Thyroid hormone receptor: A new player in epinephrine-induced larval metamorphosis of the hard-shelled mussel.

Many marine invertebrate larvae undergo a dramatic morphological and physiological transition from a planktonic larva to a benthic juvenile. The mechanisms of this metamorphosis in bivalves are mainly unknown. The recent identification in bivalves of a thyroid hormone receptor (TR) gene raises the possibility that as occurs in vertebrate metamorphosis, TRs regulate this developmental process. An evolutionary study of TR receptors revealed they are ubiquitous in the molluscs. Knock-down of the TR gene in pediveliger larvae of the hard-shelled mussel, Mytilus coruscus (Mc), using electroporation of siRNA significantly (p < 0.01) reduced TR gene expression. TR gene knock-down decreased pediveliger larval metamorphosis by 54% and was associated with a significant (p < 0.01) reduction in viability compared to control larvae. The TR in the hard-shelled mussel appears to be an essential regulatory factor for the successful epinephrine-induced metamorphosis of the pediveliger larvae to post-larvae. It is hypothesised that the knock-down of TR by siRNA transfection affects the "competence" of pediveliger larvae for the metamorphic transition by reducing their ability to respond to the inducer. The involvement of TR in the epinephrine-induced metamorphosis of a mollusc, the hard-shelled mussel, suggests the role of TR in this process probably emerged early during evolution.

Carotenoid biosynthesis genes LcLCYB, LcLCYE, and LcBCH from wolfberry confer increased carotenoid content and improved salt tolerance in tobacco.

Carotenoids play essential roles in plant growth and development and provide plants with a tolerance to a series of abiotic stresses. In this study, the function and biological significance of lycopene ß-cyclase, lycopene ε-cyclase, and ß-carotene hydroxylase, which are responsible for the modification of the tetraterpene skeleton procedure, were isolated from Lycium chinense and analyzed. The overexpression of lycopene ß-cyclase, lycopene ε-cyclase, and ß-carotene hydroxylase promoted the accumulation of total carotenoids and photosynthesis enhancement, reactive oxygen species scavenging activity, and proline content of tobacco seedlings after exposure to the salt stress. Furthermore, the expression of the carotenoid biosynthesis genes and stress-related genes (ascorbate peroxidase, catalase, peroxidase, superoxide dismutase, and pyrroline-5-carboxylate reductase) were detected and showed increased gene expression level, which were strongly associated with the carotenoid content and reactive oxygen species scavenging activity. After exposure to salt stress, the endogenous abscisic acid content was significantly increased and much higher than those in control plants. This research contributes to the development of new breeding aimed at obtaining stronger salt tolerance plants with increased total carotenoids and vitamin A content.

Quantification of Cardiomyocyte Contraction In Vitro and Drug Screening by MyocytoBeats.

Cardiomyocyte contractility is the crucial feature of heart function. Quantifying cardiomyocyte contraction in vitro is essential for disease phenotype characterization, mechanism illumination, and drug screening. Although many experimental methods have been employed to determine contraction dynamics in vitro, a time-saving and easy-to-use software is still needed to be developed. We presented a reliable tool, named MyocytoBeats, to measure cardiomyocyte contraction by processing recorded videos. Analysis results by MyocytoBeats of various experimental models have shown a significant linear relationship with another validated software. We also performed pharmacology screen in the platform, and astragaloside IV was identified to stabilize the frequency and amplitude of cardiomyocyte in the arrhythmia model. MyocytoBeats is a high-performance tool for generating cardiomyocyte contraction data of vitro study and shows a great potential in cardiac pharmacology study.

A transient wave of Bhlhe41+ resident macrophages enables remodeling of the developing infarcted myocardium.

The immune system plays a critical role during myocardial injury, contributing to repair and remodeling post myocardial infarction (MI). The myocardial infarct and border zone exhibit high heterogeneity, in turn leading to reconstructing macrophage subsets and specific functions. Here we use a combination of single-cell RNA sequencing, spatial transcriptomes, and reporter mice to characterize temporal-spatial dynamics of cardiac macrophage subtype in response to MI. We identify that transient appearance of monocyte-derived Bhlhe41+ Mφs in the "developing" infarct zone peaked at day 7, while other monocyte-derived macrophages are identified in "old" infarct zone. Functional characterization by co-culture of Bhlhe41+ Mφs with cardiomyocytes and fibroblasts or depletion of Bhlhe41+ Mφs unveils a crucial contribution of Bhlhe41+ Mφs in suppression of myofibroblast activation. This work highlights the importance of Bhlhe41+ Mφ phenotype and plasticity in preventing excessive fibrosis and limiting the expansion of developing infarct area.

Anti-inflammatory effect of Danhong injection through inhibition of GSDMD-mediated pyroptosis.

BACKGROUND: Pyroptosis is an inflammatory form of cell death that has been implicated in various infectious and non-infectious diseases. Gasdermin family proteins are the key executors of pyroptotic cell death, thus they are considered as novel therapeutic targets for inflammatory diseases. However, only limited gasdermin specific inhibitors have been identified to date. Traditional Chinese medicines have been applied in clinic for centuries and exhibit potential in anti-inflammation and anti-pyroptosis. We attempted to find candidate Chinese botanical drugs which specifically target gasdermin D (GSDMD) and inhibit pyroptosis. METHODS: In this study, we performed high-throughput screening using a botanical drug library to identify pyroptosis specific inhibitors. The assay was based on a cell pyroptosis model induced by lipopolysaccharides (LPS) and nigericin. Cell pyroptosis levels were then evaluated by cell cytotoxicity assay, propidium iodide (PI) staining and immunoblotting. We then overexpressed GSDMD-N in cell lines to investigate the direct inhibitory effect of the drug to GSDMD-N oligomerization. Mass spectrometry studies were applied to identify the active components of the botanical drug. Finally, a mouse model of sepsis and a mouse model of diabetic myocardial infarction were constructed to verify the protective effect of the drug in disease models of inflammation. RESULTS: High-throughput screening identified Danhong injection (DHI) as a pyroptosis inhibitor. DHI remarkably inhibited pyroptotic cell death in a murine macrophage cell line and bone marrow-derived macrophages. Molecular assays demonstrated the direct blockade of GSDMD-N oligomerization and pore formation by DHI. Mass spectrometry studies identified the major active components of DHI, and further activity assays revealed salvianolic acid E (SAE) as the most potent molecule among these components, and SAE has a strong binding affinity to mouse GSDMD Cys192. We further demonstrated the protective effects of DHI in mouse sepsis and mouse myocardial infarction with type 2 diabetes. CONCLUSION: These findings provide new insights for drug development from Chinese herbal medicine like DHI against diabetic myocardial injury and sepsis through blocking GSDMD-mediated macrophage pyroptosis.

Structural repurposing of SGLT2 inhibitor empagliflozin for strengthening anti-heart failure activity with lower glycosuria.

Sodium-glucose cotransporter 2 (SGLT2) inhibitors have been reapproved for heart failure (HF) therapy in patients with and without diabetes. However, the initial glucose-lowering indication of SGLT2i has impeded their uses in cardiovascular clinical practice. A challenge of SGLT2i then becomes how to separate their anti-HF activity from glucose-lowering side-effect. To address this issue, we conducted structural repurposing of EMPA, a representative SGLT2 inhibitor, to strengthen anti-HF activity and reduce the SGLT2-inhibitory activity according to structural basis of inhibition of SGLT2. Compared to EMPA, the optimal derivative JX01, which was produced by methylation of C2-OH of the glucose ring, exhibited weaker SGLT2-inhibitory activity (IC50 > 100 nmol/L), and lower glycosuria and glucose-lowering side-effect, better NHE1-inhibitory activity and cardioprotective effect in HF mice. Furthermore, JX01 showed good safety profiles in respect of single-dose/repeat-dose toxicity and hERG activity, and good pharmacokinetic properties in both mouse and rat species. Collectively, the present study provided a paradigm of drug repurposing to discover novel anti-HF drugs, and indirectly demonstrated that SGLT2-independent molecular mechanisms play an important role in cardioprotective effects of SGLT2 inhibitors.

Mechanistic insight of weak magnetic field trigger transformation of amorphous Fe(III)-(oxy)hydroxide for enhanced ferrate (VI) towards selective removal of natural organic matter.

It was important to regulate the formation of Fe-hydroxyl during ferrate (Fe(VI)) oxidation and hydrolysis which was beneficial for interfacial adsorption of natural organic matter (NOM). Based on the influence of weak magnetic field (WMF) on the physical and chemical characteristics of particles in chemistry. This study investigated the effect of WMF on Fe(VI) oxidation and Fe(III) flocculation performance by regulating iron species during hydrolysis, for NOM removal. Results indicated WMF efficiently accelerate the removal of NOM that the reactions rate constants in magnetization system was twice as much as the control group. With the structure and electrochemical analysis, WMF enhanced Hydrogen-bond and caused much polar hydroxyl groups combined with iron ions, further triggered Fe(III) transformed to amorphous Fe-hydroxide and ferrihydrite with large specific surface area and high surface activity which removed the pollutants by adsorption and co-precipitation, instead of crystalloid Fe2O3 and Fe3O4. In addition, the nucleation aggregation behavior and interaction energy of Fe-(oxy)hydroxide revealed that the lower free energy obtained in magnetization system, could lead to higher nucleation rate, and promoted the aggregation. WMF increased hydrophobicity of Fe-(oxy)hydroxides, further more easily adsorbed with humic acid (HA) and bovine serum albumin (BSA) with lower interaction energies than in control group. The selective removal mechanism of Fe-(oxy)hydroxide hardly to aggregate with pollutants which caused by the difference of electrostatic interaction, was illustrated that electronegativity HA and SA were liable to electrostatically attract with Fe-(oxy)hydroxide and removed while the low electronegativity BSA was difficult to remove which its attraction was weakened.

Hypoxia-Activated Albumin-Binding Exatecan Prodrug for Cancer Therapy.

As an effective drug delivery strategy for traditional antitumor drugs, the stimulus-responsive albumin-based prodrugs are getting more and more attention. These prodrugs only release drugs in specific tumor microenvironments, which can prevent premature release of the drug in the circulation. Tumor hypoxia is a fundamental feature of the solid tumor microenvironment. As a hypoxia-activated linker, the 5-position branched linker of 1-methyl-2-nitro-5-hydroxymethylimidazole can be a trigger for albumin-based prodrugs. In this study, we report the synthesis and biological evaluation of the hypoxia-activated albumin-binding prodrug Mal-azo-Exatecan. After intravenous administration, the maleimide on the side chain can rapidly bind to endogenous albumin, enabling the prodrugs to accumulate in tumors, where tumor-associated hypoxia microenvironments trigger the selective release of Exatecan. The 5-position branched linker of 1-methyl-2-nitro-5-hydroxymethylimidazole as a cleavable linker has high plasma stability and does not cause Exatecan release from HSA-azo-Exatecan during circulation in vivo, avoiding systemic side effects caused by Exatecan.

Tracing PRX1+ cells during molar formation and periodontal ligament reconstruction.

Neural crest-derived mesenchymal stem cells (MSCs) are known to play an essential function during tooth and skeletal development. PRX1+ cells constitute an important MSC subtype that is implicated in osteogenesis. However, their potential function in tooth development and regeneration remains elusive. In the present study, we first assessed the cell fate of PRX1+ cells during molar development and periodontal ligament (PDL) formation in mice. Furthermore, single-cell RNA sequencing analysis was performed to study the distribution of PRX1+ cells in PDL cells. The behavior of PRX1+ cells during PDL reconstruction was investigated using an allogeneic transplanted tooth model. Although PRX1+ cells are spatial specific and can differentiate into almost all types of mesenchymal cells in first molars, their distribution in third molars is highly limited. The PDL formation is associated with a high number of PRX1+ cells; during transplanted teeth PDL reconstruction, PRX1+ cells from the recipient alveolar bone participate in angiogenesis as pericytes. Overall, PRX1+ cells are a key subtype of dental MSCs involved in the formation of mouse molar and PDL and participate in angiogenesis as pericytes during PDL reconstruction after tooth transplantation.

The characteristic of electrode of degradation of bio-electrochemical system based on in-situ ultrasonic monitoring: Biofilm and ion precipitation.

Electrode interface behavior is a decisive factor affecting the performance of bio-electrochemical systems, and traditional monitoring methods cannot provide real-time feedback. Therefore, in situ ultrasonic monitoring was performed to continuously monitor the formation process of electroactive biofilm and salt precipitation on the cathode surface. The results showed that biofilm was attached to the cathode surface first. Then, Ca2+ and Mg2+ precipitation gradually invaded the biofilm and accumulated between the cathode and the biofilm. The electrochemical performance of the biofilm adhesion and initial ion invasion process was improved. However, the electrochemical performance of the precipitation layer was decreased, while the operation time increases. In this paper, based on the air cathode scaling analyzing a new method for monitoring the electrode interface of bio-electrochemical system was provided, and the performance was recovered by using reverse electric field.

SN38-based albumin-binding prodrug for efficient targeted cancer chemotherapy.

Developing new therapeutic strategies that damage tumour cells without harming normal tissues is among the primary obstacles in chemotherapy. In this study, a novel ß-glucuronidase-sensitive albumin-binding prodrug was designed and synthesized to selectively deliver the drug SN38 to tumour sites and maximize its efficacy. After intravenous administration, the prodrug Mal-glu-SN38 covalently bound to plasma albumin through the Michael addition, enabling it to accumulate in the tumour and release SN38 when triggered by extracellular ß-glucuronidase. Compared to irinotecan, Mal-glu-SN38 displayed a slower plasma clearance and increased drug exposure over time. Moreover, Mal-glu-SN38 caused an increase in tumour-site accumulation of both the albumin-prodrug conjugate and free SN38 released from albumin conjugate when compared with irinotecan. After administration of multiple doses, Mal-glu-SN38 also significantly delayed the tumour growth, resulting in an impressive reduction or even disappearance of tumours (67% of mice cured) without causing any observable side effects.

A camptothecin-based, albumin-binding prodrug enhances efficacy and safety in vivo.

The albumin-based drug delivery system is an effective drug delivery strategy for traditional chemotherapeutic drugs that can improve their antitumour efficacies and reduce systemic toxicities. The camptothecin derivative CPTS0001 has excellent antitumour activity in vitro, but it shows toxicity and side effects in vivo. In this study, we report the synthesis and biological evaluation of the ß-glucuronidase-reactive albumin-binding prodrug Mal-glu-CPTS0001 based on quaternary ammonium. After intravenous administration, the compound covalently binds to plasma albumin through Michael addition, enabling it to accumulate in tumours, where tumour-associated ß-glucuronidase triggers the selective release of CPTS0001. This prodrug significantly reduced the toxicity of the parent drug, and the maximum tolerated dose was increased by 2.5 times. At the same time, this prodrug enhanced the selectivity in vivo and improved the preferential accumulation of prodrug in tumours. Notably, this prodrug exhibited excellent in vivo antitumour effects in a murine breast cancer xenograft model without visible pathological toxicity.

Near-future levels of ocean temperature weaken the byssus production and performance of the mussel Mytilus coruscus.

Marine mussels are key ecological engineers that form dense aggregations to maintain the vital habitat in benthic systems. It is essential to understand the consequences of mussel byssus attachment in elevated temperatures associated with ocean warming. We evaluated byssus production and the mechanical performance of threads in the mussel Mytilus coruscus at 21° (control), 27 °C (average temperature in the M. coruscus habitat during the summer season) and 31 °C (4 °C raised) for 72 h. We quantified byssus secretion and shedding number, measured byssal breaking force, byssal polyphenol oxidase (PPO) activity, byssal thread length and diameter. Expression of byssus foot protein genes was analyzed by quantitative real-time PCR in foot tissue. High seawater temperature decreased the number of newly secreted byssus and the diameter of byssal threads, leading to the reduction of byssal breaking force and the alteration of the weakest part of the thread. Increased breakpoints in the upper part of the thread (proximal region) were higher at 27 °C than at 21 °C. High-temperature stress significantly reduced the PPO activity in byssus at 31 °C in comparison to 21 °C. The expression of mussel foot protein genes was affected by elevated temperature. The increased gene expression of byssus collagen-like protein 2 (Mccol2) at 31 °C conflicted with the number of byssuses produced. Suggesting the reduction of mussel foot protein abundance is not the cause of decreased byssus production at 31 °C. These results show that byssus, as an extracellular structure of mussels, may be highly susceptible to the adverse effects of ocean warming.

Hysteresis effect on backwashing process in a submerged hollow fiber membrane bioreactor (MBR) applied to membrane fouling mitigation.

Hysteresis effect on backwashing in a submerged MBR was investigated with dead-end hollow fiber membranes. The out-of-step changes in TMP and flux is the real hysteresis effect which is common but easily overlooked. Methods of visualization and ultrasonic spectrum analysis were implemented. The results showed that fouling layer is just the culprit of hysteresis effect. Fouling level and fiber length were determined as two key factors that affect hysteresis effect by data and model derivation. Moreover, a hysteresis evaluation index "τbw" is proposed to quantify the result of TMP vs time. The relationship between influence factors and "τbw" is interactive. A linear relationship between fouling level and "τbw" was found as well as an extreme value between fiber length and "τbw". A lower fouling level (lower backwashing flow) and optimal backwashing duration will be helpful for an effective backwashing no matter for membrane fouling control or energy cost reduce.