Maternal nutrition during pregnancy and lactation is crucial for pup brain development. A low-protein isocaloric diet (LPID) has been associated with cognitive deficits, but its sex-specific effects on early hippocampal maturation remain unclear. This study examined whether maternal LPID alters astrocytic activation, measured by glial fibrillary acidic protein (GFAP) expression and cell density, in hippocampal subregions of male and female pups on postnatal day 10 (PND10). Pregnant Wistar rats were fed either a control diet (18% protein) or an LPID (6% protein) throughout gestation and lactation. The pups' brains were analyzed via immunohistochemistry using GFAP and 4',6-diamidino-2-phenylindole (DAPI) staining. Maternal LPID caused sex-specific changes limited to male pups. These showed marked GFAP upregulation in the CA1 pyramidal and radiatum layers, with increases of up to 225% compared to females and 124% compared to controls. DAPI staining revealed higher cell density in CA1 (up to +34%), CA3 (+34%), and the dentate gyrus (+45%), respectively. The hilus showed a 71% increase compared to females. No significant differences were observed in LPID females across hippocampal regions. These findings suggest that maternal LPID during early development induces hippocampal astrogliosis, evidenced by increased GFAP expression, in a sex-specific manner. This process disrupts cellular organization exclusively in males, which highlights their increased vulnerability to early-life nutritional stress.
Phenylketonuria (PKU) is a rare, autosomal recessive inborn error of metabolism caused by genetic variants of the PAH gene, which encodes phenylalanine hydroxylase (PAH). In PKU, the PAH enzyme has reduced affinity for its substrate [phenylalanine (Phe)] or cofactor [tetrahydrobiopterin (BH4)], shows decreased substrate activation or becomes destabilized owing to protein misfolding. Approximately 3500 PAH variants have been identified that vary in residual PAH activity and thus disease severity and BH4 responsiveness. Burdensome, lifelong dietary restriction remains the standard of care for PKU. Until recently, only two pharmacological therapies were approved for PKU: sapropterin and pegvaliase; however, the need for additional treatment options remained. In 2025, sepiapterin (Sephience™), a novel oral treatment for children and adults with PKU, was approved in several countries, including the USA. Sepiapterin has pharmacological and clinical properties unique from existing therapies. A key differentiating factor is that it has two distinct mechanisms of action: it acts as an independent chaperone for PAH, stabilizing misfolded enzyme variants, and it serves as a bioavailable precursor to BH4, increasing intracellular BH4 levels. Preclinical studies demonstrate that sepiapterin increases PAH activity across a broad range of genetic variants, including those associated with severe phenotypes and BH4-nonresponsiveness. Clinical trials (e.g., phase 3 APHENITY, AMPLIPHY) show that sepiapterin can significantly lower blood Phe versus placebo and sapropterin, with efficacy observed in both classic PKU and BH4-nonresponsive participants. In clinical studies, sepiapterin is consistently well tolerated, with limited side effects and a convenient oral formulation. On the basis of its dual mechanism of action and favorable safety and tolerability profile, sepiapterin has the potential to treat a broad range of individuals with PKU across the spectrum of disease severities and ages. Thus, all individuals should be considered for an initial treatment trial with sepiapterin to establish responsiveness, regardless of genetic variant or phenotypic severity. Sepiapterin: how it works and its potential benefits for people with phenylketonuria Phenylketonuria is a rare genetic condition that causes changes in the enzyme phenylalanine hydroxylase, stopping it from working. As a result, the body cannot break down the amino acid phenylalanine, leading to a buildup of phenylalanine in the blood and brain. Without treatment, phenylketonuria can affect the way the brain develops and how people think or behave. Currently, the main treatment is a specialized, restrictive, low-protein diet. Two medicines, sapropterin and pegvaliase, are also available; however, they are not beneficial for many individuals with phenylketonuria because of age restrictions, side effects, or lack of effectiveness. So, new treatment options are needed. Sepiapterin (Sephience™) is a new treatment approved in 2025 in several countries for children and adults with phenylketonuria. Sepiapterin works differently from existing treatments. It works in two ways: it interacts directly with phenylalanine hydroxylase to make the enzyme work better, and it turns into a substance called tetrahydrobiopterin in the body, which also helps phenylalanine hydroxylase work better. Here, clinicians summarize how sepiapterin works and its demonstrated benefits across the full range of phenylketonuria disease severities. Clinical trials show that sepiapterin lowers phenylalanine levels, including in many individuals with “classic” phenylketonuria (the most severe type). In addition, sepiapterin is well tolerated with minimal potential side effects. Overall, these data suggest that all individuals with phenylketonuria should be considered for treatment with sepiapterin to assess potential benefits.
To clarify sex-specific differences in the risk profiles of anastomotic leakage (AL) after rectal cancer surgery, for which sex is widely recognized as an important risk factor. This retrospective multicenter study included 1,854 patients (1,196 men and 658 women) who underwent rectal cancer surgery with anastomosis between 2015 and 2025 at our institution. Patients were stratified by sex, and risk factors for AL (grade ≥ B) were analyzed separately using logistic regression. Interaction analyses between sex and perioperative factors were also conducted. AL occurred more frequently in men than in women (9.9% vs. 4.1%, respectively). In sex-stratified multivariate analyses, an anastomotic site ≤ 5 cm from the anal verge and transanal tube placement were identified as independent risk factors for AL in men, whereas minimally invasive surgery and the use of indocyanine green fluorescence imaging were associated with a reduced risk of AL. In women, a preoperative C-reactive protein-to-albumin ratio ≥ 0.07 and operative period were independent risk factors for AL. A significant interaction was observed between sex and low anastomosis rates. Sex-specific differences in AL risk profiles have been suggested. Sex-specific risk assessment may contribute to more individualized strategies for diverting stoma decision-making.
Glutamine-fructose-6-phosphate amidotransferase 2 (GFPT2) is a rate-limiting enzyme of the hexosamine biosynthetic pathway, but its protein-level expression and functional relevance in lung adenocarcinoma (LUAD) remain unclear. GFPT2 protein abundance was evaluated in Clinical Proteomic Tumor Analysis Consortium (CPTAC) LUAD proteomics and validated by Western blotting in paired LUAD and adjacent tissues. The Cancer Genome Atlas Lung Adenocarcinoma (TCGA-LUAD) transcriptomes were analyzed for differential expression and pathway enrichment, with survival assessed by Kaplan-Meier and Cox regression. Stable GFPT2 knockdown/overexpression in A549 cells was used to examine proliferation, migration, invasion, and transforming growth factor-β (TGF-β)/Smad2/3 signaling by Western blotting. GFPT2 protein was significantly upregulated in LUAD compared with adjacent tissues. TCGA-based analyses showed distinct transcriptional profiles between GFPT2 mRNA-high and GFPT2-low tumors, with enrichment of extracellular matrix organization, cell adhesion, and TGF-β-related signaling. High GFPT2 mRNA expression was associated with poorer overall survival and remained an independent adverse prognostic factor in multivariable Cox models. In A549 cells, GFPT2 overexpression was accompanied by enhanced proliferation, migration, and invasion, whereas GFPT2 knockdown showed the opposite effects. Consistently, GFPT2 upregulation was associated with increased TGF-β1 levels and Smad2/3 phosphorylation, suggesting a positive association with TGF-β/Smad signaling activity. GFPT2 is upregulated at the protein level in LUAD and is associates with aggressive cellular phenotypes and unfavorable prognosis. Integrated multi-omics analyses and cell-based experiments support a link between GFPT2 and TGF-β/Smad2/3 pathway activation, indicating GFPT2 as a potential prognostic biomarker and therapeutic target.
Protein phosphorylation regulates diverse cellular processes, yet its analysis at the single-cell level remains challenging due to the low abundance of phosphoproteins. Here, we present a highly sensitive system for profiling phosphorylation of kinase substrates in individual cells. The method integrates fluorescence labeling of single-cell proteomes, immunoprecipitation using antibodies recognizing phosphorylation within specific amino acid motifs, miniaturized SDS-PAGE, and single-molecule detection using a custom-built light-sheet fluorescence microscope. We applied this approach to analyze substrates of casein kinase 2 (CK2) in HeLa cells treated with the phosphatase inhibitor calyculin A. Bulk and pseudo-single-cell analyses confirmed treatment-induced accumulation of phosphorylated CK2 substrates and demonstrated quantitative performance over biologically relevant input ranges. Importantly, true single-cell measurements revealed heterogeneous phosphorylation patterns across molecular weight regions, highlighting cell-to-cell variability in CK2 signaling that is obscured in bulk analyses. This platform enables profiling of the phosphorylation states of a wide range of kinase substrates in individual cells and provides a foundation for dissecting heterogeneous signaling dynamics.
RNA interference (RNAi) serves as a crucial tool for gene function research and pest control. Nevertheless, its application in lepidopteran insects is restricted by the low efficiency of gene silencing. Spodoptera litura (Lepidoptera: Noctuidae), a globally-distributed polyphagous agricultural pest, has developed multiple strategies to resist control measures. This situation highlights the urgent need for innovative strategies to promote functional genomics and molecular target characterization studies. In this study, a novel cuticular protein SlCP was identified, which is vital for epidermal development and larval survival. Silencing of SlCP gave rise to epidermal contraction, a reduction in body size, a decrease in feed intake, and larval mortality. To address the problem of low RNAi efficiency, imidazole-modified graphene quantum dots (IGQDs) were utilized as nanocarriers for the delivery of dsSlCP. IGQD mediated delivery led to a 4.65-fold greater reduction in SlCP transcript levels compared with naked double-stranded RNA and elevated larval mortality from 38.00% to 56.00%. These findings establish that SlCP functions in epidermal development and present a nanomaterial-assisted strategy that enhances RNAi delivery efficacy in S. litura.
Antimicrobial peptides (AMPs) are promising alternatives to conventional antibiotics against multidrug-resistant pathogens, but their application is limited by low yield, structural instability, and potential toxicity. In this study, we designed a hybrid peptide, Aur-Defb3, by fusing a bioinformatically optimized rainbow trout β-defensin-derived sequence with a conserved aurein motif. To improve predicted physicochemical properties, we introduced a Leucine-to-Phenylalanine (L-to-F) substitution into the defensin-derived segment, hypothesizing that the aromatic side chain may contribute to hydrophobic-core stability and membrane interaction. Recombinant Aur-Defb3 was expressed in Komagataella phaffii GS115 and purified by Ni-IDA affinity chromatography. HPLC analysis showed that the major purified peak accounted for 97.08% of the integrated peak area, and the concentration of the purified fraction was estimated by A280 measurement. Antimicrobial assays revealed antibacterial activity against both Gram-positive and Gram-negative bacteria, with MIC values ranging from 31.25 to 250 μg/mL (average 97.7 μg/mL). Hemolysis assays showed low hemolytic activity toward rainbow trout erythrocytes under the tested conditions, with approximately 10% hemolysis observed at 256 μg/mL. Scanning electron microscopy (SEM) showed concentration-dependent morphological damage to bacterial cells, including surface roughening, deformation, and disruption, suggesting that membrane damage may contribute to the antibacterial activity of Aur-Defb3. These results demonstrate the successful expression of Aur-Defb3, supporting its potential for further structural characterization, precise yield quantification, and in vivo mechanistic evaluation.
Pulmonary arterial hypertension (PAH) has responded favorably to the new fusion protein named sotatercept. This study aimed to evaluate the safety and efficacy of sotatercept in PAH. The online databases like PubMed/Medline, Cochrane Library, and ClinicalTrials.gov were searched until 15 September 2024 for studies comparing sotatercept to placebo using keywords such as "sotatercept, pulmonary arterial hypertension, and 'fusion protein.' Of the 213 titles searched, after removing duplicates, 142 studies were evaluated for eligibility. Two RCTs were finally included. Subgroup analysis was conducted for the two doses(0.7 mg/kg and 0.3 mg/kg). In the safety analysis, there were no significant differences in total adverse events[RR = 1.03(0.95, 1.12),I2 = 0,p = 0.50], serious adverse events[RR = 0.96(0.38, 2.42), I2 = 56, p = 0.93] and mortality[RR = 0.39(0.01, 11.81),I2 = 61%,p = 0.59]. Whereas, a significant difference was observed with non-serious adverse events[RR = 1.15(1.03-1.30),I2 = 0%,p = 0.02], telangiectasia[RR = 3.34(1.26-8.83),p = 0.02], thrombocytopenia[RR = 3.69(1.23-11.0),I2 = 0%,p = 0.02] and epistaxis[RR = 5.58(2.20-14.13), p = 0.0003]. No significant difference was observed with headache, nasopharyngitis, hypotension, neutropenia, nausea, diarrhea, rash, anemia, and dizziness. A significant change was observed with the majority of efficacy parameters, like Pro BNP[MD = -1150.38(-1846.44, -454.32),I2 = 79,p = 0.001], pulmonary vascular resistance[MD = -234.40(-325.30,-143.51), I2 = 72, p < 0.00001], pulmonary arterial wedge pressure(p = 0.007) and improvement in WHO functional class[OR = 3.23(1.25, 8.34), I2 = 61, p = 0.02]. This study demonstrates that sotatercept is more effective than placebo in treating PAH and has a fair safety profile.ClinicalTrial.gov Identifier numbers: NCT04576988 and NCT03496207 PROSPERO: CRD42024534695. Based on current research, the pooled analysis of the studies demonstrates that sotatercept is a well-tolerated and helpful treatment option for managing pulmonary arterial hypertension (PAH). Sotatercept does not seem to worsen or increase the risk of overall side effects or serious health issues. The more common side effects of headache, sore throat, hypotension, nausea, diarrhea, skin rash, anemia, and dizziness did not differ to a clinically meaningful extent. Physicians, however, must pay attention to low platelet counts in some patients owing to the potential for bleeding and elevated hemoglobin levels, which may require dose modification.Sotatercept has demonstrated promising results in enhancing several important health metrics, including decreasing pulmonary blood pressure, enhancing physical functioning, and reducing levels of pro-BNP, a sign of cardiac strain. Sotatercept was used along with other standard treatments of PAH in the study. Since the available evidence stems from a limited number of studies, further research involving larger cohorts and extended observational periods is essential for clarifying long-term safety.
Carrimycin is a macrolide antibiotic widely used in the pharmaceutical and clinical fields， with its primary composition consisting of three distinct derivatives of spiramycin. For decades， macrolides like carrimycin have been valued for their efficacy against Gram-positive bacteria and certain atypical pathogens. However， as research has progressed， recent studies have revealed that carrimycin， along with various other structurally modified spiramycin derivatives， exhibits significant antitumor activity in multiple experimental models. This finding suggests that the spiramycin molecular scaffold possesses intrinsic potential beyond its conventional antibacterial role. By strategically modifying the substituents attached to this core structure， it may be feasible to develop novel compounds with enhanced and more selective antitumor properties. Despite this promising outlook， research into the specific targets and comprehensive mechanisms underlying the antitumor effects of spiramycin derivatives remains notably insufficient. A major gap exists in the precise identification of their molecular targets within cancer cells and the detailed signaling pathways they modulate. This lack of mechanistic understanding poses a substantial barrier to the rational design， optimization， and clinical translation of spiramycin-based antitumor therapeutics. To systematically address this knowledge gap and elucidate the antitumor mechanism of this compound class， the present study adopted an activity-based protein profiling （ABPP） strategy. ABPP is a chemoproteomic approach that enables the direct identification of functionally active proteins that interact with small-molecule probes in a native biological context. As a first step， we designed and synthesized a novel， high-activity spiramycin derivative termed n-hexanoyl spiramycin （h-SPM）. Building upon the structure of h-SPM， we subsequently engineered and synthesized a structurally analogous activity-based probe. This probe was specifically functionalized with chemical handles （such as an alkyne group） compatible with ABPP methodologies， allowing for downstream bioorthogonal conjugation and enrichment steps. The experimental workflow began by incubating this active probe with live cancer cells. During this co-incubation period， the probe engaged with and covalently bound to its potential protein targets within the complex cellular environment. Following the interaction， cells were lysed， and the probe-labeled proteins were efficiently isolated and purified using affinity-based enrichment techniques-specifically via copper-catalyzed azide-alkyne cycloaddition （click chemistry） to immobilize them onto a solid support. The enriched protein pool was then subjected to in-depth analysis using liquid chromatography-mass spectrometry （LC-MS）. This analytical phase yielded detailed， proteome-wide information on the identities of proteins that interact with the h-SPM-based probe. To extract biological insights from the list of identified proteins， we performed comprehensive bioinformatic analysis using Gene Ontology （GO） enrichment. This systematic classification provided crucial information regarding the biological processes， molecular functions， and cellular components associated with the captured proteins. The functional annotations derived from GO analysis allowed us to evaluate and prioritize several promising candidate target proteins for further experimental validation. Through this integrated proteomic and bioinformatic approach， we successfully identified multiple potential cellular targets of h-SPM. Notable among these were amyloid precursor protein （APP） and low-density lipoprotein receptor （LDLR）， both of which are implicated in diverse cellular processes such as cell adhesion， signal transduction， and metabolic regulation. To confirm the biological relevance of these interactions， we employed Western Blotting experiments. These studies verified that the expression or post-translational modification states of APP and LDLR were altered in response to h-SPM treatment， thereby confirming their status as responsive molecular targets. After identifying and preliminarily validating APP as a key interactor， we proceeded to investigate its functional role in the drug's mechanism of action. Using short hairpin RNA （shRNA）-mediated protein knockdown， we generated cancer cell lines with significantly reduced APP expression. Comparative analysis of drug sensitivity between these knockdown cells and their wild-type counterparts revealed a marked attenuation of h-SPM's antitumor effects in the absence of APP. Complementary to this， cell staining assays， including immunofluorescence， were conducted to visualize morphological changes， alterations in cell viability， and the subcellular localization of relevant biomarkers following h-SPM treatment. Collectively， these functional experiments provided compelling evidence that APP plays a critical and indispensable role in mediating the antitumor activity of h-SPM. Our findings thus outline a preliminary model of the drug's mechanism， likely involving cellular pathways regulated or influenced by APP. In summary， this study achieves two significant objectives. First， it establishes and validates a robust， generalizable target-screening platform based on ABPP， specifically tailored for the investigation of spiramycin-class compounds. Second， and more importantly， it delivers novel biological insights by pinpointing specific protein targets such as APP. The identification of these targets provides invaluable mechanistic clues and a solid theoretical foundation for the future development of this family of compounds. This work effectively advances spiramycin derivatives from compounds with observed phenotypic activity toward agents with an emerging mechanistic understanding， thereby paving the way for more targeted drug design and informed combination therapy strategies in oncology. Future studies will focus on delineating the detailed downstream consequences of APP engagement by h-SPM and exploring the therapeutic potential of other identified targets in preclinical models. 可利霉素是一种在医药领域广泛使用的大环内酯类抗生素，主要成分为3种不同的螺旋霉素衍生物。然而随着相关研究的深入，近期发现可利霉素以及多种不同结构的螺旋霉素衍生物都具有一定的抗肿瘤活性，说明利用螺旋霉素作为母核，进行不同的取代基修饰，可能会得到更好的抗肿瘤活性药物。虽然螺旋霉素有着成为抗肿瘤药物的潜力，但是关于其作用靶点及完整机制的研究依然欠缺。为了揭示其抗肿瘤作用机制，本研究采用了基于活性的蛋白质组分析（ABPP）策略，设计并合成了一种高活性螺旋霉素衍生物正己酰螺旋霉素（h-SPM），进而依据其结构合成了一种结构相似且带有适用于ABPP实验官能团的活性分子探针。利用该探针与细胞蛋白质共孵育，使两者结合，从而捕获h-SPM潜在的药物靶点，随后对探针结合的蛋白质进行分离纯化与质谱分析，获得详细的靶点蛋白质信息。进一步通过基因本体（GO）分析挖掘这些蛋白质的性质和功能，得到明确的功能信息，并据此筛选出若干潜在靶点蛋白质。通过上述方法，从蛋白质谱的结果中鉴定出包括淀粉样前体蛋白（APP）、低密度脂蛋白受体（LDLR）在内的多个h-SPM潜在作用靶点，并采用免疫印迹（Western Blotting）方法确定这些蛋白质对h-SPM产生了响应。随后，通过短发夹RNA（shRNA）介导的蛋白质敲低及细胞染色等实验，证明蛋白质APP在该药物发挥作用的过程中扮演了关键角色，初步揭示了该类药物的抗肿瘤作用机制。本研究不仅建立了适用于螺旋霉素类化合物的靶点筛选方法，也为该类药物的后续开发提供了关键靶点线索与理论依据。
Glycosylation is a critical post-translational modification (PTM) in biotherapeutics that influences protein structure, stability, pharmacokinetics, biological activity, and immunogenicity. Comprehensive characterization of glycosylation and other PTMs is therefore important for biopharmaceutical development and quality assessment. Aflibercept is a recombinant fusion protein containing five N-glycosylation sites and multiple potential O-glycosylation sites. Although site-specific N-glycosylation of aflibercept has been previously investigated, integrated characterization of glycosylation heterogeneity and aspartic acid (Asp) isomerization using an electron-transfer/higher-energy collision dissociation (EThcD) based workflow remains limited. In this study, EThcD data-dependent MS2 (ddMS2) peptide mapping was performed on a Thermo Scientific Orbitrap Excedion Pro mass spectrometer equipped with the EASY-ETD option for detailed characterization of aflibercept. Site-specific N-glycosylation profiles at all five glycosylation sites were identified and relatively quantified. In addition, an O-glycopeptide localized at S12 was detected, providing direct experimental evidence supporting site-specific O-glycosylation in aflibercept. The EThcD fragmentation approach also enabled differentiation and localization of isoAsp residues through characteristic diagnostic fragment ions, including low-abundance peptides containing multiple Asp residues. Several additional PTMs, including deamidation and oxidation, were simultaneously characterized within the same analytical workflow. The results also indicate that EThcD peptide mapping can support the integrated multi-attribute method (MAM) concept for complex glycoproteins by combining glycosylation analysis, Asp/isoAsp differentiation, and PTM profiling in a single experiment. This workflow may be useful for detailed structural characterization and analytical comparability assessment of therapeutic glycoproteins and biosimilars.
As Southeast Asian countries advance toward malaria elimination, challenges such as residual transmission reservoirs and zoonotic spill-over require high-resolution micro-surveillance. While conventional diagnostics often miss hidden transmission foci in low transmission settings, serological assays provide an essential supplement by measuring both historical and recent exposure. This scoping review assesses the implementation of antibody surveillance across Southeast Asian countries and highlights its potential in the concerted effort towards regional malaria elimination. This review adhered to the PRISMA-ScR guidelines to identify peer-reviewed studies on malaria serology in Southeast Asian countries. Systematic searches of PubMed, Web of Science and Scopus were conducted for articles published between January 2000 and May 2025. Multiple independent reviewers screened titles, abstracts and full texts against predefined eligibility criteria, focusing on English-language primary studies involving human samples. Extracted data including study location, population, serological assay type, serological markers and key findings were synthesised to provide a comprehensive overview of malaria antibody surveillance. Of 6,849 records identified, 33 studies across seven Southeast Asian countries met the inclusion criteria. Study designs consisted of 26 cross-sectional, five longitudinal and one each of a prospective cohort and a control trial. Over time, research focuses have shifted from high-transmission assessments to low-transmission and elimination contexts, increasingly utilising multiplex assays and protein microarrays. While Plasmodium falciparum and Plasmodium vivax markers predominated, recent studies have incorporated Plasmodium knowlesi and vector exposure antigens. Furthermore, seroconversion rate estimation and machine learning approaches effectively identified historical transmission declines and residual hotspots missed by conventional microscopy or molecular methods. Serological surveillance offers a transformative supplement to the diagnostic gaps prevalent in Southeast Asia's low-transmission settings. By generating a high-resolution exposure profile, advanced seroepidemiological tools enable precise micro-stratification of transmission foci, the differentiation of recent from historical exposure and the targeted management of zoonotic spill-over. Ultimately, integrating these methods provides a robust, cost-effective framework for identifying high-risk populations, verifying transmission interruption and achieving regional malaria-free certification.
The epigenetic control of immune responses plays a crucial role in the development and progression of cancer. The need to identify biomarkers and create new predictive models is crucial to reliably estimate response rates in patients receiving tumor immunotherapy, which are currently low. We screened for variably methylated loci associated with immunotherapy response, focusing on relevant pathways. The expression of immunotherapy-related methylation loci was analyzed in tissues, and quantitative trait locus (QTL) features were summarized. Relationships among tumor mutational Burden (TMB), neoantigen (NeoAg), programmed death-ligand 1 (PD-L1), and immunotherapy efficacy were examined. Motif analysis identified sequence preferences linked to methylation. Six machine learning models were constructed and compared to select the optimal.Due to the lack of external validation datasets, subgroup analysis was conducted. The five CpG loci showing the strongest response to immunotherapy were cg00045061, cg00107488, cg00056433, cg00090974, and cg00072957. Differentially methylated sites enriched the ubiquitin-proteasome pathway, with most loci located in the N shore region of CpG islands. GO enrichment highlighted microvillus length modulation and CXCR4 chemokine receptor binding. The support vector machine (SVM) model exhibited optimal performance, with precision of 0.796, accuracy of 0.833, F1-score of 0.663, recall of 0.55 and AUC of 0.894.The optimal SVM model achieved an AUC of 0.85 for melanoma dataset and 0.75 for lung cancer dataset. Tumor methylation sites hold promise as predictive biomarkers for immunotherapy efficacy. The SVM model is the optimal machine learning approach for predicting methylation sites in immunotherapy. Immunotherapy is a treatment that helps the body’s own immune system fight cancer. However, it does not work well for everyone, and doctors need better ways to predict which patients will benefit. This study looked at DNA methylation, which can turn genes on or off without changing the DNA sequence itself. We found five specific methylation sites that are linked to how well patients respond to immunotherapy. These markers are involved in important cell processes, including protein breakdown and immune cell signaling. We also built a computer model using a method called support vector machine that can predict treatment outcomes based on these methylation patterns. This model performed very well and also worked on different types of cancer, including melanoma and lung cancer. Our findings suggest that these DNA methylation markers could help doctors choose the right treatment for each patient and improve the success of immunotherapy.This study provides new ideas for predicting future immunotherapy.
ObjectivePost-lumbar puncture headache is a common complication following lumbar puncture. Although several risk factors have been established, the role of low body mass index remains controversial. This study aimed to evaluate the association between body mass index and post-lumbar puncture headache in Chinese adults undergoing lumbar puncture.MethodsIn total, 499 patients who underwent lumbar puncture at the Department of Neurology between January 2022 and March 2024 were enrolled. Data on demographic characteristics, blood pressure, lumbar puncture history, and cerebrospinal fluid parameters were collected. Univariate and multivariate logistic regression models were employed to assess the relationship between body mass index and post-lumbar puncture headache. Subgroup analyses were conducted to evaluate the consistency of this association across sex, age, blood pressure, cerebrospinal fluid volume, and cerebrospinal fluid protein levels.ResultsPost-lumbar puncture headache occurred in 13% (65/499) of patients; among these, 63.1% were female. Compared with those without post-lumbar puncture headache, those with a headache were younger (p < 0.001), had lower body mass index (p = 0.003), lower systolic and diastolic blood pressures (p = 0.001 and p = 0.001), lower cerebrospinal fluid protein level (p < 0.001), and greater cerebrospinal fluid collection volume (p < 0.001). No significant association was found between primary disease classification and post-lumbar puncture headache (p = 0.164). In univariate logistic regression analysis, the risk of post-lumbar puncture headache significantly decreased with increasing body mass index values (odds ratio = 0.9, 95% confidence interval: 0.84-0.97, p = 0.003). After multivariable adjustment for sex, age, blood pressure, cerebrospinal fluid protein level, and cerebrospinal fluid volume, the association remained significant (odds ratio = 0.92, 95% confidence interval: 0.85-0.99, p = 0.033). Further analysis, in which patients were grouped by body mass index levels and assessed using two multivariate regression models, revealed that individuals with a low body mass index had a significantly higher risk of post-lumbar puncture headache. In Model I, adjusted for sex and age, the odds ratio was 3.2 (95% confidence interval: 1.41-7.23, p = 0.005); in Model II, further adjusted for systolic pressure, diastolic pressure, cerebrospinal fluid protein level, and cerebrospinal fluid collection volume, the odds ratio was 3.14 (95% confidence interval: 1.36-7.25, p = 0.007). The findings were robust across both models. The findings were consistent in the subgroup analyses.ConclusionLower body mass index is significantly associated with an increased risk of post-lumbar puncture headache. These findings underscore the importance of considering body mass index in preoperative risk assessment and postoperative management for patients undergoing lumbar puncture. However, due to the cross-sectional study design, causal inference cannot be established.
ObjectiveTo examine associations between systemic inflammation, sex hormones, and vitamin D levels in U.S. women.MethodsA cross-sectional study was conducted using NHANES 2021-2023 data from 3179 women aged 18-80 years. Participants were grouped by high-sensitivity C-reactive protein (hs-CRP) quartiles. Demographic, anthropometric, lifestyle, dietary, and clinical variables were analyzed. Serum biomarkers included hs-CRP, sex hormones, lipid parameters, and 25-hydroxyvitamin D3 (25OHD3). Multivariable linear regression assessed independent associations between hs-CRP and endocrine markers after adjustment for confounders.ResultsWomen in the highest hs-CRP quartile had higher age, body weight, waist circumference, body mass index, and prevalence of hypertension and diabetes than those in the lowest quartile. Higher hs-CRP levels were associated with lower concentrations of 17α-hydroxyprogesterone, androstenedione, anti-Müllerian hormone (AMH), dehydroepiandrosterone sulfate, follicle-stimulating hormone (FSH), sex hormone-binding globulin, and 25OHD3. After adjustment, hs-CRP remained positively associated with AMH (β = 0.212, p = 0.007), estrone (β = 0.142, p = 0.048), low-density lipoprotein cholesterol (β = 0.241, p = 0.002), and total cholesterol (β = 0.224, p = 0.001), and inversely associated with FSH (β = -0.194, p = 0.039) and luteinizing hormone (β = -0.191, p = 0.019).ConclusionSystemic inflammation was associated with reproductive hormones and lipid profiles, while many associations weakened after adjustment for adiposity. These findings highlight complex links between inflammation, endocrine function, and cardiometabolic health, influenced by obesity-related factors.
Current licensed influenza vaccines primarily protect by eliciting antibodies against the viral hemagglutinin (HA) glycoprotein, thereby blocking viral attachment and fusion with host cells. Unlike most vaccines, influenza vaccines must be administered annually because circulating viruses undergo continuous antigenic drift and population antibody titers wane over time. Despite yearly reformulation, influenza vaccine effectiveness remains highly variable, often below 45%, largely due to antigenic mismatches. These mismatches arise from ongoing HA evolution following strain selection and from egg-adaptation during production or propagation in animal cell cultures, which can alter key HA epitopes relative to circulating strains. Even when an antigenic match is favorable, repeated annual vaccination may elicit immunological phenomena that attenuate protective responses. Serial vaccination in young and older adults can increase regulatory T-cell activation, reducing vaccine-induced antibody titers. In older adults, this may be compounded by age-associated CD4+ T-cell memory populations that have reduced capacity to activate HA-specific B cells. While natural influenza infection induces durable memory B cells, conventional vaccination does not reliably generate such long-lived memory, suggesting a fundamental limitation of current vaccine platforms. Collectively, these observations underscore the need to re-evaluate influenza vaccination strategies, particularly to improve protection in high-risk groups such as older adults. Reducing antigenic mismatch remains essential and can be facilitated by improving the vaccine selection process and expanding the use of recombinant protein and mRNA vaccine production platforms that do not rely on egg- or animal-cell culture technologies. In parallel, the rational selection and development of adjuvants that minimize T-regulatory cell induction while enhancing durable memory B-cell formation and long-lived plasma cells may help overcome the immunological constraints associated with repeated annual vaccination. Beyond active immunization, complementary countermeasures are critical for mitigating severe outcomes, including hospitalizations and deaths. Antiviral drugs and monoclonal antibodies, especially those engineered for extended in vivo half-life, represent important adjuncts for protecting vulnerable populations such as the elderly, young children, and immunocompromised individuals. Strengthening and advancing these modalities should be prioritized as part of an integrated strategy to improve influenza control and reduce the global burden of disease.
Chitosan (CS) is widely used as a biodegradable packaging material in the food industry; however, integration of ultrasound-generated nanoemulsions into a functionally stratified CS architecture, has rarely been reported. Herein, bilayer films were fabricated consisting of a chitosan support layer and an ultrasound-assisted chitosan/lecithin/cinnamon essential oil (C/L/CEO) nanoemulsion top layer, with CEO incorporated at 2-6 minimum inhibitory concentrations (MIC). Among the formulations, the C/L/CEO1-C bilayer film (0.25 mg CEO per 100 mL nanoemulsion) exhibited optimal properties, including high tensile strength (45.6 MPa), elongation at break (33%), low water vapor permeability (1.78 × 10-10 g·m-1·s-1·Pa-1), low solubility (14.4%), excellent UV-blocking capacity, and enhanced hydrophobicity (water contact angle 101.33°). The films effectively inhibited Staphylococcus aureus and Escherichia coli O157:H7, and extended the refrigerated shelf life of salmon to 12 days. Increasing CEO concentration reduced lipid oxidation but promoted protein oxidation, highlighting the need to balance antimicrobial efficacy and oxidative stability.
Poor nutritional status is a modifiable risk factor that has been shown to have adverse outcomes in spine surgery, including higher rates of complications, poorer functional outcomes, longer hospital stays, and increased healthcare costs. As such, interest has emerged in clinical practice regarding the use of tools and important nutrient profiles that can be used to assess nutritional status both pre- and postoperatively. The purpose of this review is to synthesize the current evidence surrounding the clinical utility of various nutritional screening tools, as well as to evaluate the importance of targeted interventions. These strategies include protein and amino acid supplementation, vitamin D optimization, and preoperative carbohydrate loading; prior studies have associated these interventions with improved fusion rates, lower infection risk, and accelerated recovery. Furthermore, economic analyses in the setting of nutritional optimization are explored. Despite these benefits, substantial barriers like inconsistent protocols and patient nonadherence remain. In addition, reliance on single markers like albumin may be misleading due to inflammatory confounding, highlighting the need for multifactorial assessment that incorporates surrogates of bone quality and baseline health status. As the spine surgical population ages and procedures grow more complex, preoperative nutritional optimization represents a low-risk strategy with potential for substantial advancements in patient recovery. This review advocates for the standardization of multidisciplinary nutritional protocols as a key component of comprehensive perioperative spine care.
The Complete Ammonium and Nitrate removal via Denitratation-Anammox over Nitrite (CANDAN) process is a promising low-carbon biological nitrogen removal technology. Coupling CANDAN with hydroxyapatite (HAP) precipitation can enhance granular sludge strength and biomass retention. However, prolonged operation under calcium-rich conditions may induce excessive mineral accumulation, resulting in granule calcification, mass transfer limitation, and deterioration of nitrogen removal performance. This study systematically evaluated low-intensity ultrasound (LIU) as a recovery strategy for calcified CANDAN granular sludge and elucidated the underlying structural mechanisms. A sequencing batch reactor seeded with calcified CANDAN-HAP granules was operated under LIU-assisted recovery conditions. After 133 days, complete functional restoration was achieved, with NH₄⁺-N removal efficiency increasing from 21.08 ± 3.8% to 83.02 ± 2.4%, TN removal efficiency reaching 90.21 ± 3.9%, and negligible NO₂⁻-N accumulation during stable operation. Multi-scale structural analyses revealed that calcification was dominated by the formation of a dense extracellular polymeric substances (EPS)-inorganic "wall" on the granule surface. This structure, enriched with protein-dominated tightly bound EPS (T-EPS), occluded surface pores and restricted substrate diffusion into the granule interior. LIU progressively disrupted this diffusion barrier, transforming granule surfaces from compact and impermeable to porous and heterogeneous, while reconstructing internal pore networks. Anaerobic ammonium-oxidizing bacteria (AnAOB) preserved within granule cores rapidly regained activity after mass transfer was restored. Microbial community analysis further demonstrated that LIU caused only transient community restructuring, with functional populations stabilizing during long-term operation. Overall, this study demonstrates that LIU is a low-damage method restoring calcified CANDAN granules primarily through structural reconfiguration and mass transfer recovery, providing a practical strategy for mitigating calcification risks and maintaining stable long-term operation of granular nitrogen removal systems.
To meet the rising food demands of our growing population, the Food and Agriculture Organization proposed edible insects as sustainable sources of animal protein. Although hundreds of million people already consume insects around the tropics, western societies remain averse to entomophagy. To trace whether ancient Europeans consumed insects, we here apply two complementary genomic approaches. Metagenomic screening on 745 ancient anatomically modern human dental calculus returned limited insect DNA traces, with read abundances well below those observed in Neanderthals, western chimpanzees, and gorillas. In addition, genes encoding stomach-expressed chitinases show two of the most significant signatures of latitudinal differentiation genome-wide. Clines are consistent with evolutionary benefits of entomophagy in tropical regions and with expression quantitative trait locus data supporting low chitin digestibility in present-day Europeans. Ancient genomes confirm that both clines already existed at the onset of agriculture and persisted despite massive migrations. Together, our findings support occasional and possibly incidental insect consumption in Europe over the past ~9000 years.
Enzymes play a crucial role in metabolism. The rate at which enzyme sequence data is emerging far exceeds the rate at which we can measure the associated catalytic constants, posing a significant problem for the advancement of the fields of metabolic engineering and synthetic biology. Here, we present KcatNeuroCortex, an interpretable and novel deep learning framework for enzyme catalytic efficiency prediction. The proposed architecture combines Bi-directional Gated Recurrent Units (Bi-GRU) with a multi-attention mechanism designed to mirror how enzymes work. It first captures local functional motifs through a segmentation-based strategy, and then integrates these into a global representation of long-range interactions that shape the catalytic landscape, leading to improved interpretability and prediction accuracy. KcatNeuroCortex achieves good results, with R² values of 0.74 and RMSE values of 0.77, representing a 57% improvement compared to DLKcat. We demonstrate that our segmentation-aware, attention-enhanced approach achieves competitive performance compared to conventional sequence-based models, especially on diverse and low-similarity sequences. The framework is robust, scalable, and interpretable, making it a valuable tool for enzyme engineering and large-scale kinetic parameter estimation. Also, the work establishes that deep learning can move beyond prediction to provide a biologically meaningful understanding of enzyme catalysis, positioning KcatNeuroCortex as a very valuable tool for the enzyme engineering community.