Fucoidan from Cladosiphon okamuranus enhances antioxidant activity and prevents reproductive dysfunction in polystyrene microplastic-induced male rats.

Plastic pollution, including microplastic, has emerged as a severe environmental and public health problem. The health risks, especially in the case of reproductive damage caused by polystyrene microplastic (PS-MP) exposure, are emerging problems that need to be solved. This study aimed to investigate the effects of fucoidan extracted from Cladosiphon okamuranus on the polystyrene microplastic-induced oxidative stress of the Leydig (LC540) cells and reproductive damage in male rats. The oxidative stress of the LC540 cells and reproductive damage in the rats were induced by PS-MP. The fucoidan treatment reduces nitric oxide (NO) and reactive oxygen species generation in the LC540 cells. In the animal study, fucoidan treatment enhanced enzymatic antioxidant activities (glutathione peroxidase, superoxide dismutase, glucose-6-phosphate dehydrogenase, and glutathione reductase) and reduced malondialdehyde and nitric oxide production. Fucoidan supplementation also downregulates tumor necrosis factor-alpha, interleukin-6, and caspase-3 expression. Additionally, fucoidan upregulates testosterone levels, prevents the reduction of epithelium thickness, and reduces the area of the seminiferous tubule lumen. According to these conditions, fucoidan from Cladosiphon okamuranus prevents reproductive damage by downregulating oxidative stress and pro-inflammatory cytokines. Therefore, fucoidan can be used as a source of food supplements or functional food ingredients for reproductive or testicular damage management.

Effects of taurine against benzo[α]pyrene-induced cell cycle arrest and reactive oxygen species-mediated nuclear factor-kappa B apoptosis via reduction of mitochondrial stress in A549 cells.

Taurine is a free amino acid that prevents reactive oxygen species (ROS) formation. ROS production is associated with oxidative stress, cell proliferation, apoptosis, inflammation, and DNA alterations in benzo[α]pyrene (BaP)-induced lung cells. Here, we assessed the effect of adding of 25 mM taurine on human pulmonary alveolar epithelial A549 cells treated with different concentrations of BaP. After culturing for 24 h, the cells were tested for biomarkers including cell viability, cellular morphology, Annexin V-FITC/propidium iodide, cell cycle regulation, ROS accumulation, mitochondrial membrane potential (MMP), and expression of related signaling genes and proteins. BaP induced cell cycle arrest and decreased cell viability in a dose-dependent manner. In addition, 50 µM BaP induced a 52.2% increase in ROS levels and inhibited MMP by up to 80%; however, taurine decreased BaP-induced ROS production by 19.5% and restored MMP. The expression of nuclear factor-kappa B (NF-κB), B-cell lymphoma-2 (BCL-2) homologous antagonist killer (Bak), BCL-2-associated X protein (Bax), and cytochrome c at both the mRNA and protein levels were increased, and the expression of BCL-2 and BCL-x1 was decreased by BaP treatment. Furthermore, BaP activated caspase-3/7 expression by up to 25%. However, taurine decreased the expression of NF-κB, Bak, Bax and cytochrome c levels, reduced caspase-3/7 activities, and increased the expression of BCL-2 and BCL-x1. Hence, taurine attenuates BaP-induced oxidative stress and mitochondrial dysfunction by inhibiting the NF-κB-mediated intrinsic apoptosis pathway in A549 cells. Taurine can be considered as a preventive molecule to prevent lung damage.

Comprehensive detection of 120 additives in food using nontargeted MS data acquisition.

The compliance assessment on the labeling of food additives is a hard job, because there are nearly thousand legal food additives can be used in food, and countless illegal additives must also deal with. This study developed a non-targeted data acquisition screening method based on liquid chromatography high resolution mass spectrometry (HRMS) in which a precursor ion and two product ions of each analyte are able to be recorded. The high throughput screening method worked as foodomics that characterized and identified every food components as long as they were ionized in terms of theory. The data acquisition method called data independent acquisition (DIA) was achieved by a full scan form m/z 70-1050, and then followed wide window fragmentations of product ions recording. A full scan and the followed fragmentations generated 21 spectra in 2.6 s contributed about 6 data points for a typical 0.2-0.3 min width peak in HPLC. A detection database list of 120 additives included 79 colorants, 13 sweeteners, 12 preservatives and 7 antioxidants was established. Thirty-three commercial samples including beverages, candies, and sauces were surveyed for testing additives. Sweeteners (rebaudioside A) and flavoring agents (malic acid and fumaric acid) were found the most under declared additives. HPLC column often do not provide adequate retention for highly polar compounds such as organic acids (flavoring agents). In this study they were coeluted, but were able to be separated and determined by HRMS worked as the secondary separation tool. The surveillance results showed there is still room for food manufacturers to improve the connection between their product information and consumers.

The complete mitochondrial genome of Conus quercinus (Neogastropoda: Conidae).

The complete mitochondrial genome sequence of cone snail Conus quercinus a kind of worm-hunting sea snails, was performed by next-generation sequencing. The mitogenome is 16,439 bp in length, including 13 protein-coding genes, 22 tRNA genes, two ribosomal RNA genes (12S and 16S rRNA), and one control region. It has overall base composition of A (28.1%), T (38.2%), C (14.7%), and G (18.6%). It shows 75.9% identity with C. capitaneus, which also belongs to worm-hunting sea snail. The phylogenetic analysis was conducted with 22 closely related species to assess their phylogenetic relationship. The complete mitogenome of the C. quercinus provides important DNA molecular data for further phylogeography.

Mitochondrial DNA sequence of Conus textile (Neogastropoda: Conidae).

The cone snail Conus textile belongs to the family Conidae. It is a kind of molluscivorous species. The complete mitochondrial DNA sequence was constructed by next-generation sequencing in this study. The mitogenome of C. textile is 15,765 bp in length, including 13 protein-coding genes, 22 tRNA genes, 2 ribosomal RNA genes and 1 control region. The base composition was 27.3% A, 37.9% T, 15.7% C and 19.1% G. The phylogenetic tree of C. textile with the other 6 Conus species and 15 Neogastropoda sea snails was built. It provides fundamental data for further research of phylogeny and biogeography with this genus.

The complete mitochondrial genome of Conus capitaneus (Neogastropoda: Conidae).

The complete mitochondrial genome sequence of cone snail Conus capitaneus, a kind of worm-hunting sea snails, was performed by next-generation sequencing. The mitogenome is 15,829 bp in length, including 13 protein-coding genes, 22 tRNA genes, 2 ribosomal RNA genes (12S and 16S rRNA) and 1 control region. It has an overall base composition of A (25.6%), T (36.6%), C (16.3%) and G (21.5%). It shows 79.8% identity with C. tribblei, which also belongs to worm-hunting sea snail. The phylogenetic analysis was conducted with 21 closely related species to assess their phylogenetic relationship. The complete mitogenome of the C. capitaneus provides important DNA molecular data for further phylogeography.

The complete mitochondrial genome of Conus striatus (Neogastropoda: Conidae).

Conus striatus is a kind of piscivorous cone snail. We have sequenced it by next generation sequencing method. We used de novo assembly and reference mapping methods to assemble mitogenome. The mitochondrial genome is 15,738 bp, containing 13 protein coding genes, 22 transfer RNAs and 2 ribosomal RNAs genes. The overall base composition of C. striatus is 25.9% for A, 16.3% for C, 20.8% for G and 38.6% for T. The phylogenetic analysis was conducted with 18 related species and confirmed the classification status. The complete mitogenome of the C. striatus provides an essential and important DNA molecular data for further phylogeography and evolutionary analysis for cone snail phylogeny.

Development of standardized methodology for identifying toxins in clinical samples and fish species associated with tetrodotoxin-borne poisoning incidents.

Tetrodotoxin (TTX) is a naturally occurring toxin in food, especially in puffer fish. TTX poisoning is observed frequently in South East Asian regions. In TTX-derived food poisoning outbreaks, the amount of TTX recovered from suspicious fish samples or leftovers, and residual levels from biological fluids of victims are typically trace. However, liquid chromatography-mass spectrometry and liquid chromatography-tandem mass spectrometry methods have been demonstrated to qualitatively and quantitatively determine TTX in clinical samples from victims. Identification and validation of the TTX-originating seafood species responsible for a food poisoning incident is needed. A polymerase chain reaction-based method on mitochondrial DNA analysis is useful for identification of fish species. This review aims to collect pertinent information available on TTX-borne food poisoning incidents with a special emphasis on the analytical methods employed for TTX detection in clinical laboratories as well as for the identification of TTX-bearing species.

The complete mitochondrial genome of Conus tulipa (Neogastropoda: Conidae).

The complete mitogenome sequence of the cone snail Conus tulipa (Linnaeus, 1758) has been sequenced by next-generation sequencing method. The assembled mitogenome is 16,599 bp in length, including 13 protein-coding genes, 22 transfer RNA genes and 2 ribosomal RNA genes. The overall base composition of C. tulipa is 28.7% A, 15.2% C, 18.4% G and 37.7% T. It shows 81.1% identity to the cone snail C. consors, 78.5% to C. borgesi and 77.5% to C. textile. Using the 13 protein-coding genes and 2 ribosomal RNA genes of C. tulipa in this study, together with 18 other closely species, we constructed the species phylogenetic tree to verify the accuracy and utility of new determined mitogenome sequence. The complete mitogenome of the C. tulipa provides an essential and important DNA molecular data for further phylogeography and evolutionary analysis for cone snail phylogeny.

Differential proteomics to explore the inhibitory effects of acidic, slightly acidic electrolysed water and sodium hypochlorite solution on Vibrio parahaemolyticus.

Slightly acidic electrolysed water (SlAEW) and acidic electrolysed water (AEW) have been demonstrated to effectively inactivate food-borne pathogens. However, the underlying mechanism of inactivation remains unknown. Therefore, in this study, a differential proteomic platform was used to investigate the bactericidal mechanism of SlAEW, AEW, and sodium hypochlorite (NaOCl) solutions against Vibrio parahaemolyticus. The upregulated proteins after SlAEW, AEW, and NaOCl treatments were identified as outer membrane proteins K and U. The downregulated proteins after the SlAEW, AEW, and NaOCl treatments were identified as adenylate kinase, phosphoglycerate kinase, glyceraldehyde-3-phosphate dehydrogenase (GAPDH), and enolase, all of which are responsible for energy metabolism. Protein synthesis-associated proteins were downregulated and identified as elongation factor Tu and GAPDH. The inhibitory effects of SlAEW and AEW solutions against V. parahaemolyticus may be attributed to the changes in cell membrane permeability, protein synthesis activity, and adenosine triphosphate (ATP) biosynthesis pathways such as glycolysis and ATP replenishment.

Plancitoxin I from the venom of crown-of-thorns starfish (Acanthaster planci) induces oxidative and endoplasmic reticulum stress associated cytotoxicity in A375.S2 cells.

The crown-of-thorns starfish Acanthaster planci is a venomous starfish whose venom provokes strong cytotoxicity. In the present study, the purified cytotoxic toxin of A. planci venom (CAV) was identified as plancitoxin I protein by mass spectrum analyses. This study aims to investigate the molecular mechanism underlying the cytotoxicity function of plancitoxin I by focusing on the oxidative stress, mitochondrial dysfunction and endoplasmic reticulum (ER) stress pathway in human melanoma A375.S2 cells. The results indicated that after being treated with CAV toxin, A375.S2 cells significantly decreased viability in a dose-dependent manner. The CAV was found to reduce the cellular antioxidant enzymes such as SOD and CAT, and there was a significant decrease in total thiol level and mtDNA integrity, and it enhanced the lipid peroxidation. In addition, CAV increased cytosolic Ca(2+) concentration, and enhanced the expression of the ER molecular chaperones GRP78 and CHOP in a dose-dependent manner. CAV significantly elevated the activity of caspase-3, -8 and -9, and reduced the ratio of Bcl-2/Bax. The cells exhibited apoptosis were determined by using propidium iodide (PI) staining of DNA fragmentation (sub-G1 peak). In summary, the results demonstrated that plancitoxin I inhibits the proliferation of A375.S2 cells through induction of oxidative stress, mitochondrial dysfunction and ER stress associated apoptosis.

Taurine resumed neuronal differentiation in arsenite-treated N2a cells through reducing oxidative stress, endoplasmic reticulum stress, and mitochondrial dysfunction.

The goal of the study is to investigate the preventive effect of taurine against arsenite-induced arrest of neuronal differentiation in N2a cells. Our results revealed that taurine reinstated the neurite outgrowth in arsenite-treated N2a cells. Meanwhile, arsenite-induced oxidative stress and mitochondrial dysfunction as well as degradation of mitochondria DNA (mtDNA) were also inhibited by co-treatment of taurine. Since oxidative stress and mitochondrial dysfunction is closely associated with endoplasmic reticulum (ER) stress, we further examined indicators of ER stress, 78 kDa glucose-regulated protein (GRP78), and C/EBP-homologous protein (CHOP) protein expression. The results demonstrated that taurine significantly reduced arsenite-induced ER stress in N2a cells. In the parallel experiment, arsenite-induced disruption of intracellular calcium homeostasis was also ameliorated by taurine. The proven bio-function of taurine preserved a preventive effect against deleteriously cross-talking between oxidative stress, mitochondria, and ER. Overall, the results of the study suggested that taurine reinstated neuronal differentiation by inhibiting oxidative stress, ER stress, and mitochondrial dysfunction in arsenite-treated N2a cells.

Cytotoxic and apoptotic activities of the plancitoxin I from the venom of crown-of-thorns starfish (Acanthaster planci) on A375.S2 cells.

This study reports on a cytotoxic toxin derived from the venom of the crown-of-thorns starfish Acanthaster planci (CAV). The protein toxin was isolated through both ion-exchange and gel-filtration chromatography, and characterized by sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) and mass spectrum analyzes. The CAV was identified as plancitoxin I protein. The mechanistic role of the CAV toxin was explored in human malignant melanoma A375.S2 cell death. The results indicated that after incubation with CAV toxin, cells significantly decreased in A375.S2 cell viability and increased in the lactate dehydrogenase (LDH) level in a dose-dependent manner. The assays indicated that CAV toxin promoted reactive oxygen species (ROS) production, induced nitric oxide (NO) formation, lost mitochondrial membrane potential (ΔΨm) and induced inter-nucleosomal DNA fragmentation in A375.S2 cells. The molecular cytotoxicity of the CAV toxin was tested through evaluation of the apoptosis/necrosis ratio by double staining with annexin V-FITC and a propidium iodide (PI) assay. The results suggested that CAV toxin induced a cytotoxic effect in A375.S2 cells via the apoptotic procedure, and may be associated with the regulation of the p38 pathways.

Antioxidative and anticancer activities of various ethanolic extract fractions from crown-of-thorns starfish (Acanthaster planci).

Many studies currently researching marine invertebrates to determine the therapeutic potential of their bioactive materials have been showing very promising results. The crown-of-thorns starfish Acanthaster planci, an Echinodermata of the class Asteroidea, is infamous as the unique venomous starfish and as a destroyer of coral reefs. Starfish possesses many useful pharmacological and biological characteristics. In this study, A. planci was extracted with 70% ethanol and lyophilized to obtain an ethanol fraction. The ethanol fraction was dissolved with water and defatted with petroleum ether to obtain a non-polar fraction. The residual solution was successively partitioned with ethylacetate and butanol to obtain an ethylacetate fraction and butanol fraction, respectively. Four fractions were used to examine the antioxidant and anticancer properties. The ethanol fraction of A. planci contained the highest antioxidant effects such as ABTS, DPPH, Fe(2+) chelating activity and reducing power when compared with four fractions. Among the four fractions, the butanol fraction was especially shown to inhibit human malignant melanoma A375.S2 cells' proliferation, which is involved in the apoptotic progression. This fraction could induce apoptosis and even necrosis in A375.S2 cells as evidenced by double staining with an Annexin V-FITC and PI assay and DNA fragmentation analysis. These results indicated that the starfish A. planci is a good resource for obtaining the biologically active substances for antioxidant and anticancer effects.

Toxin and species identification of toxic octopus implicated into food poisoning in Taiwan.

A food poisoning incident due to ingestion of unknown octopus occurred in Taipei in December, 2010. The serum and urine from victims (male 38 and 43 years old) were collected, determined the toxicity, and identified tetrodotoxin (TTX) by high-performance liquid chromatography with tandem mass spectrometry (LC-MS/MS). It was found that only urine contained the trace of TTX. Then, two retained specimen (one without blue ring in the skin and another with small blue ring in the skin) were collected from victims and examined for the toxicity and toxin. Meanwhile, 6 specimens of octopus without blue ring in the skin and 4 specimens of octopus with blue ring in the skin were re-collected from the market. Both retained octopus samples were found to contain TTX. However, re-collected market's octopus without blue ring in the skin did not show to contain TTX the and was identified as Octopus aegina by using the analysis of cytochrome b gene (Cyt b) and cytochrome c oxidase subunit I gene (COI). Only octopus with blue ring in the skin contained TTX and was identified as Hapalochlaena fasciata by using the analysis of Cyt b and COI. Therefore, this octopus food poisoning was caused by toxic octopus H. fasciata and the causative agent was TTX.

Spine venom of crown-of-thorns starfish (Acanthaster planci) induces antiproliferation and apoptosis of human melanoma cells (A375.S2).

The crown-of-thorns starfish (Acanthaster planci) is a venomous starfish. In this study, the extraction of A. planci spine venom (ASV) was performed by phosphate saline buffer, followed by assaying the cytotoxicity on human normal and tumor cells. It was found that human melanoma cells (A375.S2) were the most sensitive to the ASV solution. The cells, after incubation with ASV, significantly appeared to decrease cell viability and increase lactate dehydrogenase (LDH) release with a dose-dependent relationship. The extract of spine promoted loss of mitochondrial membrane potential (ΔΨm) and induced inter-nucleosomal DNA fragmentation in human melanoma cells. The cells exhibited apoptosis by using propidium iodide (PI) staining of DNA fragmentation; it was then determined by flow cytometry (sub-G1 peak). The molecular cytotoxicity of ASV was tested through evaluation of the apoptosis/necrosis ratio by double staining with annexin V and PI assay. The A. planci spine venom showed significant antiproliferation. The human melanoma cells revealed apoptosis at low dose (1.25 µg/ml), and necrosis occurred at high dose (5 µg/ml).

Hemolytic activity of venom from crown-of-thorns starfish Acanthaster planci spines.

BACKGROUND: The crown-of-thorns starfish Acanthaster planci is a venomous species from Taiwan whose venom provokes strong hemolytic activity. To understand the hemolytic properties of A. planci venom, samples were collected from A. planci spines in the Penghu Islands, dialyzed with distilled water, and lyophilized into A. planci spine venom (ASV) powder. RESULTS: Both crude venom and ASV cause 50% hemolysis at a concentration of 20 µg/mL. The highest hemolytic activity of ASV was measured at pH 7.0-7.4; ASV-dependent hemolysis was sharply reduced when the pH was lower than 3 or greater than 8. There was almost no hemolytic activity when the Cu2+ concentration was increased to 10 mM. Furthermore, incubation at 100°C for 30 to 60 minutes sharply decreased the hemolytic activity of ASV. After treatment with the protease α-chymotrypsin, the glycoside hydrolase cellulase, and the membrane component cholesterin, the hemolytic activity of ASV was significantly inhibited. CONCLUSIONS: The results of this study provide fundamental information about A. planci spine venom. The hemolytic activity was affected by pH, temperature, metal ions, EDTA, cholesterin, proteases, and glycoside hydrolases. ASV hemolysis was inhibited by Cu2+, cholesterin, α-chymotrypsin, and cellulose, factors that might prevent the hemolytic activity of venom and provide the medical treatment for sting.

Taurine prevented cell cycle arrest and restored neurotrophic gene expression in arsenite-treated SH-SY5Y cells.

The study investigated the effect of taurine on cell viability and neurotrophic gene expression in arsenite-treated human neuroblastoma SH-SY5Y cells. Arsenite-induced intracellular reactive oxygen species (ROS) and interrupted cell cycle in SH-SY5Y cells. In addition, arsenite reduced mitochondria membrane potential (MMP) and decreased neurotrophic gene expressions such as n-myc downstream-regulated gene 4 (NDRG-4), brain-derived neurotrophic factor (BDNF) and sirtuin-1 (SIRT-1) in SH-SY5Y cells. In parallel, taurine prevented cell cycle, restored MMP and reduced the intracellular ROS level, and taurine recovered NDRG-4, BDNF and SIRT-1 gene expressions in arsenite-treated SH-SY5Y cells while taurine alone has no effect on these parameters.

Hemolytic activity of venom from crown-of-thorns starfish Acanthaster planci spines

Background : The crown-of-thorns starfish Acanthaster planci is a venomous species from Taiwan whose venom provokes strong hemolytic activity. To understand the hemolytic properties of A. planci venom, samples were collected from A. planci spines in the Penghu Islands, dialyzed with distilled water, and lyophilized into A. planci spine venom (ASV) powder. Results : Both crude venom and ASV cause 50% hemolysis at a concentration of 20 μg/mL. The highest hemolytic activity of ASV was measured at pH 7.0-7.4; ASV-dependent hemolysis was sharply reduced when the pH was lower than 3 or greater than 8. There was almost no hemolytic activity when the Cu2+ concentration was increased to 10 mM. Furthermore, incubation at 100°C for 30 to 60 minutes sharply decreased the hemolytic activity of ASV. After treatment with the protease α-chymotrypsin, the glycoside hydrolase cellulase, and the membrane component cholesterin, the hemolytic activity of ASV was significantly inhibited. Conclusions : The results of this study provide fundamental information about A. planci spine venom. The hemolytic activity was affected by pH, temperature, metal ions, EDTA, cholesterin, proteases, and glycoside hydrolases. ASV hemolysis was inhibited by Cu2+, cholesterin, α-chymotrypsin, and cellulose, factors that might prevent the hemolytic activity of venom and provide the medical treatment for sting.(AU)

Hemolytic activity of venom from crown-of-thorns starfish Acanthaster planci spines

Background : The crown-of-thorns starfish Acanthaster planci is a venomous species from Taiwan whose venom provokes strong hemolytic activity. To understand the hemolytic properties of A. planci venom, samples were collected from A. planci spines in the Penghu Islands, dialyzed with distilled water, and lyophilized into A. planci spine venom (ASV) powder. Results : Both crude venom and ASV cause 50% hemolysis at a concentration of 20 g/mL. The highest hemolytic activity of ASV was measured at pH 7.0-7.4; ASV-dependent hemolysis was sharply reduced when the pH was lower than 3 or greater than 8. There was almost no hemolytic activity when the Cu2+ concentration was increased to 10 mM. Furthermore, incubation at 100°C for 30 to 60 minutes sharply decreased the hemolytic activity of ASV. After treatment with the protease -chymotrypsin, the glycoside hydrolase cellulase, and the membrane component cholesterin, the hemolytic activity of ASV was significantly inhibited. Conclusions : The results of this study provide fundamental information about A. planci spine venom. The hemolytic activity was affected by pH, temperature, metal ions, EDTA, cholesterin, proteases, and glycoside hydrolases. ASV hemolysis was inhibited by Cu2+, cholesterin, -chymotrypsin, and cellulose, factors that might prevent the hemolytic activity of venom and provide the medical treatment for sting.