SlRBP1 promotes translational efficiency via SleIF4A2 to maintain chloroplast function in tomato.

Many glycine-rich RNA-binding proteins (GR-RBPs) have critical functions in RNA processing and metabolism. Here, we describe a role for the tomato (Solanum lycopersicum) GR-RBP SlRBP1 in regulating mRNA translation. We found that SlRBP1 knockdown mutants (slrbp1) displayed reduced accumulation of total chlorophyll and impaired chloroplast ultrastructure. These phenotypes were accompanied by deregulation of the levels of numerous key transcripts associated with chloroplast functions in slrbp1. Furthermore, native RNA immunoprecipitation-sequencing (nRIP-seq) recovered 61 SlRBP1-associated RNAs, most of which are involved in photosynthesis. SlRBP1 binding to selected target RNAs was validated by nRIP-qPCR. Intriguingly, the accumulation of proteins encoded by SlRBP1-bound transcripts, but not the mRNAs themselves, was reduced in slrbp1 mutants. Polysome profiling followed by RT-qPCR assays indicated that the polysome occupancy of target RNAs was lower in slrbp1 plants than in wild-type. Furthermore, SlRBP1 interacted with the eukaryotic translation initiation factor SleIF4A2. Silencing of SlRBP1 significantly reduced SleIF4A2 binding to SlRBP1-target RNAs. Taking these observations together, we propose that SlRBP1 binds to and channels RNAs onto the SleIF4A2 translation initiation complex and promotes the translation of its target RNAs to regulate chloroplast functions.

Dnttip2 is essential for 18S rRNA processing and digestive organ development in zebrafish.

Dnttip2 is one of the components of the small subunit (SSU) processome. In yeast, depletion of dnttip2 leads to an inefficient processing of pre-rRNA and a decrease in synthesis of the mature 18S rRNA. However, the biological roles of Dnttip2 in higher organisms are poorly defined. In this study, we demonstrate that dnttip2 is a maternal gene in zebrafish. Depletion of Dnttip2 leads to embryonic lethal with severe digestive organs hypoplasia. The loss of function of Dnttip2 also leads to partial defects in cleavage at the A0-site and E-site during 18S rRNA processing. In conclusion, Dnttip2 is essential for 18S rRNA processing and digestive organ development in zebrafish.

Dual-Modality Imaging-Guided Manganese-Based Nanotransformer for Enhanced Gas-Photothermal Therapy Combined Immunotherapeutic Strategy Against Triple-Negative Breast Cancer.

Activating the stimulator of the interferon gene (STING) is a promising immunotherapeutic strategy for converting "cold" tumor microenvironment into "hot" one to achieve better immunotherapy for malignant tumors. Herein, a manganese-based nanotransformer is presented, consisting of manganese carbonyl and cyanine dye, for MRI/NIR-II dual-modality imaging-guided multifunctional carbon monoxide (CO) gas treatment and photothermal therapy, along with triggering cGAS-STING immune pathway against triple-negative breast cancer. This nanosystem is able to transfer its amorphous morphology into a crystallographic-like formation in response to the tumor microenvironment, achieved by breaking metal-carbon bonds and forming coordination bonds, which enhances the sensitivity of magnetic resonance imaging. Moreover, the generated CO and photothermal effect under irradiation of this nanotransformer induce immunogenic death of tumor cells and release damage-associated molecular patterns. Simultaneously, the Mn acts as an immunoactivator, potentially stimulating the cGAS-STING pathway to augment adaptive immunity, resulting in promoting the secretion of type I interferon, the proliferation of cytotoxic T lymphocytes and M2-macrophages repolarization. This nanosystem-based gas-photothermal treatment and immunoactivating therapy synergistic effect exhibit excellent antitumor efficacy both in vitro and in vivo, reducing the risk of triple-negative breast cancer recurrence and metastasis; thus, this strategy presents great potential as multifunctional immunotherapeutic agents for cancer treatment.

DJ-1 depletion prevents immunoaging in T-cell compartments.

Decline in immune function during aging increases susceptibility to different aging-related diseases. However, the underlying molecular mechanisms, especially the genetic factors contributing to imbalance of naïve/memory T-cell subpopulations, still remain largely elusive. Here, we show that loss of DJ-1 encoded by PARK7/DJ-1, causing early-onset familial Parkinson's disease (PD), unexpectedly diminished signs of immunoaging in T-cell compartments of both human and mice. Compared with two gender-matched unaffected siblings of similar ages, the index PD patient with DJ-1 deficiency showed a decline in many critical immunoaging features, including almost doubled non-senescent T cells. The observation was further consolidated by the results in 45-week-old DJ-1 knockout mice. Our data demonstrated that DJ-1 regulates several immunoaging features via hematopoietic-intrinsic and naïve-CD8-intrinsic mechanisms. Mechanistically, DJ-1 depletion reduced oxidative phosphorylation (OXPHOS) and impaired TCR sensitivity in naïve CD8 T cells at a young age, accumulatively leading to a reduced aging process in T-cell compartments in older mice. Our finding suggests an unrecognized critical role of DJ-1 in regulating immunoaging, discovering a potent target to interfere with immunoaging- and aging-associated diseases.

Three-dimensional Assessment of Longitudinal Surgical Outcome in Patients with Unilateral Cleft Lip and Palate: A Modified Rotation Advancement Technique.

BACKGROUND: Optimization of a modified rotation advancement technique is hampered by lack of objective measures to quantify the longitudinal surgical outcome. METHODS: We collected and assessed facial 3D images of 115 consecutive patients who underwent primary repair between 2017 and 2019. Photogrammetry was performed preoperatively, immediately postoperatively and at a first and second follow-up interval, occurring at an average year of 0.6 and 5.3 years, respectively. 10 additional age-matched noncleft control subjects were also included. RESULTS: Growth lag in cleft side lateral lip and gradual elongation of medial lip height on the cleft side caused continuous deviation of philtrum towards the cleft side. The columellar length on the cleft side continued to grow slower, accompanied by a persistent widening of alar base width on the cleft side, leading to in the gradual deviation of columella towards the cleft side. The pre-operative and post-operative nasolabial asymmetry would increase with greater degree of postoperative deficiencies. Right clefts presented with greater degrees of deficiencies in lateral lip height in preoperative measurement, but this discrepancy of the laterality of clefts was not observed in the two follow-up periods. CONCLUSION: The surgical outcome of this modified rotational advancement technique in unilateral cleft lip primary repair is promising. Growth lag in lateral lip and lateral displacement of alar base cause continuous deviation of philtrum towards the cleft side. Pre-operative severity does predict post-operative outcomes. Laterality of oral clefts does not significantly affect the long-term outcomes of surgery. PRACTICAL IMPLICATION: This surgical technique meets the current trend of cleft lip and palate primary repair and is worth promoting.

The effects and significance of Dicyclopentadiene on the expression of Intersectin-1 after cerebral ischemia-reperfusion in rats.

OBJECTIVE: This study mainly observed the changes in Intersectin-1 (ITSN-1) expression in rat brain tissue after ischemia-reperfusion intervened by Dicyclopentadiene. METHODS: SD rats were randomly divided into non-middle Cerebral Artery Occlusion model group (normal group, sham operation group) and Middle Cerebral Artery Occlusion (MCAO) model group [Ischemia reperfusion (cerebral ischemia reperfusion)] reperfusion,IR) (6h, 24h, 72h, 1w, 2w) group, butylphthalein intervention group], First of all, Use Western The expression of ITSN-1 in the cerebral tissue of infarction side after ischemia-reperfusion injury in each group was measured by blotting, and then the loss and degree of nerve function after ischemia-reperfusion injury in each group was evaluated by Zea-Longa scoring method. The morphological changes of cells in the ischemic penumbra region in the normal group and the MCAO model group for 24h were observed by HE staining. Next, 24h was selected as the reperfusion point for intervention with butylphthalein sodium chloride injection. Finally, Zea-Longa scoring method was used to evaluate whether the rats had neurological impairment and its degree, TTC (Triphenyltetrazolium chloride) staining was used to determine whether the rats had cerebral infarction and its extent, and Western The expression of ITSN-1 in the cerebral tissue of infarcted rats after ischemia-reperfusion injury was measured by blotting. RESULTS: 1. Zea-Longa scoring: Scores, except for the normal group and sham operation group (which scored 0), ranged between 2.75 ± 0.46 in the ischemia-reperfusion 24h group and 1.88 ± 0.35 in the Dicyclopentadiene intervention group, showing statistically significant decreases (P<0.05). 2. HE staining results: The cell structures in the brain tissues of normal group rats were normal with regular nuclear shapes and sizes. There were no obvious abnormal changes. Rats in the ischemia-reperfusion 24h group showed obviously swollen cells, reduced and aggregated nucleus, and cell necrosis in the ischemic penumbra. 3. TTC staining results: Except for the normal group and the sham operation group, which had no infarcts, the ischemia-reperfusion 24h group had the largest volume ratio of cerebral infarction. The volume ratio of cerebral infarction in the Dicyclopentadiene intervention group relatively reduced, making a difference with statistical significance (P<0.05). 4. Western blotting results: After cerebral ischemia-reperfusion in rats, ITSN-1 expression in the infarction-side brain tissue dynamically changed. ITSN-1 expression in the ischemia-reperfusion 24h group was significantly lower among other groups compared to the normal group (P<0.05). After 24 hours, the expression gradually increased after using Dicyclopentadiene intervention, the difference was statistically significant (P<0.05). CONCLUSION: After cerebral ischemia-reperfusion in rats, ITSN-1 expression dynamically changed in the infarction-side brain tissue. Dicyclopentadiene can alleviate ischemia-reperfusion injuries in rats, which might be related to the regulation of ITSN-1 expression.

[Chemical constituents from roots of Kadsura heteroclita].

The dichloromethane fraction of Kadsura heteroclita roots was separated and purified by chromatographic techniques(e.g., silica gel, Sephadex LH-20, ODS, MCI column chromatography) and semi-preparative HPLC. Twenty compounds were isolated from K. heteroclita, and their structures were identified by NMR, MS, UV, and X-ray single crystal diffraction techniques. Twenty compounds were isolated from K. heteroclita, which were identified as xuetongdilactone G(1), mallomacrostin C(2), 3,4-seco(24Z)-cychmrt-4(28),24-diene-3,26-dioic acid 3-methyl ester(3), nigranoic acid(4), methyl ester schizanlactone E(5), schisandronic acid(6), heteroclic acid(7), wogonin(8),(2R,3R)-4'-O-methyldihydroquercetin(9), 15,16-bisnor-13-oxo-8(17),11E-labdadien-19-oic acid(10), stigmast-4-ene-6ß-ol-3-one(11), psoralen(12),(1R,2R,4R)-trihydroxy-p-menthane(13), homovanillyl alcohol(14), 2-(4-hydroxyphenyl)-ethanol(15), coniferaldehyde(16),(E)-7-(4-hydroxy-3-methoxyphenyl)-7-methylbut-8-en-9-one(17), acetovanillone(18), vanillic acid(19) and vanillin(20). Compound 1 is a new compound named xuetongdilactone G. Compounds 2-3 and 8-20 are isolated from K. heteroclita for the first time.

Chronic Exposure to Palmitic Acid Down-Regulates AKT in Beta-Cells through Activation of mTOR.

High circulating lipids occurring in obese individuals and insulin-resistant patients are considered a contributing factor to type 2 diabetes. Exposure to high lipid concentration is proposed to both protect and damage beta-cells under different circumstances. Here, by feeding mice a high-fat diet (HFD) for 2 weeks to up to 14 months, the study showed that HFD initially causes the beta-cells to expand in population, whereas long-term exposure to HFD is associated with failure of beta-cells and the inability of animals to respond to glucose challenge. To prevent the failure of beta-cells and the development of type 2 diabetes, the molecular mechanisms that underlie this biphasic response of beta-cells to lipid exposure were explored. Using palmitic acid (PA) in cultured beta-cells and islets, the study demonstrated that chronic exposure to lipids leads to reduced viability and inhibition of cell cycle progression concurrent with down-regulation of a pro-growth/survival kinase AKT, independent of glucose. This AKT down-regulation by PA is correlated with the induction of mTOR/S6K activity. Inhibiting mTOR activity with rapamycin induced Raptor and restored AKT activity, allowing beta-cells to gain proliferation capacity that was lost after HFD exposure. In summary, a novel mechanism in which lipid exposure may cause the dipole effects on beta-cell growth was elucidated, where mTOR acts as a lipid sensor. These mechanisms can be novel targets for future therapeutic developments.

KLF13 overexpression protects sepsis-induced myocardial injury and LPS-induced inflammation and apoptosis.

Sepsis remains a worldwide public health problem. This study aims to explore the role and mechanism of transcriptional factors (TFs) in sepsis-induced myocardial injury. Firstly, TF KLF13 was selected to explore its role in sepsis-induced myocardial injury. The caecal ligation and puncture (CLP) -induced sepsis mouse model was established and the septic mice were examined using standard histopathological methods. KLF13 expression was detected in the septic mouse heart and was also seen in a lipoploysaccharide (LPS) -induced cellular inflammation model. To explore this further both pro-apoptotic cleaved-caspase3/caspase3 and Bax levels and anti-apoptotic Bcl2 levels were examined, also in both models, In addition inflammatory cytokine (IL-1ß, TNF-α, IL-8 and MCP-1) production and IκB-α protein level and p65 phosphorylation were examined in both septic mice and LPS-induced cells. Thus three parameters - cardiomyocyte apoptosis, inflammatory response and NF-κB pathway activation were evaluated under similar conditions. The septic mice showed significant oedema, disordered myofilament arrangement and degradation and necrosis to varying degrees in the myocardial cells. KLF13 was downregulated in both the septic mouse heart and the LPS-induced cellular inflammation model. Furthermore, both models showed abnormally increased cardiomyocyte apoptosis (increased cleaved-caspase3/caspase and Bax protein levels and decreased Bcl2 level), elevated inflammation (increased production of inflammatory cytokines) and the activated NF-κB pathway (increased p65 phosphorylation and decreased IκB-α protein level). KLF13 overexpression notably ameliorated sepsis-induced myocardial injury in vivo and in vitro. KLF13 overexpression protected against sepsis-induced myocardial injury and LPS-induced cellular inflammation and apoptosis via inhibiting the inflammatory pathways (especially NF-κB signalling) and cardiomyocyte apoptosis.

Project-based learning course on metabolic network modelling in computational systems biology.

Project-based learning (PBL) is a dynamic student-centred teaching method that encourages students to solve real-life problems while fostering engagement and critical thinking. Here, we report on a PBL course on metabolic network modelling that has been running for several years within the Master in Integrated Systems Biology (MISB) at the University of Luxembourg. This 2-week full-time block course comprises an introduction into the core concepts and methods of constraint-based modelling (CBM), applied to toy models and large-scale networks alongside the preparation of individual student projects in week 1 and, in week 2, the presentation and execution of these projects. We describe in detail the schedule and content of the course, exemplary student projects, and reflect on outcomes and lessons learned. PBL requires the full engagement of students and teachers and gives a rewarding teaching experience. The presented course can serve as a role model and inspiration for other similar courses.