an acute abdomen, especially in patients with acute lymphoblastic leukemia.