Dietary effects of vitamin C on antioxidant capacity, intestinal microbiota and the resistance of pathogenic bacteria in cultured Silver pomfret (Pampus argenteus).

As most teleosts are unable to synthesize vitamin C, supplemental diets containing vitamin C diets play a crucial role in fish health. The aim of this study was to investigate the effect of dietary vitamin C on the intestinal enzyme activity and intestinal microbiota of silver pomfre (Pampus argenteus). Four experimental diets were supplemented with basic diets containing 300 mg of vitamin C/kg (group tjl3), 600 mg of vitamin C/kg (group tjl6), and 1200 mg of vitamin C/kg (group tjl12), as well as vitamin C-free supplemental basic diet (group tjl0), respectively. The four diets were fed to juvenile P. argenteus (average initial weight: 4.68 ± 0.93 g) for 6 weeks. The results showed that the activity of SOD (superoxide dismutase) and CAT (catalase) increased significantly while that of MDA (malondialdehyde) decreased significantly in group tjl3 compared to vitamin group tjl0. At the genus level, groups tjl0, tjl6, and tjl12 contained the same dominant microbial community, Stenotrophomonas, Photobacterium, and Vibrio, whereas group tjl3 was dominated by Stenotrophomonas, Delftia, and Bacteroides. Among the fish fed with a basic diet containing 300 mg of vitamin C/kg, the intestines exhibited a notable abundance of probiotic bacteria, including lactic acid bacteria (Lactobacillus) and Bacillus. The abundance of Aeromonas in groups tjl3 and tjl6 was lower than that of the vitamin C-free supplemental basic diet group, whereas Aeromonas was not detected in group tjl12. In addition, a causative agent of the disease outbreak in cultured P. argenteus, Photobacterium damselae subsp. Damselae (PDD) was the dominant microbiota community in groups tjl0, tjl6 and tjl12, whereas the abundance of PDD in group tjl3 was the lowest among the diets. Taken together, the diets supplied with vitamin C could influence the composition microbial community of P. argenteus. The low level of vitamin C (300 mg of vitamin C/kg per basic diet) supplementation could not only improve the antioxidant capacity but also resist the invasion of pathogenic bacteria.

Effects of different conditions on the artificial incubation effect and physiological indexes of redclaw crayfish eggs.

We explored the effects of different conditions on the artificial incubation of redclaw crayfish eggs in an effort to improve this process. Samples at the egg and juvenile stages were selected. The samples at different stages were separated from the pleopods, then they were placed in incubator boxes and sterilized with different disinfectant solutions. The density was 300,400 and 500 eggs/incubator box, the vibration frequency was 11,16 and 26 vibrations/min, and the water circulation cycle was 2.1, 4.8 and 7.1 cycles/h. The results showed the eggs disinfected with 3000 ppm formaldehyde for 15 min had stronger antioxidant capacity. The hatching and survival rates of five pairs of appendage stage group were significantly lower than those of other groups. In the egg stage, acid phosphatase (ACP) level of compound eye pigmentation stage group was significantly higher than those of other groups. In the juvenile stage, malondialdehyde (MDA) content of five pairs of appendage stage group was significantly higher than those of other groups. The survival rate of 500 eggs/box group was significantly higher than that of other groups. In the egg stage, alkaline phosphatase (AKP) level of 400 eggs/box group was significantly higher than that of other groups. The survival rate of 11 vibrations/min group was significantly higher than that of other groups. In the egg stage, ACP and AKP levels of 11 vibrations/min group were significantly higher than those of 26 vibrations/min group. In the juvenile stage, superoxide dismutase (SOD), ACP and AKP levels of 11 vibrations/min group was significantly higher than those of 26 vibrations/min group. In the juvenile stage, AKP level of 4.8 cycles/h group was significantly lower than that of other groups. In conclusion, egg development at the stage after seven pairs of appendages, with a density of 400 eggs/box, vibration frequencies set at 11 vibrations/min achieved high hatching rates (93.58 %) and survival rates (75.67 %). Moreover, bronopol or hydrogen peroxide might have a better choice to replace formaldehyde if further exploration was conducted to reduce stimulation of the in vitro-grown egg. These conditions could be used on a large scale to optimize the production of redclaw crayfish.

Quantitative Crotonylome Analysis Expands the Roles of p300 in the Regulation of Lysine Crotonylation Pathway.

Lysine crotonylation (Kcr) is a recently identified post-translational modification (PTM) that is regulated by an acetyltransferase, p300. The p300-catalyzed histone Kcr is able to stimulate transcription to a greater degree than the well-studied histone lysine acetylation (Kac). Despite these progresses, the global Kcr substrates regulated by p300 remain largely unknown, hindering efforts to establish mechanistic links between Kcr and p300-mediated phenotypes. Here, a quantitative proteomics study to characterize the p300-regulated lysine crotonylome is reported. A total of 816 unique endogenous crotonylation sites are identified across 392 proteins, with 88 sites from 69 proteins being decreased by more than 0.7-fold (log2 < 0.5) and 31 sites from 17 proteins being increased by more than 1.4-fold (log2 > 0.5) in response to p300 knockout (KO). The most downregulated crotonylome alterations under p300 deficiency concern components of the nonsense-mediated decay, infectious disease, and viral/eukaryotic translation pathways. Moreover, some p300-targeted Kcr substrates are potentially linked to diseases such as cancer. Taken together, this study reveals the lysine crotonylome in response to p300, which sheds light on the role for lysine crotonylation in regulation of diverse cellular processes and provides new insights into mechanisms of p300 functions.

Impact of Admission Glucose on Non-Diabetic Patients with ST-Segment Elevation Myocardial Infarction Treated with Percutaneous Coronary Intervention: A Meta-Analysis.

BACKGROUND: Impaired admission glucose (AG) is thought to significantly increase the risk of both early and late death with ST-segment elevation myocardial infarction (STEMI), especially for non-diabetic patients. However, several earlier studies contradict these relationships. Through our meta-analysis, we aimed to evaluate such a relation between impaired AG, the risk of death and STEMI. METHODS: We accessed PubMed, EMBASE, Web of Science, and the Cochrane Library and systematically searched their databases to identify all related prospective cohort studies. The relative risks (RRs) with their 95% confidence interval (CI) were pooled quantitatively. RESULTS: The pooled, unadjusted relative risks of early outcome events indicated that patients who had glucose concentrations ≥ the range of 6.1-11.1 mmol/L, had a 4.38-fold (95% CI, 3.23-5.94) higher early mortality. For late outcome events, the pooled unadjusted RR indicated patients who had glucose concentrations ≥ the range 7.8-11.1 mmol/L, and had a 2.69-fold (95% CI, 2.16-3.34) higher late mortality based on full participants, whereas patients had a 1.65-fold (95% CI, 1.33-2.04) higher late mortality based on based on in-hospital or 30-day survivors. CONCLUSIONS: In conclusion, the present meta-analysis demonstrated that impaired admission glucose may be an effective prognostic marker for significantly increased risk of early death. Regarding the long-term outcomes based on full population or early survival, high admission glucose also has a distinct but poorer prognostic impact on long-term mortality than early mortality. KEY WORDS: Admission glucose • Meta-analysis • Myocardial infarction • Non-diabetic.

Identification and expression analysis of a doublesex1 gene in Daphnia pulex during different reproductive stages.

The gene doublesex (dsx) has shown deep conservation in the sex determination in many organisms. Environmental stimuli initiate a switch in the reproductive strategy of Daphnia pulex from asexual to sexual reproduction; however, occasionally, changes in environmental conditions will not lead to this transition. So study genetic responses to environmental stimuli and the molecular basis for the switch of reproductive stages are urgently needed. Therefore, we isolated and sequenced a D. pulex doublesex1 gene (Dpdsx1) and analyzed its expression and location by quantitative polymerase chain reaction (qPCR) and whole-mount in situ hybridization in D. pulex during different stages of reproduction. The predicted amino acid sequence has 335 amino acids that contained one DM domain and one dimerization domain, which is characteristic of insect orthologs of Dsx. Real-time PCR showed that Dpdsx1 expression decreased significantly (P < 0.05) in different reproductive stages in the following order: male, parthenogenetic female, ephippial female, resting egg, and juvenile female. Whole-mount in situ hybridization revealed that Dpdsx1 is expressed in the first antennae, first thoracic limb and compound eye in males, whereas expression levels in the corresponding sites of parthenogenetic and ephippial females were relatively weak. Dpdsx1 could not be detected in the gonads of males or ephippial and parthenogenetic females. Taken together, these different reproductive stages' and sex specific expression patterns are regulated temporally and spatially. We speculate that Dpdsx1 may involve in switching different stages of reproduction and in sexual differentiation in D. pulex.

Cloning and expression analysis of a transformer gene in Daphnia pulex during different reproduction stages.

The full-length cDNA of a transformer gene (Dptra) was cloned from the cladoceran Daphnia pulex using RACE. Dptra expression was assessed by qPCR and whole-mount in situ hybridization in different reproductive stages. The Dptra cDNA, 1652bp in length, has a 1158-bp open reading frame that encodes a 385 amino acid polypeptide containing one Sex determination protein N terminal (SDP_N) superfamily, eight putative phosphorylation sites, and an arginine-serine (RS)-rich domain at the N-terminus. Dptra showed 81%, 53%, 51% and 45% identity to orthologous genes in Daphnia magna, Apis mellifera, Apis cerana and Bombus terrestris, respectively. Phylogenetic analysis based on deduced amino acid sequences revealed that Dptra clustered in the hymenopteran clade and was most closely related to D. magna and A. mellifera. qPCR showed that Dptra expression increased significantly (P<0.05) in different reproductive stages in the following order: male, ephippial female, parthenogenetic female, resting egg and juvenile female. Dptra expression is significantly different between males and females and it is significantly greater in ephippial females and males than in parthenogenetic D. pulex (with summer eggs). Whole-mount in situ hybridization revealed that Dptra was expressed at different levels between males and females. In males, hybridization signals were found in the first antennae, second antennae and thoracic limb, whereas expression levels in the corresponding sites of parthenogenetic and ephippial females were relatively weak. This suggests that the Dptra gene plays significant roles in switching modes of reproduction and in sexual differentiation.

Pathological analysis of hemolymphs of Charybdis japonica infected with Vibrio alginolyticus.

We explored the pathogenic mechanism of Vibrio alginolyticus in the stone crab Charybdis japonica by studying the hemolymph of C. japonica artificially infected by V. alginolyticus. To this end, Wright-Geimsa staining and electron microscopy were used, and phenoloxidase (PO) activity and the immune protection rate of C. japonica injected with immune polysaccharide during infection were analyzed. The results indicated that the total hemocyte and hyaline hemocyte (HH) counts in diseased crabs were significantly lower than those in healthy crabs (P < 0.05), whereas the large granule hemocytes (LGHs) were significantly higher in diseased crabs than in healthy crabs (P < 0.05). The cellular sizes of HHs and LGHs showed an increasing trend after V. alginolyticus infection, while the nuclear/cytoplasmic ratio (NP) of these cells showed a sharp decline after V. alginolyticus infection (P < 0.05). Micro-pathological analysis of hemocytes revealed fewer hemocytes in the hemolymph of diseased crabs and the presence of disintegrated cells. Ultrastructural and micro-pathological analyses showed damage in all types of hemocytes. The mitochondria were damaged and incomplete in structure, parts of the nuclear membrane were anamorphic and parts of the nuclei had shrunk, hematocyte nuclei exhibited heterochromatinization, hemocyte granules were increased in the polysaccharide-treated group infected with V. alginolyticus, and the numbers of mitochondria and rough endoplasmic reticulum were also increased. PO activity in the two Vibrio-infected groups peaked at 6 h and 24 h after infection, respectively, and PO activity increased in the hemolymph of infected crabs but sharply decreased with prolonged infection. Finally, the PO activities in the two Vibrio-infected groups were significantly lower than controls at 120 h post-infection (P < 0.05). Interestingly, PO activity was higher in polysaccharide-treated crabs than non-polysaccharide-challenged infected crabs, resulting in an immunoprotective rate of 69.64% at 7 days post-infection. This phenomenon suggests that polysaccharides could enhance the organism's antibacterial defenses by improving immune-related enzyme activity.

Inhibition of glycogen synthase kinase-3 reverses tau hyperphosphorylation induced by Pin1 down-regulation.

One of the neuropathological hallmarks of Alzheimer's disease (AD) is the occurrence of neurofibrillary tangles (NFTs) that are composed of abnormally hyperphosphorylated microtubule-associated protein tau. Abnormal tau hyperphosphorylation is mainly induced due to the imbalance between protein kinases and phosphatases. In the tanglerich subregions of the hippocampus and parietal cortex in the brain of AD patients, the levels of the phosphorylationdependent protein peptidyl-prolyl cis-trans isomerase (Pin1) were found to be low. Although Pin1 can regulate tau phosphorylation, it is not clear whether the inhibition of glycogen synthase kinase 3 (GSK-3), the primary mediator of tau phosphorylation in AD, could reverse tau hyperphosphorylation induced due to the down-regulation of Pin1. We found that while suppression of Pin1, either by using its inhibitor Juglone or a shRNA plasmid against Pin1, induces tau hyperphosphorylation and GSK-3ß activation both in vivo and in vitro, inhibition of GSK-3ß by SB216763 or LiCl reverses tau hyperphosphorylation. Our data suggest that GSK-3ß activation plays an important role in tau hyperphosphorylation induced by the down-regulation of Pin1, and the inhibition of GSK-3ß might be a potential therapeutic approach for AD pathology.

Characterization of Cdc2 kinase in the red claw crayfish (Cherax quadricarinatus): evidence for its role in regulating oogenesis.

Cdc2 kinase is a catalytic subunit of the maturation-promoting factor (MPF), a central factor for inducing the meiotic maturation of oocytes. MPF has been studied in a wide variety of animal species; however, its expression in crustaceans is poorly characterized. In this study, a complete cDNA sequence of Cdc2 kinase was cloned from the red claw crayfish, Cherax quadricarinatus, and its spatiotemporal expression profiles were analyzed. The Cdc2 cDNA (1,769 bp) encodes for a 299 amino acid protein with a calculated molecular weight of 34.7 kDa. Quantitative real-time PCR demonstrated that Cdc2 mRNA was expressed mainly in the ovary tissue and the expression decreased as the ovaries developed. Immunohistochemistry analysis revealed that the Cdc2 protein relocated from the cytoplasm to the nucleus during oogenesis. These findings suggest that Cdc2 kinase may play an important role in the gametogenesis and gonad development in C. quadricarinatus.

[Study on adsorption properties of organic vapor on activated carbons].

Adsorption technology is widely used in oil vapor recovery, and adsorbents have decisive effect on separation. Three kinds of activated carbon (AC) were chosen to study their adsorption properties and adsorption energy, where n-hexane and n-heptane acted as adsorbate and adsorption experiments were conducted at 293.15 K. At the same time, regression formula of Logistic model was used to fit the throughout curves of active carbons. The results showed that: surface area and pore volume of activated carbon were the main factors affecting its adsorption properties; the adsorption behavior of n-hexane and n-heptane were corresponding to Langmuir adsorption isotherm model; adsorption energy of these three kinds of activated carbon became greater with increasing specific surface area. Fitting curve of Logistic model had high similarity with the experimental results, which could be used in the prediction of breakthrough curves of activated carbons.

Polymorphisms of the IL-18 promoter and bronchial asthma.

To study the association of polymorphisms of the IL-18 promoter (-607C/A and -137G/C) with bronchial asthma, 120 subjects with bronchial asthma were selected as the experimental group (49 cases with bronchial asthma and allergic rhinitis as experimental group 1 and 71 cases with bronchial asthma without allergic rhinitis as experimental group 2) and 120 healthy individuals were selected as the control group. A polymerase chain reaction sequence-specific primer (PCR-SSP) was used to identify the genotypic polymorphisms between the experimental and control groups. The results revealed that the frequencies of the CC, CA and AA genotypes at -607C/A in experimental group 1 were 18.37, 40.82 and 40.82%, respectively; these frequencies were 32.39, 54.92 and 12.68%, respectively, in experimental group 2. There was a statistically significant difference in the genotype distribution between experimental groups 1 and 2 (χ²=12.81; P<0.05). The allele frequencies of genotypes C and A were 38.78 and 61.22%, respectively, in experimental group 1 and 59.86 and 40.14%, respectively, in experimental group 2. The allele frequencies at -607C/A were significantly different between experimental groups 1 and 2 (χ²=10.32; P<0.05). Frequencies of genotypes GG, GC and CC at -137C/A in experimental group 1 were 48.98, 46.94 and 4.08%, respectively, and 80.28, 14.08 and 4.08% in experimental group 2 with significant difference (χ²=15.73, P<0.05). Allele frequencies of G and C in experimental group 1 were 72.45 and 27.55%, and 87.32 and 12.68% in experimental group 2 with statistically significant difference (χ²=8.42, P<0.05). In conclusion, polymorphisms of the IL-18 gene promoter at -607C/A and -137G/C were associated with allergic rhinitis in bronchial asthma, indicating that polymorphisms of the IL-18 gene promoter at -607C/A and -137G/C differentially affect the pathogenesis of asthma and allergic rhinitis.

Effects of white spot syndrome virus infection on immuno-enzyme activities and ultrastructure in gills of Cherax quadricarinatus.

In this study, we explored the pathogenic mechanism of white spot syndrome virus (WSSV) in crayfish, Cherax quadricarinatus, by investigating activities of enzymes related to innate immune function during infection. After 6-12 h of exposure to WSSV, the activities of four enzymes, phenoloxidase (PO), peroxidase (POD), superoxide dismutase (SOD) and lysozyme (LSZ), increased in the gills of C. quadricarinatus but then sharply decreased during longer infection times. Except for PO, the activities of other enzymes in the WSSV-infected crayfish (Group II) were significantly lower than those of the controls at 72 h post-exposure (P < 0.01). Interestingly, the enzyme activities in the group treated with polysaccharides before challenge with WSSV (Group III) were higher than those in Group II. This phenomenon demonstrated that the polysaccharides could improve the immuno-enzyme activities and enhance the organism's antiviral defenses. Morphological examination by transmission electron microscopy revealed abundant WSSV particles and significant damage in the gills of infected crayfish. WSSV infection caused parts of the gill epithelium and microvilli to be reduced in number and size or damaged; meanwhile, the mitochondria morphology changed, with parts of the cristae diminished leaving large vacuoles. Moreover, electron dense deposits appeared and heterochromatinized nuclei could be seen in blood cells with ruptured nuclear membranes and outflow of nucleoplasm. The findings of this study furthers our understanding of the biochemical alterations induced by viral infections, including changes in the antioxidant status, oxidative stress and lysozyme activity, which could help to advance strategies for control of WSSV in crayfish.