Synthesis and Hypoglycemic Effect of Insulin from the Venom of Sea Anemone Exaiptasia diaphana.

Sea anemone venom, abundant in protein and peptide toxins, serves primarily for predatory defense and competition. This study delves into the insulin-like peptides (ILPs) present in sea anemones, particularly focusing on their role in potentially inducing hypoglycemic shock in prey. We identified five distinct ILPs in Exaiptasia diaphana, exhibiting varied sequences. Among these, ILP-Ap04 was successfully synthesized using solid phase peptide synthesis (SPPS) to evaluate its hypoglycemic activity. When tested in zebrafish, ILP-Ap04 significantly reduced blood glucose levels in a model of diabetes induced by streptozotocin (STZ) and glucose, concurrently affecting the normal locomotor behavior of zebrafish larvae. Furthermore, molecular docking studies revealed ILP-Ap04's unique interaction with the human insulin receptor, characterized by a detailed hydrogen-bonding network, which supports a unique mechanism for its hypoglycemic effects. Our findings suggest that sea anemones have evolved sophisticated strategies to activate insulin receptors in vertebrates, providing innovative insights into the design of novel drugs for the treatment of diabetes.

Genome-wide identification of resistance genes and cellular analysis of key gene knockout strain under 5-hydroxymethylfurfural stress in Saccharomyces cerevisiae.

In bioethanol production, the main by-product, 5-hydroxymethylfurfural (HMF), significantly hinders microbial fermentation. Therefore, it is crucial to explore genes related to HMF tolerance in Saccharomyces cerevisiae for enhancing the tolerance of ethanol fermentation strains. A comprehensive analysis was conducted using genome-wide deletion library scanning and SGAtools, resulting in the identification of 294 genes associated with HMF tolerance in S. cerevisiae. Further KEGG and GO enrichment analysis revealed the involvement of genes OCA1 and SIW14 in the protein phosphorylation pathway, underscoring their role in HMF tolerance. Spot test validation and subcellular structure observation demonstrated that, following a 3-h treatment with 60 mM HMF, the SIW14 gene knockout strain exhibited a 12.68% increase in cells with abnormal endoplasmic reticulum (ER) and a 22.41% increase in the accumulation of reactive oxygen species compared to the BY4741 strain. These findings indicate that the SIW14 gene contributes to the protection of the ER structure within the cell and facilitates the clearance of reactive oxygen species, thereby confirming its significance as a key gene for HMF tolerance in S. cerevisiae.

A Chromosome-Level Genome Assembly of the Reef Stonefish (Synanceia verrucosa) Provides Novel Insights into Stonustoxin (sntx) Genes.

Reef stonefish (Synanceia verrucosa) is one of the most venomous fishes, but its biomedical study has been restricted to molecular cloning and purification of its toxins, instead of high-throughput genetic research on related toxin genes. In this study, we constructed a chromosome-level haplotypic genome assembly for the reef stonefish. The genome was assembled into 24 pseudo-chromosomes, and the length totaled 689.74 Mb, reaching a contig N50 of 11.97 Mb and containing 97.8% of complete BUSCOs. A total of 24,050 protein-coding genes were annotated, of which metalloproteinases, C-type lectins, and stonustoxins (sntx) were the most abundant putative toxin genes. Multitissue transcriptomic and venom proteomic data showed that sntx genes, especially those clustered within a 50-kb region on the chromosome 2, had higher transcription levels than other types of toxins as well as those sntx genes scatteringly distributed on other chromosomes. Further comparative genomic analysis predicted an expansion of sntx-like genes in the Percomorpha lineage including nonvenomous fishes, but Scorpaenoidei species experienced extra independent sntx duplication events, marking the clear-cut origin of authentic toxic stonustoxins. In summary, this high-quality genome assembly and related comparative analysis of toxin genes highlight valuable genetic differences for potential involvement in the evolution of venoms among Scorpaeniformes fishes.

Discovery of new strains for furfural degradation using adaptive laboratory evolution in Saccharomyces cerevisiae.

In industrial production, the excessive discharge of furfural can pose harm to soil microorganisms, aquatic animals and plants, as well as humans. Therefore, it is crucial to develop efficient and cost-effective methods for degrading furfural in the environment. Currently, the use of Saccharomyces cerevisiae for furfural degradation in water has shown effectiveness, but there is a need to explore improved efficiency and tolerance in S. cerevisiae for this purpose. In this study, we isolated and evolved highly efficient furfural degradation strains, namely YBA_08 and F60C. These strains exhibited remarkable capabilities, degrading 59% and 99% furfural in the YPD medium after 72 h of incubation, significantly higher than the 31% achieved by the model strain S288C. Through analysis of the efficient degradation mechanism in the evolutionary strain F60C, we discovered a 326% increase in the total amount of NADH and NADPH. This increase likely promotes faster furfural degradation through intracellular aldehyde reductases. Moreover, the decrease in NADPH content led to a 406% increase in glutathione content at the background level, which protects cells from damage caused by reactive oxygen species. Mutations and differential expression related to cell cycle and cell wall synthesis were observed, enabling cell survival in the presence of furfural and facilitating rapid furfural degradation and growth recovery. Based on these findings, it is speculated that strains YBA_08 and F60C have the potential to contribute to furfural degradation in water and the production of furfuryl alcohol, ethanol, and FDCA in biorefinery processes.

Effects of Polystyrene Diet on the Growth and Development of Tenebrio molitor.

In recent years, the role of Tenebrio molitor in degrading polystyrene foam through its gut microbes has become the focus of research. However, little literature has reported the effect of feeding on polystyrene foam on the growth and development of Tenebrio molitor. In this study, we investigated the impacts of different polystyrene by evaluating the vital signs of Tenebrio molitor fed in the intestines and excrement fluids using RNA-Seq t.echnology and then verifying the transcriptome sequencing findings using qRT-PCR technology. The average weight of Tenebrio molitor larvae in the wheat bran group increased significantly. Tenebrio molitor larvae in the PS group, on the other hand, didn't grow as much and had a much lower average weight than those in the wheat bran group. Compared to the bran group, the excrement of Tenebrio molitor fed only on polystyrene foam was flaky and coarse, increased nitrogen and phosphorus atomic concentration ratios by about 50%, decreased potassium atomic concentration ratios by 63%, with the enterocytes and circular muscle of Tenebrio molitor falling as well. Kyoto Encyclopedia of Genes and Genomes enrichment indicated that the differential genes were mainly related to metabolic pathways. There was an agreement between qRT-PCR and RNA-Seq analyses for the growth and development genes chitinase, heat shock protein 70, and cytochrome P450. Only feeding polystyrene foam shall lead to the growth and development retardation of Tenebrio molitor.

Preparation and photocatalytic application of terbium and sulfur co-doped titanium nanomaterials.

Titanium-based nanomaterials co-doped with terbium (Tb) and sulfur (S) were synthesized by sol-gel method via a facile step. Physicochemical properties of the resulting composites were characterized by X-ray diffraction (XRD), Scanning electron microscope (SEM), X-ray photo-electron spectroscopy (XPS) and UV-Vis diffused reflectance spectroscopy (DRS). Methylene blue (MB) was used as a degradation target for evaluating the photocatalytic performance. The factors which influence the photocatalytic activity were investigated, including calcined temperatures and S doping amount. Tb, S (2 wt%) co-doped TiO2 composite calcined at 500 °C exhibited the highest photocatalytic activity with a degradation rate of 72.4% in 3 h. The reaction constant was 0.11529, 0.26025, 0.35038 and 0.41462 h-1 for undoped TiO2, Tb-doped TiO2, S-doped TiO2 and Tb, S co-doped TiO2, respectively. Importantly, the synergistic effect of terbium and sulfur dopants was profoundly discussed. Furthermore, recycling tests and acute toxicity experiments were carried out to confirm the reusability and biosafety of Tb, S co-doped TiO2.

The complete mitochondrial DNA genome of a cone snail, Conus betulinus (Neogastropoda: Conidae), from the South China sea.

The complete mitochondrial genome of the tubular cone snail Conus betulinus is presented in this study. The C. betulinus mitochondrial genome was 16,240 bp with 13 protein-coding genes (PCGs), 22 transfer RNA (tRNA) genes, 2 ribosomal RNA (rRNA) genes, and a non-coding AT-rich region (D-loop). The overall base composition was estimated to be 25.67% for A, 38.26% for T, 21.38% for G, and 14.69% for C, with a high A + T content of 63.93%. Phylogenetic analyses based on 13 PCGs showed the close relationship of vermivorous C. betulinus with the common ancestor of molluscivorous Conus textile and Conus gloriamaris, providing a basis for further studies on the phylogenetics of cone snails according to their dietary type.

Jiaolong capsule protects SD rats against 2,4,6-trinitrobenzene sulfonic acid induced colitis.

ETHNOPHARMACOLOGICAL RELEVANCE: Jiaolong capsule (JLC) was approved for the therapy of gastrointestinal diseases by the State Food and Drug Administration (SFDA) of China. It has a satisfactory curative effect in the treatment of patients with inflammatory bowel disease, however, the mechanism remains to be elucidated. AIM OF THE STUDY: In current study, the effects and possible mechanisms of JLC on 2,4,6-trinitrobenzene sulfonic acid (TNBS)-induced colitis were investigated. MATERIALS AND METHODS: Sulfasalazine and JLC were administrated orally and initialized 6 h after TNBS enema, once a day for seven consecutive days. The effect of JLC on intestinal microbial populations and LPS/TLR-4/NF-κB pathway was observed and assessed. Thirty female SD rats were distributed into six groups randomly and equally, namely, control, TNBS, TNBS + sulfasalazine (625 mg/kg), and TNBS + three different doses of JLC (25, 50, and 100 mg/kg) groups. RESULTS: The effect of JLC on restoring normal structures of colorectum and repairing colonic damage were superior to that of sulfasalazine. JLC showed a positive effect in re-balancing intestinal bacteria population of colitis, and suppressed the activation of LPS/TLR-4/NF-κB pathway. CONCLUSION: The results suggest that JLC demonstrated a beneficial effect on treating colitis in a rat model. The possible mechanisms may be through the regulatory effect of intestinal commensal bacteria and down-regulation of LPS/TLR-4/NF-κB pathway.

Rapid preparation of terbium-doped titanium dioxide nanoparticles and their enhanced photocatalytic performance.

Further applications of photocatalysis were limited by the high recombination probability of photo-induced electron-hole pairs in traditional titanium dioxide nanoparticles (TiO2 NPs). Herein, we modified them with rare earth metal via a facile sol-gel method, using tetrabutyl titanate as a precursor and terbium (III) nitrate hexahydrate as terbium (Tb) source. The resulting samples with different Tb doping amounts (from 0 to 2%) have been characterized by X-ray diffraction, UV-visible diffuse reflectance spectroscopy, X-ray photo-electron spectroscopy and a scanning electron microscope. The photocatalytic performance of Tb-doped TiO2 was evaluated by the degradation of methylene blue. The effects of Tb doping amount and initial pH value of solution were investigated in detail. The composite with Tb doping amount of 1.0 wt% showed the highest photocatalytic performance. It exhibited approximately three times enhancement in photocatalytic activity with a reaction rate constant of 0.2314 h-1 when compared with that of commercial P25 (0.0827 h-1). In addition, it presented low toxicity on zebrafishes with 96 h-LC50 of 23.2 mg l-1, and has been proved to be reusable for at least four cycles without significant loss of photocatalytic activity. A probable photocatalytic mechanism of Tb-doped TiO2 was proposed according to the active species trapping experiments. The high photocatalytic performance, excellent reusability and low toxicity of Tb-doped TiO2 indicated that it is a promising candidate material in the future treatment of dye wastewater.

The complete chloroplast genome sequence of Alpinia oxyphylla Miq. and comparison analysis within the Zingiberaceae family.

Alpinia oxyphylla Miq. (A. oxyphylla) is an important edible and traditional herbal medicine. In this study, the complete chloroplast genome of A. oxyphylla was sequenced, analysed, and compared to five species in the Zingiberaceae family. The size of the A. oxyphylla chloroplast genome was 161351 bp, which consisted of a large single-copy (LSC, 87248 bp) and small single-copy (SSC, 16175 bp) region separated by a pair of inverted repeats (IRa and IRb, 28964 bp each). The genome encoded 132 unique genes, including 87 protein-coding genes, 37 tRNAs and four rRNAs. The GC content of the genome was 36.17%. A total of 53 simple sequence repeats (SSRs) and 80 long repeats were identified in the A. oxyphylla chloroplast genome. The chloroplast genome of A. oxyphylla shared the highest sequence similarity of >90% with the chloroplast genome of A. zerumbet, and six chloroplast genomes in the Zingiberaceae family were compared by using CGView Comparison Tool (CCT). According to the phylogenetic tree, the Zingiberaceae family is divided into two categories, which coincide with the classification of the characteristics of sun-like and shade-like in plants. Our results reveal the phototrophic component of NADH-dehydrogenase (ndhB and ndhC), photosystem II (psbZ) and ATP synthase (atpE, atpF) exhibit adaptive evolution under different environments, and the strength of light is an important trigger for the adaptations at the chloroplast level.

Echinacoside alleviates hypobaric hypoxia-induced memory impairment in C57 mice.

Acute hypobaric hypoxia (HH) gives rise to persistent cognitive impairment, influencing memory function specifically. Echinacoside (ECH), one of the phenylethanoids isolated from the stems of Cistanche salsa, has been reported to prevent ischemia induced by neuronal injury traditionally. This study then tried to investigate whether ECH could alleviate HH-induced memory deficit. Ten C57 mice were used as control, and 50 were exposed to HH equivalent to 6,100 m for 7 days in a decompression chamber and were given ECH daily (50, 75, or 100 mg/kg) through gavage during the period of exposure. Cognitive performance was evaluated by the Morris water maze test. ECH, especially at 100 mg/kg, significantly reduced HH-induced memory decline. Furthermore, ECH increased the expression of nuclear factor E2 p45-related factor 2, heme oxygenase-1, NAD(P)H:quinone oxidoreductase 1, and γ-glutamyl cysteine synthetase in mRNA and protein levels, suggesting that the Keap1-Nrf2-ARE signaling pathway might be involved in neuronal adaptation. The results indicate that ECH could prevent HH-induced memory impairment, which is associated with antioxidant effect of ECH in the hippocampus.

Toxic Effects of TiO2 NPs on Zebrafish.

Titanium dioxide nanoparticles (TiO2 NPs) have become a widely used nanomaterial due to the photocatalytic activity and absorption of ultraviolet light of specific wavelengths. This study investigated the toxic effects of rutile TiO2 NPs on zebrafish by examining its embryos and adults. In the embryo acute toxicity test, exposure to 100 mg/L TiO2 NPs didn't affect the hatching rate of zebrafish embryos, and there was no sign of deformity. In the adult toxicity test, the effects of TiO2 NPs on oxidative damage in liver, intestine and gill tissue were studied. Enzyme linked immunosorbent assay (ELISA) and fluorescence-based quantitative real-time reverse transcription PCR (qRT-PCR) were used to detect the three antioxidant enzymes: superoxide dismutase (SOD), catalase (CAT) and glutathione S transferase (GSTs) in the above mentioned zebrafish organs at protein and gene levels. The results showed that long-term exposure to TiO2 NPs can cause oxidative damage to organisms; and compared with the control group, the activity of the three kinds of enzyme declined somewhat at the protein level. In addition, long-term exposure to TiO2 NPs could cause high expression of CAT, SOD and GSTs in three organs of adult zebrafish in order to counter the adverse reaction. The effects of long-term exposure to TiO2 NPs to adult zebrafish were more obvious in the liver and gill.

Paris saponin VII reverses chemoresistance in breast MCF-7/ADR cells.

ETHNOPHARMACOLOGICAL RELEVANCE: The development of a multidrug-resistant (MDR) phenotype is a main obstacle to the successful treatment of breast cancer. Saponins of several herbs are considered as promising candidates for drug resistance treatment. We extracted Paris saponin VII (PS VII) from Trillium tschonoskii Maxim. and investigated whether it could sensitize chemoresistant breast cancer cells MCF-7/ADR to the cytotoxic effects of adriamycin. MATERIALS AND METHODS: MCF-7/ADR cells were exposed to 0.5 µM PSVII plus different concentrations of adriamycin (0-100 µM). Then, MTT assay and adriamycin accumulation assay were used to assess cell proliferation and intracellular adriamycin retention. P glycoprotein levels and intracellular rhodamine 123 (Rh-123) accumulations were investigated to measure the expression and activity of P-glycoprotein. A xenograft model of nude mouse was utilized to observe the effect of PSVII in vivo. RESULTS: Treatment with PSVII influenced cell viability of MCF-7/ADR cells, as well as sensitized MCF-7/ADR cells to the cytotoxic effects of adriamycin. Moreover, PSVII significantly downregulated MDR1 expression in MCF-7/ADR cells. Intravenous administration of PSVII significantly enhanced anticancer efficacy of adriamycin to MCF-7/ADR xenograft model in nude mice. CONCLUSION: These findings suggested a possible application of PSVII in combination with chemotherapy and/or as neo-adjuvant therapy in the treatment of MDR breast cancer.

A Novel Yeast Surface Display Method for Large-Scale Screen Inhibitors of Sortase A.

Fluorescence resonance energy transfer substrates of sortase A are too expensive to be used to roughly screen high-throughput sortase A inhibitors. This makes therapeutic strategies difficult to realize in a clinical therapeutic use. Instead, we design here an LPETG-EGFP (leucine, proline, glutamic, threonine and glycine-enhanced green fluorescence) protein displayed on a yeast surface as a substrate by adaptively reducing the cost. We do this by optimizing the induction conditions of sortase A expression in Escherichia coli DE3(BL21) and catalyzing LPETG proteins, which are displayed on surface of Pichia pastoris. Different expression conditions of sortase A include: induction temperature (22 °C, 28 °C, 37 °C and 40 °C), induction time (4 h, 5 h, 6 h and 7 h) and induction concentration of isopropyl ß-d-thiogalactoside IPTG (0.25 mmol/L, 0.5 mmol/L, 1 mmol/L, and 2 mmol/L). The fluorescence change of the LPETG-EGFP protein on the surface of P. pastoris over time was detected by flow cytometry and fluorescence spectrophotometry, and then the sensitivities of the two methods were compared. Using berberine chloride as an inhibitor, the activity of sortase A was investigated with the substrates of LPETG-EGFP protein, and compared to Dabcyl-QALPETGEE-Edans. A high yield of sortase A was achieved by inducing 1.0 mmol/L IPTG at 28 °C for 6 h. The intensity of green fluorescence of substrates displayed on the yeast surface was increased over time, while the stability was decreased slightly. Both fluorescence spectrophotometery and flow cytometry were fit for detection because of their high sensitivity. We utilized two different substrates of sortase A to investigate sortase A activity, which resulted in the increase of fluorescence intensity with respect to the increased time of growth. However, the method with Dabcyl-QALPETGEE-Edans as its substrate was more robust. Thus, the method described in this paper is a simple and cheap method which is very suitable for high-throughput analysis, but the conventional method is much more sensitive. The method described in this paper is expected to lead to large-scale screening of sortase A inhibitors which can be used to decrease the risk of drug resistance development.

Modified apple polysaccharide prevents colitis through modulating IL-22 and IL-22BP expression.

Chronic intestinal inflammation enhances cell proliferation, angiogenesis, and migration, then promotes the development of colorectal cancer (CRC). Many ingredients of apples have been proven to have anti-inflammatory properties, and show benefits for colitis treatment. In our previous studies, we found modified apple polysaccharide (MAP) could prevent colitis associated colorectal carcinogenesis effectively. Herein, we further our study to observe the effect of MAP on dextran sodium sulfate (DSS)-induced colitis and to investigate the possible mechanisms. IL-22 has both pathogenic and protective effects during intestinal tissue damage. It could be neutralized by the soluble IL-22 receptor, known as the IL-22 binding protein (IL-22BP). A DSS-induced colitis mouse model, a mouse CRC cell line MCA-38 and a mouse dendritic cell line DC2.4 were treated with MAP. Western blot, ELISA, BrdU staining and a co-culture system were used to detect the expression of IL-22 and IL-22BP. MAP significantly protected ICR mice against DSS-induced colitis, and inhibited the growth of MCA-38 cells. The mechanisms may be that MAP down-regulated IL-22 level and up-regulated expression of IL-22BP. These data may provide another molecular basis for understanding how apples act to prevent colitis and suggest that MAP has a potential to treat colitis and prevent CRC.

Differential gene responses in the embryo of the green mussel Perna viridis exposed to dichlorodiphenyltrichloroethane (DDT).

The green-lipped mussel, Perna viridis, is considered to be an ideal indicator for marine environmental pollution. Dichlorodiphenyltrichloroethane (DDT), a typical persistent organic pollutant, is extensively distributed in marine environments. However, little is known about the toxic effects of DDT on the embryo of marine animals, particularly in marine bivalves. Using next-generation sequencing technology, we studied P. viridis embryo after DDT stress at the transcriptome level. A total of 99 202 unigenes were obtained based on the 2383 bp of unigene N50. These differentially expressed genes (DEGs) participated in the various molecular pathways of biological effects, including oxidative stress, detoxification, innate immunity and neurobehavioral disease. Quantitative real-time PCR was performed to verify the mRNA expression of several genes identified by differential gene expression (DGE) analysis. The results indicated that DDT was in induced a dose-dependent manner in the embryo of P. viridis, and most genes involved in oxidative stress and detoxification were up-regulated by DDT exposure; however, the immunity-related genes were down-regulated, except the genes involved in phagocytosis. Gene expression changes in embryo from P. viridis provide a preliminary basis to better understand the molecular toxic response mechanisms of embryo to DDT.

Remediation of acid mine drainage using microbial fuel cell based on sludge anaerobic fermentation.

The aim of this work is to utilize the microbial fuel cell for removing metals and sulfate from acid mine drainage using sewage sludge organics and simultaneous electricity generation. The enriched sulfate-reducing mixed culture was used as the cathodic biofilm and the sludge as the substrate. Under anaerobic conditions, 71.2% sulfate, 99.7% heavy metals, and 51.6% total chemical oxygen demand are removed at an electrode spacing of 4 cm and a sludge concentration of 30% (v/v) after 10-day treatment. A maximum power density of 51.3 mW/m2 is obtained. Approximately 79.5% of the dissipated sulfate is converted to element sulfur or polysulfides. The sulfide concentration is kept at below 20 mg-S/L. The concentrations of heavy metals are in the range of 0.02-0.06 mg/L in the effluent, which are far below the levels required by Chinese legislation. Microbial community analysis reveals that sulfate-reducing bacteria in Desulfuromonadales are dominant on the cathodic biofilm at the end of experiments. This study shows the potential of synchronous degradation of residual sludge and treatment of AMD with electricity harvesting.

Thyroid Disruption in Zebrafish Larvae by Short-Term Exposure to Bisphenol AF.

Bisphenol AF (BPAF) is extensively used as a raw material in industry, resulting in its widespread distribution in the aqueous environment. However, the effect of BPAF on the hypothalamic-pituitary-thyroidal (HPT) axis remains unknown. For elucidating the disruptive effects of BPAF on thyroid function and expression of the representative genes along the HPT axis in zebrafish (Danio rerio) embryos, whole-body total 3,3',5-triiodothyronine (TT3), total 3,5,3',5'-tetraiodothyronine (TT4), free 3,3',5-triiodothyronine (FT3) and free 3,5,3',5'-tetraiodothyronine (FT4) levels were examined following 168 h post-fertilization exposure to different BPAF concentrations (0, 5, 50 and 500 µg/L). The results showed that whole-body TT3, TT4, FT3 and FT4 contents decreased significantly with the BPAF treatment, indicating an endocrine disruption of thyroid. The expression of thyroid-stimulating hormone-ß and thyroglobulin genes increased after exposing to 50 µg/L BPAF in seven-day-old larvae. The expressions of thyronine deiodinases type 1, type 2 and transthyretin mRNAs were also significantly up-regulated, which were possibly associated with a deterioration of thyroid function. However, slc5a5 gene transcription was significantly down-regulated at 50 µg/L and 500 µg/L BPAF exposure. Furthermore, trα and trß genes were down-regulated transcriptionally after BPAF exposure. It demonstrates that BPAF exposure triggered thyroid endocrine toxicity by altering the whole-body contents of thyroid hormones and changing the transcription of the genes involved in the HPT axis in zebrafish larvae.