Wiley最新文献

Romidepsin (RMD), an FDA-approved histone deacetylase inhibitor for treating cancer, demonstrates potent latency-reversing activity in vitro but has yielded suboptimal outcomes in kick-and-kill HIV-cure trials when administered as a 5 mg/m² infusion. We hypothesized that distinct pharmacokinetic targets exist between the pharmacodynamic goals of cytotoxicity for cancer therapy and latency reversal for HIV cure. To address this question, we designed an empirical modeling framework incorporating patient pharmacokinetics as a New Approach Methodology. Using clonal- and primary cell models of HIV-1 latency with readouts for different stages of latency-reversing activity, we performed exposure-response and dose-fractionation studies to define pharmacokinetic drivers of efficacy in latency, then designed tailored dosing strategies to meet these targets in static- and dynamic-culture systems, including a hollow-fiber model. Across all systems, the only consistent pharmacokinetic parameter driving latency-reversing activity, was RMD exposure time, with maximal activity achieved in cells exposed to ~4-6 ng/mL RMD for ≥ 20 hours (AUCs of ~80-120 ng*hour/mL). With current clinical-dosing strategies achieving AUCs of 240-600 ng*hour/mL, the same outcome occurred only when RMD concentrations greatly exceeded a cytotoxicity threshold, suggesting response was a byproduct of cytotoxic processes rather than targeted latency reversal. These findings indicate that conventional oncologic dosing strategies for RMD might not be directly applicable to HIV-1 latency reversal, offering a mechanistic explanation for suboptimal clinical outcomes. We provide proof-of-concept for RMD's limited clinical efficacy and propose a "low and slow" dosing strategy targeting time-driven, toxicity-minimized drug exposure, demonstrating how in vitro pharmacokinetic/pharmacodynamic investigations applying clinical pharmacokinetic data may inform dose optimization for clinical trials of latency reversal.

Background and aims: Dual use of cigarettes and e-cigarettes is often employed by smokers as a harm reduction strategy. This study evaluated the harm reduction potential of e-cigarettes by investigating changes in urinary biomarker of exposure (BOE) to tobacco-specific nitrosamines (TSNAs) and nicotine among smokers transitioning to dual use.

Methods: Data from 8688 adult smokers in waves 1 to 5 of the Population Assessment of Tobacco and Health Study (United States) were analyzed. 'Transitioned dual users' were defined as those changing from exclusive cigarette smoking to dual use across two consecutive waves. BOE levels were assessed before and after this transition and compared with those who continued smoking cigarettes exclusively ('remained smokers') across the same time frame. Propensity score weighting was used to adjust for baseline confounders. Further analyses were stratified by baseline cigarette consumption, dichotomized at the cohort median into heavy and light smoking groups.

Results: Observational analysis revealed that transitioned dual users experienced a statistically significant reduction in TSNAs without a statistically significant change in nicotine. The regression analysis comparing transitioned dual users with 'remained smokers' showed that transitioning to dual use was statistically significantly associated with a 13% decrease in 4-(methylnitrosamino)-1-(3-pyridyl)-1-butanol (NNAL) [95% confidence interval (CI) = -20% to -6%] and a 10% decrease in N'-nitrosonornicotine (NNNT) (95% CI = -15% to -4%). In contrast, dual use was statistically significantly associated with a 17% increase in total nicotine equivalents-2 (95% CI = 12%-23%) and 8% increase in total nicotine equivalents-6 (95% CI = 5%-12%). Stratified analysis further showed that the association between dual use and TSNAs reduction was attenuated among the baseline heavy smoking group (>13 cigarettes/day), with one-fourth of the effect estimates and no statistical significance.

Conclusions: Smokers transitioning to dual use alongside e-cigarettes appear to show reductions in tobacco-specific nitrosamines exposure, particularly among lighter smokers, but do not appear to show reductions in nicotine exposure.

Aims: To quantify changes in carbohydrate intake, insulin delivery and glycaemia across menstrual cycle phases in women with type 1 diabetes using the MiniMed 780G automated insulin delivery system (MM780G).

Materials and methods: Women on MM780G provided menstrual cycle data. Continuous glucose monitoring and insulin delivery data were retrieved from the CareLink portal. Daily values for time in range (TIR, 3.9-10 mmol/L), total carbohydrates, total basal, total bolus and total insulin were calculated. Menstrual phases were classified using two methods: Method 1 compared the pre-menses week with the menses week; Method 2 compared five physiological phases with mid-follicular as reference. Linear mixed-effects models with random intercepts for participant and cycle were fitted.

Results: Fifteen women (mean age 35.9 years, BMI 23.3 kg/m², HbA1c 50.8 mmol/mol [6.8%]) contributed 118 cycles. In the pre-menses week, carbohydrate intake (β = +0.164 g kg^-1, adjusted p = 0.003), total daily bolus (β = +0.018 U kg^-1, adjusted p = 0.001), total daily basal (β = +0.011 U kg^-1, adjusted p = 0.003) and total daily dose (β = +0.029 U kg^-1, adjusted p < 0.001) were significantly higher than the menses week, yet TIR was lower (β = -3.487 percentage points, adjusted p < 0.001). In Method 2, total daily basal and total daily dose were significantly higher in the late luteal phase, and TIR was lower in the mid luteal phase (β = -3.34 percentage points, adjusted p = 0.007).

Conclusions: Menstrual cycle phase is associated with significant changes in carbohydrate intake, insulin delivery and TIR in women using MiniMed 780G. Despite increased insulin delivery, TIR was lower in the luteal phase, suggesting partial adaptation to menstrual cycle-related physiological changes.

Excessive nitrogen (N) application can affect soil health and crop performance, yet the multi-generational effects of prolonged high-N exposure on wheat seed germination remain unclear. We profiled seed metabolomes and endophytic bacterial communities across seven consecutive wheat generations (F1-F7) under normal-N and high-N regimes, and assessed germination of F7 seeds. Spatial metabolomics, spatial transcriptomics and 16S rRNA sequencing were used to resolve embryo- and endosperm-specific responses. Seven generations of high-N treatment delayed progeny seed germination and decreased the germination index by 12.3%, whereas the vigor index was not significantly affected. Spatial omics revealed strong embryo-endosperm heterogeneity and a high-N-associated shift in endosperm metabolism, including a 73.9% decrease in endosperm L-aspartic acid. Compartment-resolved microbiome analysis showed enrichment of Bacillus under high-N exposure. Functional assays showed that L-aspartic acid promoted germination/early outgrowth, whereas Bacillus grew with L-aspartic acid as the sole C/N source and reduced germination under L-aspartic acid-supplemented conditions. These results support a model in which multi-generational high N reduces wheat seed germination through endosperm amino-acid depletion and altered endophytic bacterial assembly.

Botrytis cinerea is a devastating necrotrophic pathogen threatening global agriculture. Although modulating light intensity holds promise for sustainable disease management, the regulatory strategies and underlying mechanisms remain largely undefined. Three light intensity levels (200, 500 and 800 µmol·m^-2·s^-1) were applied in this study. By integrating multilevel pathological and cytological observations with transcriptional and metabolic pathway analyses, we systematically investigated how light intensity regulates defence responses against B. cinerea in tomato (Solanum lycopersicum). The results revealed that 200 µmol·m^-2·s^-1 light most effectively attenuated disease progression by orchestrating a multilayered defence network. This network encompasses stomatal closure to restrict pathogen invasion, phenylalanine-derived lignin biosynthesis for cell wall reinforcement, upregulation of pathogenesis-related proteins to facilitate pathogen degradation and maintenance of redox homeostasis to mitigate oxidative damage. These coordinated responses, rather than direct pathogen inhibition by light intensity, collectively restrict B. cinerea invasion. Application of exogenous phenylalanine and the PAL inhibitor 2-aminoindan-2-phosphonic acid (AIP) further validated that light intensity dictates metabolic flux partitioning within the phenylpropanoid pathway, a core mechanism for enhanced resistance. This study provides the first evidence that low-light-induced resistance to B. cinerea stems from phenylpropanoid metabolic reprogramming and ROS homeostasis, offering novel insights into light-modulated plant defence against necrotrophic pathogens.

Loquat (Eriobotrya japonica) regeneration and transformation are difficult, hindering gene function studies. To overcome this limitation, we developed a rapid and stable regeneration and transformation system for hairy roots in loquat by optimising key conditions, in order to explore the potential functions of key genes involved in root growth and development. LEAFY COTYLEDON 2 (LEC2) is a central hub linking embryonic development, lipid synthesis, root system development, and hormonal regulation. However, the role of the LEC2 protein in regulating root growth and development in loquat remains unclear. This study demonstrates a specific interaction between LEC2 and FUSCA3 (FUS3) in loquat, with FUS3 acting downstream of LEC2. Overexpression of LEC2 or FUS3 showed longer root length and increased root numbers, while loss-of-function mutants of either LEC2 or FUS3 exhibited shorter root lengths and reduced root numbers, and overexpression of FUS3 effectively compensated for the root growth defects in LEC2 knockout mutants. In summary, the stable Agrobacterium rhizogenes-mediated transformation system provides an effective approach for gene function analysis and genetic engineering research in loquat, and the LEC2-FUS3 complex functions as a key regulator in auxin-regulated root growth and development.

Here, we evaluated foliar-applied silicon quantum dots (SiQDs) with different surface charges on soybean salt tolerance. Positively charged SiQDs (P-SiQDs) exhibited stronger foliar retention and penetration than negatively charged SiQDs (N-SiQDs), resulting in a 35.3% higher silicon accumulation in leaves. Under 200 mM NaCl stress, foliar application of P-SiQDs increased shoot dry weight and reduced the Na⁺/K⁺ ratio. Compared with N-SiQDs, P-SiQDs more effectively alleviated salt-induced damage to thylakoid ultrastructure and improved photosynthetic performance. A life cycle field pot trial further showed that SiQDs increased the 100-seed weight by 20.1%-25.9% under salt stress. Leaf metabolomics showed significant alterations in lipid metabolite pathways associated with redox homeostasis, accompanied by shifts in the phyllosphere microbiome. P-SiQDs increased the abundance of Chloroflexota, Actinomycetota, and genera such as Paenarthrobacter, Variovorax, and Xanthobacter. Meanwhile, the phyllosphere microbiome shifted toward life-history strategies related to growth, resource acquisition, and salt stress tolerance. In addition, P-SiQDs promoted root growth, nodulation, and nitrogenase activity, leghemoglobin content, and total nitrogen accumulation, together with changes in root exudate composition and the enrichment of salt-tolerant rhizosphere bacteria, including Sphingomonas and Novosphingobium. These findings indicate that surface charge modification is an effective strategy to enhance the efficiency of foliar nano-fertilizer application.

Vegetable cultivation is heavily reliant on an ample water supply, with water deficiency limiting seedling growth and final yield. The phytohormone abscisic acid (ABA) is recognised for its role in drought tolerance by promoting stomatal closure to reduce water loss. This study identifies that drought treatment upregulated the C2H2-type zinc finger protein SlZAT5 and the bHLH transcription factor SlJIG, leading to enhanced ABA accumulation and drought tolerance. Immunoprecipitation-Mass Spectrometry (IP-MS) analysis revealed the interaction between SlZAT5 and SlJIG. Both transcription factors can bind to the promoter of the ABA catabolism gene SlCYP707A3. However, SlJIG alone was insufficient for transcriptional repression of SlCYP707A3, which instead required SlZAT5's transrepression activity. Additionally, SlJIG strengthened the inhibitory effect of SlZAT5 on SlCYP707A3. Overexpression of either SlZAT5 or SlJIG improved tomato drought tolerance, whereas knockout of either gene reduced drought tolerance, with the double mutant exhibiting a further decline. These findings elucidate the role of the SlZAT5-SlJIG module in regulating tomato drought responses via ABA homoeostasis.