Key Points: METTL3 was upregulated in CKD, leading to increased m ⁶ A modification levels. M ⁶ A modifications were enriched in genes related to the cyclic guanosine monophosphate–AMP synthase-stimulator of IFN genes pathway in CKD. Normalizing heightened METTL3 and m ⁶ A modification levels showed promise as a strategy to combat kidney disease. Background: Chemical modifications on RNA profoundly affect RNA function and regulation. m ⁶ A, the most abundant RNA modification in eukaryotes, plays a pivotal role in diverse cellular processes and disease mechanisms. However, its importance is understudied in human CKD samples regarding its influence on pathological mechanisms. Methods: Liquid chromatography–tandem mass spectrometry and methylated RNA immunoprecipitation sequencing were used to examine alterations in m ⁶ A levels and patterns in CKD samples. Overexpression of the m ⁶ A writer METTL3 in cultured kidney tubular cells was performed to confirm the effect of m ⁶ A in tubular cells and explore the biological functions of m ⁶ A modification on target genes. In addition, tubule-specific deletion of Mettl3 (Ksp-Cre Mettl3 ^f/f ) mice and antisense oligonucleotides inhibiting Mettl3 expression were used to reduce m ⁶ A modification in an animal kidney disease model. Results: By examining 127 human CKD samples, we observed a significant increase in m ⁶ A modification and METTL3 expression in diseased kidneys. Epitranscriptomic analysis unveiled an enrichment of m ⁶ A modifications in transcripts associated with the activation of inflammatory signaling pathways, particularly the cyclic guanosine monophosphate–AMP synthase (cGAS)-stimulator of IFN genes (STING) pathway. m ⁶ A hypermethylation increased mRNA stability in cGAS and STING1 as well as elevated the expression of key proteins within the cGAS-STING pathway. Both the tubule-specific deletion of Mettl3 and the use of antisense oligonucleotides to inhibit Mettl3 expression protected mice from inflammation, reduced cytokine expression, decreased immune cell recruitment, and attenuated kidney fibrosis. Conclusions: Our research revealed heightened METTL3-mediated m ⁶ A modification in fibrotic kidneys, particularly enriching the cGAS-STING pathway. This hypermethylation increased mRNA stability for cGAS and STING1, leading to sterile inflammation and fibrosis.

Prostate cancer (PC) develops in a microenvironment where the stromal cells modulate adjacent tumor growth and progression. Here, we demonstrated elevated levels of monoamine oxidase B (MAOB), a mitochondrial enzyme that degrades biogenic and dietary monoamines, in human PC stroma, which was associated with poor clinical outcomes of PC patients. Knockdown or overexpression of MAOB in human prostate stromal fibroblasts indicated that MAOB promotes cocultured PC cell proliferation, migration, and invasion and co-inoculated prostate tumor growth in mice. Mechanistically, MAOB induces a reactive stroma with activated marker expression, increased extracellular matrix remodeling, and acquisition of a protumorigenic phenotype through enhanced production of reactive oxygen species. Moreover, MAOB transcriptionally activates CXCL12 through Twist1 synergizing with TGFβ1-dependent Smads in prostate stroma, which stimulates tumor-expressed CXCR4-Src/JNK signaling in a paracrine manner. Pharmacological inhibition of stromal MAOB restricted PC xenograft growth in mice. Collectively, these findings characterize the contribution of MAOB to PC and suggest MAOB as a potential stroma-based therapeutic target.

This study aimed to characterize the plasmid-mediated quinolone resistance (PMQR) in fluoroquinolone nonsusceptible E. coli (FQNSEC) isolated from patients with urinary tract infections (UTIs) in 2019-2010 and 2020. A total of 844 E. coli isolates were collected from UTI patients at National Cheng Kung University Hospital. The antimicrobial susceptibility of E. coli isolates to 21 antibiotics was determined by disk diffusion tests. The distribution of phylogenetic groups, virulence factor, and PMQR genes was determined by PCR. Conjugation assays were performed to investigate the transferability of qnr genes from FQNSEC isolates to E. coli C600. We found 211 (41.9%) and 152 (44.7%) E. coli isolates were FQNSEC in 2009-2010 and 2020, respectively. Phylogenetic group B2 was dominant in FQNSEC isolates (52.34%), followed by group F (10.47%), group B1 (9.64%), and group D (9.64%). FQNSEC isolates were more resistant to 17 of 19 tested antimicrobial agents, compared to the fluoroquinolone susceptible E. coli. PMQR screening results showed 34, 22, and 10 FQNSEC isolates containing aac(6')-Ib-cr, qnr genes, and efflux pump genes (qepA or oqxAB), respectively. PMQR E. coli isolates were more nonsusceptible to gentamicin, amoxicillin, ampicillin/sulbactam, imipenem, cefazolin, cefuroxime, cefmetazole, ceftriaxone, ceftazidime, and cefepime compared to non-PMQR FQNSEC. Moreover, 16 of 22 qnr-carrying plasmids were transferrable to the recipient C600. Here, we reported the high prevalence of MDR- and XDR-E. coli in FQNSEC isolates. Moreover, qnr-carrying plasmids were highly transferable and led to the resistance to other classes of antibiotics in the transconjugants.

Purpose: To characterize gut microbiota composition in men with non-obstructive azoospermia (NOA) and evaluate associations with sperm retrieval outcomes following microdissection testicular sperm extraction (mTESE). Methods: In this prospective cohort, fecal samples from 39 azoospermic men (obstructive azoospermia [OA], n = 14; NOA, n = 25) were analyzed. NOA patients were stratified into successful (NOA ( +)) and unsuccessful (NOA ( −)) sperm retrieval groups. Full-length 16S rRNA sequencing (PacBio) was performed. Microbial composition, diversity, and differential taxa were analyzed using QIIME2, DADA2, and linear discriminant analysis effect size (LEfSe). Receiver operating characteristic (ROC) analysis was used to assess the predictive performance of candidate markers. Results: Compared with OA, NOA patients showed reduced Bacteroidota and increased Proteobacteria, indicating gut dysbiosis. Alpha diversity was similar between groups, whereas beta-dispersion revealed greater microbial variability in NOA. Within the NOA cohort, the NOA ( +) group exhibited a higher Firmicutes-to-Bacteroidota ratio and distinct microbial profiles. Finegoldia magna and other Firmicutes taxa were enriched in NOA ( +), whereas Bacteroidota-associated taxa predominated in NOA ( −). The Firmicutes-to-Bacteroidota ratio predicted successful retrieval with an area under the curve of 0.76, and Finegoldia magna abundance showed high specificity, although these findings were exploratory. Conclusion: Gut microbiota composition differs by azoospermia etiology and is associated with mTESE outcomes. Enrichment of Finegoldia magna and increased Firmicutes in NOA ( +) suggest a potential role of the gut–testis axis. Gut microbiota profiling may represent a non-invasive correlate of sperm retrieval success, although these exploratory findings require validation in larger independent cohorts.

Objectives: Examine the understanding of terminologies and management patterns of bacillus Calmette‐Guérin (BCG)‐unresponsive nonmuscle invasive bladder cancer (NMIBC) in six territories in Asia‐Pacific.Methods: This study involved two phases: (1) a survey with 32 urologists and 7 medical oncologists (MOs) and (2) a factorial experiment and in‐depth interviews with 23 urologists and 2 MOs. All clinicians had ≥8 years' experience managing NMIBC patients in Australia, Hong Kong, Japan, South Korea, Singapore, and Taiwan. Data from Phase 1 were summarized using descriptive statistics; content and thematic analyses applied in Phase 2.Results: In phase 1, 35% of clinicians defined BCG‐unresponsive as BCG‐refractory, ‐relapse and ‐resistant, 6% defined it as BCG‐refractory and ‐relapse; 22% classified BCG‐failure as BCG‐refractory, ‐relapse, ‐resistant, and when muscle‐invasive bladder cancer is detected. If eligible and willing, 50% (interquartile range [IQR], 50%–80%) of BCG‐unresponsive patients would undergo radical cystectomy (RC), and 50% (IQR 20%–50%) of RC‐eligible patients would receive bladder‐sparing treatment or surveillance. In phase 2, we found that 32%, 88%, and 48% of clinicians, respectively, used “BCG‐unresponsive,” “BCG‐refractory,” and “BCG‐relapse” in clinical practice but with no consistent interpretation of the terms. Compared with EAU definitions, 8%–60% of clinicians appropriately classified 9 tumor types that are persistent or recurrent after adequate BCG. Fifty percent of clinicians mentioned a lack of bladder‐preserving treatment that outperforms RC in quality of life as a reason to retreat BCG‐unresponsive patients with BCG.Conclusions: Our study revealed varied understanding and application of BCG‐unresponsive terminologies in practice. There is a need for a uniform and simple definition of BCG‐unresponsive disease in Asia‐Pacific.

Background: Triplet therapy with androgen deprivation therapy (ADT), docetaxel, and androgen receptor pathway inhibitors (ARPIs), either darolutamide or abiraterone, is one of the standard treatments for metastatic hormone-sensitive prostate cancer (mHSPC); however, no randomized trial has directly compared both ARPIs within this regimen. Given practical and financial constraints, a head-to-head randomized trial directly comparing these ARPIs may be unlikely. Objective: To compare the real-world effectiveness of darolutamide- versus abiraterone-based triplet therapy in patients with mHSPC. Design: Retrospective cohort study using a multinational electronic health record network. Methods: We conducted a retrospective cohort study using the TriNetX research network. Patients with mHSPC treated with darolutamide- or abiraterone-based triplet therapy were identified and balanced using propensity-score matching. The primary outcome was overall survival (OS), and the secondary outcome was time to next treatment (TTNT), defined by treatment switch. Prespecified subgroup analyses focused on age, cardiometabolic comorbidities, and polypharmacy-related medication use. Results: Among 1607 eligible patients, 1252 were included after propensity score matching (626 per group). Darolutamide-based triplet therapy was associated with longer OS compared with abiraterone (hazard ratio (HR) 0.42). A favorable difference in TTNT was also observed (HR 0.66). These associations were consistent across clinically relevant subgroups, including patients aged ⩾65 years and those with ischemic heart disease or diabetes mellitus. In polypharmacy-enriched subgroups, darolutamide-based therapy was consistently associated with more favorable OS and TTNT. Conclusion: In this large real-world analysis, darolutamide-based triplet therapy was associated with longer OS and TTNT compared with abiraterone-based triplet therapy in patients with mHSPC, particularly among older individuals and those with comorbidities. These findings are hypothesis-generating, suggesting that ARPI selection within triplet therapy may have clinically relevant implications, supporting the need for prospective studies.