Influence of dietary fibre-enriched foods and of blueberry-rich diet on saliva composition in humans

The main objective of the thesis is to explore the possible “cross-talk” between the brain, intestine and specific foods components (i.e. dietary fibres and blueberry polyphenols). This objective will be achieved through a short-term study and a long-term intervention in humans, with the non-invasive analysis of saliva samples and breath compositions. The short-term study aimed at evaluating the impact of dietary fibres on salivary NAEs during mastication and in the post-prandial phase. Three types of biscuits enriched with 3% barley β-glucan (βGB) or whole-wheat bran (WWBB) or without dietary fibre (control, CB) were developed. A crossover randomized human study with eighteen healthy and fasting participants was carried out. Saliva samples from subjects in a resting condition, upon mastication of parafilm and one of the three biscuits were collected. Subsequently, the amount of biscuits consumed in an ad-libitum breakfast was measured and post-prandial saliva samples, blood glucose, and questionnaires of appetite and food liking were collected over the following two hours. Data demonstrated that salivary NAEs concentration increased only upon food mastication instead of mastication itself, independently from dietary fibre composition of the food. The type of biscuits did not influence individual appetite nor post- prandial blood glucose; on the contrary it influenced the persistence of NAEs in saliva over 30 min after consumption. Future studies will clarify the mechanisms behind this finding and the role of salivary NAEs in food liking and appetite cues after food consumption. The aim of long-term intervention study was to test the influence of a one week-consumption of blueberries on the volatile organic compounds (VOCs) of breath and saliva in humans. Fourteen healthy volunteers participated in this three-week single blind study with a two- week cross over design. After a one-week of baseline period with a low polyphenol-diet (BL), subjects were grouped randomly to continue the same diet (control diet, CT) or to add 200 g/day of fresh blueberries (intervention diet, INT) for one week. In the following week they switched to the other arm. At the end of each week, fasting subjects reached the laboratory to collect saliva samples and to have on-line analysis of breath by PTR-ToF-MS. After INT and CT, difference was found neither in VOC fingerprints nor in single VOC of breath and saliva samples. Nevertheless, a significant correlation was shown between saliva and breath composition for methanol, formaldehyde, ethanol, acetone and propanol. Numerous previous studies focused on VOCs composition of breath of patients and of healthy subjects, but very few studies focused on salivary VOCs. Therefore the link found in this study between saliva and breath VOCs may open a new research path to clarify the mechanisms behind the metabolic effect of the dietary intervention in humans

Molecular cloning, sequence analysis and structure prediction of the related to b0,+ amino acid transporter (rBAT) in Cyprinus carpio L.

In this study, the full-length cDNA of basic amino acid transporter gene rBAT was cloned from intestinal cells of Cyprinus carpio L. using reverse transcription polymerase chain reaction (RT-PCR) and rapid-amplification of cDNA ends (RACE) methods. The amplified product was 2370 bp, including a 42 bp 5'-untranslated region, a 288 bp 3'-untranslated region, and a 2040 bp open reading frame (ORF), which encoded 679 amino acids. The predicted amino acid sequence showed high similarity with that of zebrafish (83.5%), and low similarity with that of rat (50.90%). The 3-D protein models were predicted by the comparative protein modeling program SWISS-MODEL. The prediction result displayed that the Cyprinus carpio L. rBAT had a hydrophilic cytoplasmic N terminus, a single membrane-spanning domain, and an extracellular C terminus. The structural core was a β-sheet at the N terminus. The rBAT associates with the light subunit b0,+AT by a disulfide bridge with conserved cysteine residues (residues 109). A better understanding of the functional roles and regulation mechanism of rBAT would provide unique opportunities to investigate the biochemical processes underlying amino acid metabolism in C. carpio L., and support the foundation for improving aquaculture culture of C. carpio L.Keywords: rBAT gene, cDNA sequence analysis, protein tertiary structure, Cyprinus carpio

Characterization of Shewanella sp. Isolated from Cultured Loach Misgurnus anguillicaudatus

Shewanella infection of fish has become a significant problem in aquaculture. In September 2014, a disease was seen in cultured loach (Misgurnus anguillicaudatus) in Xuzhou, central China. A gram-negative bacillus was isolated from the diseased loaches and was tentatively named strain MS1, which was then identified as Shewanella sp. by physiological and biochemical characteristics analysis. The strain MS1 showed highest 16S rRNA sequence identities (98.93%, 98.87%) with the latest two species listed (Shewanella sp. MR7, Shewanella sp. MR4). The phylogenetic tree constructed on the basis of 16S rRNA gene sequences strongly indicated that the strain MS1 is most closely related to the new Shewanella strains MR7 and MR4. The isolate MS1 was confirmed as the pathogen of the infected loaches by experimental reinoculation. The strain was susceptible to most antimicrobial agents tested, but resistant to glycopeptides (vancomycin, teicoplanin) and lincosamide (lincomycin, clindamycin). This is the second report on Shewanella sp. isolated from the diseased loach

A 550,000-year record of East Asian monsoon rainfall from Be-10 in loess

Cosmogenic Be-10 flux from the atmosphere is a proxy for rainfall. Using this proxy, we derived a 550,000-year-long record of East Asian summer monsoon (EASM) rainfall from Chinese loess. This record is forced at orbital precession frequencies, with higher rainfall observed during Northern Hemisphere summer insolation maxima, although this response is damped during cold interstadials. The Be-10 monsoon rainfall proxy is also highly correlated with global ice-volume variations, which differs from Chinese cave delta O-18, which is only weakly correlated. We argue that both EASM intensity and Chinese cave delta O-18 are not governed by high-northern-latitude insolation, as suggested by others, but rather by low-latitude interhemispheric insolation gradients, which may also strongly influence global ice volume via monsoon dynamics

Seasonal variations of ATPase activity and antioxidant defenses in gills of the mud crab Scylla serrata (Crustacea, Decapoda)

【英文摘要】The mud crab Scylla serrata is an important commercial crustacean being widely distributed along the southeastern coast of China. This crab and particularly its gills are subjected to seasonal fluctuations of environmental factors due to direct exposure to seawater. The investigation of seasonal variations of ATPase and antioxidant defenses in gills could be helpful for the understanding of physiological regulation mechanism of seasonal adaptations. In this study, mud crabs were collected from the subtropical waters near Xiamen island, Southeast China (24°26′46′′N, 118°04′04′′E), in August and November, 2002, and February and May, 2003, respectively, being considered as specimens from summer, autumn, winter and spring, respectively. Only healthy intermoult male crabs without carapace or appendage damage were used, having a carapace width of 7.05 ± 0.52 cm and a wet weight of 130 ± 20 g. The activities of four ATPases and three antioxidation enzymes (superoxide dismutase, catalase, glutathione peroxidase) and the content of malondialdehyde in gills were measured. The results showed that the activities of Na+, K+-ATPase, Mg2+-ATPase, Ca2+-ATPase and Ca2+, Mg2+-ATPase reached maximum levels similarly in summer and decreased in winter. The activities of superoxide dismutase and catalase were higher in summer and lower in winter, with a significant decrease in winter compared to the other seasons (p < 0.01). Glutathione peroxidase activity was higher in summer, and lower in autumn, being significantly higher in summer or winter than in spring or autumn (p < 0.01). Malondialdehyde content was higher in summer and lower in spring with significant differences among the different seasons (p < 0.01). In summary, the obvious seasonal activity variations of four ATPases and antioxidant enzymes and the content of malondialdehyde reflect a seasonal regulation of the physiological metabolising enzyme and the antioxidant capacity to cope with seasonal alterations of environment factors such as fluctuating salinities and temperatures. Communicated by K. Yin

Antioxidation and ATPase activity in the gill of mud crab Scylla serrata under cold stress

【英文摘要】 Mud crab (Scylla serrata) is an important commercial crustacean in China. An experiment was designed to study the effect of cold stress on S. serrata. After a one-week adaptation at 28oC, the temperature is suddenly reduced to 4oC. The crabs were sampled every 2 h for 10 h and dissected immediately to measure the enzyme activity. The crabs at room temperature (28oC) were used as the control group. The activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPX), the content of mal...the National High-Tech Research and Development Program of China (863 Program) (No. 2002AA603013

CcGSDMEa functions the pore-formation in cytomembrane and the regulation on the secretion of IL-lβ in common carp (Cyprinus carpio haematopterus)

GSDME is the only direct executor of caspase-dependent pyroptosis in both canonical and non-canonical inflammasomes known to date in fish, and plays an important role in anti-bacterial infection and inflammatory response. In order to determine the regulation of GSDMEa on antibacterial infection in innate immune response, the CcGSDMEa gene in common carp (Cyprinus carpio haematopterus) was first identified and characterized, and then its function related to immune defense was investigated. Our results showed that the expressions of CcGSDMEa at the mRNA and protein levels were both significantly increased after Aeromonas hydrophila intraperitoneal infection at the early stage than that in the control group. We found that CcGSDMEa could be cleaved by inflammatory caspase (CcCaspase-1b) and apoptotic caspases (CcCaspase-3a/b and CcCaspase-7a/b). Interestingly, only the CcGSDMEa-NT (1-252 aa) displayed bactericidal activity to Escherichia coli and could punch holes in the membrane of HEK293T cells, whereas CcGSDMEa-FL (1-532 aa) and CcGSDMEa-CT (257-532 aa) showed no above activity and pore-forming ability. Overexpression of CcGSDMEa increased the secretion of CcIL-1β and the release of LDH, and could reduce the A. hydrophila burdens in fish. On the contrary, knockdown of CcGSDMEa reduced the secretion of CcIL-1β and the release of LDH, and could increase the A. hydrophila burdens in fish. Taken together, the elevated expression of CcGSDMEa was a positive immune response to A. hydrophila challenge in fish. CcGSDMEa could perform the pore-formation in cell membrane and the regulation on the secretion of IL-lβ, and further regulate the bacterial clearance in vivo. These results suggested that CcGSDMEa played an important role in immune defense against A. hydrophila and could provide a new insight into understanding the immune mechanism to resist pathogen invasion in teleost

The 10Be record as a proxy of paleomagnetic reversals and excursions: A Mediterranean perspective

The 10Be/9Be ratio is acknowledged as an effective tool for establishing the stratigraphic position of paleomagnetic excursions. Still, our data suggest that, in particular depositional settings, the interplay between climate, sedimentation and oceanography may jeopardize a realistic depiction of the natural 10Be/9Be record

Differential proteomic profiles and characterizations between hyalinocytes and granulocytes in ivory shell Babylonia areolata

Abstract(#br)The haemocytes of the ivory shell, Babylonia areolata are classified by morphologic observation into the following types: hyalinocytes (H) and granulocytes (G). Haemocytes comprise diverse cell types with morphological and functional heterogene and play indispensable roles in immunological homeostasis of invertebrates. In the present study, two types of haemocytes were morphologically identified and separated as H and G by Percoll density gradient centrifugation. The differentially expressed proteins were investigated between H and G using mass spectrometry. The results showed that total quantitative proteins between H and G samples were 1644, the number of up-regulated proteins in G was 215, and the number of down-regulated proteins in G was 378. Among them, cathepsin, p38 MAPK, toll-interacting protein-like and beta-adrenergic receptor kinase 2-like were up-regulated in G; alpha-2-macroglobulin-like protein, C-type lectin, galectin-2-1, galectin-3, β-1,3-glucan-binding protein, ferritin, mega-hemocyanin, mucin-17-like, mucin-5AC-like and catalytic subunit of protein kinase A were down-regulated in G. The results showed that the most significantly enriched KEGG pathways were the pathways related to ribosome, phagosome, endocytosis, carbon metabolism, protein processing in endoplasmic reticulum and oxidative phosphorylation. For phagosome and endocytosis pathway, the number of down-regulation proteins in G was more than that of up-regulation proteins. For lysosome pathway, the number of up-regulation proteins in G was more than that of down-regulation proteins. These results suggested that two sub-population haemocytes perform the different immune functions in B. areolata