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Neurogenesis in adult mammalian brain persists in restricted areas, especially the subgranular zone (SGZ) of the hippocampus and the ventricular-subventricular zone (V-SVz), where neural stem cells (NSCs) occupy neurogenic niches. These NSC niches provide signals that regulate stem cell behavior. Among extrinsic modulators, Growth Differentiation Factor 11 (GDF11 or BMP11) which is a transforming growth factor-β (TGF-β) superfamily member, was shown to play key role in the NSC biology and brain aging. In this review, the most recent molecular mechanisms of GDF11 signaling in the regulation of NSC will be addressed. GDF11 plays mainly through activin type II receptors (ActRIIA/B) and ALK4/ALK5, activating classical Smad2/3 pathways that impact transcriptional networks controlling neural cell behavior. Moreover, GDF11 stimulates non-Smad signaling pathways - including ERK, p38, JNK, and PI3K/AKT - providing context-dependent integration of proliferative and anti-proliferative signals. Furthermore, GDF11 functions as a feedback regulator limiting the number of progenitor cells and organizing neurogenic timing. In the adult brain, GDF11 plays important role in neurovascular remodeling, glial inflammatory states, and extracellular matrix interactions. Despite its recognized roles, the effect of GDF11 on aging remains a subject of intense debate, characterized by conflicting reports regarding its circulating levels, tissue-specific dynamics, and dose-dependent effects. Recent evidence suggests that GDF11 acts as a context-dependent modulator, integrating systemic, vascular, and cellular cues to maintain NSC homeostasis and neurogenic potential. Therefore, elucidating the exact cellular and molecular mechanisms by which GDF11 controls NSC behavior is vital to advancing novel therapeutic strategies for neurodegenerative disorders and age-related cognitive decline.

Plants rely on complex signaling networks to interact with beneficial plant growth-promoting rhizobacteria (PGPR). Among these microbes, Caulobacter RHG1 is recognized by Arabidopsis thaliana roots via specific receptor-like kinases (RLKs), but the molecular mechanisms underlying the plant growth-promotion effect remain largely unknown. To investigate these mechanisms, we used an integrative approach combining proteomics and phosphoproteomics. Our phosphoproteome analysis revealed that RHG1 triggers a range of signaling events, involving two RLKs (LysM RLK1-INTERACTING KINASE 1 [LIK1] and an uncharacterized RLK, AT5G49770). Notably, via the proteome analysis, we revealed that upon RHG1 treatment, defense proteins are downregulated early. Later on, developmental processes, such as ribosome biogenesis and maturation, might lead to enhanced cell division and/or activity to promote growth. Our findings suggest that RHG1 triggered phosphorylation-based signaling pathways, resulting in the timely modulation of defense and developmental processes, which eventually lead to plant growth promotion. Our data, combined with that of previous reports, suggest that RHG1 activates a broad network of regulatory mechanisms that extend beyond a single pathway. Understanding the mechanisms behind such beneficial plant-microbe interactions can pave the way for more effective commercialization of PGPR-based agricultural products.

Lenalidomide (Len) is the standard of care for red blood cell transfusion-dependent (TD) patients with myelodysplastic syndromes (MDS) and del(5q). A Phase III clinical trial (SintraRev) demonstrated better efficacy of early treatment with Len in anemic del(5q) MDS patients prior to TD, but clonal evolution with this approach remained unexplored. We evaluated changes in mutational profiles of non-TD del(5q) MDS patients treated with Len to assess whether early administration of Len could reduce or worsen the mutational burden in those patients. The molecular profile of patients included in the SintraRev trial was analyzed by targeted sequencing. Median follow up of patients was 60.6 months (interquartile range [IQR] 32.3-73.9). Next-generation sequencing (NGS) was available in a total of 51 patients. SF3B1 (25.5%) and TP53 (21.3%) were the most frequently mutated genes. Clonal evolution during treatment among patients receiving Len or placebo (Pcb) was markedly different, with a 41.2% reduction in predominating clones in the Len arm, while remaining stable/increase in 100% of Pcb patients after 2 years. Mutations in SF3B1 and TP53 decreased/remained stable in the Len cohort, while only DNMT3A mutations increased under cytogenetic response. Regarding disease progression, only 20% of Len patients carrying TP53 mutations developed acute myeloid leukemia (AML), similar to the Pcb group (20%). Treatment with low-dose Len in transfusion-independent del(5q) MDS reduced the mutational burden of most genes and did not promote the expansion of preexisting clones or AML progression, especially TP53-mutated clones. Early administration of Len in del(5q) MDS patients without TD may be an effective therapeutic approach with a manageable safety profile regarding clonal evolution.

Atypical endometrial hyperplasia and endometrial adenocarcinoma are successive stages of malignant transformation of the uterine epithelium. Usually genomic instability and tumor mutation burden progressively increase during carcinogenesis. In case of endometrial cancer, the presence of a mutation in the exonuclease domain of the POLE gene and/or a DNA mismatch repair deficiency (dMMR) leads to genomic instability at early stages of tumor development. This paper describes a clinical case of a 49-year-old female patient with synchronous presence of several stages of the endometrium malignant transformation: glandular polyp without signs of atypia, atypical hyperplasia and highly differentiated endometrioid adenocarcinoma. In all substrates, the POLE p.Pro286Arg mutation was detected. In the glandular polyp sample, it was detected with a minimal variant allele frequency (1%). At the same time, in the tissue of atypical hyperplasia and adenocarcinoma, this mutation led to an explosive accumulation of oncogenic genetic variants. This observation indicates role of mutation in the POLE gene as an early driver of carcinogenesis and emphasizes the importance of its determination in atypical endometrial proliferations. Атипическая гиперплазия эндометрия и эндометриальная аденокарцинома представляют собой последовательные этапы злокачественного перерождения эпителия тела матки. В процессе канцерогенеза геномная нестабильность и мутационная нагрузка прогредиентно нарастают, однако в случае рака эндометрия наличие мутации в экзонуклеазном домене гена POLE и/или дефицита системы репарации ошибочно спаренных нуклеотидов ДНК (dMMR) приводит к геномной нестабильности на ранних этапах развития опухоли. В данной работе описывается клиническое наблюдение пациентки 49 лет, в гистологическом материале которой были выявлены одновременно 3 стадии злокачественной трансформации эндометрия: железистый полип без признаков атипии, атипическая гиперплазия и высокодифференцированная эндометриоидная аденокарцинома. Во всех субстратах была выявлена мутация POLE p.Pro286Arg, которая в образце железистого полипа обнаруживалась с минимальным значением доли альтернативного аллеля (1%). При этом в ткани атипической гиперплазии и аденокарциномы такая мутация приводила к взрывному накоплению онкогенных генетических вариантов. Данное наблюдение указывает на роль мутации в гене POLE как раннего драйвера канцерогенеза и подчеркивает важность ее определения в атипичных эндометриальных пролиферациях.

Nanoparticles (NPs) have attracted increasing attention because of their antimicrobial properties. This study aimed to evaluate the antibacterial and antibiofilm activities of nickel oxide (NiO) nanoparticles against representative uropathogenic Escherichia coli (UPEC) O-serotypes isolated from patients with urinary tract infections (UTIs) and characterized by ERIC-PCR typing. A total of 153 UPEC isolates were characterized by PCR-based O-serogrouping and ERIC-PCR typing. NiO nanoparticles were synthesized using the sol-gel method. The antibacterial and antibiofilm activities of NiO nanoparticles were evaluated against one representative biofilm-forming isolate from each ERIC type using the broth microdilution and microtiter plate methods, respectively. Among the 153 UPEC isolates, seven O-serogroups, O2, O4, O6, O8, O15, O16, and O25, were detected, with O25 being the most prevalent. ERIC-PCR analysis classified the isolates into 19 distinct clusters (E1-E19) at an 80% similarity threshold, comprising 13 common types (each containing ≥ 2 isolates) and 6 unique (singleton) types. No significant association was observed between O-serogroups and ERIC-PCR clusters (p > 0.05). NiO nanoparticles exhibited antibacterial and antibiofilm activity against 19 representative UPEC isolates, with MIC values ranging from 250 to 500 µg/mL and significant inhibition of biofilm formation at concentrations of 125-500 µg/mL (p = 0.001). Biofilm inhibition was also observed at sub-MIC concentrations. NiO nanoparticles demonstrated in vitro antibacterial and antibiofilm activity against 19 representative UPEC isolates. However, broader isolate screening and comprehensive toxicity and in vivo studies are required to further evaluate the antimicrobial activity and potential applicability of NiO nanoparticles.

Brain metastases occur in 30-55% of patients with HER2-positive metastatic breast cancer, presenting significant therapeutic challenges. Circulating tumor DNA (ctDNA) monitoring has emerged as a potentially useful tool for earlier molecular detection of disease activity. We report a 35-year-old woman with HER2-positive metastatic breast cancer diagnosed during pregnancy who achieved pathological complete response after standard therapy. Given her young age, de novo stage IV disease, and atypical pregnancy-associated presentation, baseline comprehensive genomic profiling was performed at diagnosis. Serial ctDNA monitoring detected positivity approximately three months prior to radiologic evidence of brain metastases. Genomic profiling revealed newly emergent alterations potentially contributory to evolving disease biology, including MET amplification and HER2 V777L mutation. Treatment with trastuzumab deruxtecan plus pyrotinib, informed by ctDNA dynamics and the molecular profile, was associated with intracranial disease control over 17 months of follow-up. Drug-induced hepatotoxicity was managed through dose modifications and supportive care without treatment interruption. This case illustrates the feasibility of incorporating ctDNA monitoring into treatment decisions and provides a hypothesis-generating observation regarding combination therapy in managing HER2-positive brain metastases. The temporal relationship between molecular and radiologic findings observed here suggests potential value for earlier detection of disease activity, although whether such lead time translates into improved clinical outcomes requires prospective validation. The findings support prospective evaluation of this approach, including the ongoing TROPHY trial investigating this therapeutic approach.

This study reports a 50-year-old male case of Thelazia callipaeda infection without a history of pet contact. The patient exhibited chronic ocular surface inflammation after exposure to wild insects and was misdiagnosed multiple times with bacterial conjunctivitis before definitive diagnosis by an ophthalmic specialist. This report emphasizes the core steps of the T. callipaeda extraction procedure. These steps include preoperative preparation, topical ocular anesthesia using 0.4% oxybuprocaine hydrochloride eye drops, sufficient exposure of the surgical field with an eyelid speculum, precise clamping and complete removal of nematodes under ophthalmic visualization, and immediate postoperative management. The entire procedure was recorded in a supplementary operative video. Additionally, the procedure can be conducted under conventional outpatient clinic conditions, making it suitable for primary medical institutions. Postoperatively, the patient received anti-infective therapy with 0.5% levofloxacin eye drops, administered four times daily, and ocular surface moisturization with 0.1% sodium hyaluronate eye drops, administered four times daily, for 1 week, with standard follow-up at 1, 3, and 6 months postoperatively. The clinical manifestations, including conjunctival hyperemia and foreign body sensation, were completely alleviated without recurrence, and ocular surface function returned to normal. Etiological confirmation was verified through morphological observation, characterized by serrated cuticular striations, a hexagonal buccal capsule, and a gravid female reproductive system, and recombinase polymerase amplification-CRISPR-Cas12a molecular detection targeting the 18S ribosomal ribonucleic acid gene. Critical procedural precautions to prevent nematode fragmentation and residual inflammation are also underscored. This case enriches the clinical reports of T. callipaeda. infection without a pet contact history and offers a practical diagnosis and treatment protocol for ophthalmologists.

Fibroblast growth factor 21 (FGF21) is an endocrine hormone central to metabolic regulation under nutritional and cellular stress. Predominantly synthesized by the liver, FGF21 exerts pleiotropic effects through fibroblast growth factor receptor 1c (FGFR1c) and its co receptor β-Klotho, targeting adipose tissue, skeletal muscle, pancreas, and the central nervous system. In diabetes mellitus, FGF21 enhances insulin sensitivity, stimulates adiponectin secretion, alleviates hepatic steatosis, and protects pancreatic β-cells from oxidative and endoplasmic reticulum stress. These effects are mediated via key molecular pathways, including AMPK, SIRT1, PGC-1α, and mTOR signaling. Interestingly, despite elevated circulating FGF21 levels in obesity and type 2 diabetes, biological responsiveness is diminished-a phenomenon termed FGF21 resistance, characterised by impaired receptor expression, inflammatory signaling interference, and downstream signaling deficits. This review presents an in depth mechanistic overview of FGF21 signaling, its regulatory networks, and implications for glucose and lipid homeostasis, inflammation, mitochondrial health, and autophagy. Additionally, it discusses the development of pharmacologically optimized FGF21 analogs and multi agonists designed to overcome resistance and enhance therapeutic efficacy. As our comprehension of FGF21 biology deepens, elucidation of its molecular processes, resistance phenomena, and pharmacological refinement will be crucial in establishing FGF21 as a fundamental element of precision medicine for metabolic illnesses. The incorporation of FGF21 based strategies with combinatorial pharmacotherapy and tissue targeted delivery methods offers a potential approach to tackle the complicated pathophysiology of diabetes and improve clinical outcomes in affected populations.

Understanding how transcription factors (TFs) regulate target promoters is central to molecular biology. However, widely used reporter-based assays typically require specialized instrumentation, costly substrates, and labor-intensive procedures. The recently developed RUBY reporter gene, which enables visible betalain accumulation, offers a simple and equipment-free alternative for monitoring gene expression. Here, we established a pEAQ-RUBY reporter system for efficient analysis of TF-mediated promoter regulation. We first optimized a quantitative method for betalain extraction in tobacco and identified 40% ethanol as a safe and effective solvent. We then constructed pGreenII 0800-RUBY reporter vectors by replacing the LUC coding sequence with RUBY while preserving the multiple cloning site for flexible promoter insertion. Four commonly used effector vectors were evaluated, and pEAQ exhibited the highest expression efficiency. Using this system, we confirmed the positive regulation of SARD1 on the SA biosynthetic gene promoter ICS1 and MYC2 on the JA biosynthetic gene promoters AOS, LOX2, and OPR3. We also validated the negative regulation of TCP20 and TCX8 on the LOX2 promoter. All results were consistent with previous reports. Collectively, the pEAQ-RUBY reporter system provides a simple, visible, and safe alternative to conventional luciferase-based assays for studying transcriptional regulation in plants.

SLC6A8 encodes the creatine transporter (CRT), which mediates creatine transport across the plasma membrane in the brain, including the blood-brain barrier and neurons. Creatine transporter deficiency (CTD), caused by pathogenic variants in SLC6A8, leads to cerebral creatine depletion and cognitive impairment. Here, we investigated the developmental molecular mechanisms underlying CTD using the pathogenic c.1681G>C (G561R) variant of Slc6a8, which corresponds to a variant identified in SLC6A8 in a patient with CTD. In vitro analyses using HEK293 cells expressing mutant mouse CRT carrying the G561R variant demonstrated impaired N-glycan maturation and plasma membrane localization of the transporter, resulting in markedly reduced creatine uptake, consistent with previous reports on the corresponding human CRT variant. To investigate the in vivo effects of this pathogenic variant, we generated CRT-G561R knock-in mice by introducing the c.1681G>C point mutation into the mouse Slc6a8 gene using the CRISPR/Cas9 system. These male mice exhibited severe reductions in brain creatine levels, postnatal growth retardation, and impaired spatial memory, despite preserved gross brain morphology. Quantitative proteomic analyses of the hippocampus and cerebral cortex during postnatal development revealed region-dependent protein alterations in CTD. The hippocampus showed pronounced early postnatal remodeling involving proteins related to actin cytoskeleton organization and vesicle-mediated membrane trafficking, whereas the cerebral cortex exhibited a more gradual response involving creatine biosynthesis-related enzymes and later-emerging mitochondrial pathways, including the mitochondrial translation machinery. These findings demonstrate stage- and region-dependent proteomic remodeling during postnatal brain development in CTD.Significance Statement Creatine transporter deficiency (CTD) causes cerebral creatine depletion and intellectual disability; however, the developmental mechanisms linking creatine loss to brain dysfunction remain unclear. We performed developmental proteomic profiling of the hippocampus and cerebral cortex using a mouse model carrying a pathogenic Slc6a8 variant identified in patients with CTD. Creatine transporter dysfunction induces distinct region- and stage-dependent molecular responses during postnatal brain maturation. The hippocampus shows early alterations in cytoskeleton-dependent membrane trafficking pathways, consistent with impaired synaptic and circuit maturation, whereas the cerebral cortex exhibits progressive metabolic and mitochondrial adaptations. These findings suggest that impaired creatine-dependent energy buffering disrupts distinct developmental programs across brain regions, potentially contributing to cognitive dysfunction by hindering early hippocampal circuit maturation.

Pancreatic ductal adenocarcinoma (PDAC) is among the deadliest cancers, with a 5-year survival rate of 3.2% in metastatic cases. Cisplatin, a DNA-damaging agent, offers minimal benefit due to PDAC's altered DNA repair and protective stroma. Tumor Treating Fields (TTFields), low intensity alternating electric fields that disrupt mitosis, have shown promise in other solid tumors. We investigated whether TTFields enhance cisplatin efficacy in PDAC and explored the underlying biology. PDAC cell lines were treated with TTFields and cisplatin alone or in combination. Assays assessed viability, migration, and clonogenic survival. Orthotopic mouse models evaluated tumor volume. Mechanistic studies included KRAS signaling analysis and RNA sequencing. TTFields significantly reduced PDAC cell viability, migration, and clonogenic survival, with the strongest effects in combination with cisplatin. In vivo, combined therapy achieved ∼85% tumor volume reduction. Molecular profiling revealed suppression of KRAS-driven signaling, stromal activation, and survival pathways beyond single-agent effects. Transcriptomic analysis showed alteration of immunogenic tumor microenvironment, downregulation of inflammatory survival signaling, and upregulation of genes linked to cytoskeletal and mitochondrial remodeling, changes predicted to limit tumor spread and promote apoptosis. TTFields potentiate cisplatin's therapeutic impact in PDAC and reveal a novel biology where electrical and chemical stress converge to reprogram tumor survival. This well tolerated multimodal strategy offers a promising avenue for improving outcomes in PDAC, a cancer urgently needing more effective treatments.

Prostate cancer (PCa) ranks as the fifth most prevalent cancer type globally, with its incidence rising due to lifestyle changes. The prognosis significantly worsens at the metastatic stage, with a 5-year survival rate dropping to 30%. This study focuses on the androgen signaling pathway's role in PCa etiology and aims to identify molecular mechanisms that could serve as potential biomarkers for early diagnosis of the disease in a population of Iranian men. We analyzed the expression levels of long non-coding RNAs (lncRNAs) PCAT18, CTBP1 antisense RNA (CTBP1-AS), and HOTAIR in blood samples from 30 PCa patients and 30 individuals with benign prostatic hyperplasia (BPH). RT-PCR and specific primers were used for quantification. We observed a significant downregulation of PCAT18, HOTAIR and CTBP1-AS in the PCa cohort compared to BPH. ROC curve analysis revealed HOTAIR as the most promising biomarker, with an area under the curve of 0.6897. Further analysis indicated that PCAT18 and HOTAIR demonstrate considerable sensitivity and specificity for distinguishing PCa from BPH, suggesting their potential utility in clinical diagnosis and prognosis evaluation. Significant correlation was found between LNCRNAs expression and demographic information for CTBP1-AS expression vs. size of tumor, PCAT18 expression vs. PSA and PCAT18 expression vs. grade. These results implying that lncRNAs dysregulation may be more indicative of cancer biology. These findings highlight the importance of CTBP1, PCAT18 and HOTAIR as potential biomarkers in prostate cancer and underscore the necessity for further investigation into their roles in cancer progression and therapeutic strategies.

Yam is a medicinal plant and previous studies have demonstrated that the yam glycoprotein (GLP1) possessed immunomodulatory bioactivity. However, how GLP1 exerted immunomodulatory effect and the molecular mechanism is unclear. To investigate the underlying mechanisms, the immunoregulation of GLP1 on macrophages are examined. The composition of GLP1 was investigated using high-performance liquid chromatography (HPLC) and UV visible spectrophotometer (UV-Vis). The activity of macrophages was detected, and the effects of GLP1 on macrophages were studied using immunofluorescence, flow cytometry, Enzyme-linked immunosorbent assay (ELISA), and Western blotting. Results indicated that GLP1, as a biomolecule, contained abundant monosaccharides and possessed O-glycopeptide bonds. GLP1 bound to pattern recognition receptors (PRRs) and was internalized by macrophages via toll-like receptor 4 (TLR4), thereby exerting its immunomodulatory effects. Further experimental studies revealed that GLP1 promoted RAW264.7 cells proliferation by advancing cells from G0/G1 phase to S phase. Additionally, GLP1 stimulated the secretion of reactive oxygen species (ROS) from mitochondria and induced macrophage polarization toward the M1 phenotype. GLP1 is a functional protein and has the capacity to regulate macrophages immune responses via the TLR4/NF-κB pathway. The findings highlighted the potential of GLP1 might as a dietary immunostimulant at the molecular level.

Mitochondrial neurogastrointestinal encephalomyopathy (MNGIE) is an autosomal recessive disorder caused by mutations in TYMP, which disrupt thymidine metabolism. This study aimed to characterize a novel homozygous TYMP variant and provide insights into its potential structural and functional consequences through bioinformatic analyses. We identified a homozygous TYMP variant (c.131G>C, p.R44P) in a proband with MNGIE using whole-exome sequencing and Sanger sequencing. Computational structural analyses and molecular modeling were performed to predict the impact of the R44P substitution on thymidine phosphorylase (TP) stability, homodimerization, catalytic activity, and substrate binding. The homozygous TYMP c.131G>C variant was confirmed in the proband. Computational analyses suggested that the p.R44P substitution may destabilize TP and potentially impair homodimerization. Molecular modeling further predicted altered thymidine binding and disrupted active-site geometry. These predicted perturbations are hypothesized to contribute to defective nucleotide metabolism, thymidine accumulation, and deoxynucleotide triphosphate pool imbalance, which may ultimately result in mitochondrial genomic instability manifesting as mitochondrial DNA deletions and depletion. Our findings report the TYMP c.131G>C variant in a homozygous configuration, extending beyond a recently described compound heterozygous case. The bioinformatic predictions support the classification of this variant as likely pathogenic in MNGIE, though functional studies are warranted to validate these findings.

The emergence of highly virulent and multidrug-resistant Salmonella strains continues to pose a major global public health concern. Building on our previous investigation of 80 Salmonella isolates obtained from avian and human sources in northeastern Algeria (Annaba and Constantine) over a three-year period (2017-2019), in which their resistance profiles to 15 antimicrobial agents were assessed, the present study further explores the molecular features of virulence by targeting two additional plasmid-encoded genes, spvB and spvR. All isolates were subjected to polymerase chain reaction (PCR) amplification using gene-specific primers. The results revealed that spvB was detected in 86.25% of isolates, while spvR was identified in 73.75%, suggesting a strong association with strains capable of causing severe systemic infections. When compared with previous reports, notable geographical variations were observed, which may reflect differences in host reservoirs, ecological conditions, and selective pressures arising from antimicrobial use. The coexistence of spv virulence genes and multidrug-resistant (MDR) phenotypes in our isolates underscores the substantial risk of dissemination of highly pathogenic and resistant Salmonella strains between animals and humans. These findings highlight the need to integrate virulence gene surveillance into existing antimicrobial resistance monitoring programs to better understand and manage the zoonotic transmission dynamics of Salmonella in Algeria.

This study reports the complete mitochondrial genome (15,610 bp) of Stauroderus scalaris, a grasshopper species from Xinjiang, China, which comprises 13 protein-coding genes, 2 rRNA genes, 22 tRNA genes, and a control region. Phylogenetic analysis confirms its close evolutionary relationship with the genera Gomphocerus and Gomphocerippus, supporting its placement within the tribe Gomphocerini. These findings enrich the available mitochondrial genome resources for grasshoppers and provide valuable molecular data for subsequent research on the phylogeny and evolutionary history of the subfamily Gomphocerinae.

Rearranged during transfection (RET) fusion is a driver aberration in non-small cell lung cancer (NSCLC). Approval of RET inhibitors selpercatinib and pralsetinib prompts adequate molecular testing for RET fusions in NSCLC. The aim of this study was to determine changes in the detection of RET fusions and the effect of various testing strategies in stage IV NSCLC patients using real-world data. Patients diagnosed with metastatic non-squamous NSCLC in 2017 and 2019 were selected from Netherlands Cancer Registry (NCR) database and linked to their pathology reports form the Dutch nationwide pathology databank (Palga). Based on these data, RET fusion testing rates, prevalence and laboratory variation were evaluated. A total of 3651 patients and 3934 patients were included in 2017 and 2019, respectively. The prevalence of RET fusion in non-squamous NSCLC was 1.3% and 0.7%, in 2017 and 2019, respectively. The overall RET fusion testing rate increased from 22.6% (2017) to 31.8% (2019). Between 2017 and 2019, a shift from FISH (91% to 47%) to RNA-based testing (6% to 46%) for the detection of RET fusions was observed. The prevalence calculated from real-world data in the Netherlands with nation-wide coverage was 1.3% in 2017 and 0.7% in 2019. The decrease in detected RET fusions between 2017 and 2019 was associated with decreased use of FISH testing The prevalence of RET fusions in non-squamous NSCLC published in the literature (1-2%) is therefore likely overestimated.

Gliomas are molecularly heterogeneous central nervous system tumors with variable treatment responsiveness and survival outcomes. O6-methylguanine-DNA methyltransferase (MGMT) promoter methylation is an important biomarker because it is associated with reduced DNA repair capacity and increased sensitivity to alkylating agents, particularly temozolomide (TMZ). However, its clinical meaning is not uniform across glioma subtypes and should not be interpreted as a universal prognostic marker. In IDH-wildtype glioblastoma, MGMT promoter methylation has the strongest evidence as a predictive biomarker for benefit from TMZ-containing therapy, although survival advantages in treated cohorts should be interpreted mainly as treatment-associated effects unless treatment-independent prognostic value is demonstrated. In IDH-mutant astrocytoma, MGMT methylation may partly reflect IDH-associated global hypermethylation and appears to have limited independent clinical value. In IDH-mutant, 1p/19q-codeleted oligodendroglioma, MGMT methylation may provide supportive, treatment-context-dependent information, particularly in patients receiving alkylating chemotherapy. In pediatric-type and rare molecularly defined gliomas, including histone-altered tumors, MGMT status remains exploratory and should be interpreted alongside methylation class, lineage-defining alterations, tumor location, and treatment history. Technical factors, including assay platform, CpG-site selection, cutoff definition, tissue quality, tumor-cell content, and intratumoral heterogeneity, further complicate interpretation. Because MGMT methylation is biologically continuous, this review further argues that borderline results should be reported as gray-zone or intermediate categories when validated, and that quantitative methylation values should be integrated into subtype-aware multivariable models rather than being reduced exclusively to binary calls. This review summarizes the biological and clinical relevance of MGMT promoter methylation across glioma subtypes and proposes a subtype-aware, treatment-conditional, and assay-aware framework for interpreting its predictive and survival-related significance.

To investigate the human milk oligosaccharide (HMO) composition, longitudinal change and the influence of maternal factors in a multi-ethnic Asian cohort. In the prospective GUSTO mother-offspring cohort, maternal sociodemographic, genetic, and obstetric characteristics were related to the concentrations of the 19 most-abundant HMOs (quantified by HPLC, n = 248 mothers) at 3-weeks (n = 205) and 3-months (n = 114) postpartum (71 matched cases, 28.6%). Mothers providing samples were Chinese (73.8%), Malay (14.5%) and Indian (11.7%). Across ethnicities, individual HMO concentrations and proportions of secretors, were comparable. Approximately 70% were secretors, determined by distinctly higher 2'-fucosyllactose (2'-FL, 3-weeks: 16.0% of total HMO; 3-months: 10.4%) and lacto-N-fucopentaose-I (LNFP-I), compared with non-secretors (2'-FL, 3-weeks, 0.3%; 3-months: 0.2%). 3-fucosyllactose (3-FL) comprised 9.5% and 21.7% of total HMO concentration in secretors and non-secretors, respectively, at 3-weeks, and 30.4% and 42.9%, respectively, at 3-months. Compared with term, preterm cases had higher 3'-sialyllactose (3'SL) at 3-weeks [adjusted mean difference 1.23 SDs (95%CI 0.44-2.01); p = 0.002]. Longitudinal changes in individual HMO concentrations from 3-weeks to 3-months were generally consistent across secretor groups. HMO-secretor-phenotype showed 97% concordance with status predicted by the FUT2 single nucleotide polymorphism (SNP) rs1047781. Chinese, Malay and Indian women exhibited similar HMO compositions, with the FUT2 rs1047781 SNP being a strong determinant of secretor status. Preterm delivery may influence specific early postpartum HMO concentrations, including a higher 3'SL concentration. The concentration of 2'-FL was lower, while 3-FL was higher, compared with published reports on non-Asians, suggesting multi-ethnic studies of infant nutrition and health should consider Asian HMO biology.