Relationship between intracellular Ca²âº and ROS during fluoride-induced injury in SH-SY5Y cells.

The mechanisms underlying the neurotoxicology of endemic fluorosis still remain obscure. To explore lactate dehydrogenase (LDH) leakage, intracellular Ca²âº concentration ([Ca²âº]i ) and reactive oxygen species (ROS) production induced by fluoride, human neuroblastoma (SH-SY5Y) cells were incubated with sodium fluoride (NaF, 20, 40, 80 mg/L) for 24 h, with 40 mg/L NaF for 3, 6, 12, 18, 24 h, and N-acetyl-L-cysteine (NAC), ethyleneglycol-bis-(ß-aminoethyl ether)-N,N,N',N'-tetraacetic acid (EGTA), 1,2-bis(O-aminophenoxy)ethane-N,N,N',N'-tetraacetic acid tetra(acetoxymethyl) ester (BAPTA-AM) alone or combined with fluoride (40 mg/L) respectively for 12 h in vitro. The results showed that the LDH levels in the 40 and 80 mg/L fluoride-treated groups were significantly higher than that of the control group (in the test level of 0.05, the difference were statistical significance). [Ca²âº]i and ROS reached a peak at 3 h and 12 h respectively after exposure to 40 mg/L fluoride. Fluoride coincubated with NAC (antioxidant) dramatically decreased ROS and LDH levels compared with the fluoride only group (in the test level of 0.05, the difference were statistical significance). However, fluoride-induced increase in [Ca²âº]i was not affected by NAC. BAPTA-AM (intracellular calcium chelator) markedly lowered fluoride-induced increase of [Ca²âº]i , ROS and LDH levels while EGTA (extracellular calcium chelator) have no effects on them. These results indicate that fluoride-related Ca²âº release from the site of intracellular calcium storage causes the elevation of ROS contributing to the cytotoxicity in SH-SY5Y cells.

Impacts of conventional and biodegradable microplastics on juvenile Lates calcarifer: Bioaccumulation, antioxidant response, microbiome, and proteome alteration.

Discarded plastic bag is a main component of marine debris, posing potential threats to marine biota. This study was conducted to assess the potential effects of microplastics on juvenile Lates calcarifer. Fish were exposed via diet to two microplastic types from conventional polyethylene (PE) and biodegradable (Bio) plastic bags for 21 days. Antioxidative enzymes activity, intestinal microbiome and proteome were determined. PE and Bio microplastics were found to accumulate in gastrointestinal tracts, and no mortality was observed. Microplastics exposure did not induce significant antioxidant response except for the glutathione reductase (GR) modulation. Intestinal microbiome diversity decreased significantly in PE group based on Simpson index. Both types of microplastics induced proteome modulation by down-regulating proteins associated with immune homeostasis. Bio microplastics maintained higher intestinal microbial diversity and induced more proteins alteration than PE microplastics. This study provides toxicological insights into the impacts of conventional and biodegradable microplastics on juvenile L. calcarifer.

Effects of the Fas/Fas-L pathway on fluoride-induced apoptosis in SH-SY5Y cells.

The mechanisms underlying fluoride-induced apoptosis in neurons still remain unknown. To investigate apoptosis, caspase-3 activity, and mRNA expression of Fas, Fas-L, and caspases (-3 and -8) induced by fluoride, human neuroblastoma (SH-SY5Y) cells were incubated with 0, 20, 40, and 80 mg/L sodium fluoride (NaF) for 24 h in vitro. The data show that cell viability in the 40 and 80 mg/L fluoride groups were significantly lower than that of the control group. The percentages of apoptosis in the 40 and 80 mg/L fluoride groups were markedly higher than those in the control group, and they increased with the increase in fluoride concentration. The activity of caspase-3 and mRNA expression levels for Fas, Fas-L, and caspases (-3 and -8) in the 40 and 80 mg/L fluoride groups were significantly higher than those in the control group. An agonistic anti-Fas monoclonal antibody (CH-11) significantly augmented apoptosis induction by fluoride, showing a synergistic effect, while a Fas-blocking antibody (ZB4) partly inhibited fluoride-induced apoptosis of SH-SY5Y cells. The results indicate that fluoride exposure could induce apoptosis in SH-SY5Y cells, and the Fas/Fas-L signaling pathway may play an important role in the process.

Microplastic accumulation in fish from Zhanjiang mangrove wetland, South China.

Microplastics (MPs) are widespread in marine and estuarine environments, but the contamination of MPs in mangrove wetlands is relatively unknown. Here, we quantify the presence of MPs in fish collected from Zhanjiang mangrove wetland, the largest mangrove in South China, which provide baseline data on MPs accumulation in fish in mangrove environment as the first evidence in China. MPs were found in 30 out of 32 fish species at an average abundance of 2.83 ±â€¯1.84 items individual-1, ranged from 0.6 to 8.0 items individual-1 in each species. MPs were detected in gills, stomach and intestine, and not found in muscles and livers. Positive relationship was found between MPs abundance and body length or weight of mangrove fish. The dominant polymers identified by micro-FTIR were polyethylene, polyethylene terephthalate, polypropylene and cellophane. MPs consisted primarily of fibers and with the prominent size range of 0.02-1 mm. The body sizes, living habitats and feeding habits of fish are important factors affecting MPs accumulation in different fish species. This study revealed the wide presences of MPs in fish species within a mangrove wetland.

Low level of microplastic contamination in wild fish from an urban estuary.

Microplastic accumulation in estuarine environments is considered the dominant input of land-based plastics into the oceans. In this study, the level of microplastic contamination was evaluated in 26 species of wild fish from the Pearl River Estuary, South China. Results showed that microplastics abundance ranged from 0.17 items individual-1 (Boleophthalmus pectinirostris & Acanthogobius flavimanus) to 1.33 items individual-1 (Plectorhynchus cinctus) among different species. The distribution of microplastic abundance in the gills and gastrointestinal tracts was not significantly different. Microplastics in gills are strongly related to the filtration area of gills in 15 fish species. Fibers were the dominant shapes accounting for 93.45% of the total shapes. The majority of microplastics were <3 mm in size. The most common polymer composition was polyethylene terephthalate (38.2%) and the most common color was black (30.36%). The findings of this study provide baseline data for microplastic contamination in wild fish from an urban estuary.

Effects of fluoride on the expression of NCAM, oxidative stress, and apoptosis in primary cultured hippocampal neurons.

The mechanisms underlying the neurotoxicity of endemic fluorosis still remain unknown. To investigate the expression level of neural cell adhesion molecules (NCAM), oxidative stress, and apoptosis induced by fluoride, the primary rat hippocampal neurons were incubated with 20, 40, and 80 mg/l sodium fluoride for 24 h in vitro. The results showed that the cell survival rate in the 80 mg/l fluoride-treated group was significantly lower than that of the control group. Forty and 80 mg/l of fluoride induced significantly increased lactate dehydrogenase release, intracellular reactive oxygen species, and the percentage of apoptosis. Compared with control group, the malondialdehyde levels were significantly elevated while glutathione levels and glutathione peroxidase activities were decreased in all fluoride-treated groups, accompanied by the markedly reduced superoxide dismutase activity in 80 mg/l fluoride-treated group. With respect to NCAM mRNA expression levels, a significant dose-dependent decrease was observed in 40 and 80 mg/l fluoride-treated groups against the control group. In addition, as compared to the control group, the protein expression levels of NCAM-180 in 40 and 80 mg/l fluoride-treated groups, NCAM-140 in all fluoride-treated groups, and NCAM-120 in the 80 mg/l fluoride-treated group were significantly decreased. Our study herein suggested that fluoride could cause oxidative stress, apoptosis, and decreased mRNA and protein expression levels of NCAM in rat hippocampal neurons, contributing to the neurotoxicity induced by fluoride.

[The dose-effect relationship of water fluoride levels and renal damage in children].

OBJECTIVE: To explore the dose-effect relationship of water fluoride levels and renal damage in children and observe the difference of renal function between high-loaded fluoride people and dental fluorosis people in the same water fluoride level region. METHODS: 210 children were divided into seven groups in term of drinking water fluoride levels and whether they suffered from dental fluorosis. Fluoride concentrations in urine and serum and activities of urine NAG and gamma-GT were determined. RESULTS: The urine and serum fluoride of high-loaded fluoride people and dental fluorosis people increased compared with control, moreover fluoride contents in urine and serum increased gradually with the increase of fluoride level in drinking water. Urine NAG and gamma-GT activities significantly increased in dental fluorosis people from area of 2.58 mg/L fluoride in drinking water and in those two groups from area of 4.51 mg/L fluoride in drinking water. Moreover, there existed an obvious dose-effect relationship between the drinking water fluoride concentration and NAG and gamma-GT activity. CONCLUSION: Over 2.0 mg/L fluoride in drinking water can cause renal damage in children, and the damage degree increases with the drinking water fluoride content. Renal damage degree is not related to whether the children suffered from dental fluorosis and mainly due to water fluoride concentration.

Mameliella alba gen. nov., sp. nov., a marine bacterium of the Roseobacter clade in the order Rhodobacterales.

A Gram-negative, non-motile, rod-shaped bacterial strain, JLT354-W(T), that accumulates poly-beta-hydroxybutyrate granules was isolated from the South China Sea. Phylogenetic analysis based on 16S rRNA gene sequences showed that the strain was related to members of the genera Antarctobacter, Sagittula, Oceanicola and Loktanella; levels of similarity between strain JLT354-W(T) and members of the above genera were less than 92.0 %. The predominant fatty acid of strain JLT354-W(T) was C(18 : 1)omega7c (83.1 %); significant amounts of C(18 : 0) (7.9 %) and C(12 : 1) 3-OH (3.7 %) were also present. The predominant respiratory ubiquinone was Q-10. The DNA G+C content of strain JLT354-W(T) was 63.7 mol%. The isolate was distinguishable from members of the order Rhodobacterales based on phenotypic and biochemical characteristics. On the basis of the taxonomic data presented, strain JLT354-W(T) is considered to represent a novel species of a new genus, for which the name Mameliella alba gen. nov., sp. nov. is proposed. The type strain of Mameliella alba is JLT354-W(T) (=LMG 24665(T)=CGMCC 1.7290(T)).

Dose-effect relationship between drinking water fluoride levels and damage to liver and kidney functions in children.

Although a dose-effect relationship between water fluoride levels and damage to liver and kidney functions in animals has been reported, it was not demonstrated in humans. To evaluate the effects of drinking water fluoride levels on the liver and kidney functions in children with and without dental fluorosis, we identified 210 children who were divided into seven groups with 30 each based on different drinking water fluoride levels in the same residential area. We found that the fluoride levels in serum and urine of these children increased as the levels of drinking water fluoride increased. There were no significant differences in the levels of total protein (TP), albumin (ALB), aspartate transamine (AST), and alanine transamine (ALT) in serum among these groups. However, the activities of serum lactic dehydrogenase (LDH), urine N-acetyl-beta-glucosaminidase (NAG), and urine gamma-glutamyl transpeptidase (gamma-GT) in children with dental fluorosis and having water fluoride of 2.15-2.96 mg/L and in children having water fluoride of 3.15-5.69 mg/L regardless of dental fluorosis were significantly higher than children exposed to water fluoride of 0.61-0.87 mg/L in a dose-response manner. In contrast to children with dental fluorosis and having water fluoride of 2.15-2.96 and 3.10-5.69 mg/L, serum LDH activity of children without dental fluorosis but exposed to the same levels of water fluoride as those with dental fluorosis were also markedly lower, but the activities of NAG and gamma-GT in their urine were not. Therefore, our results suggest that drinking water fluoride levels over 2.0mg/L can cause damage to liver and kidney functions in children and that the dental fluorosis was independent of damage to the liver but not the kidney. Further studies on the mechanisms and significance underlying damage to the liver without dental fluorosis in the exposed children are warranted.