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The aggregation of Amyloid- β (A β) peptides is associated with neurodegeneration in Alzheimer's disease (AD). We previously identified novel naphtalene derivatives, including the lead compound Amylovis-201, able to form thermodynamically stable complexes with A β species, peptides and fibrils. As the drug showed a chemical scaffold coherent for an effective interaction with the σ 1 receptor chaperone and as σ 1 agonists are currently developed as potent neuroprotectants in AD, we investigated the pharmacological action of Amylovis-201 on the σ 1 receptor. We report that Amylovis-201 is a potent σ 1 agonist by several in silico , in vitro and in vivo assays and that its anti-amnesic and neuroprotective effects involve a pharmacological action at σ 1 receptors. Furthermore, we show for the first time that classical σ 1 receptor agonist (PRE-084), and antagonist (NE-100) are able to interact and disaggregate A β 25–35 fibrils. Interestingly, Amylovis-201 was the only compound inhibiting A β 25–35 aggregates formation. Our results therefore highlight a dual action of Amylovis-201 as anti-aggregating agent and σ 1 receptor agonist that could be highly effective in long-term treatment against neurodegeneration in AD. Amylovis-201, known to interfere with Amyloid- β aggregation, also acts as a sigma-1 agonist in vitro and in vivo. The drug is therefore a dual-acting compound with synergic extracellular and intracellular modes of action. [Display omitted] [ABSTRACT FROM AUTHOR]

Multiple oxaliplatin-resistance mechanisms have been proposed such as increase of anti-inflammatory M2 macrophages and lack of cytotoxic T-cells. Thereby oxaliplatin chemotherapy promotes an immunosuppressive tumor microenvironment and inhibits anti-tumor efficacy. It has been shown that toll-like receptor (TLR) agonists are capable of triggering broad inflammatory responses, which may potentially reduce oxaliplatin-resistance and improve the efficacy of chemotherapy. In this study, we established colorectal tumor-bearing zebrafish and mice, and investigated the effects of TLR agonists and oxaliplatin in macrophage function and anti-tumor T cell immunity as well as tumor growth control in vivo. To increase the potential of this strategy as well minimize side effects, neutral liposomes carrying oxaliplatin and cationic liposomes co -loaded with TLR agonists Poly I:C and R848 were employed for maximum immune activation. Both of two liposomal systems exhibited good physicochemical properties and excellent biological activities in vitro. The combination strategy delivered by liposomes showed more pronounced tumor regression and correlated with decreased M2 macrophage numbers in both zebrafish and mice. Increasing numbers of dendritic cells, DC maturation and T cell infiltration mediated via immunogenic cell death were observed in treated mice. Our study offers valuable insights into the potential of liposomal combination therapy to improve cancer treatment by reprogramming the tumor microenvironment and enhancing immune responses. [Display omitted] [ABSTRACT FROM AUTHOR]

• CA is the most abundant bioactive compound found in Vaccinium dunalianum Wight. • A COX-2/1 zebrafish model was set up to illustrate CA's anti-inflammatory effects. • CA's anti-inflammatory mechanism was analyzed with systematic network pharmacology. 6′- O -Caffeoylarbutin (CA) is the most abundant compound found within Vaccinium dunalianum Wight, which is used as a traditional Chinese herbal tea for its effects in lowering uric acid and anti-rheumatoid arthritis. No reports to date, however, have explored the pharmacological impact of CA on inflammatory activity. In this study, the anti-inflammatory targets and molecular mechanisms of CA were evaluated through zebrafish inflammation model, network pharmacology and molecular docking. The results showed that CA was able to inhibit cyclooxygenase (COX-2/1) at a dose-dependent fashion in 12- O -tetradecanoylphorbol-13-acetate (TPA)-treated zebrafish. Network pharmacology analyses was employed to derive 156 putative CA-related anti-inflammatory targets, which were subjected to functional enrichment analyses to obtain 511 associated Gene Ontology (GO) terms and 180 Kyoto Encyclopedia of Genes and Genomes (KEGG) pathways. Moreover, 9 core targets (HRAS, SRC, MMP9, HSP90AA, CASP, ANXA5, EGFR, ESR1, and AKT1) associated with the anti-inflammatory effect of CA were further selected and identified based on network topology analyses and molecular docking. Overall, the results of this study suggest that CA exerts its anti-inflammatory effects through a complex mechanism of action, and these findings offer a robust foundation for future in-depth analyses exploring the pharmacological pathways through which CA can prevent rheumatoid arthritis-associated damage. [ABSTRACT FROM AUTHOR]

Bioactive glass nanoparticles (BGNs) are well-recognized multifunctional biomaterials for bone tissue regeneration due to their capability to stimulate various cellular processes through released biologically active ions. Understanding the correlation between BGN composition and cellular responses is key to developing clinically usable BGN-based medical devices. This study investigated the influence of CaO content of binary SiO 2 -CaO BGNs (CaO ranging from 0 to 10 mol%) on osteogenic differentiation of rat bone marrow mesenchymal stem cells (rBMSCs) and in vivo bone regeneration in zebrafish osteoporosis model. The results showed that BGNs could promote osteogenic differentiation of rBMSCs by indirectly releasing active ions or directly interacting with rBMSCs by internalization. In both situations, BGNs of a higher CaO content could promote the osteogenic differentiation of rBMSCs to a greater extent. The internalized BGNs could activate the transcription factors RUNX2 and OSX, leading to the expression of osteogenesis-related genes. The results in the zebrafish osteoporosis model indicated that the presence of BGNs of higher CaO contents could enhance bone regeneration and rescue dexamethasone-induced osteoporosis to a greater extent. These findings demonstrate that BGNs can stimulate osteogenic differentiation of rBMSCs by releasing active ions or internalization. A higher CaO content facilitates osteogenesis and bone regeneration of zebrafish as well as relieving dexamethasone-induced osteoporosis. The zebrafish osteoporosis model can be a potent tool for evaluating the in vivo bone regeneration effects of bioactive materials. Bioactive glass nanoparticles (BGNs) are increasingly used as fillers of nanocomposites or as delivery platforms of active ions to regenerate bone tissue. Various studies have shown that BGNs can enhance osteogenic differentiation of bone marrow mesenchymal stem cells (BMSCs) by releasing active ions. However, the correlation between BGN composition and cellular responses and in vivo bone regeneration effect has still not been well investigated. Establishment of a suitable in vivo animal model for investigating this correlation is also challenging. The present study reports the influence of CaO content in binary SiO 2 -CaO BGNs on osteogenic differentiation of BMSCs extracellularly and intracellularly. This study also demonstrates the suitability of zebrafish osteoporosis model to investigate in vivo bone regeneration effect of BGNs. [Display omitted] [ABSTRACT FROM AUTHOR]

The current research sheds light on the biological toxicity of organophosphate flame retardants (OPFRs), yet it overlooks the neurotoxicity and potential molecular mechanisms of tris(1,3-dichloro-2-propyl) phosphate (TiBP), a prominent constituent of the OPFRs. To address this, we utilized zebrafish larvae as a model to investigate TiBP's acute toxicity and neurotoxic effects, along with the associated molecular pathways. Our findings revealed that the 96 h and 120 h LC50 values for TiBP were 56.51 mg/L and 48.85 mg/L, respectively. Gradient exposure based on the 120 h LC50 demonstrated that TiBP induced developmental toxicity, characterized by elevated heart rate, reduced body length, and diminished eye distance. Additionally, a decrease in swimming activity was observed in the light test, along with the inhibition of the neuro crest cell development in Tg (HuC:eGFP) and Tg (sox10: eGFP) zebrafish larvae following TiBP exposure, as well as the alterations of neurogenesis and ACh-related genes. Expression of key neurodevelopment genes, including mbpa , gap43 , nestin , ngfra , was significantly downregulated. Furthermore, heightened anxiety-like behaviors in open field and phototaxis tests were observed, concomitant with neurotransmitter imbalances. Specifically, there was an increase in DA levels, a decrease in GABA, and an upregulation of AChE activity. These disruptions were primarily mediated through transcriptional dysregulation of neurotransmitter synthesis, transport, and reception. Upon exposure to TiBP, zebrafish larvae exhibited a concentration-dependent increase in both ROS level and apoptosis. An upregulation of antioxidant enzymes and their transcription levels indicated the presence of oxidative stress in the larvae. The induction of ddit3 was congruent with the observed apoptosis, suggesting that it may be triggered by oxidative stress via the ERs-CHOP pathway. In summary, our study indicates that oxidative stress is a pivotal molecular event in the neurotoxicity induced by TiBP, implicating the disruption of the GABAergic, dopaminergic, and cholinergic systems, as well as triggering apoptosis. [Display omitted] • TiBP causes developmental toxicity in zebrafish with decreased interocular distance. • Acute TiBP toxicity in larvae indicated by 96 h/120 h LC50 of 56.51 and 48.85 mg/L. • Neurogenesis inhibition & neuro crest cell alloplasia linked to swimming activity. • TiBP induces anxiety-like behaviors via neurotransmitter imbalances. • TiBP triggers oxidative stress, affecting neurotransmitters via ERs-CHOP pathway. [ABSTRACT FROM AUTHOR]

Gastrodia elata (GE) has been widely used in clinical practice for many years with the functions of relieving stroke, suppressing liver Yang, dispelling wind and clearing collaterals. Our group's previous experimental studies have proved that GE has therapeutic effect on cerebral ischemia reperfusion injury (CIRI) (Ding et al., 2022). However, the active components of GE in treating CIRI remain unclear and require further research. The purpose of this paper was to explore the potential effective components of GE improving CIRI based on the "Spectrum-effect" correlation. Zebrafish model was used for verification in vivo experimental. First, the absorption components and metabolites of GE in rat serum were identified using ultra-high performance liquid chromatography-quadrupole time-of-flight tandem mass spectrometry (UHPLC-Q-TOF/MS). Second, pharmacodynamic indexes were determined by ELISA kit method, and the effect-time curve of each pharmacodynamic indexes was established. The potential compounds were screened using the statistical method of grey correlation between pharmacodynamic indicator and component response. Finally, the zebrafish CIRI model was successfully established, and the in vivo effect of the active components of GE was verified intuitively. 45 chemical components were detected in GE. A total of 87 active components in serum of GE were identified including 25 prototype components and 62 metabolites. GE can improve CIRI by regulating the levels of interleukin-6 (IL-6), interleukin-1β (IL-1β), and tumor necrosis factor-α (TNF-α), MDA levels and SOD levels. It was found that p‑hydroxy benzaldehyde (PHB), p-hydroxybenzyl alcohol (PHBA) and gastrodin (GA) of GE were the possibly main active components by grey correlation statistics. The in vivo experiments of zebrafish model showed that PHB, PHBA, and GA have the ability to ameliorate cerebral thrombosis by regulation of oxidative stress and apoptosis. The potential active components of GE on CIRI were initially excavated using UHPLC-Q-TOF-MS/MS, pharmacodynamics, and in vivo experiments of zebrafish model. It makes up for the disadvantages of separate research on chemical components and pharmacodynamics, and reflects the material basis of pharmacodynamics more objectively. It has provided theoretical basis for further quality evaluation and scientific foundation for rational drug using of GE in clinical. [Display omitted] [ABSTRACT FROM AUTHOR]

Advanced probes for imaging viscous lipids microenvironment in vitro and in vivo are desirable for the study of membranous organelles and lipids traffic. Herein, a reaction-based dihydroquinoline probe (DCQ) was prepared via linking a diethylamino coumarin fluorophore with a N -methylquinoline moiety. DCQ is stable in low viscous aqueous mediums and exhibits green fluorescence, which undergoes fast autoxidation in high viscous mediums to form a fluorescent product with deep-red to near-infrared (NIR) emission, rendering the ability for dual-color imaging. Living cell imaging indicated that DCQ can effectively stain lysosomal membranes with deep-red fluorescence. Super-resolution imaging of lysosome vesicles has been achieved by DCQ and stimulated emission depletion (STED) microscopy. In addition, DCQ realizes multiple organs imaging in zebrafish, whose dual-color emission can perfectly discriminate zebrafish's yolk sac, digestive tract and gallbladder. Most importantly, DCQ has been successfully used to establish a gallbladder-visualizable zebrafish model for the evaluation of drug stress. Super resolution imaging of lysosome membrane in living cells and visualizing multiple organs in living zebrafishes have been achieved by a reaction-based probe DCQ. DCQ realizes the development of a gallbladder-visualizable zebrafish model for the evaluation of drug stress. [Display omitted] • The proposed probe DCQ undergoes oxidation in viscosity-dependent manners. • Green to NIR fluorescence conversion realizes dual-color imaging. • STED super-resolution imaging of lysosome membranes has been achieved. • A high-quality gallbladder-visualizable zebrafish model has been developed using DCQ. [ABSTRACT FROM AUTHOR]

Here, we designed, synthesized and characterized three new cyclometalated Ru(II) complexes, [Ru(phen) 2 (1-(4-Ph-Ph)-IQ)]+ (phen = 1,10-phenanthroline, IQ = isoquinoline, RuIQ9), [Ru(phen) 2 (1-(4-Ph-Ph)-7-OCH 3 -IQ)]+ (RuIQ10), and [Ru(phen) 2 (1-(4-Ph-Ph)-6,7-(OCH 3) 2 -IQ)]+ (RuIQ11). The cytotoxicity experiments conducted on both 2D and 3D multicellular tumor spheroids (MCTSs) indicated that complexes RuIQ9–11 exhibited notably higher cytotoxicity against A549 and A549/DDP cells when compared to the ligands and precursor compounds as well as clinical cisplatin. Moreover, the Ru(II) complexes displayed low toxicity when tested on normal HBE cells in vitro and exposed to zebrafish embryos in vivo. In addition, complexes RuIQ9–11 could inhibit A549 and A549/DDP cell migration and proliferation by causing cell cycle arrest, mitochondrial dysfunction, and elevating ROS levels to induce apoptosis in these cells. Mechanistic studies revealed that RuIQ9–11 could suppress the expression of Nrf2 and its downstream antioxidant protein HO-1 by inhibiting Nrf2 gene transcription in drug-resistant A549/DDP cells. Simultaneously, they inhibited the expression of efflux proteins MRP1 and p-gp in drug-resistant cells, ensuring the accumulation of the complexes within the cells. This led to an increase in intracellular ROS levels in drug-resistant cells, ultimately causing damage and cell death, thus overcoming cisplatin resistance. More importantly, RuIQ11 could effectively inhibit the migration and proliferation of drug-resistant cells within zebrafish, addressing the issue of cisplatin resistance. Accordingly, the prepared Ru(II) complexes possess significant potential for development as highly effective and low-toxicity lung cancer therapeutic agents to overcome cisplatin resistance. Three novel cyclometalated Ru(II) complexes RuIQ9–11 could suppress the expression of Nrf2 and its downstream antioxidant protein HO-1 by inhibiting Nrf2 gene transcription in A549/DDP cells, ultimately causing cell death. RuIQ9–11 also exerted a prominent inhibitory effect on 3D MCTSs of A549 and A549/DPP cells. RuIQ11 could effectively inhibit the migration and proliferation of A549/DPP cells within zebrafish. [Display omitted] • Three new cyclometalated Ru(II) complexes were synthesized and characterized. • RuIQ9–11 exhibited relative low toxicity on both normal HBE cells in vitro and zebrafish embryos in vivo. • RuIQ9–11 could suppress the expression of Nrf2 and its downstream antioxidant protein HO-1, ultimately causing cell death. • RuIQ9–11 exerted a prominent inhibitory effect on 3D multicellular tumor spheroid model. • RuIQ11 could effectively inhibit the migration and proliferation of drug-resistant cells within zebrafish. [ABSTRACT FROM AUTHOR]

[Display omitted] • Homologous gene was analyzed between DEGs of NAFLD patients and zebrafish model. • DAG feeding can alleviate NAFLD symptoms both in larval and grown zebrafish. • DAG diet can prevent the occurrence of NAFLD in zebrafish model. • Mechanism of DAG in ameliorating NAFLD was investigated through transcriptomic analysis. The escalating prevalence of non-alcoholic fatty liver disease (NAFLD), driven by inadequate dietary habits and metabolic imbalances, underscores the urgent need for effective dietary interventions. This study investigated the potential of 80 % peanut diacylglycerol (DAG) as a functional lipid component in preventing NAFLD and associated intestinal microenvironment based on a high-cholesterol diet (HCD)-induced zebrafish model. We first found that the zebrafish fatty liver model shared multiple similar pathways like lipid-induced atherosclerosis, insulin resistance with NAFLD patient in terms of lipid metabolism. Employing larvae and grown zebrafish, we aimed to elucidate the preventive and alleviating effect of a DAG diet on NAFLD respectively. Results showed that DAG post-intervention significantly decreased hepatic lipid accumulation and diminished intestinal inflammatory in fatty liver zebrafish model. Notably, transcriptomic sequencing illuminated upregulation in genes related to lipid metabolism (e.g., fasn, mogat2, cd36), suggesting that peanut DAG accelerated lipid transportation and fostered a metabolic environment less conducive to NAFLD symptoms. Besides, reduced liver lipid droplets and hepatic inflammatory suggested DAG feeding could also prevent the formation of NAFLD. The study provided compelling evidence supporting the incorporation of DAG into the diet as a strategy for the prevention and management of fatty liver disease, proposing a novel approach to functional food development aimed at combating metabolic diseases. [ABSTRACT FROM AUTHOR]

Perillaldehyde (PAE), a prevalent flavoring agent, has raised safety concerns due to conflicting evidence regarding its toxicity. This study provides a comprehensive assessment of the developmental and neurotoxic effects of PAE in zebrafish, elucidating the underlying mechanisms of its toxicity. Results showed that PAE affected the viability and hatching rate of zebrafish at 96 h postfertilization with the 50 % lethal concentration (LC50) of 7.975 mg/L. Furthermore, exposed‌ to a non-lethal concentration of 4 mg/L PAE induced a spectrum of morphological abnormalities, such as pericardial edema, delayed yolk sac absorption, reduced body length, and microphthalmia. Behavioral observations revealed that PAE reduced motor ability, and was accompanied by an increase in spontaneous turning angle and angular velocity. Using the TG(elav13:EGFP) transgenic model, we observed the number of newborn neurons was reduced, indicating that PAE induced obvious neurotoxic effects. Additionally, this concentration facilitated the accumulation of reactive oxygen species (ROS) and malondialdehyde (MDA), concomitantly decreasing the activity of antioxidant enzymes. QRT-PCR analysis revealed that PAE down-regulated Nestin and Neurogenin1 gene expression, up-regulated Glipr1a and Nox1 gene expression, and inhibited the Nrf2/HO-1 pathway. Notably, co-administration of N -acetylcysteine (NAC), an inhibitor of oxidative stress, mitigated oxidative stress levels and partially ameliorated the neurotoxicity. These findings suggest that oxidative stress is the primary mediator of PAE-induced neurotoxicity. This study provides crucial insights for the safe application of PAE. • PAE led to the developmental toxicity in zebrafish embryos. • PAE influenced the locomotor behavior of zebrafish larvae. • PAE disrupted gene expression related to neurodevelopment and oxidative stress. • The neurotoxicity induced by PAE can be reversed by NAC. • Neurotoxicity induced by PAE was linked to the inhibition of the NRF2 pathway. [ABSTRACT FROM AUTHOR]

• Acetyl quinolone thiosemicarbazone and its derivatives were synthesized by simple work up and column free products. • Phenyl ring substituted thiosemicarbazide 5b and 6b shows the good in-vitro antioxidant activity. • From the In-vivo embryo toxicity analysis, all the synthesized compounds had a less toxic. • The compound 5b and 6b shows the good binding energy for the protein 2C0U, 3NM8. In this study, we successfully synthesized a series of acetyl quinolone thiosemicarbazones (5a-d, 6a-d) using a simple work-up procedure yielding column-free products. The structures of the synthesized compounds were confirmed by various spectroscopic techniques, including 1H, 13C, and 2D NMR, along with FT-IR. Detailed 2D NMR analyses such as COSY, HSQC, and HMBC were also performed. The biological activities were evaluated through in-vitro, in-vivo , and in-silico studies. In-vitro antioxidant properties were assessed using radical scavenging assays, revealing significant activity for compounds 5b and 6b , likely due to the aromatic ring substitution in the thiosemicarbazide side chain. In-vivo zebrafish embryo toxicity studies indicated minimal toxicity for these compounds. Additionally, in-silico studies showed that compounds 5b and 6b have good binding energies for proteins 2C0U, 3NM8 and 2X08. Further the DFT study of HOMO and LUMO energy gap is small for 5b and 6b. These findings suggest that acetyl quinolone thiosemicarbazones are promising candidates for various biological applications due to their potent antioxidant activity and low toxicity. [Display omitted] [ABSTRACT FROM AUTHOR]

• Carbon dots are synthesized by carbonization of chitosan. • Carbon dots exhibit excitation dependent luminescent properties. • Chitosan and gelatin are crosslinked to improve the drug encapsulation and solubility of the resultant crosslinked polymer. • Carbon dots embedded in the crosslinked chitosan/gelatin polymer can act as an imaging probe for bioimaging applications. • Antibacterial, antioxidant and cytotoxicity of the nanocomposite is examined. The present work elucidates the synthesis and properties of chitosan/gelatin/carbon dot nanocomposite. The synthesized carbon dots are incorporated into the chitosan-gelatin matrix in the ratio of 1:1. This ratio was chosen as it showed better release, anti-bacterial and antioxidant properties. The integration of carbon dots and dopamine into the chitosan-gelatin matrix was confirmed by XRD, UV, IR analysis and TEM micrographs. Toxicity studies of the composite were examined with the help of the IC-21 macrophage cell line. The release of the drug was tabulated in-vitro , and Korsmeyer - Peppas Case II transport release kinetics was observed over a period of 30 h in pH 7. After 30 h, 90 % of the encapsulated dopamine was released from the matrix. The addition of carbon dots enhances the controlled and extended release of the drug. The system is also tested for antibacterial and antioxidant properties. The highlight of this work is the facile synthesis and luminescent property of chitosan-derived carbon dots. The embryos and larvae of zebrafish were treated with the synthesized carbon dots, which provided them with an incubation period of 24 h. They were then observed under a fluorescent microscope with DAPI, FTIC and TRITC filters. Excitation-dependant emission, i.e., blue, green, and red emission, was seen in the sample when excited with UV, blue and green wavelengths, respectively. Its fluorescence properties help in its use as a tracer in monitoring the flow and movement of the drug-loaded composite. [Display omitted] [ABSTRACT FROM AUTHOR]

[Display omitted] • Emoxypine Succinate mitigates iron overload-induced neurodegeneration in a zebrafish model. • Improved behavioral performance observed in treated zebrafish, evidenced by enhanced exploration and memory functions. • Significant reduction in oxidative stress markers following emoxypine succinate treatment. • Suppression of neuroinflammatory markers, including IL-1β, TNF-α, CDK-5, GSK-3β, and NLRP3, highlights its potential in modulating neuroinflammation associated with neurodegenerative disorders. Excessive iron accumulation in the brain plays a significant role in neurodegenerative processes, contributing to the pathogenesis of Alzheimer's disease (AD). AD, a prominent neurological disorder affecting the central nervous system, is characterized by the accumulation of beta-amyloid (Aβ) and tau phosphorylation. This accumulation leads to the subsequent development of cognitive impairments, particularly in learning and memory functions. This study investigates the neuroprotective effects of emoxypine succinate in a zebrafish model of iron overload-induced neurodegeneration. Iron was administered to the zebrafish for 28 days to induce neurodegeneration. Following induction, Emoxypine succinate was employed as a treatment intervention for 14 days (concentrations of 4 mg/L, 8 mg/L, and 12 mg/L). Following the end of the treatment, behavioral tests (Y maze test, Novel tank test) were conducted on the zebrafish, and the biochemical (MDA, Catalase, SOD, GSH) and molecular parameters (AchE, Iron levels, IL-1β, TNF-α, CDK-5, GSK-3β, and NLRP3) of the zebrafish brain were also assessed. In the novel tank test, emoxypine succinate-treated groups exhibited significantly increased time in the upper zone (p < 0.001), higher distance travelled (p < 0.001), and shorter latency to the top (p < 0.001) compared to the negative control. Similarly, the Y-maze test revealed improved time in the novel arm (p < 0.001) and total distance travelled (p < 0.001) in treated groups versus the negative control. Assessment of oxidative stress parameters demonstrated significant reductions in oxidative stress in emoxypine succinate-treated groups. Furthermore, AChE activity decreased significantly (p < 0.001), and brain iron levels decreased substantially (p < 0.001) in treated groups, indicating positive therapeutic outcomes. Molecular analysis showed a significant reduction in pro-inflammatory markers like IL-1β, TNF-α, CDK-5, GSK-3β, and NLRP3 (p < 0.001). This comprehensive study highlights the potential efficacy of emoxypine succinate in mitigating neurodegeneration associated with iron dysregulation. [ABSTRACT FROM AUTHOR]

• Novel compound heterozygous mutations causing TNNT1 myopathy. • Combination of missense and splicing variants resulting in an intermediate phenotype. • Impact of a missense variant validated using a zebrafish loss-of-function model. Nemaline myopathies are clinically and genetically heterogeneous disorders caused by several different genes. One of them is TNNT1 , which was initially described in Amish families and has not been reported in Asian populations. Although most TNNT1 myopathies are caused by loss-of-function mutations, several recent studies have shown that missense mutations can also be pathogenic. A 16-year-old Korean boy with progressive muscle weakness visited the Seoul National University Hospital. He showed generalized myopathy, which was predominant in the paraspinal and neck muscles. Moreover, nemaline rods were observed in a muscle biopsy. Whole-exome sequencing of DNA samples of the patient and his younger brother, who had a similar phenotype, revealed novel compound heterozygous mutations in TNNT1 (c.724G>C (p.Ala242Pro) and c.611+1G>A). Sanger sequencing of cDNA extracted from muscle samples of the patient confirmed partial or total skipping of exon 11 in the splicing variant. The impact of the missense variant on muscle integrity and locomotor activity was verified using a zebrafish loss-of-function model. Here, we reported novel familial cases of TNNT1 myopathy with intermediate clinical presentations caused by compound heterozygous mutations and demonstrated their functional defects using an animal model. [ABSTRACT FROM AUTHOR]

Phloretin loaded porous starch (Ph-PS) were prepared for its application in food. The effects of Ph-PS in vitro release and its ability against AAPH-induced oxidative stress in vivo zebrafish model were investigated. Ph-PS was prepared by absorption method, the physical and chemical characterization showed that PS decreased the crystallinity of Ph obviously. Ph-PS exhibited higher release amount and faster release rate of Ph compared to free Ph in vitro release study. What's more, the effect of Ph-PS reduced ROS generation and lipid peroxidation was better than that of free Ph in zebrafish model. These findings suggest Ph-PS is a new and simple strategy to improve dissolution rate and antioxidant ability of Ph. In this paper: • Phloretin loaded porous starch were prepared by physical absorption method, the crystallinity of Ph decreased obviously. • Ph-PS exhibited higher release amount and faster release rate of Ph in vitro release study compared to free Ph. • The effect of Ph-PS reduced ROS generation and lipid peroxidation was better than that of free Ph in zebrafish model. [ABSTRACT FROM AUTHOR]

Selenium nanoparticles (SeNPs), a potential cancer therapeutic agent, have attracted widespread attention owing to their high bioavailability and remarkable anticancer activity. Nevertheless, the poor water solubility and dispersibility of SeNPs seriously limit their applications. In the present study, we synthesized stable and individual spherical selenium nanoparticles (CL90-Tw-SeNP2) with an average diameter of approximately 79 nm using a polysaccharide extracted from Citrus limon (CL90) and Tween-80 as the decorator and stabilizers. The proportion of selenium in CL90-Tw-SeNP2 was 10.6%. CL90-Tw-SeNP2 possessed high stability and good dispersion in water for more than three months. The subsequent biological assay revealed that CL90-Tw-SeNP2 showed remarkable antitumor effects against HepG2 cells, with an IC 50 value of 49.13 μg/mL, by inducing cell apoptosis. Furthermore, an in vivo zebrafish assay to explore possible applications indicated that CL90-Tw-SeNP2 could inhibit the proliferation and migration of tumors and the zebrafish angiogenesis. These results indicated that CL90-Tw-SeNP2 could be a potential agent for cancer treatment, especially against human liver hepatoma cancer. [Display omitted] • High stability and selenium content of CL90-Tw-SeNP2 were synthesized successfully. • Citrus limon polysaccharides and Tween-80 acted as the stabilizers in the reaction. • CL90-Tw-SeNP2 exerted remarkable antitumor effects by inducing apoptosis. • CL90-Tw-SeNP2 inhibited the proliferation and migration of tumors. • CL90-Tw-SeNP2 can block the angiogenesis of zebrafish. [ABSTRACT FROM AUTHOR]

A water-soluble polysaccharide identified here as ADP80-2 was acquired from Angelica dahurica. ADP80-2 was a gluco-arabinan composed of arabinose and a trace of glucose with a molecular weight of 9950 g/mol. The backbone of ADP80-2 comprised →5)- α -L-Ara f -(1→, →3, 5)- α -L-Ara f -(1→, →6)- α -D-Glc p -(1→, with a terminal branch α -L-Ara f -(1 → residue. In terms of immunoregulatory activity, ADP80-2 can significantly promote the phagocytosis, the production of nitric oxide (NO), and the secretion of cytokines (IL-6, IL-1 β , and TNF- α) of macrophage. In addition to the cellular immunomodulatory activities, the chemokines related to immunoregulation were significantly increased in the zebrafish model after treated with ADP80-2. These biological results indicated that ADP80-2 with immunomodulatory effects was expected to be useful for the development of new immunomodulatory agents. Simultaneously, the discovery of ADP80-2 further revealed the chemical composition of A. dahurica used as a traditional Chinese medicine and spice. [Display omitted] • An active polysaccharide ADP80-2 was isolated from Angelica dahurica. • ADP80-2 was a gluco-arabinan without triple helix conformation in solution. • ADP80-2 showed prominent immunomodulatory activity both in vivo (zebrafish embryo) and in vitro (RAW264.7). [ABSTRACT FROM AUTHOR]

Polygala tenuifilia Willd (Polygalaceae), a traditional Chinese medicine, has been used for a long time to treat various illnesses with serious adverse reactions. Glycyrrhizae radix et rhizoma processing is generally used to reduce the adverse reactions. The aim of this study was to validate the irritation caused by raw Polygalaceae (RPA), to investigate whether processed Polygalaceae (PGA) was less irritating, and to screen and validate irritant properties of virgaureagenin G (polygala acid, PA), 3,6′-disinapoylsucrose (DSS), Tenuifolia (TEN) and polygalaxanthone III (POL), which had pharmacologically active in Polygalaceae. Zebrafish model, Draize test and High-Performance Liquid Chromatography (HPLC) were utilized to achieve the aim. Scanning Electron Microscopy (SEM) and optical microscope were used to determine the presence of calcium oxalate needle crystal in RPA and PGA. Zebrafish egg spinning changes and zebrafish embryo behavior were used for irritation validation, irritation comparison and irritant screening. For additional evidence, the Draize test, HE staining of rabbit eyes and ELISA kit were used. Finally, changes in the composition of RPA and PGA were investigated using HPLC. SEM and optical microscopy revealed no calcium oxalate needle crystals in Polygalaceae. RPA, PGA, PA and DSS were able to accelerate the spinning of zebrafish eggs and the movement of embryos, while TEN and POL were not. RPA, PGA, DSS and PA may cause rabbit eyes to become hyperemic and swollen, resulting in damage to the iris, cornea and conjunctiva and increased levels of interleukin-6 (IL-6) and interleukin-10 (IL-10). Comparatively, the effects caused by PGA were less severe than those caused by RPA. In addition, compared to RPA, PGA had lower levels of DSS and PA. RPA, PGA, DSS, and PA were irritating. However, processing and curing could reduce the irritation by reducing the levels of DSS and PA. DSS and PA could be two potential irritants of Polygalaceae. [Display omitted] • Polygala tenuifilia Willd is irritating. • Polygala tenuifilia Willd processed by Glycyrrhizae Radix et Rhizoma is less irritating. • 3,6′-disinapoylsucrose and virgaureagenin G are two of the irritating ingredients of Polygala tenuifilia Willd. • Processed Polygala tenuifilia Willd has lower content of 3,6′-disinapoylsucrose and virgaureagenin G. [ABSTRACT FROM AUTHOR]

Four new undescribed halimane- and labdane-type diterpenoids, named zeylleucapenoids E–H (1–4), along with four known analogues (5–8), were isolated from the aerial parts of Leucas zeylanica (L.) R. Br. Their structures were determined by comprehensive spectroscopic analysis and computational calculations. Compounds 1 and 2 are the highly modified halimane diterpenoids featuring a 6/6/6-fused tricyclic system with an unusual six-membered 6,11-ether ring. Compound 8 exhibits nontoxic effects for zebrafish embryo, while it displays efficient reduction against NO production in a dose-dependent manner and strongly suppresses the secretion of LPS-induced TNF- α and IL-6 cytokines in RAW264.7 macrophages. In addition, marked reductions of iNOS and COX-2 expression were observed. Molecular docking analysis indicated that 8 has high affinities with the target amino acid residues on protein-binding sites, which may be a possible mechanism contributing to the anti-inflammatory potential of this molecule. [Display omitted] • Four new diterpenoids were purified from Leucas zeylanica. • Two structurally unique 6/6/6-fused tricyclic halimane-type structures were purified from Leucas zeylanica. • ECD and VCD calculations were employed to define the absolute configuration of new diterpenoids. • The in vitro anti-inflammatory effects of all the isolates were evaluated. • The anti-inflammatory mechanism was preliminarily investigated using molecular docking and Western blotting analysis. • The nontoxic effects were conducted with a zebrafish model. [ABSTRACT FROM AUTHOR]

Redox control seems to be indispensable for proper embryonic development. The ratio between glutathione (GSH) and its oxidized disulfide (GSSG) is the most abundant cellular redox circuit. We used zebrafish harboring the glutaredoxin 1-redox sensitive green fluorescent protein (Grx1-roGFP) probe either in mitochondria or cytosol to test the hypothesis that the GSH:GSSG ratio is strictly regulated through zebrafish embryogenesis to sustain the different developmental processes of the embryo. Following the GSSG:GSH ratio as a proxy for the GSH-dependent reduction potential (E h GSH) revealed increasing mitochondrial and cytosolic E h GSH during cleavage and gastrulation. During organogenesis, cytosolic E h GSH decreased, while that of mitochondria remained high. The similarity between E h GSH in brain and muscle suggests a central regulation. Modulation of GSH metabolism had only modest effects on the GSSG:GSH ratios of newly hatched larvae. However, inhibition of GSH reductase directly after fertilization led to dead embryos already 10 h later. Exposure to the emerging environmental pollutant Perfluorooctane Sulfonate (PFOS) disturbed the apparent regulated E h GSH as well. Mitochondrial and cytosolic GSSG:GSH ratios are almost identical in different organs during zebrafish development indicating that the E h GSH might follow H 2 O 2 levels and rather indirectly affect specific enzymatic activities needed for proper embryogenesis. Our data confirm that vertebrate embryogenesis depends on strictly regulated redox homeostasis. Disturbance of the GSSG:GSH circuit, e.g. induced by environmental pollution, leads to malformation and death. [Display omitted] • No organ specific mitochondrial and cytosolic GSH:GSSG ratios in zebrafish larvae. • Almost no effects of manipulated GSH metabolism on E h GSH in older zebrafish larvae. • Inhibition of GSH reductase in young larvae leads to malformation and death. • The environmental pollutant Perfluorooctane Sulfonate affects E h GSH in larvae. [ABSTRACT FROM AUTHOR]

Oxidative stress disorders and diseases caused by drug-resistant bacteria have emerged as significant public health concerns. Plant-based medications like protease inhibitors are growing despite adverse effects therapies. Consecutively, in this study, trypsin inhibitors from Dioscorea bulbifera L. (DbGTi trypsin inhibitor) ground tubers were isolated, purified, characterized, and evaluated for their potential cytotoxicity, antibacterial, and antioxidant activities. DbGTi protein was purified by Q-Sepharose matrix, followed by trypsin inhibitory activity. The molecular weight of the DbGTi protein was found to be approximately 31 kDa by SDS-PAGE electrophoresis. The secondary structure analysis by circular dichroism (CD) spectroscopy revealed that the DbGTi protein predominantly comprises β sheets followed by α helix. DbGTi protein showed competitive type of inhibition with V max = 2.1372 × 10−1 μM/min, K m = 1.1805 × 102 μM, & K i = 8.4 × 10−9 M and was stable up to 70 °C. DbGTi protein exhibited 58 % similarity with Dioscorin protein isolated from Dioscorea alata L. as revealed by LC-MS/MS analysis. DbGTi protein showed a non-toxic effect, analyzed by MTT, Haemolytic assay and in vivo studies on zebrafish model. DbGTi protein significantly inhibited K. pneumoniae and has excellent antioxidant properties, confirmed by various antioxidant assays. The results of anti-microbial, cytotoxicity and antioxidant assays demonstrate its bioactive potential and non-toxic nature. • Isolated & characterized DbGTi protein from Dioscorea bulbifera. L tubers. • DbGTi exhibits strong thermal stability and competitive trypsin inhibition. • DbGTi exhibits potent antimicrobial efficacy against Klebsiella pneumoniae. • DbGTi displays exceptional antioxidant efficacy across diverse radicals. • Non-cytotoxicity against humans, shown by cell viability and in vitro assays. [ABSTRACT FROM AUTHOR]

Pectins are highly sought after as food additives, functional ingredients, and structural polymers. So far, no study has been performed on the extraction, characterization, and application of pectins from garlic pomace. In this study, a medium-sized low-methoxyl pectin rich in rhamnogalacturonan-I (RG-I) and galactose was prepared from garlic pomace (termed "GP"; molecular weight, Mw, 324.08 kDa; degree of methyl esterification, DM, 48.10%; RG-I, 35.20%), via citric acid-water extraction at 65 °C for 120 min and ethanol precipitation. GP contained six neutral monosaccharides (fucose, rhamnose, arabinose, galactose, glucose, and xylose) and two acidic monosaccharides (galacturonic acid and glucuronic acid), and could protect zebrafish against metronidazole-induced oxidative damage. Compared with commercial citrus pectin, GP had a comparable Mw, much more RG-Is and homogalacturonans, higher protein and phenolic contents, higher degree of branching, lower crystallinity, higher thermal stability (over 30–377 °C), higher in vitro and in vivo antioxidant capacities, higher viscosity, and higher emulsifying and emulsion-stabilizing abilities. Accordingly, garlic pomace was a good source of RG-I-rich and galactose-rich pectins with high emulsifying and antioxidant capacities. GP can be used for making foods and pharmaceuticals possessing simultaneously high emulsifying, emulsion-stabilizing, and antioxidant capacities. This study demonstrates a sustainable garlic waste valorization approach. [Display omitted] • Pectins rich in RG-I and galactose (GP) were prepared from garlic pomace. • GP was a medium-sized low-methoxyl pectin with a high degree of branching. • GP showed more excellent emulsifying abilities than commercial citrus pectin. • GP effectively protected zebrafish against metronidazole-induced oxidative damage. • GP is a good ingredient/additive with high emulsifying and antioxidant capacities. [ABSTRACT FROM AUTHOR]

Citrobacter infections are becoming an increasingly significant health problem in aquaculture in South-Eastern countries. The objective of this study was to isolate and evaluate the potential of lytic bacteriophages against Citrobacter infections. TEM analysis revealed that the isolated phages Citrophage MRM19 and Citrophage MRM57 were identified to be Siphovirus and Podovirus family of the order Caudovirales. The phage life-cycle studies showed that Citrophage MRM19 had an adsorption time of 18 ± 1 min and a latency period of 25 ± 3 min with burst size of 110 ± 20 phages/infected cell and Citrophage MRM57 had an adsorption time of 15 ± 1 min and a latency period of 25 ± 2 min with burst size of 50 ± 5 phages/infected cell. In vitro studies indicated that the bacterial load was reduced by 5 and 7 log units within 12 h by Citrophage MRM19 and Citrophage MRM57. The in vivo efficacy of the phages was studied using zebrafish (Danio rerio) as a model organism in low-scale tanks. The study unveiled that the use of phages increased the survival up to 17%, 23%, and 26% in the case of Citrophage MRM19, Citrophage MRM57, and phage cocktail treatment, respectively. Our study indicated that bacteriophages are suitable biocontrol agents against Citrobacter spp. especially in aquaculture industry. [ABSTRACT FROM AUTHOR]

• AS-48 is a 70-residue circular bacteriocin produced by Enterococcus strains. • Toxicity of AS-48 in in vitro and in vivo models has been evaluated. • The haemolytic activity and lacks of pro-inflammatory effects of AS-48 are minimal. • AS-48 has scarce ability to cause loss of cell viability at therapeutic concentrations. • AS-48 is a promising therapeutic agent against a vast array of microbial infections. The in vitro antimicrobial potency of the bacteriocin AS-48 is well documented, but its clinical application requires investigation, as its toxicity could be different in in vitro (haemolytic and antibacterial activity in blood and cytotoxicity towards normal human cell lines) and in vivo (e.g. mice and zebrafish embryos) models. Overall, the results obtained are promising. They reveal the negligible propensity of AS-48 to cause cell death or impede cell growth at therapeutic concentrations (up to 27 μM) and support the suitability of this peptide as a potential therapeutic agent against several microbial infections, due to its selectivity and potency at low concentrations (in the range of 0.3–8.9 μM). In addition, AS-48 exhibits low haemolytic activity in whole blood and does not induce nitrite accumulation in non-stimulated RAW macrophages, indicating a lack of pro-inflammatory effects. The unexpected heightened sensitivity of zebrafish embryos to AS-48 could be due to the low differentiation state of these cells. The low cytotoxicity of AS-48, the absence of lymphocyte proliferation in vivo after skin sensitization in mice, and the lack of toxicity in a murine model support the consideration of the broad spectrum antimicrobial peptide AS-48 as a promising therapeutic agent for the control of a vast array of microbial infections, in particular, those involved in skin and soft tissue diseases. [ABSTRACT FROM AUTHOR]

Graphical abstract Highlights • HFD enhances HCC progression and modulates the immune response in the liver microenvironment. • HFD induced changes in macrophage polarization with increased numbers of TNFα-positive macrophages in the liver. • HFD reduces T cell infiltration to liver area in NASH-associated HCC larvae. • Ablation of macrophages reduces disease progression in NASH-associated HCC larvae, but not in HCC alone. • Metformin specifically affects the progression induced by diet in NASH-associated HCC in zebrafish. Background & Aims Non-alcoholic fatty liver disease/non-alcoholic steatohepatitis (NAFLD/NASH) is an increasing clinical problem associated with progression to hepatocellular carcinoma (HCC). The effect of a high-fat diet on the early immune response in HCC is poorly understood, while the role of metformin in treating NAFLD and HCC remains controversial. Herein, we visualized the early immune responses in the liver and the effect of metformin on progression of HCC using optically transparent zebrafish. Methods We used live imaging to visualize liver inflammation and disease progression in a NAFLD/NASH-HCC zebrafish model. We combined a high-fat diet with a transgenic zebrafish HCC model induced by hepatocyte-specific activated beta-catenin and assessed liver size, angiogenesis, micronuclei formation and inflammation in the liver. In addition, we probed the effects of metformin on immune cell composition and early HCC progression. Results We found that a high-fat diet induced an increase in liver size, enhanced angiogenesis, micronuclei formation and neutrophil infiltration in the liver. Although macrophage number was not affected by diet, a high-fat diet induced changes in macrophage morphology and polarization with an increase in liver associated TNFα-positive macrophages. Treatment with metformin altered macrophage polarization, reduced liver size and reduced micronuclei formation in NAFLD/NASH-associated HCC larvae. Moreover, a high-fat diet reduced T cell density in the liver, which was reversed by treatment with metformin. Conclusions These findings suggest that diet alters macrophage polarization and exacerbates the liver inflammatory microenvironment and cancer progression in a zebrafish model of NAFLD/NASH-associated HCC. Metformin specifically affects the progression induced by diet and modulates the immune response by affecting macrophage polarization and T cell infiltration, suggesting possible effects of metformin on tumor surveillance. Lay summary This paper reports a new zebrafish model that can be used to study the effects of diet on liver cancer. We found that a high-fat diet promotes non-resolving inflammation in the liver and enhances cancer progression. In addition, we found that metformin, a drug used to treat diabetes, inhibits high-fat diet-induced cancer progression in this model, by reducing diet-induced non-resolving inflammation and potentially restoring tumor surveillance. [ABSTRACT FROM AUTHOR]

Lead (Pb) contamination is a worldwide public health threaten. Besides close restraint of lead exposure, it's emergency to discover compounds that could help to cue toxicities caused by lead. Zebrafish embryos and early larvae can serve as valuable screening tools in early pre-clinical phase of drug screening and research. In order to establish a zebrafish lead poisoning model that could be used for drug screening and research, zebrafish embryos at 6 h post-fertilization (hpf) were treated with lead at different concentrations by soaking intermittently, raised in lead work solution at night while in fish water during the day. After treated for 90 h, death and severe trunk curvature were observed on zebrafish in 640 μM group, obvious dysplasia, blood toxicity, excessive reactive oxygen species (ROS), severe neurotoxicity, such as shorter length of peripheral motor neurons, neuronal apoptosis, and axonal injury, and neurobehavior impairment were induced by lead at 80, 160 and 320 μM, similar to phenotypes reported in rodent. Moreover, the mRNA level of genes related to neurodevelopment, memory, and antioxidation were significantly down regulated, and apoptosis-related genes were up regulated, consistent to zebrafish phenotypic change. Finally, zebrafish were intermittently exposed to 80 μM lead solution to establish the lead poisoning model, and the efficacy of a safe chelating agent Meso-2,3-Dimercaptosuccinic acid (DMSA) was tested at a series of concentrations to validate the zebrafish model. The result showed concentration-dependent decrease of lead content in zebrafish in DMSA treated groups compared with model group. The above data fully demonstrated a zebrafish model of lead poisoning suitable for drug screening was successfully developed, which was expected to provide a rapid and economic tools for discovering antidotes of lead and drugs of neuroprotection. [Display omitted] • Exposed zebrafish to high concentrations of lead intermittently in acid condition. • Validated lead neurotoxicity in zebrafish. • Revealed the blood toxicity of lead in larval zebrafish. • Developed a acute neurobehavior impairment model for rapid drug screening. [ABSTRACT FROM AUTHOR]

Cigarette smoke exposure poses significant health risks, including oxidative stress, inflammation, tissue damage, and neurodegenerative diseases. Luteolin, a natural flavonoid known for its antioxidant and anti-inflammatory properties, is of interest in countering these effects. This study aims to assess luteolin's protective potential against cigarette smoke extract (CSE) in adult zebrafish. Adult zebrafish were exposed to CSE for 15 days, inducing smoke-related damage. Subsequent luteolin treatment assessed its impact. Evaluations included antioxidant enzymes (SOD, CAT), nitric oxide (NO), LDH activity (cellular damage), tissue integrity, fibrosis, amyloid plaque accumulation, and CSE component analysis via HPLC. CSE exposure heightened oxidative stress, reducing SOD and CAT activity and elevating NO levels, leading to cellular damage and tissue disruption, notably fibrosis and amyloid plaque accumulation. Inflammatory markers TNF-α and IL-1β also increased. Luteolin treatment restored SOD and CAT activity, reduced LDH and NO activity, counteracting oxidative damage. It also mitigated fibrosis and reduced amyloid plaque deposition, preserving tissue integrity. Luteolin reduced TNF-α and IL-1β levels and CSE components, displaying anti-inflammatory effects. This study underscores luteolin's potential as a protective agent against cigarette smoke-induced harm in a zebrafish model. • Cigarette smoke promotes oxidative damage, inflammation, fibrosis and accumulation of amyloid plaques in in-vivo zebrafish. • Luteolin reduces such stress by restoring antioxidant enzymes and protecting cellular integrity. • Cigarette smoke induced inflammation and tissue damage in gills and brain was neutralised by luteolin. • The overexpression of pro-inflammatory cytokines such as TNF-α and IL-1β was suppressed due to luteolin. [ABSTRACT FROM AUTHOR]

Dimethenamid, one of the acetamide herbicides, is widely used on soybeans and corns to inhibit weed growth. Although other acetamide herbicides have been reported to have several toxicities in non-target organisms including developmental toxicity, the toxicity of dimethenamid has not yet been studied. In this research, we utilized the zebrafish animal model to verify the developmental toxicity of dimethenamid. It not only led to morphological abnormalities in zebrafish larvae but also reduced their viability. ROS production and inflammation responses were promoted in zebrafish larvae. Also, uncontrolled apoptosis occurred when the gene expression level related to the cell cycle and apoptosis was altered by dimethenamid. These changes resulted in toxicities in the cardiovascular system, liver, and pancreas are observed in transgenic zebrafish models including fli1a:EGFP and L-fabp:dsRed;elastase:GFP. Dimethenamid triggered morphological defects in the heart and vasculature by altering the mRNA levels related to cardiovascular development. The liver and pancreas were also damaged through not only the changes of their morphology but also through the dysregulation in their function related to metabolic activity. This study shows the developmental defects induced by dimethenamid in zebrafish larvae and the possibility of toxicity in other non-target organisms. [Display omitted] • Dimethenamid causes developmental and morphological abnormalities in zebrafish. • Dimethenamid induces oxidative stress and inflammatory response in embryos. • Dimethenamid increases cardiovascular damage in zebrafish larvae during development. • Dimethenamid triggers hepatotoxicity and pancreatic toxicity in zebrafish larvae. • Dimethenamid may cause the detrimental effects on aquatic ecosystems. [ABSTRACT FROM AUTHOR]

Ten spirocyclic polycyclic polyprenylated acylphloroglucinols (PPAP), hunascynols A–J (1 – 10), and 12 known analogs were isolated from the aerial parts of Hypericum ascyron Linn. Compounds 1 and 2 , which share a 1,2- seco -spirocyclic PPAP skeleton, could be derived from spirocyclic PPAP, with a common octahydrospiro[cyclohexan-1,5′-indene]-2,4,6-trione core, through a cascade of Retro-Claisen, keto-enol tautomerism, and esterification reactions. Aldolization of normal spirocyclic PPAP yielded 3 , which has a caged framework with a 6/5/6/5/6 ring system. The structures of these compounds were determined using spectroscopy and X-ray diffraction. The inhibitory activities of all isolates were tested in three human cancer cell lines and a zebrafish model. Compounds 1 and 2 displayed moderate cytotoxicity against HCT116 cells (IC 50 6.87 and 9.86 μM, respectively). The mechanisms of these compounds were evaluated using Western blot assays. Compounds 3 and 5 inhibited the growth of sub-intestinal vessels in zebrafish embryos. Further, the target genes were screened using real-time PCR. Ten undescribed spirocyclic polycyclic polyprenylated acylphloroglucinols, including two 1,2- seco and one with a cage-like framework, were isolated from Hypericum ascyron Linn. Some of these compounds exhibit anti-tumor activity. [Display omitted] • 10 undescribed and 12 known spirocyclic PPAP were isolated from Hypericum ascyron. • 1 and 2 are 1,2- seco -spirocyclic PPAP and 3 has a cage-like framework. • 1 and 2 affected the growth of HCT116 in vitro. • 3 and 5 reduced the length of sub-intestinal vessels in zebrafish embryos. [ABSTRACT FROM AUTHOR]

In noise-induced hearing loss (NIHL), noise exposure damages cochlear sensory hair cells, which lack the capacity to regenerate. Following noise insult, intense metabolic activity occurs, resulting in a cochlear free radical imbalance. Oxidative stress and antioxidant enzyme alterations, including lipoxygenase upregulation, have been linked to chronic inflammation, which contributes to hearing impairment. We previously proposed Scutellaria baicalensis (SB) extract as an alternative therapeutic for preventing NIHL and attributed its pharmacological effects to baicalein. Although baicalein was most effective, its concentration in SB extract is much lower compared to baicalin. In this study, we performed enzymatic bioconversion using an Sumizyme (SM) enzyme to increase baicalein concentration in SB extract and consequently improve its therapeutic efficacy. HPLC analysis revealed that baicalein concentration in SB extract after bioconversion (BSB) was significantly increased. Moreover, BSB-treated mice exhibited significantly improved auditory function compared with control mice and tended to have improved auditory function compared with SB-treated mice. We also demonstrated that BSB effectively stimulates hair cell regeneration compared to SB that did not achieve the same effect in a zebrafish model. Finally, when compared the abilities of SB and BSB to inhibit lipoxygenase (LOX), BSB showed a greater efficacy. Cumulatively, our data suggest that BSB exhibits improved pharmacological properties for treating NIHL compared with SB. [ABSTRACT FROM AUTHOR]

Detecting and measuring the dynamic redox events that occur in vivo is a prerequisite for understanding the impact of oxidants and redox events in normal and pathological conditions. These aspects are particularly relevant in cardiovascular tissues wherein alterations of the redox balance are associated with stroke, aging, and pharmacological intervention. An ambiguous aspect of redox biology is how redox events occur in subcellular organelles including mitochondria, and nuclei. Genetically-encoded Rogfp2 fluorescent probes have become powerful tools for real-time detection of redox events. These probes detect hydrogen peroxide (H 2 O 2 ) levels and glutathione redox potential (E GSH ), both with high spatiotemporal resolution. By generating novel transgenic (Tg) zebrafish lines that express compartment-specific Rogfp2-Orp1 and Grx1-Rogfp2 sensors we analyzed cytosolic, mitochondrial, and the nuclear redox state of endothelial cells and cardiomyocytes of living zebrafish embryos. We provide evidence for the usefulness of these Tg lines for pharmacological compounds screening by addressing the blocking of pentose phosphate pathways (PPP) and glutathione synthesis, thus altering subcellular redox state in vivo. Rogfp2-based transgenic zebrafish lines represent valuable tools to characterize the impact of redox changes in living tissues and offer new opportunities for studying metabolic driven antioxidant response in biomedical research. [ABSTRACT FROM AUTHOR]

Summary Vascular mural cells (vMCs) are essential components of the vertebrate vascular system, controlling blood vessel maturation and homeostasis. Discrete molecular mechanisms have been associated with vMC development and differentiation. The function of hemodynamic forces in controlling vMC recruitment is unclear. Using transgenic lines marking developing vMCs in zebrafish embryos, we find that vMCs are recruited by arterial-fated vessels and that the process is flow dependent. We take advantage of tissue-specific CRISPR gene targeting to demonstrate that hemodynamic-dependent Notch activation and the ensuing arterial genetic program is driven by endothelial primary cilia. We also identify zebrafish foxc1b as a cilia-dependent Notch-specific target that is required within endothelial cells to drive vMC recruitment. In summary, we have identified a hemodynamic-dependent mechanism in the developing vasculature that controls vMC recruitment. [ABSTRACT FROM AUTHOR]

[Display omitted] • Thiabendazole induces morphological abnormalities in zebrafish larvae. • Thiabendazole triggers apoptosis and cell cycle arrest in zebrafish embryos. • Thiabendazole causes cardiovascular and motor neuronal toxicity in zebrafish larvae. • Thiabendazole stimulates ROS production and inflammatory response in zebrafish. • Thiabendazole may cause the detrimental effects on the aquatic environment. Thiabendazole, a benzimidazole fungicide, is widely used to prevent yield loss in agricultural land by inhibiting plant diseases derived from fungi. As thiabendazole has a stable benzimidazole ring structure, it remains in the environment for an extended period, and its toxic effects on non-target organisms have been reported, indicating the possibility that it could threaten public health. However, little research has been conducted to elucidate the comprehensive mechanisms of its developmental toxicity. Therefore, we used zebrafish, a representative toxicological model that can predict toxicity in aquatic organisms and mammals, to demonstrate the developmental toxicity of thiabendazole. Various morphological malformations were observed, including decreased body length, eye size, and increased heart and yolk sac edema. Apoptosis, reactive oxygen species (ROS) production, and inflammatory response were also triggered by thiabendazole exposure in zebrafish larvae. Furthermore, PI3K/Akt and MAPK signaling pathways important for appropriate organogenesis were significantly changed by thiabendazole. These results led to toxicity in various organs and a reduction in the expression of related genes, including cardiovascular toxicity, neurotoxicity, and hepatic and pancreatic toxicity, which were detected in flk1:eGFP , olig2:dsRED , and L-fabp:dsRed;elastase:GFP transgenic zebrafish models, respectively. Overall, this study partly determined the developmental toxicity of thiabendazole in zebrafish and provided evidence of the environmental hazards of this fungicide. [ABSTRACT FROM AUTHOR]

Cardiovascular pharmaceuticals (CVPs) are globally present in inland waters and have also been found in the sediment and plasma of fish from the Sava River, Croatia. Based on the previous research, CVPs amiodarone (AMI), ramipril (RAM), simvastatin (SIM), and verapamil (VER) have been selected for this study. Their effect has been investigated, individually and in a mixture, on the development of the zebrafish embryo Danio rerio (Hamilton, 1822) within the first 96 h of development. Upon exposure to environmentally relevant concentrations of tested CVPs (0.1, 1, and 10 μg/L) zebrafish survival and development as apparent from observed morphological abnormalities, heartbeat rates and changes in behavior, hatching success, larval length and oxidative stress level were monitored. The CVP causing the highest mortality and pathological changes was SIM (1 and 10 μg/L), which corresponds well with the observed effects during zebrafish exposure to CVPs' mixtures (4 and 40 μg/L). All pharmaceuticals affected cardiac function and decreased heart rate. SIM (1 μg/L), VER and RAM (10 μg/L) decreased larval length, while induced oxidative stress was recorded in the SIM- and VER-exposed specimens. Behavioral alterations of zebrafish were observed only in AMI-treated group (10 μg/L). Our amino acid sequence comparison and structural and docking analysis showed a highly conserved binding site between human and zebrafish HMG-CoA reductase for SIM and its main metabolite simvastatin acid. Using these ecotoxicological bioassays on a zebrafish model with particular emphasis on sublethal endpoints, the risk of CVPs, especially statins, for fish in inland waters has been identified. [Display omitted] • Cardiovascular drugs are widespread in the aquatic environment. • Cardiovascular drugs affect cardiac physiology and growth, induced oxidative stress. • Simvastatin caused severe adverse effects on zebrafish embryonal development. • Mixture toxicity analysis highlights interaction of cardiovascular drugs. [ABSTRACT FROM AUTHOR]

In this study, a mandarin fish (Siniperca chuatsi) rhabdovirus (SCRV) S3 was isolated, purified, and identified from live mandarin fish collected from a commercial market. The full-length genome of S3 is 11,582 bp and has a 36 bp insertion in the 5'-UTR of the genome. The genome homology between S3 and the representative species in the genera of Perhabdovirus , Spriviviruses , and Novirhabdoviruses is 29.6% ∼ 31.0%, 25.7% ∼ 25.8, and 0.3% respectively. The cell tropism study showed that S3 could not only infect fish-derived cells but also mammalian and human-derived cells at 28 °C. However, S3 had no infectivity to all the tested cells at 37 °C. The temperature was an important barrier to the infection of SCRV S3. The upper limit of temperature for S3 growth in cells is 32 °C and the lowest measurable virus replication temperature is 10 °C. The optimum growth temperature is 28 °C. The animal study showed that SCRV S3 exhibited strong pathogenicity to zebrafish. Mortality in blue zebrafish (Danio rerio) and red zebrafish (Danio rerio) at 28 °C was 53% and 73%, respectively. Zebrafish can be used as an infection model for SCRV. This study will aid in understanding the biological characterizations of SCRV and has important guiding significance for the effective prevention and control strategies of SCRV. • The full genome of SCRV S3, which has a 36 bp insertion in the 5'-UTR, is 11582 bp. • SCRV S3 could infect mammalian and human-derived cells at 28 °C, but not at 37 °C. • The growth temperature range of SCRV S3 is 10–32 °C. The optimum temperature is 28 °C. • Zebrafish can be used as an infection model for SCRV. [ABSTRACT FROM AUTHOR]

Tyrosinase is an oxidase that is the rate-limiting enzyme for controlling the production of melanin in the human body. Overproduction of melanin could lead to a variety of skin disorders. Glabridin, an isoflavan, isolated from the root of Glycyrrhiza glabra Linn, has exhibited several pharmacological activities, including excellent inhibitory effects on tyrosinase. In this paper, the inhibitory kinetics of glabridin on tyrosinase and their binding mechanisms were determined using spectroscopic, zebrafish model and molecular docking techniques. The results indicate that glabridin reversibly inhibits tyrosinase in a noncompetitive manner through a multiphase kinetic process with the IC 50 of 0.43 μmol/L. It has been shown that glabridin had a strong ability to quench the intrinsic fluorescence of tyrosinase mainly through a static quenching procedure, suggesting a stable glabridin-tyrosinase complex may be generated. The results of molecular docking suggest that glabridin did not directly bind to the active site of tyrosinase. Moreover, according to the results of zebrafish model system, glabridin shows no effects on melanin synthesis in zebrafish but presents toxicity to zebrafish embryo. The possible inhibitory mechanisms, which will help to design and search for tyrosinase inhibitors especially for glabridin analogues, were proposed. [ABSTRACT FROM AUTHOR]

Human mitochondrial DNA (mtDNA) mutations have been associated with a wide spectrum of clinical abnormalities. However, nuclear modifier gene(s) modulate the phenotypic expression of pathogenic mtDNA mutations. In our previous investigation, we identified the human GTPBP3 related to mitochondrial tRNA modification, acting as a modifier to influence of deafness-associated mtDNA mutation. Mutations in GTPBP3 have been found to be associated with other human diseases. However, the pathophysiology of GTPBP3-associated disorders is still not fully understood. Here, we reported the generation and characterization of Gtpbp3 depletion zebrafish model using antisense morpholinos. Zebrafish gtpbp3 has three isoforms localized at mitochondria. Zebrafish gtpbp3 is expressed at various embryonic stages and in multiple tissues. In particular, the gtpbp3 was expressed more abundantly in adult zebrafish ovary and testis. The expression of zebrafish gtpbp3 can functionally restore the growth defects caused by the mss1 / gtpbp3 mutation in yeast. A marked decrease of mitochondrial ATP generation accompanied by increased levels of apoptosis and reactive oxygen species were observed in gtpbp3 knockdown zebrafish embryos. The Gtpbp3 morphants exhibited defective in embryonic development including bleeding, melenin, oedema and curved tails within 5 days post fertilization, as compared with uninjected controls. The co-injection of wild type gtpbp3 mRNA partially rescued these defects in Gtpbp3 morphants. These data suggest that zebrafish Gtpbp3 is a structural and functional homolog of human and yeast GTPBP3. The mitochondrial dysfunction caused by defective Gtpbp3 may alter the embryonic development in the zebrafish. In addition, this zebrafish model of mitochondrial disease may provide unique opportunities for studying defective tRNA modification, mitochondrial biogenesis, and pathophysiology of mitochondrial disorders. [ABSTRACT FROM AUTHOR]

Aims Paraquat (PQ) is a pesticide highly toxic to human beings, and a well-known trigger of oxidative stress. Although several animal models of PQ poisoning have been developed, some disadvantages limit their application in vivo. A zebrafish model was used in the present study to better define mechanisms of oxidative stress injury induced by PQ. Main methods The toxicity of PQ was evaluated in the AB strain of zebrafish, and apoptosis was assessed by acridine orange staining. Macrophage migration was identified using the TG ( zlyz : EGFP ) transgenic strain, and angiogenesis was observed using the fli1a-EGFP casper strain. Following the validation of gene changes by zebrafish-based in vivo quantitative real-time PCR, network analysis was performed using the Ingenuity Pathway Analysis software. Key findings We first established the LC 50 of PQ in the zebrafish model, and then found that robust oxidative stress and antioxidant genes were activated after PQ exposure. Moreover, apoptosis and distinct macrophage activation and migration were identified for the first time in PQ-exposed zebrafish. Utilizing this model, both extrinsic and intrinsic pathways involved in PQ-induced apoptosis were elucidated. We further demonstrated that macrophage migration was specifically induced by PQ, and that Rho family members and JNK-MMP13 signaling participated in this process. Significance Zebrafish is a promising tool for investigating the mechanisms of oxidative stress injury induced by PQ, and for screening effective anti-oxidant drugs. [ABSTRACT FROM AUTHOR]

GLE1 mutations cause lethal congenital contracture syndrome 1 (LCCS1), a severe autosomal recessive fetal motor neuron disease, and more recently have been associated with amyotrophic lateral sclerosis (ALS). The gene encodes a highly conserved protein with an essential role in mRNA export. The mechanism linking Gle1 function to motor neuron degeneration in humans has not been elucidated, but increasing evidence implicates abnormal RNA processing as a key event in the pathogenesis of several motor neuron diseases. Homozygous gle1 −/− mutant zebrafish display various aspects of LCCS, showing severe developmental abnormalities including motor neuron arborization defects and embryonic lethality. A previous gene expression study on spinal cord from LCCS fetuses indicated that oligodendrocyte dysfunction may be an important factor in LCCS. We therefore set out to investigate the development of myelinating glia in gle1 −/− mutant zebrafish embryos. While expression of myelin basic protein ( mbp ) in hindbrain oligodendrocytes appeared relatively normal, our studies revealed a prominent defect in Schwann cell precursor proliferation and differentiation in the posterior lateral line nerve. Other genes mutated in LCCS have important roles in Schwann cell development, thereby suggesting that Schwann cell deficits may be a common factor in LCCS pathogenesis. These findings illustrate the potential importance of glial cells such as myelinating Schwann cells in motor neuron diseases linked to RNA processing defects. [ABSTRACT FROM AUTHOR]

[Display omitted] • Abundant Lewis- and Brønsted-acid sites on H-OMS-2 in favor of C C cleavage of PVA molecule. • H-OMS-2 could act as both effective PMS activator and adsorbent with superior catalytic stability. • Quenching experiments and EPR confirmed that non-radical mechanism played the key role in H-OMS-2/PMS system for PVA removal. • The toxicity of PVA wastewater has been reduced after adsorption and degradation. The present work introduced the use of acidified rod-like manganese dioxide (H-OMS-2) as effective peroxymonosulfate (PMS) activator and adsorbent for the removal of polyvinyl alcohol (PVA). The adsorption efficiency for PVA solution (80 mg/L) was 78 % after 60 min when H-OMS-2 (0.6 g/L) was used as adsorbent and the maximum adsorption capacity was 25.5 mg/g. After the addition of PMS (7 mM), PVA was entirely degraded within 40 min after adsorption/desorption equilibrium, and 64 % mineralization rate was reached after 5 h via the synergistic adsorption and oxidation process. The apparent activation energy ( Ea ) of H-OMS-2/PMS system was 11.14 kJ/mol that was lower than the value in OMS-2/PMS system (24.87 kJ/mol). And, H-OMS-2 could be recycled for 4 times with 90 % of degradation rate at 40 min and 56 % of mineralization rate after 24 h. Quenching test and electron paramagnetic resonance (EPR) results demonstrated that 1O 2 was the primary reactive oxygen species rather than ·OH, SO 4 ̇- and O 2 ̇-. In addition, non-radical pathway via the direct electron transfer existed. The complex between Mn3+ and PMS (electron acceptor) acted as main active species to oxidize adsorbed PVA (electron donor). The characterization and degradation performance of catalyst indicated that abundant Lewis- and Brønsted-acid sites on H-OMS-2 are beneficial to C C cleavage of PVA, along with the void-confinement effects of H-OMS-2 due to its highly structured mesoporous pore size, which can result in the promotion of the degradation. Finally, study on the degradation pathway of PVA along with the aquatic toxicity experiment via zebrafish model system indicated that intermediates containing carbonyl groups are harmful for the survival and development of zebrafish. [ABSTRACT FROM AUTHOR]

[Display omitted] • A purified mannose-containing polysaccharide fractions was obtained from Jiuzao. • The backbone of LWZ-P' was comprised of → 3)-α- d -Manp-(1 → and → 2)-α- d -Manp-(1→. • LWZ-P' showed antioxidant activities toward scavenging different radicals in vitro. • LWZ-P' registered the best protective effects on hydrogen peroxide-induced zebrafish. Recovery of valuable compounds from Baijiu by-products is one of the important challenges currently faced by the brewing industry. In recent years, the research on Baijiu has shifted from Baijiu flavour to functional compounds, especially the extraction and activity of Jiuzao polysaccharides. The polysaccharide from Laowuzeng Jiuzao (LWZ-P') was extracted by water extraction and alcohol precipitation, and then purified by column chromatography to obtain pure polysaccharide. The physical and chemical properties of LWZ-P' were analyzed by UV, FT-IR, GPC, NMR, and HPAEC. As a result, LWZ-P' was mainly composed of mannose (37.26 %), with a relative molecular weight of 32 402 g/mol, possessing the primary backbone of → 3)-α - d - Man p -(1 → and → 2)-α - d - Man p -(1 → with branches consisting of α - d - Man p -(1 → residue. In addition, LWZ-P' had significant activities of DPPH radical, ABTS free radical, superoxide anion free radical and hydroxyl free radical in a dose-dependent manner. Meanwhile, LWZ-P' presented a protective effect against the generation of H 2 O 2 -induced reactive oxygen species, effectively inhibiting cell death in zebrafish. The current study aims to recover polysaccharides from brewed products, better understand the functional value of Jiuzao , and lay a theoretical foundation for the development of Jiuzao polysaccharide-related foods. [ABSTRACT FROM AUTHOR]

Gu Sui Bu (GSB), the dried rhizome of Drynaria fortunei J. Sm., is widely used in traditional Chinese medicine for treating fractures and osteoporosis. Although glucocorticoids are widely prescribed in modern medicine, the efficacy of GSB in treating glucocorticoid-induced osteoporosis (GIOP) remains unclear. GIOP is one of the most prevalent forms of osteoporosis and increases the risk of fracture, which can cause severe complications in elderly people. Safe, efficacious, and cost-effective treatment options for GIOP are thus warranted. The present study investigated the efficacy and mechanism of GSB for treating GIOP. We established an efficient and robust in vivo GIOP model by optimizing zebrafish larvae rearing conditions and the dose and duration of dexamethasone treatment. Bone calcification was evaluated through calcein staining. To quantify the degree of vertebral mineralization in the larvae, we developed a scoring system based on the rate of vertebral calcification; this system reduced quantification errors among individual zebrafish caused by inconsistencies in staining or imaging parameters. Quantitative real-time polymerase chain reaction was used to access the expression levels of genes essential to the differentiation and function of bone cells. High-performance liquid chromatography was employed to identify naringin in the GSB extract. GSB significantly reversed the dexamethasone-induced calcification delay in zebrafish larvae. GSB enhanced osteoblast activity by increasing the expression of collagen I , osteopontin , and osteonectin and repressed bone resorption by decreasing the expression of matrix metalloproteinases (mmps), including mmp9 and mmp13a. We also identified naringin as one of the constituents of GSB responsible for the herbal extract's anti-GIOP activity. Using the in vivo zebrafish GIOP model that we established, the efficacy of traditional Chinese medicines in treating GIOP could be systematically investigated. GSB has an osteogenic effect and may thus be an efficacious and cost-effective treatment option for GIOP. Notably, bone resorption activity was found to be retained after GSB treatment, which would be beneficial for maintaining normal bone remodeling. [Display omitted] [ABSTRACT FROM AUTHOR]

In this work four vanadium complexes (compounds 1 , 2 , 3 and 4) and one molybdenum complex (compound 5) with hydrazone ligands derived from pyridoxal were synthesized and characterized. All compounds are mononuclear species, two of them (compounds 3 and 5) are dioxide complexes and the other three (compounds 1 , 2 and 4) monoxide complexes. The vanadium atom of the compound 3 is five-coordinated and all the other compounds have a six coordinated environment polyhedron. The poses for the potential intercalation of the compounds 2 and 3 with DNA were obtained by using AutoDock software. Optimizations were also performed at PM6-D3H4 semi-empirical level whereas the study of the nature of the interaction was carried out by means of the Energy Decomposition Analysis and the Non-Covalent Interaction index by using in both cases Density Functional Theory computations. The cytotoxicity in lung cancer cells (A549 cell line) of all the compounds was also evaluated. After 24 h of treatment, vanadium complexes showed high values of IC 50 , between 419.93 ± 22.58 and 685.88 ± 46.55 μM. After 48 h, the results showed that the compound 3 had the lowest IC 50 value, 65.32 ± 9.95 μM, and the compound 2 the highest value, 375.28 ± 32.09 μM. The molybdenum complex showed the lowest IC 50 value at 48 h (11.22 ± 1.34 μM). The toxicity of the compounds 3 , 4 and 5 was tested in vivo , using zebrafish model, and the molybdenum complex showed higher toxic effects than the studied vanadium complexes. [Display omitted] [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance The root of Croton crassifolius Geisel is traditionally used in China for the treatment of snake bites, stomach ache, sternalgia, joint pain, pharyngitis, jaundice and rheumatoid arthritis, while in Thailand, it has been used as an anticancer herbal medicine by the indigenous people. Yet, its pharmacological studies are still limited, especially towards its anticancer property. Anti-angiogenesis is a promising therapeutic strategy in the anti-cancer treatment. Previous studies have shown strong anti-angiogenic activity in the low polar fraction of the herb. Nevertheless, the potent compound which is responsible for the anti-angiogenesis, and its molecular mechanism have never been reported. Aim of the study To determine the potent anti-angiogenic component in C. crassifolius and its molecular mechanism of action. Materials and methods C. crassifolius was extracted using supercritical fluid extraction and steam distillation. The anti-angiogenic activities of the two extracts were evaluated in the zebrafish model by quantitative endogenous alkaline phosphatase assay. The chemical compounds in the active extract were isolated using chromatographic methods, and their structures were elucidated using different spectroscopic techniques. The content/quantity of the active compounds in this extract was determined with HPLC analysis. The molecular mechanism of the most active compound was further studied using the real-time PCR assay. Besides, its cytotoxicity on various cancer and normal cell lines was evaluated using the cell-counting kit. Results Supercritical fluid extract (SFE) of C. crassifolius showed better anti-angiogenic activity than that of steam distillation extract (SDE). Three sesquiterpenes, namely, cyperenoic acid, 8-hydroxy-α-guaiene and (+)-guaia-l(10),ll-dien-9-one, were isolated and identified in the SFE. Among them, cyperenoic acid displayed the strongest anti-angiogenic activity by 51.7% of the control at 10 μM, while the others showed little effect. HPLC results showed that cyperenoic acid was the major component in the SFE with 9.97% (w/w). Results of the real-time PCR assay suggested that the cyperenoic acid affected multiple molecular targets related to angiogenesis including vascular endothelial growth factor (Vegfa), angpiopoietin (Angpt), and their receptors. Cytotoxicity assay showed cyperenoic acid possessed little toxicity toward cancer and normal cells. Conclusions Cyperenoic acid is an important anti-angiogenic component present in C. crassifolius and serve as a potent inhibitor in the angiogenesis in the zebrafish embryo model. The anti-angiogenic property, but not the cytotoxicity, of C. crassifolius provides a scientific basis for its traditional use in cancer treatment. [ABSTRACT FROM AUTHOR]

Ethnopharmacological relevance Piper wallichii (Miq.) Hand.-Mazz. is a medicinal plant used widely for the treatment of rheumatoid arthritis, inflammatory diseases, cerebral infarction and angina in China. Previous study showed that lignans and neolignans from Piper spp. had potential inhibitory activities on platelet aggregation. In the present study, we investigated the chemical constituents of Piper wallichii and their antithrombotic activities, to support its traditional uses. Materials and methods The methanolic extract of the air-dried stems of Piper wallichii was separated and purified using various chromatographic methods, including semi-preparative HPLC. The chemical structures of the isolates were determined by detailed spectroscopic analysis, and acidic hydrolysis in case of the new glycoside 2 . Determination of absolute configurations of the new compound 1 was facilitated by calculated electronic circular dichroism using time-dependent density-functional theory. All compounds were tested for their inhibitory effects on platelet aggregation induced by platelet activating factor (PAF) in rabbits׳ blood model, from which the active ones were further evaluated the in vivo antithrombotic activity in zebrafish model. Results A new neolignan, piperwalliol A ( 1 ), and four new aromatic glycosides, piperwalliosides A–D ( 2 – 5 ) were isolated from the stems of Piper wallichii , along with 25 known compounds, including 13 lignans, six aromatic glycosides, two phenylpropyl aldehydes, and four biphenyls. Five known compounds ( 6 – 10 ) showed in vitro antiplatelet aggregation activities. Among them, (–)-syringaresinol ( 6) was the most active compound with an IC 50 value of 0.52 mM. It is noted that in zebrafish model, the known lignan 6 showed good in vivo antithrombotic effect with a value of 37% at a concentration of 30 μM, compared with the positive control aspirin with the inhibitory value of 74% at a concentration of 125 μM. Conclusion This study demonstrated that lignans, phenylpropanoid and biphenyl found in Piper wallichii may be responsible for antithrombotic effect of the titled plant. [ABSTRACT FROM AUTHOR]

We investigated the protective effect of octaphlorethol A (OPA) isolated from Ishige foliacea against high-glucose-induced oxidative damage, as indicated by reactive oxygen species (ROS) generation, lipid peroxidation, and cell death, in human umbilical vein endothelial cells (HUVECs) and in a zebrafish model. OPA treatment significantly and dose-dependently decreased ROS generation, lipid peroxidation, and cell death in HUVECs and in a zebrafish model. Our results show that OPA can protect HUVECs and zebrafish against high glucose by suppressing intracellular ROS generation, lipid peroxidation, and cell death. [ABSTRACT FROM AUTHOR]

[Display omitted] • Hypolipidaemic effect of various dark teas were examined using a zebrafish model. • Microbial fermentation significantly improved the hypolipidaemic effect of CDTEs. • The mechanisms underlying hyperlipidaemia regulation by CDTEs were proposed. In order to investigate the hypolipidaemic and antioxidant effects of various dark teas produced from different post-fermentation using the same raw material, a hyperlipidaemia zebrafish model combined with binding bile salts assay and antioxidant assays were performed in this study. Results showed that the hypolipidaemic effect of dark tea extracts increased significantly (p < 0.05) while the antioxidant ability decreased sharply compared with raw material. Particularly, Liupao tea (50%) and Pu-erh tea (48%) showed promising hypolipidaemic potential; however, the antioxidant capacity of Pu-erh tea decreased (31–49%) most dramatically. Besides, the levels of total polyphenols and catechins decreased sharply, but theabrownin, gallic acid, and caffeine increased significantly after post-fermentation. Moreover, the potential mechanisms of regulating hyperlipidaemia by dark tea extracts were discussed. These results suggest that microbial fermentation significantly affects the bioactivity of dark teas, and provide theoretical basis for processing and improving of dark tea products for hyperlipidaemia therapy. [ABSTRACT FROM AUTHOR]

Rare deletions and duplications on the long arm of Chromosome 21 have previously been reported in many patients with craniofacial and developmental phenotypes. However, this Down Syndrome Critical Region (DSCR) contains multiple genes, making identifying a single causative gene difficult. Here, we report a case of a boy with bicoronal craniosynostosis, facial dysmorphism, developmental delay, and intellectual impairment who was found by whole genome sequencing to have a homozygous missense mutation in the Single-Minded Homolog 2 (SIM2) gene (c.461 A > G, p.Tyr154Cys) within the DSCR. SIM2 encodes an essential bHLH and PAS domain transcription factor expressed during fetal brain development and acts as a master regulator of neurogenesis. This variant is globally very rare, segregates in the family, and is predicted to be highly deleterious by in silico analysis, 3D molecular modeling of protein structure, and functional analysis of zebrafish models. Zebrafish expressing the human SIM2p.Y154C variant displayed a progressed microcephaly-like phenotype and head shape abnormalities. When combined with careful phenotyping of the patient vis-à-vis previously reported cases harboring structural variants in this critical 21q22 region, the data support a pathogenic role of SIM2 in this complex syndrome and demonstrates the utility of next-generation sequencing in prioritizing genes in contiguous deletions/duplications syndromes and diagnosing microarray-negative patients in the craniofacial clinic. [ABSTRACT FROM AUTHOR]

Abstract: Ethnopharmacological relevance: “Qin-Jiao” is a well-known traditional Chinese medicinal (TCM) herb having been used generally for fighting rheumatoid arthritis (RA) since ancient times. The root of Gentiana dahurica Fisch (Gentianaceae) is one of the four officially validated “Qin-Jiao” as listed in the Chinese Pharmacopoeia. In addition, it is a common Tibetan medicinal herb used for the treatment of tonsillitis, urticaria, and RA, while the flowers have been used as a Mongolian herb for curing cough sore throat and eliminating the phlegm due to its anti-inflammatory effect. Aim of the study: The aim of the study was to characterize the anti-inflammatory compounds in “Qin-Jiao”, on the basis of detailed investigation on not only the phytochemical study of Gentiana dahurica, but also the bioactive evaluation on compounds obtained presently and previously from different “Qin-Jiao” origins and Gentiana species. Materials and methods: The ethanol extract of air-dried roots of Gentiana dahurica was suspended into H2O and extracted with EtOAc and n-BuOH, successively. Repeated column chromatography (CC) and semi-preparative HPLC were carried out on each of the fractions. The isolated compounds were determined by detailed spectroscopic analysis and acidic hydrolysis. Anti-inflammatory activities of 18 isolates, together with 12 typical compounds obtained previously by our group from the other “Qin-Jiao” origins (Gentiana crassicaulis, Gentiana straminea) and Gentiana rigescens, were tested by inhibitory effects on LPS-induced NO production in macrophage RAW264.7 cells and TPA-induced cyclooxygenases-2 and -1 (COXs-2/1) production on zebrafish model. Results: A new lignan glycoside (1) was identified, together with 20 known compounds, including 10 iridoid glycosides (2–11), three steroids (12–14), four lignans (15–18), one phenylpropanoid (19) and two triterpenes (20–21). Anti-inflammatory bioassay showed that only compound 21 displayed potential inhibitory effect on NO production (IC50=16.85μM), while 20 tested compounds had inhibitory activities on COXs-2/1. Among them, the triterpenoid 21 was the most active compound with an inhibitory value of 78% at a concentration of 30μM. All the tested compounds showed no cytotoxicity on five human cancer cell lines (40μM) and zebrafish (30μM), except for 21 displaying weak cytotoxicity on human myeloid leukemia HL-60 (IC50=16.43μM). Conclusion: Most of compounds particularly iridoid glycosides from “Qin-Jiao” display potential inhibitory effect on COXs-2/1. The results support the historical importance of the well-known TCM herb, “Qin-Jiao”, having been commonly used for fighting RA. As major components, the bioactive iridoid glycosides should play important role in the anti-inflammatory effect of “Qin-Jiao”. Although further research will be required to evaluate the selective activities of the COXs-2/1 inhibitors, this work validates the medicinal use of “Qin-Jiao” and provides information for different “Qin-Jiao” origins having different treating effects on RA. [Copyright &y& Elsevier]