[Analysis of 14 cases of melanosis caused by 1, 8-dinitronaphthalene and 1, 8-diaminonaphthalene].

14 workers in the 1, 8-diaminonaphthalene workshop of a chemical company in Nantong City had symptoms or signs of varying degrees of pruritus and pigmentation of the face, neck and waist. Pathological examination of skin biopsies showed hyperkeratosis, the basal cells were liquefied and denatured. Seven workers were eventually diagnosed with occupational melanosis. To explore the causes of occupational melanosis caused by exposure to 1, 8-dinitronaphthalene and 1, 8-diaminonaphthalene, and to provide reference for the prevention and treatment of occupational melanosis in the future, this paper reported 14 cases of melanosis in the skin of workers in chemical industry.

Development of antioxidant gliadin particle stabilized Pickering high internal phase emulsions (HIPEs) as oral delivery systems and the in vitro digestion fate.

In this paper, we demonstrate for the first time the use of gliadin particles to structure algal oil (rich in DHA) and to exert chemical stability against lipid oxidation via the Pickering high internal phase emulsion (HIPE) strategy. The gliadin/chitosan colloid particles (GCCPs) were effectively adsorbed and anchored at the algal oil-water interface. Concomitantly, the particle-coated droplets as building blocks constructed a percolating 3D-network framework, endowing Pickering HIPEs with viscoelastic and self-supporting attributes. In addition, Pickering HIPEs loaded with shell (HIP-curEs) or core curcumin (HIPEs-cur) were constructed to depress the oxidation of algal oil. The content of primary (lipid hydroperoxides) and secondary (malondialdehyde and hexanal) oxidation products in HIPEs was lower than that in bulk oil. The oxidative stability of HIPEs was further improved in shell and core curcumin. An in vitro gastrointestinal (GI) model was constructed to characterize the lipid digestion, lipid oxidation as well as curcumin bioaccessibility of the ingested Pickering HIPEs. Lipid oxidation in the Pickering HIPEs was retarded under GI fluids, especially in the presence of core curcumin. The free fatty acid (FFA) fraction released was below 30% for all HIPEs, reflecting that the Pickering HIPEs formed restrict the digestion of fat or oil and potentially help to fight obesity. Interestingly, this route enhanced the bioaccessibility of curcumin from only 2.13% (bulk algal oil) to 53.61% (core curcumin); in particular, it reached 76.82% for shell curcumin. These results help to fill the gap between the physicochemical performance of the gliadin particle stabilized Pickering HIPEs and their potential applications as oral delivery systems of nutraceuticals. This work opens concomitantly an attractive strategy to convert liquid oils into antioxidant soft solids without artificial trans fats, as a potential alternative for PHOs.

Characterization and development of 56 EST-SSR markers derived from the transcriptome of Odontobutis potamophila.

Expressed sequence tags (ETSs) are the sources of microsatellite development. In this study, we isolated and characterized microsatellite markers for Odontobutis potamophila by using Illumina RNA-sequencing. We sequenced a large number of ESTs and screened 200 potential microsatellites. Consequently, a total of 56 novel polymorphic microsatellite repeat markers were identified in thirty-two individuals from a wild population area (Jiande, Zhejiang Province, China). The number of alleles per locus varied from two to eight, the observed heterozygosity (HO) ranged from 0.03571 to 0.9375, and the expected heterozygosity (HE) ranged from 0.14326 to 0.81549. The average number of alleles, HO, and HE were 5.0, 0.4467, and 0.5518, respectively. By the calculation, the range of polymorphism information content (PIC) was 0.1177-0.8492. Most of the loci showed moderate or high polymorphism. These newly developed EST-simple sequence repeat (EST-SSR) markers would serve as an efficient tool for analyzing population connectivity and provide sufficient information for genetic diversity research, parentage, and molecular breeding of O. potamophila and other fishes with similar genetic relationship.

Development of stable Pickering emulsions/oil powders and Pickering HIPEs stabilized by gliadin/chitosan complex particles.

In this paper, we demonstrate the use of gliadin/chitosan complex particles (GCCPs) as particulate stabilizers of oil-in-water emulsions of natural oils and water. For this purpose, we fabricated GCCPs through a facile anti-solvent procedure and demonstrated their usage in the formation of Pickering emulsions and Pickering high internal phase emulsions (HIPEs). The GCCPs can be used to produce surfactant-free o/w Pickering emulsions and Pickering HIPEs; unfortunately these emulsions were labile to coalescence. NaCl addition and/or pH regulation, and the combination were used to modify the surface wettability of the complex particles to achieve stable emulsions. The microstructures, e.g., interfacial frameworks, GCCP partition between the continuous phase and interfacial region, and the state of the droplets, of Pickering emulsions were visualized by confocal laser scanning microscopy (CLSM), confirming that the inclusion of NaCl and slightly adjusting pH toward 4.0 and/or 5.0 benefited the adsorption and accumulation of colloid particles at the droplet surface to form an engineered interfacial structure, bridging droplets together through a percolating layer of colloidal particles at the oil/water interface. A schematic representation for the formation route of the emulsions is proposed to relate the physical performance and rheological property with the interfacial structures and aggregate behaviors in the Pickering system stabilized by the complex particles. Interestingly, direct freeze-drying of the emulsions transformed unstable Pickering emulsions into stable oil powders. This study opens a promising route based on Pickering HIPEs or oil powders to structure liquid oils into solid-like fats without artificial trans-fat, which outlines new directions for future fundamental research.

Phylogenetic relationships of twenty Gymnothorax species based on cytochrome b sequence data.

To study the phylogenetic relationships of the genus Gymnothorax (moray eels) distributed in South China Sea, polymerase chain reactions were performed, and the amplification products were sequenced by cloning into the PMD18T-vector (TaKaRa). The entire gene sequences encoding cytochrome b (1140 bp) for 16 Gymnothorax (G. flavimarginatus, G. meleagris, G. undulates, G. reticularis, G. reevesi, G. melanospilus, G. rueppeliae, G. javanicus, G. chilospilus, G. pseudothyrsoideus, G. fimbriatus, G. hepaticus, G. berndti, G. curostus, G. favagineus, and G. margaritophorus) were obtained. Four additional Gymnothorax sequences from GenBank were also included. The nucleotide composition, genetic distances, and base substitution saturation analysis were calculated using the MEGA 5.0 Software. Phylogenetic analysis was performed using maximum-parsimony, maximum-likelihood (ML), and neighbor-joining (NJ). The results were as follows: 1) base-substitution saturation analysis suggested that both in third codon positions, and the full-length cytochrome b data set, Ts are not saturated, but Tv substitutions may be saturated, 2) the genus Gymnothorax, native to the South China Sea, is divided into four distinct clades, with two clades in the NJ and ML trees, and 3) according to our experimental data, G. melanospilus (Bleeker, 1855) and G. favagineus (Bloch and Schneider, 1801) are the same species.

Molecular cloning, characterization, and expression of vacuolar-type-H⁺-ATPase B1 (VHAB1) gene in the gill of Anguilla marmorata.

We explored the molecular mechanism of the regulation of vacuolar-type-H+-ATPase B1 (VHAB1) in elvers in the response to salinity. The full-length cDNA of VHAB1 in Anguilla marmorata (designated as AmVHAB1), which was 1741 base pairs (bp) in length, was found to encompass a 1512-bp open reading frame encoding a polypeptide with 503 amino acids (55.9 kDa), an 83-bp 5'-untranslated region, and a 146-bp 3'-untranslated region. The mRNA and protein expression levels of AmVHAB1 in the gill were evaluated at different time points (0, 1, 3, 6, 12, 24, 48, 72, and 96 h, and 15 days) during the exposure to various salinity levels (0, 10, and 25‰). The results indicated that the expression levels of AmVHAB1 mRNA in the gill significantly increased and reached the highest level at 1 h exposure in the brackish water (BW, 10‰) group and at 6 h exposure in the seawater (SW, 25‰) group. The salinity level affected the relative expression level of AmVHAB1 mRNA in the gill, which was increased by approximately 44-fold in the SW group when compared with that in fresh water. Immunoblotting analysis showed that VHA expression was significantly higher in the BW and SW groups, with the highest expression level was detected at 96 h exposure. We found that the AmVHAB1 gene in elvers from A. marmorata plays an important role in the adaptation to seawater.

Development and validation of single nucleotide polymorphism markers in Odontobutis potamophila from transcriptomic sequencing.

Transcriptome sequencing technology has been applied in the development and discovery of single nucleotide polymorphism (SNP) markers in fish. In this study, a panel of 120 expressed sequence tag (EST)-derived SNPs was selected by several selection filters from the resultant EST library of Odontobutis potamophila using Illumina Sequencing. In total, 37 SNPs from 120 putative SNPs were considered as the true SNPs using Sanger sequencing. For each SNP locus of 30 individuals of one wild population of O. potamophila that was successfully calculated, the number of alleles per locus was 2 with an observed heterozygosity of 0.0000-0.9000 and an expected heterozygosity of 0.1000-0.5263. A total of 33 loci conformed to Hardy-Weinberg equilibrium (HWE), and 4 loci deviated from HWE after Bonferroni correction. These 33 SNP markers will benefit the studies of population genetic structure, population evolution analysis, and construction of a high-density linkage map of O. potamophila.

Neutrophil extracellular trap formation is associated with autophagy-related signalling in ANCA-associated vasculitis.

Increasing evidence indicates that aberrant neutrophil extracellular trap (NET) formation could contribute to the pathogenesis of anti-neutrophil cytoplasmic antibody (ANCA)-associated vasculitis (AAV). Recent research has provided evidence that a novel type of ANCA autoantibody, anti-lysosomal membrane protein-2 (LAMP-2) antibody, may have a pathogenic role in AAV. We have shown previously that anti-LAMP-2 antibody-stimulated NET formation contains autoantigens and anti-microbial peptides. The current study sought to determine whether LAMP-2, as a novel antigen of ANCA, was present on NETs in AAV patients, the influence of the anti-LAMP-2 antibody on the neutrophil apoptosis rate and the role of autophagy in anti-LAMP-2 antibody-induced NET formation. NET formation was assessed using immunofluorescence microscopy, scanning electron microscopy or live cell imaging. The neutrophil apoptosis rate was analysed using fluorescence activated cell sorting (FACS). Autophagy was detected using LC3B accumulation and transmission electron microscopy. The results showed that enhanced NET formation, which contains LAMP-2, was observed in kidney biopsies and neutrophils from AAV patients. The apoptosis rate decreased significantly in human neutrophils stimulated with anti-LAMP-2 antibody, and this effect was attenuated by the inhibitors of autophagy 3-methyladenine (3MA) and 2-morpholin-4-yl-8-phenylchromen-4-one (LY294002). The anti-LAMP-2 antibody-stimulated NET formation was unaffected by benzyloxycarbonyl-Val- Ala-Asp (OMe)-fluoromethylketone (zVAD-fmk) and necrostatin-1 (Nec-1), which are inhibitors of apoptosis and necrosis, respectively, but was inhibited by 3MA and LY294002. Moreover, the proportion of LC3BI that was converted to LC3BII increased significantly (P=0.0057), and massive vacuolizations that exhibited characteristics typical of autophagy were detected in neutrophils stimulated with anti-LAMP-2 antibody. Our results provide further evidence that autophagy is involved in ANCA-induced NET formation in human neutrophils.

Development and characterization of microsatellite loci in a threatened marine fish, Cheilinus undulatus (humphead wrasse).

Cheilinus undulatus (humphead wrasse) is a marine fish distributed widely throughout the tropical Indo-Pacific. It has been listed as vulnerable in the IUCN Red Data Book and in CITES Appendix II four times. Fifteen microsatellite loci were isolated and characterized for this species. The number of alleles ranged from 3 to 15 per locus, and the observed and expected heterozygosity ranged between 0.0323-0.7742 and 0.2597-0.8773, respectively. The polymorphism information content ranged from 0.2353-0.8520. Four microsatellite loci deviated significantly from Hardy-Weinberg expectations. No significant linkage disequilibrium was found among any of the loci. These microsatellite loci will be useful for future investigations of genetic variation in the wrasse population.

Complete mitochondrial genome sequence of the humphead wrasse, Cheilinus undulatus.

The humphead wrasse (Cheilinus undulatus) is a large coral fish that has become threatened due to habitat loss and fishing pressure. We sequenced the mitochondrial genome of C. undulatus, using a normal PCR method. The complete mtDNA sequence encoded 13 protein genes, 22 tRNA genes and 2 rRNA genes. It was found to be 16,613 bp in length and had an overall H-strand base compositions of 27.3 for A, 30.9 for C, 16.8 for G, and 25.0% for T. Compared with the sequences of 8 other members of the family Labridae, gene content, genome organization, and nucleotide compositions were similar. All tRNAs formed a typical clover-leaf structure, except tRNA(Ser) (AGY), and most of the size variations among tRNAs stemmed from variations of length in the arm and loop of TΨ, and loop of DHU.

Isolation and characterization of 16 microsatellite loci in marble goby (Oxyeleotris marmoratus).

A microsatellite-enriched genomic library for marble goby (Oxyeleotris marmoratus), a freshwater fish of considerable economic value, was obtained. A total of 16 microsatellite markers were successfully isolated and characterized in a population from the Mekong River in Vietnam. These markers had 2-20 alleles with expected heterozygosity ranging from 0.0370 to 0.8927. Linkage equilibrium was observed in most loci, and only 1 locus revealed a significant deviation from Hardy-Weinberg equilibrium. These microsatellite markers will be useful for genetic diversity and molecular marker-assisted selection studies of wild and farmed O. marmoratus.