Potentiating dual-directional immunometabolic regulation with nanomedicine to enhance anti-tumor immunotherapy following incomplete photothermal ablation.

Photothermal therapy (PTT) is a promising cancer treatment method due to its ability to induce tumor-specific T cell responses and enhance therapeutic outcomes. However, incomplete PTT can leave residual tumors that often lead to new metastases and decreased patient survival in clinical scenarios. This is primarily due to the release of ATP, a damage-associated molecular pattern that quickly transforms into the immunosuppressive metabolite adenosine by CD39, prevalent in the tumor microenvironment, thus promoting tumor immune evasion. This study presents a photothermal nanomedicine fabricated by electrostatic adsorption among the Fe-doped polydiaminopyridine (Fe-PDAP), indocyanine green (ICG), and CD39 inhibitor sodium polyoxotungstate (POM-1). The constructed Fe-PDAP@ICG@POM-1 (FIP) can induce tumor PTT and immunogenic cell death when exposed to a near-infrared laser. Significantly, it can inhibit the ATP-adenosine pathway by dual-directional immunometabolic regulation, resulting in increased ATP levels and decreased adenosine synthesis, which ultimately reverses the immunosuppressive microenvironment and increases the susceptibility of immune checkpoint blockade (aPD-1) therapy. With the aid of aPD-1, the dual-directional immunometabolic regulation strategy mediated by FIP can effectively suppress/eradicate primary and distant tumors and evoke long-term solid immunological memory. This study presents an immunometabolic control strategy to offer a salvage option for treating residual tumors following incomplete PTT.

Induction of DEPP1 by HIF Mediates Multiple Hallmarks of Ischemic Cardiomyopathy.

BACKGROUND: HIF (hypoxia inducible factor) regulates many aspects of cardiac function. We and others previously showed that chronic HIF activation in the heart in mouse models phenocopies multiple features of ischemic cardiomyopathy in humans, including mitochondrial loss, lipid accumulation, and systolic cardiac dysfunction. In some settings, HIF also causes the loss of peroxisomes. How, mechanistically, HIF promotes cardiac dysfunction is an open question. METHODS: We used mice lacking cardiac pVHL (von Hippel-Lindau protein) to investigate how chronic HIF activation causes multiple features of ischemic cardiomyopathy, such as autophagy induction and lipid accumulation. We performed immunoblot assays, RNA sequencing, mitochondrial and peroxisomal autophagy flux measurements, and live cell imaging on hearts and isolated cardiomyocytes. We used CRISPR-Cas9 gene editing in mice to validate a novel mediator of cardiac dysfunction in the setting of chronic HIF activation. RESULTS: We identify a previously unknown pathway by which cardiac HIF activation promotes the loss of mitochondria and peroxisomes. We found that DEPP1 (decidual protein induced by progesterone 1) is induced under hypoxia in a HIF-dependent manner and localizes inside mitochondria. DEPP1 is both necessary and sufficient for hypoxia-induced autophagy and triglyceride accumulation in cardiomyocytes ex vivo. DEPP1 loss increases cardiomyocyte survival in the setting of chronic HIF activation ex vivo, and whole-body Depp1 loss decreases cardiac dysfunction in hearts with chronic HIF activation caused by VHL loss in vivo. CONCLUSIONS: Our findings identify DEPP1 as a key component in the cardiac remodeling that occurs with chronic ischemia.

Association between Plant-based Diet and Risk of Chronic Diseases and All-Cause Mortality in Centenarians in China: A Cohort Study.

Background: Numerous studies have suggested the health benefits of a plant-based dietary pattern. However, whether this dietary pattern is associated with health benefits for centenarians remains unexplored. Our study aimed to investigate the correlation between 16 widely consumed Chinese food items and the incidence rates of chronic diseases and all-cause mortality among centenarians. Methods: We conducted a dietary survey on 3372 centenarians with an average age of 102.33 y in China. After rigorous screening, we identified 2675 centenarians, who underwent a 10-y follow-up study with all-cause mortality as the primary outcome. We developed 6 dietary patterns on the basis of the food consumption frequency of each participant. To model the impact of missing values, we employed multiple imputation methods, verifying the robustness of models. Results: The overall plant-based diet index (PDI), healthy plant-based diet index (hPDI), unhealthy plant-based diet index (uPDI), healthy plant-based foods index (HPF), unhealthy plant-based foods index (uHPF), and animal-based foods index (AF) scores among centenarians in China were 46.95 ± 6.29, 44.43 ± 5.76, 51.09 ± 6.26, 21.63 ± 4.79, 9.91 ± 2.41, and 14.59 ± 3.58, respectively. High scores of PDI, hPDI, and HPF were associated with a lower risk of chronic diseases. In the 10-y follow-up study, 92.90% of centenarians have died. The high scores of the PDI (HRPDI = 0.81), hPDI (HRhPDI = 0.79), and HPF (HRHPF = 0.81) scores were significantly associated with a lower risk of death compared with the low scores. Conversely, the high AF score (HRAF = 1.17) was significantly associated with a higher risk of death compared with the low scores. Conclusion: Despite the fact that a higher score in both a predominantly plant-based dietary pattern and a healthy dietary pattern can decrease the death among centenarians, not all HPFs have this effect. A higher AF predicted a higher risk of mortality, whereas higher PDI, hPDI, and HPF were associated with a lower risk of mortality among Chinese centenarians.

Decomposition analysis of the difference in depressive symptoms between urban and rural employed people in China: Unpaid work plays an important role.

OBJECTIVE: Focusing on the relationship between unpaid labor and the occurrence of depressive symptoms, this study aimed to explore the factors influencing the inequality of depressive symptoms and their contribution among Chinese urban and rural employed people. METHODS: This study utilized the 2020 China Family Panel Studies' national resampling data. Multivariate logistic regression was used to explore the factors influencing the occurrence of depressive symptoms among employed persons in urban and rural areas in China, respectively. Fairlie decomposition was used to explore the contribution of influencing factors such as unpaid labor to the difference in the occurrence of depressive symptoms between urban and rural areas. RESULTS: About 2,136 (21.70%) participants had depressive symptoms, of which 1,197 (24.75%) rural employed people had depressive symptoms and 939 (18.75%) urban employed people had depressive symptoms. The results of Fairlie decomposition analysis showed that 70.51% of the differences in depressive symptoms between urban and rural Chinese employed people could be explained by the covariates included in this study, including education level (52.44%), age (-11.91%), housework time (10.42%), self-rated health status (10.22%), self-rated income status (2.53%), exercise (2.36%), job satisfaction status (1.99%), chronic disease status (1.90%), and marital status (1.79%). CONCLUSION: This study found that the proportion of depressive symptoms was lower among urban employed residents than among rural employed residents. This difference was mainly caused by unpaid labor time, socioeconomic status, personal lifestyle, and health status. Housework, which is one of the unpaid labor, contributed to this depressive symptom difference in the third place.

Relationship between positive parenting and adolescents' depressive symptoms: Evidence from a Chinese research survey.

BACKGROUND: Adolescents often carry their depression well into their adulthood. This creates perpetual difficulties for their family and society. Research on the relationship between positive parenting and adolescent depressive symptoms is rare. The protective effect of positive parenting on adolescent depressive symptoms also remains underexplored. Parents are a vital source of feedback that shapes adolescents' self-view in crucial ways. AIMS: This study examines the latent relationships between four factors related to positive parenting and adolescent depressive symptoms. METHOD: Using data from the Chinese Family Panel Studies (CFPS), Stata MP 17.0 was used for preliminary data processing and descriptive statistics. The structural equation model (SEM) was adopted to test the seven proposed hypotheses. RESULTS: The study participants were 2,816 adolescents (52.34% male). The SEM showed that positive communication and parental praise can directly reduce depressive symptoms in adolescents (path coefficients of -0.24 and -0.13 [p < .001], respectively). Additionally, both positive communication and positive parent-adolescent interactions can reduce adolescents' depressive symptoms by heightening the intermediate factor of parental praise (path coefficients of 0.30 and 0.44 [p < .001], respectively). Conversely, positive parent-adolescent interactions did not negatively affect adolescents' depressive symptoms, as we hypothesized. CONCLUSIONS: High level of positive parenting negatively predicts the level of depressive symptoms among adolescents. Specifically, positive communication, positive parent-adolescent communication, and parental praise are the main protective factors related to positive parenting for adolescents' depressive symptoms.

HaMYBA-HabHLH1 regulatory complex and HaMYBF fine-tune red flower coloration in the corolla of sunflower (Helianthus annuus L.).

Sunflowers are well-known ornamental plants, while sunflowers with red corolla are rare and the mechanisms underlying red coloration remain unclear. Here, a comprehensive analysis of metabolomics and transcriptomics on flavonoid pathway was performed to investigate the molecular mechanisms underlying the differential color formation between red sunflower Pc103 and two yellow sunflowers (Yr17 and Y35). Targeted metabolomic analysis revealed higher anthocyanin levels but lower flavonol content in Pc103 compared to the yellow cultivars. RNA-sequencing and phylogenetic analysis identified multiple genes involved in the flavonoid pathway, including series of structural genes and three MYB and bHLH genes. Specifically, HaMYBA and HabHLH1 were up-regulated in Pc103, whereas HaMYBF exhibited reduced expression. HaMYBA was found to interact with HabHLH1 in vivo and in vitro, while HaMYBF does not. Transient expression analysis further revealed that HabHLH1 and HaMYBA cooperatively regulate increased expression of dihydroflavonol 4-reductase (DFR), leading to anthocyanin accumulation. On the other hand, ectopic expression of HaMYBF independently modulates flavonol synthase (FLS) expression, but hindered anthocyanin production. Collectively, our findings suggest that the up-regulation of HaMYBA and HabHLH1, as well as the down-regulation of HaMYBF, contribute to the red coloration in Pc103. It offers a theoretical basis for improving sunflower color through genetic engineering.

Light-Induced TaHY5-7A and TaBBX-3B Physically Interact to Promote PURPLE PERICARP-MYB 1 Expression in Purple-Grained Wheat.

Purple-grained wheat (Triticum aestivum L.) is an important germplasm source in crop breeding. Anthocyanin biosynthesis in the pericarps of purple-grained wheat is largely light-dependent; however, the regulatory mechanisms underlying light-induced anthocyanin accumulation in the wheat pericarp remain unknown. Here we determined that anthocyanins rapidly accumulate in the pericarps of the purple-grained wheat cultivar Heixiaomai 76 (H76) at 16 days after pollination under light treatment. Using transcriptome sequencing, differential gene expression analysis, and phylogenetic analysis, we identified two key genes involved in light signaling in wheat: ELONGATED HYPOCOTYL 5-7A (TaHY5-7A) and B-BOX-3B (TaBBX-3B). TaHY5-7A and TaBBX-3B were highly expressed in purple-grained wheat pericarps. The heterologous expression of TaHY5-7A partially restored the phenotype of the Arabidopsis (Arabidopsis thaliana) hy5 mutant, resulting in increased anthocyanin accumulation and a shortened hypocotyl. The heterologous expression of TaBBX-3B in wild-type Arabidopsis had similar effects. TaHY5-7A and TaBBX-3B were nucleus-localized, consistent with a function in transcription regulation. However, TaHY5-7A, which lacks a transactivation domain, was not sufficient to activate the expression of PURPLE PERICARP-MYB 1 (TaPpm1), the key anthocyanin biosynthesis regulator in purple pericarps of wheat. TaHY5-7A physically interacted with TaBBX-3B in yeast two-hybrid and bimolecular fluorescence complementation assays. Additionally, TaHY5-7A, together with TaBBX-3B, greatly enhanced the promoter activity of TaPpm1 in a dual luciferase assay. Overall, our results suggest that TaHY5-7A and TaBBX-3B collaboratively activate TaPpm1 expression to promote light-induced anthocyanin biosynthesis in purple-pericarp wheat.

Hypersensitivity Reactions Following Onabotulinum Toxin A Touch-up Injection Possibly Associated with Covid-19 Infection.

As one of the most common cosmetic procedures, botulinum toxin A (BTX-A) injections are basically safe. The typical allergic reactions include erythema, edema, pruritus, and wheal, which generally occur at the initial injection. Herein, we reported two cases of hypersensitivity reactions following cosmetic BTX-A touch-up injection. Type I allergic reactions provoked by re-exposure to the excipients such as gelatin may play a role in the event. In addition, this condition may be associated with the preceding Covid-19 infection, even with complete symptoms remission and negative tests results. Noteworthily, one case developed anaphylactic symptoms resembling purpuric drug eruption (PDE). These cases may serve as a significant complement to the botulinum toxin treatment safety profiles. LEVEL OF EVIDENCE V: This journal requires that authors assign a level of evidence to each article. For a full description of these Evidence-Based Medicine ratings, please refer to the Table of Contents or the online Instructions to Authors www.springer.com/00266 .

OsFAR1 is involved in primary fatty alcohol biosynthesis and promotes drought tolerance in rice.

MAIN CONCLUSION: OsFAR1 encodes a fatty acyl-CoA reductase involved in biosynthesis of primary alcohols and plays an important role in drought stress response in rice. Cuticular waxes cover the outermost surface of terrestrial plants and contribute to inhibiting nonstomatal water loss and improving plant drought resistance. Primary alcohols are the most abundant components in the leaf cuticular waxes of rice (Oryza sativa), but the biosynthesis and regulation of primary alcohol remain largely unknown in rice. Here, we identified and characterized an OsFAR1 gene belonging to the fatty acyl-CoA reductases (FARs) via a homology-based approach in rice. OsFAR1 was activated by abiotic stresses and abscisic acid, resulting in increased production of primary alcohol in rice. Heterologous expression of OsFAR1 enhanced the amounts of C22:0 and C24:0 primary alcohols in yeast (Saccharomyces cerevisiae) and C24:0 to C32:0 primary alcohols in Arabidopsis. Similarly, OsFAR1 overexpression significantly increased the content of C24:0 to C30:0 primary alcohols on rice leaves. Finally, OsFAR1 overexpression lines exhibited reduced cuticle permeability and enhanced drought tolerance in rice and Arabidopsis. Taken together, our results demonstrate that OsFAR1 is involved in rice primary alcohol biosynthesis and plays an important role in responding to drought and other environmental stresses.

Structural insight into the substrate-binding mode and catalytic mechanism for MlrC enzyme of Sphingomonas sp. ACM-3962 in linearized microcystin biodegradation.

Removing microcystins (MCs) safely and effectively has become an urgent global problem because of their extremely hazardous to the environment and public health. Microcystinases derived from indigenous microorganisms have received widespread attention due to their specific MC biodegradation function. However, linearized MCs are also very toxic and need to be removed from the water environment. How MlrC binds to linearized MCs and how it catalyzes the degradation process based on the actual three-dimensional structure have not been determined. In this study, the binding mode of MlrC with linearized MCs was explored using a combination of molecular docking and site-directed mutagenesis methods. A series of key substrate binding residues, including E70, W59, F67, F96, S392 and so on, were identified. Sodium dodecane sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) was used to analyze samples of these variants. The activity of MlrC variants were measured using high performance liquid chromatography (HPLC). We used fluorescence spectroscopy experiments to research the relationship between MlrC enzyme (E), zinc ion (M), and substrate (S). The results showed that MlrC enzyme, zinc ion and substrate formed E-M-S intermediates during the catalytic process. The substrate-binding cavity was made up of N and C-terminal domains and the substrate-binding site mainly included N41, E70, D341, S392, Q468, S485, R492, W59, F67, and F96. The E70 residue involved in both substrate catalysis and substrate binding. In conclusion, a possible catalytic mechanism of the MlrC enzyme was further proposed based on the experimental results and a literature survey. These findings provided new insights into the molecular mechanisms of the MlrC enzyme to degrade linearized MCs, and laid a theoretical foundation for further biodegradation studies of MCs.

QTL Mapping for Important Agronomic Traits Using a Wheat55K SNP Array-Based Genetic Map in Tetraploid Wheat.

Wheat yield is highly correlated with plant height, heading date, spike characteristics, and kernel traits. In this study, we used the wheat55K single nucleotide polymorphism array to genotype a recombinant inbred line population of 165 lines constructed by crossing two tetraploid wheat materials, Icaro and Y4. A genetic linkage map with a total length of 6244.51 cM was constructed, covering 14 chromosomes of tetraploid wheat. QTLs for 12 important agronomic traits, including plant height (PH), heading date (HD), awn color (AC), spike-branching (SB), and related traits of spike and kernel, were mapped in multiple environments, while combined QTL-by-environment interactions and epistatic effects were analyzed for each trait. A total of 52 major or stable QTLs were identified, among which may be some novel loci controlling PH, SB, and kernel length-width ratio (LWR), etc., with LOD values ranging from 2.51 to 54.49, thereby explaining 2.40-66.27% of the phenotypic variation. Based on the 'China Spring' and durum wheat reference genome annotations, candidate genes were predicted for four stable QTLs, QPH.nwafu-2B.2 (165.67-166.99 cM), QAC.nwafu-3A.1 (419.89-420.52 cM), QAC.nwafu-4A.1 (424.31-447.4 cM), and QLWR.nwafu-7A.1 (166.66-175.46 cM). Thirty-one QTL clusters and 44 segregation distortion regions were also detected, and 38 and 18 major or stable QTLs were included in these clusters and segregation distortion regions, respectively. These results provide QTLs with breeding application potential in tetraploid wheat that broadens the genetic basis of important agronomic traits such as PH, HD, AC, SB, etc., and benefits wheat breeding.

Crystal structural analysis and characterization for MlrC enzyme of Sphingomonas sp. ACM-3962 involved in linearized microcystin degradation.

Microcystinase C (MlrC), one key hydrolase of the microcystinase family, plays an important role in linearized microsystin (L-MC) degradation. However, the three-dimensional structure and structural features of MlrC are still unclear. This study obtained high specific activity and high purity of MlrC by heterologous expression, and revealed that MlrC derived from Sphingomonas sp. ACM-3962 (ACM-MlrC) can degrade linearized products of MC-LR, MC-RR and MC-YR to product 3-amino-9-methoxy-2,6,8-trimethyl-10-phenyldeca-4,6-dienoic acid (Adda), indicating the degradation function and significance in MC-detoxification. More importantly, this study reported the crystal structure of ACM-MlrC at 2.6 Å resolution for the first time, which provides a basis for further understanding the structural characteristics and functions of MlrC. MlrC had a dual-domain feature, namely N and C terminal domain respectively. The N-terminal domain contained a Glutamate-Aspartate-Histidine-Histidine catalytic quadruplex coordinated with zinc ion in each monomer. The importance of zinc ions and their coordinated residues was analyzed by dialysis and site-directed mutagenesis methods. Moreover, the important influence of the N/C-terminal flexible regions of ACM-MlrC was also analyzed by sequence truncation, and then the higher yield and total activity of variants were obtained, which was beneficial to study the better function and application of MlrC.

Associations of per- and polyfluoroalkyl substances and their alternatives with bone mineral density levels and osteoporosis prevalence: A community-based population study in Guangzhou, Southern China.

BACKGROUND: Evidence concerning associations of per- and polyfluoroalkyl substances (PFASs) exposure with bone mineral density (BMD) and osteoporosis is scarce. Additionally, no study has examined the effects of PFAS isomers and alternatives on bone health. OBJECTIVES: To evaluate the associations of PFASs and PFAS alternatives with BMD levels and osteoporosis prevalence. METHODS: A total of 1260 healthy adults from southern China were enrolled. Serum concentrations of 32 legacy PFASs, PFAS isomers, and alternatives were measured using modified liquid chromatography-tandem mass spectrometry. Logistic and linear regression models were applied to evaluate the associations of PFASs with osteoporosis prevalence and BMD levels, respectively, adjusting for confounding factors. We performed stratified analyses to assess potential effect modifications of age and sex. We further used sensitivity analyses to testify the robustness of the main findings. RESULTS: There were 204 (16.2 %) participants diagnosed with osteoporosis. Eleven of the studied PFASs (i.e., PFHpA, PFOA, PFBS, PFHpS, total-PFHxS, n-PFHxS, br-PFHxS, br-PFOS, 1m-PFOS, Σ3 + 4 + 5m-PFOS, and 6:2 Cl-PFESA) showed significant and inverse associations with BMD levels (mean differences ranged from -6.47 to -26.07 per one ln-unit increase in the PFASs). Additionally, we observed that each one ln-unit increase in PFHpA was significantly associated a 23 % (OR = 1.23, 95 % CI = 1.04, 1.45) greater odds of osteoporosis. The above associations were consistent in several sensitivity analyses we performed. Stratified analyses showed stronger associations among women and younger compared to their counterparts. CONCLUSIONS: Our findings suggested that greater PFAS exposure is associated with poorer bone health, especially in women and younger people.

Biomimetic, pH-Responsive Nanoplatforms for Cancer Multimodal Imaging and Photothermal Immunotherapy.

Photothermal therapy (PTT), by converting light to thermal energy, has become a novel and noninvasive technique for tumor thermal ablation in clinical practice. However, as a result of phagocytosis of reticuloendothelial cells, current photothermal agents (PTAs) derived from exogenous materials suffer from incompetent tumor targeting and brief internal circulation time. The resulting poor accumulation of PTAs in the target area severely reduces the efficacy of PTT. In addition, the potential toxicity of PTAs, excessive laser exposure, and possibilities of tumor recurrence and metastasis following PTT are still intractable problems that severely influence patients' quality of life. Herein, a biomimetic pH-responsive nanoprobe was prepared via cancer cell membrane coating polydopamine (PDA)-CaCO3 nanoparticles (CPCaNPs) for photoacoustic (PA)/ultrasonic (US)/thermal imaging-guided PTT. When CPCaNPs targeted and infiltrated into the tumor's acidic microenvironment, the decomposed CO2 bubbles from homologous targeting CPCaNPs enhanced ultrasonic (US) signals obviously. At the same time, the PDA of CPCaNPs not only performed efficient PTT of primary tumors but also generated photoacoustic (PA) signals. In addition, an immune checkpoint pathway blockade was combined, which inhibited tumor recurrence and metastasis significantly and improved the immunosuppressive microenvironment after PTT to a large extent. Thus, these proposed biomimetic pH-responsive CPCaNPs provide a promising strategy for precise PTT immunotherapy under the intelligent guidance of PA/US/thermal imaging and show great potential for clinical translation.

Fanconi anemia caused by biallelic inactivation of BRCA2 can present with an atypical cancer phenotype in adulthood.

Inherited biallelic pathogenic variants (PVs) in BRCA2 cause Fanconi Anemia complementation group D1 (FA-D1), a severe pediatric bone marrow failure and high-risk cancer syndrome. We identified biallelic BRCA2 PVs in a young adult with multiple basal cell carcinomas, adult-onset colorectal cancer and small cell neuroendocrine carcinoma, without bone marrow failure. No PVs were identified in any other known cancer susceptibility gene, and there was no evidence of reversion mosaicism. The proband's deceased sister had a classic FA-D1 presentation and was shown to carry the same biallelic BRCA2 PVs. A lymphoblastoid cell line derived from the proband demonstrated hypersensitivity to DNA damaging agents, and bone marrow showed aberrant RAD51 staining. Family expansion demonstrated the presence of BRCA2 related cancers in heterozygous family members. Our data highlight the striking phenotypic differences which can be observed within FA-D1 families and expands the clinical spectrum of FA-D1 to include adult presentation with a constellation of solid tumors not previously thought of as characteristic of Fanconi Anemia. Early recognition of this syndrome in a family could prevent further morbidity and mortality by implementation of hereditary breast and ovarian cancer screening and treatment strategies for heterozygous family members.

Autologous K63 deubiquitylation within the BRCA1-A complex licenses DNA damage recognition.

The BRCA1-A complex contains matching lysine-63 ubiquitin (K63-Ub) binding and deubiquitylating activities. How these functionalities are coordinated to effectively respond to DNA damage remains unknown. We generated Brcc36 deubiquitylating enzyme (DUB) inactive mice to address this gap in knowledge in a physiologic system. DUB inactivation impaired BRCA1-A complex damage localization and repair activities while causing early lethality when combined with Brca2 mutation. Damage response dysfunction in DUB-inactive cells corresponded to increased K63-Ub on RAP80 and BRCC36. Chemical cross-linking coupled with liquid chromatography-tandem mass spectrometry (LC-MS/MS) and cryogenic-electron microscopy (cryo-EM) analyses of isolated BRCA1-A complexes demonstrated the RAP80 ubiquitin interaction motifs are occupied by ubiquitin exclusively in the DUB-inactive complex, linking auto-inhibition by internal K63-Ub chains to loss of damage site ubiquitin recognition. These findings identify RAP80 and BRCC36 as autologous DUB substrates in the BRCA1-A complex, thus explaining the evolution of matching ubiquitin-binding and hydrolysis activities within a single macromolecular assembly.

ROS-responsive liposomes as an inhaled drug delivery nanoplatform for idiopathic pulmonary fibrosis treatment via Nrf2 signaling.

BACKGROUND: Idiopathic pulmonary fibrosis (IPF) is a progressive fibrotic disease with pathophysiological characteristics of transforming growth factor-ß (TGF-ß), and reactive oxygen species (ROS)-induced excessive fibroblast-to-myofibroblast transition and extracellular matrix deposition. Macrophages are closely involved in the development of fibrosis. Nuclear factor erythroid 2 related factor 2 (Nrf2) is a key molecule regulating ROS and TGF-ß expression. Therefore, Nrf2 signaling modulation might be a promising therapy for fibrosis. The inhalation-based drug delivery can reduce systemic side effects and improve therapeutic effects, and is currently receiving increasing attention, but direct inhaled drugs are easily cleared and difficult to exert their efficacy. Therefore, we aimed to design a ROS-responsive liposome for the Nrf2 agonist dimethyl fumarate (DMF) delivery in the fibrotic lung. Moreover, we explored its therapeutic effect on pulmonary fibrosis and macrophage activation. RESULTS: We synthesized DMF-loaded ROS-responsive DSPE-TK-PEG@DMF liposomes (DTP@DMF NPs). DTP@DMF NPs had suitable size and negative zeta potential and excellent capability to rapidly release DMF in a high-ROS environment. We found that macrophage accumulation and polarization were closely related to fibrosis development, while DTP@DMF NPs could attenuate macrophage activity and fibrosis in mice. RAW264.7 and NIH-3T3 cells coculture revealed that DTP@DMF NPs could promote Nrf2 and downstream heme oxygenase-1 (HO-1) expression and suppress TGF-ß and ROS production in macrophages, thereby reducing fibroblast-to-myofibroblast transition and collagen production by NIH-3T3 cells. In vivo experiments confirmed the above findings. Compared with direct DMF instillation, DTP@DMF NPs treatment presented enhanced antifibrotic effect. DTP@DMF NPs also had a prolonged residence time in the lung as well as excellent biocompatibility. CONCLUSIONS: DTP@DMF NPs can reduce macrophage-mediated fibroblast-to-myofibroblast transition and extracellular matrix deposition to attenuate lung fibrosis by upregulating Nrf2 signaling. This ROS-responsive liposome is clinically promising as an ideal delivery system for inhaled drug delivery.

Structural Insights Into the Effects of Interactions With Iron and Copper Ions on Ferritin From the Blood Clam Tegillarca granosa.

In addition to its role as an iron storage protein, ferritin can function as a major detoxification component in the innate immune defense, and Cu2+ ions can also play crucial antibacterial roles in the blood clam, Tegillarca granosa. However, the mechanism of interaction between iron and copper in recombinant Tegillarca granosa ferritin (TgFer) remains to be investigated. In this study, we investigated the crystal structure of TgFer and examined the effects of Fe2+ and Cu2+ ions on the TgFer structure and catalytic activity. The crystal structure revealed that TgFer presented a typically 4-3-2 symmetry in a cage-like, spherical shell composed of 24 identical subunits, featuring highly conserved organization in both the ferroxidase center and the 3-fold channel. Structural and biochemical analyses indicated that the 4-fold channel of TgFer could be serviced as potential binding sites of metal ions. Cu2+ ions appear to bind preferentially with the 3-fold channel as well as ferroxidase site over Fe2+ ions, possibly inhibiting the ferroxidase activity of TgFer. Our results present a structural and functional characterization of TgFer, providing mechanistic insight into the interactions between TgFer and both Fe2+ and Cu2+ ions.

Mitochondrial Glutathione Depletion Nanoshuttles for Oxygen-Irrelevant Free Radicals Generation: A Cascaded Hierarchical Targeting and Theranostic Strategy Against Hypoxic Tumor.

An oxygen-irrelevant free radicals generation strategy has shown great potential in hypoxic tumor therapy. However, insufficient tumor accumulation, nonspecific intracellular localization, and the presence of highly reductive mitochondrial glutathione (GSH) dramatically hamper the free radicals therapeutic efficacy. Herein, a hierarchical targeting system was constructed by Fe-doped polydiaminopyridine nanoshuttles, indocyanine green (ICG), and an oxygen-irrelevant radicals generator (AIPH) to possess a negative charge. An acid-specific charge-reverse capability of the shuttles was achieved to enhance cell uptake in the tumor microenvironment (TME). In addition, the iron release occurs only in the acidic TME, which can be used as acidity enhancers to strengthen the charge-reverse process, thereby leading to more efficient tumor internalization and deep penetration. Moreover, such a nanosystem has significantly improved the delivery efficiency of nanoshuttles (16.06%) in the tumor tissues at 24 h postinjection, much higher than that of naked Fe-doped polydiaminopyridine (6.59%). More importantly, the nanoshuttles enable simultaneously mitochondria targeting and corresponding GSH depleting capability to show advantages in free radicals-based therapy after charge reversion, leading to a powerful tumor inhibition rate (>95%). The prescence of iron could allow for magnetic resonance imaging, while ICG allowed for photoacoustic imaging and fluorescence imaging to guide the therapeutic process. The remarkable features of the nanoshuttles may open a new avenue to explore an oxygen-irrelevant free radicals generating system for accurate cancer theranostics.

Tomato SlCER1-1 catalyzes the synthesis of wax alkanes which increases the drought tolerance and fruit storability.

Very-long-chain (VLC) alkanes are the main wax compounds of tomato fruit and leaf. ECERIFERUM1 (CER1) and ECERIFERUM3 (CER3) are the two key genes involved in VLC alkane biosynthesis in Arabidopsis thaliana. However, the CER1 and CER3 homologous genes in tomato have not been investigated and their exact biological function remains unknown. We analyzed the wax profiles in tomato leaves and fruits at different growth stages, and characterized the CER1 and CER3 homologous genes. VLC alkanes were the predominant wax compounds both in the leaf and fruit at all developmental stages. We identified five CER1 homologs and two CER3 homologs in tomato, which were designated as SlCER1-1 to SlCER1-5 and SlCER3-1 and SlCER3-2 respectively. The genes exhibited tissue- and organ-dependent expression patterns and were induced by abiotic stresses. SlCER1-1 was localized to the endoplasmic reticulum (ER), which is also the main site of wax biosynthesis. Silencing the SlCER1-1 gene in tomato significantly reduced the amounts of n-Alkanes and branched alkanes, whereas its overexpression in Arabidopsis had the opposite effect. Under drought stress, both n-Alkanes and branched alkanes increased significantly in wild-type but not the SlCER1-1 RNAi tomato plants. Furthermore, SlCER1-1 silencing also increased the cuticular permeabilities of the leaves and fruits. In conclusion, SlCER1-1 is involved in wax alkane biosynthesis in tomato and plays an important role in the drought tolerance and fruit storability.