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.

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.

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.

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.

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.

Blue mussel (Mytilus edulis) water extract ameliorates intestinal immune response in high-fat diet-streptozotocin-induced diabetic mice.

Diabetes mellitus is a metabolic disease characterized by high blood glucose levels or hyperglycemia. Diabetes causes a decrease in immune function in the human body. Mytilus edulis has been identified as having anti-inflammatory properties and the ability to improve inflammation. Thus, this study aimed to investigate the function of Matsu M. edulis water extract (MWE) in mediating the regulation of immune responses and dysregulating the intestinal immune system in hyperglycemia mouse models. The mice were treated with MWE for seven weeks. The results showed that treatment with MWE has the ability to decrease triglyceride and total cholesterol concentrations. MWE also increases the interleukin (IL)-10 concentration and natural killer cell activation. It also improves the phagocytic capacity of monocytes in the colon and the proliferative capacity of lymphocytes in the mesentery. Furthermore, MWE also regulates the IL-6 concentration and the ratio of T helper 17 cells to regulatory T cells. Collectively, this extract can improve dyslipidemia, inflammatory responses, and dysregulation of the intestinal immune system. Therefore, M. edulis water extract can be used as an alternative treatment to reduce diabetes complications.

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.

One-step simultaneous differential scanning calorimetry-FTIR microspectroscopy to quickly detect continuous pathways in the solid-state glucose/asparagine Maillard reaction.

The stepwise reaction pathway of the solid-state Maillard reaction between glucose (Glc) and asparagine (Asn) was investigated using simultaneous differential scanning calorimetry (DSC)-FTIR microspectroscopy. The color change and FTIR spectra of Glc-Asn physical mixtures (molar ratio = 1:1) preheated to different temperatures followed by cooling were also examined. The successive reaction products such as Schiff base intermediate, Amadori product, and decarboxylated Amadori product in the solid-state Glc-Asn Maillard reaction were first simultaneously evidenced by this unique DSC-FTIR microspectroscopy. The color changed from white to yellow-brown to dark brown, and appearance of new IR peaks confirmed the formation of Maillard reaction products. The present study clearly indicates that this unique DSC-FTIR technique not only accelerates but also detects precursors and products of the Maillard reaction in real time.

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.

Species identification and vitamin A level in lutjanid fish implicated in vitamin A poisoning.

One outbreak of food poisoning associated with ingestion of the liver of a large lutjanid fish was investigated in this study. The symptoms in three patients primarily included headache, nausea, vomiting, fever, vertigo, and visual disorientation and later included peeling of the skin. The species of fish implicated in this incident was Etelis carbunculus (family Lutjanidae) as determined by direct sequence analysis and PCR plus restriction fragment length polymorphism analysis for detection of the cytochrome b gene. Subsequently, several specimens of E. carbunculus of different body weights were collected, and the level of vitamin A in the muscle and liver was determined by high-performance liquid chromatography. The average level of vitamin A in E. carbunculus muscle was 12 +/- 2 IU/g and that in the liver was 9,844 +/- 7,812 IU/g. Regression models indicate that E. carbunculus with higher body weight and liver weight will have higher levels of vitamin A levels in the liver.

Species identification of ciguatoxin-carrying grouper implicated in food poisoning.

Food poisoning due to ingestion of an unknown red grouper occurred in southern Taiwan. To identify the species of toxic red grouper implicated in food poisoning, a 475-bp sequence of the cytochrome b gene from six species of fresh red grouper meat was amplified by using a pair of primers (L14735/H15149). This fragment could be amplified when fish meat was treated with different heating processes. After sequencing, it was found that no variation in sequences was detected among individuals within each species. The species of toxic red grouper meat implicated in food poisoning was judged to be Lutjanus bohar based on sequence analysis. In addition, restriction enzyme analysis with HaeIII rapidly distinguished these six species of red grouper and the two samples implicated in food poisoning. No toxicity of viscera in 18 specimens of six red grouper species was detected, but two food poisoning samples were found to be toxic. This study indicated that DNA sequence and restriction enzyme analysis are powerful methods for identifying potentially toxic red grouper species as L. bohar.

Tetrodotoxin poisoning evidenced by solid-phase extraction combining with liquid chromatography-tandem mass spectrometry.

The toxicity and toxin component of gastropod Niotha clathrata implicated to a food paralytic poisoning incident in Kaohsiung, Taiwan in November 2006 were studied. The highest scores of average toxicity in the digestive gland and other portions from collected gastropods were 62+/-24 (mean+/-S.D.) and 32+/-16 microg/g according to tetrodotoxin (TTX) bioassay, respectively. The toxin from these gastropods was large amount and easily identified as tetrodotoxin by traditional method of HPLC-FLD. The toxin of patient's blood serum was trace amount and analyzed by a new developed method LC-MS/MS. LC-MS/MS was contracted by the LC system interfaced with the MS/MS system with a turbo ion spray interface. Positive ion detection and multiple reaction monitoring mode were used for TTX of patient serum. It was found that linearity in serum was observed within concentration ranged of 1-100 ng/ml and limit of detection was 0.1 ng/ml. The LOQ was reproducible at 1 ng/ml in serum. The blood serum showed to contain TTX of 3.30+/-0.08 ng/ml. It indicated that LC-MS/MS was more lower detectable and believable method for TTX determination than LC-MS reported previously. Furthermore, the causative agent of gastropod food poisoning was identified as TTX.

Short-term toxicity of aristolochic acid, aristolochic acid-I and aristolochic acid-II in rats.

We compared the short-term toxicity of toxic components of aristolochic acid in rats. Twenty-four female Wistar rats were divided into 4 groups and treated orally every 3-days with 10 mg/kg each of aristolochic acid, aristolochic acid-I and aristolochic acid-II for 19 days. After treatment, the relative ratio of liver and kidney weight to body weight, the concentrations of RBC, hemoglobin and hematocrit in the blood, the levels of aspartate amino transferase, alanine amino transferase, alkaline phosphatase, blood urea nitrogen and creatinine in the plasma, and the levels of urinary urea nitrogen and creatinine in the urine were significantly increased. Body weight of rats and the levels of Na(+), K(+), Ca(2+) in the urine were significantly decreased, especially for groups treated with aristolochic acid and aristolochic acid-II. Pathological examination of liver and kidney also showed cell enlargement and lesions, especially for groups treated with aristolochic acid and aristolochic acid-II. The aristolochic acid exhibited significant toxicity, and the short-term toxicity of aristolochic acid-II and aristolochic acid was similar to each other. Renal but not hepatic failure induced by aristolochic acid could be prevented by pentoxifylline.

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.

Effect of vitamin E on lipid parameters in ovariectomized rats.

The risk of cardiovascular disease drastically increases at the onset of menopause, in part, because of rise in blood cholesterol and unfavorable changes in lipid profile. This study was designed to investigate the dose-dependent effects of vitamin E supplementation on lipid parameters in ovariectomized (ovx) rats. Sixty 12-month-old female Sprague-Dawley rats were either sham-operated (sham; one group) or ovx (four groups). All rats were maintained on a semipurified caseinbased diet (AIN-93M; 75 IU vitamin E/kg of diet) for a period of 120 days. Thereafter, ovx rats were placed on one of four doses of vitamin E treatment (75, 300, 525, or 750 IU vitamin E/kg of diet), while the sham group was continued on 75 IU vitamin E/kg of diet for 100 days. Ovariectomy tended to increase (by 24%, P = 0.1) serum non?high-density lipoprotein (HDL) cholesterol and decrease (by 14%, P = 0.1) HDL cholesterol. Vitamin E did not have any significant effects on serum lipid parameters. Liver total lipids were notably increased (P < .001) in ovx animals, and supplementation with vitamin E at 525 IU/kg of diet was able to significantly reduce liver total lipids by 13%. Additionally, ovariectomy caused an increase in serum glucose and liver C18:1 fatty acid concentrations along with decreases in C18:0, C20:4, and C22:6 fatty acid concentrations. These alterations on liver fatty acid profiles were unaffected by vitamin E. The findings of this study suggest that vitamin E supplementation moderately improves lipid parameters in ovarian hormone-deficient rats.

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.

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.

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.