Acute toxicity of beta gamma-CAT, a naturally existing non-lens beta gamma-crystallin and trefoil factor complex from frog Bombina maxima skin secretions

In vertebrates, non-lens beta gamma-crystallins are widely expressed in various tissues, but their functions are unknown. The molecular mechanisms of trefoil factors, initiators of mucosal healing and being greatly involved in tumorigenesis, have remained

Apoptotic cells induce immunosuppression through dendritic cells: Critical roles of IFN-gamma and nitric oxide

Apoptotic cells induce immunosuppression through unknown mechanisms. To identify the underlying molecular mediators, we examined how apoptotic cells induce immunoregulation by dendritic cells (DC). We found that administration of DC exposed to apoptotic c

Effects of gamma-aminobutyric acid, progesterone and ionophore A23187 on acrosome reaction of tree shrew sperm in vitro: examination of acrosome reaction with an improved fluorescence microscopy

A number of acrosome reaction (AR) initiators have been found to be effective in inducing AR of human, laboratory and domestic animal sperm. Using an improved simple fluorescence microscopy, effects of gamma-aminobutyric acid (GABA), progesterone and ionophore A23187 on sperm AR of tree shrew, a useful animal model in biomedical research, have been investigated. Spontaneous AR in 4.92-7.53% of viable sperm was observed. Complete AR in 10.31-18.25% of viable tree shrew sperm was obviously induced by 5 mu M and 10 mu M calcium ionophore A23187, 1 mM GABA, and 5 mu M progesterone, and there were no significant differences between their abilities to initiate complete AR. No significant differences of AR percentages between 1- and 2-h treatments with A23187, progesterone and/or GABA were observed. These results suggested that the responses of tree shrew sperm to these AR initiators are similar to that of human and other mammalian sperm. (C) 1997 Elsevier Science B.V.

Molecular modeling on some human interleukins sharing gamma c of interleukin-2 receptor, and structure-function relationships

Five models for human interleukin-7 (HIL-7), HIL-9, HIL-13, HIL-15 and HIL-17 have been generated by SYBYL software package. The primary models were optimized using molecular dynamics and molecular mechanics methods. The final models were optimized using a steepest descent algorithm and a subsequent conjugate gradient method. The complexes with these interleukins and the common gamma chain of interleukin-2 receptor (IL-2R) were constructed and subjected to energy minimization. We found residues, such as Gln127 and Tyr103, of the common gamma chain of IL-2R are very important. Other residues, e.g. Lys70, Asn128 and Glu162, are also significant. Four hydrophobic grooves and two hydrophilic sites converge at the active site triad of the gamma chain. The binding sites of these interleukins interaction with the common gamma chain exist in the first helical and/or the fourth helical domains. Therefore, we conclude that these interleukins binds to the common gamma chain of IL-2R by the first and the fourth helix domain. Especially at the binding sites of some residues (lysine, arginine, asparagine, glutamic acid and aspartic acid), with a discontinuous region of the common gamma chain of IL-2R, termed the interleukins binding sites (103-210). The study of these sites can be important for the development of new drugs. (C) 2000 Elsevier Science B.V. All rights reserved.

Contributions of Pelagic and Benthic Dietary Sources to Freshwater Mussels: Evidence from Stable Carbon Isotope Analysis

In Gonghu Bay of Lake Taihu, tissue of five mussel species showed delta C-13 values similar to or slightly below that of pelagic suspended particulate organic matter (SPOM). This indicated that mussels in this area either fed non-selectively and so reflected available carbon in the pelagic habitat or selected for phytoplankton. The situation was the same for Anodonta woodiana woodiana and Cristaria plicata in Meiliang Bay; however, for the remaining three species, Hyriopsis cumingii, Arconaia lanceolata, and Lamprotula rochechouarti, tissue had intermediate delta C-13 values, falling between those for pelagic SPOM and benthic sediment organic matter (SOM), suggesting a possible preferential selection of phytoplankton from the pelagic SPOM but more likely reflecting local differences in pelagic SPOM and benthic SOM composition and available organic carbon sources. The mixing model showed that pelagic SPOM accounted for over 98% of carbon incorporated by all mussels in Gonghu Bay and two mussels in Meiliang Bay, suggesting the dietary importance of pelagic food sources for mussels. Less than 50% of the assimilation in H. cumingii, A. lanceolata, and L. rochechouarti came from the pelagic carbon sources in Meiliang Bay, which suggested that these species consumed a mix of benthic and pelagic derived carbon sources.

Effects of dietary cyanobacteria of two different sources on growth and recovery of hybrid tilapia (Oreochromis niloticus x O. aureus)

A 115 days feeding trial was conducted to evaluate the effect of dietary cyanobacteria on growth, microcystins (MCs) accumulation in hybrid tilapia (Oreochromis niloticus x O. aureus) and the recovery when the fish were free of cyanobacteria. Three experimental diets were formulated: the control (cyanobacteria free diet); one test diet with cyanobacteria from Lake Taihu (AMt 80.0 mu g MCs g(-1) diet) and one with cyanobacteria from Lake Dianchi (AMd, 410.0 rho g MCs g(-1) diet). Each diet was fed to fish for 60 days and then all fish were free of cyanobacteria for another 55 days. A significant increase in feeding rate (FR) was observed in fish fed AMd diet after a first 30-day exposure (1(st) EP), and in fish fed both AMt diet and AMd diet after a second 30-day exposure (2(nd) EP). Specific growth rates (SGR) of fish fed AMt diet and AMd diet were both obviously affected after the first 30-day exposure, but SGR was only significantly affected in fish fed AMt diet after the second 30-day exposure. After a 55-day recovery, there were no significant differences among diets in the indices mentioned above. Much higher concentrations of MCs were accumulated in tissues of all fish exposed to cyanobacteria. After the 55-day recovery, MC concentrations in fish tissues were significantly lower than those on day 60. (C) 2009 Elsevier Ltd. All rights reserved.

Extracellular phosphatases produced by phytoplankton and other sources in shallow eutrophic lakes (Wuhan, China): taxon-specific versus bulk activity

Extracellular phosphatases are an important part of the phosphorus cycle in aquatic environments. Phosphatase activity (PA) in plankton was studied in seven subtropical shallow lakes of different exploitation management and trophic status in the urban area of Wuhan City. Bulk PA was rather high (range 1.1-11 mu mol l(-1) h(-1)), although concentrations of soluble reactive phosphorus (SRP) were also high (range 27 mu g P l(-1) to similar to 1.5 mg P l(-1)) in all lakes. Cell-associated extracellular PA in phytoplankton was detected using the fluorescence-labelled enzyme activity technique. Phytoplankton species partly contributed to the bulk PA. We found explicit differences in the presence of cell-associated phosphatase within the main phytoplankton groups; species belonging to Chlorophyta and Dinophyta were regularly phosphatase-positive, while Cyanophyta and Bacillariophyceae were phosphatase-negative in all but one case. Furthermore, there is a certain potential of extracellular phosphatases produced by heterotrophic nanoflagellates in most of the lakes. This new finding compromises the 'traditional' interpretation of bulk phosphatase data as being due to overall phytoplankton or bacterial P regeneration.

The search for the IFN-gamma receptor in fish: Functional and expression analysis of putative binding and signalling chains in rainbow trout Oncorhynchus mykiss

Interferons (IFNs), consisting of three major subfamilies, type I, type II (gamma) and type III (lambda) IFN, activate vertebrate antiviral defences once bound to their receptors. The three IFN subfamilies bind to different receptors, IFNAR1 and IFNAR2 for type I IFNs, IFN gamma R1 and IFN gamma R2 for type II IFN, and IL-28R1 and IL-10R2 for type III IFNs. In fish, although many types I and II IFN genes have been cloned, little is known about their receptors. In this report, two putative IFN-gamma receptor chains were identified and sequenced in rainbow trout (Oncorhynchus mykiss), and found to have many common characteristics with mammalian type II IFN receptor family members. The presented gene synteny analysis, phylogenetic tree analysis and ligand binding analysis all suggest that these molecules are the authentic IFN gamma Rs in fish. They are widely expressed in tissues, with IFN gamma R1 typically more highly expressed than IFN gamma R2. Using the trout RTG-2 cell line it was possible to show that the individual chains could be differentially modulated, with rIFN-gamma and rIL-1 beta down regulating IFN gamma R1 expression but up regulating IFN gamma R2 expression. Overexpression of the two receptor chains in RTG-2 cells revealed that the level of IFN gamma R2 transcript was crucial for responsiveness to rIFN-gamma, in terms of inducing gamma IP expression. Transfection experiments showed that the two putative receptors specifically bound to rIFN-gamma. These findings are discussed in the context of how the IFN gamma R may bind IFN-gamma in fish and the importance of the individual receptor chains to signal transduction. (c) 2009 Elsevier Ltd. All rights reserved.

Comparative study and expression analysis of the interferon gamma gene locus cytokines in Xenopus tropicalis

Using bioinformatics approach, the genome locus containing interleukin (IL)-22, IL-26, and interferon gamma (IFN-gamma) genes has been identified in the amphibian, Xenopus tropicalis. Like that in other vertebrates such as fish, birds, and mammals, the Xenopus IL-22, IL-26, and IFN-gamma are clustered in the same chromosome and the adjacent genes are conserved. The genomic structures of the Xenopus IL-22, IL-26, and IFN-gamma gene were identical to that of their mammalian counterparts. The Xenopus IL-22 and IL-26 genes contained five exons and four introns while the Xenopus IFN-gamma gene consisted of four exons and three introns. The Xenopus IL-22, IL-26, and IFN-gamma share 14.1-41.6%, 14.6-31.2%, and 23.7-36.5% identity to their counterparts in other species, respectively. Reverse-transcription polymerase chain reaction (PCR) and real-time quantitative PCR analyses revealed that the expression of IL-22, IL-26, and IFN-gamma genes was significantly upregulated after simulation with bacterial polyliposaccharide and/or synthetic double-stranded poly(I:C), suggesting these cytokines like those in other vertebrates play an important role in regulating immune response in Xenopus.

Molecular cloning and characterization of common carp (Cyprinus carpio L.) TCR gamma and CD3 gamma/delta chains

Partial cDNA sequences of TCR gamma and CD3 gamma/delta were isolated from the thymus of common carp (Cyprinus carpio L.) by the method of suppression subtractive hybridization (SSH). Subsequently the full length cDNAs of carp TCR gamma and CD3 gamma/delta were obtained by means of 3' RACE and 5' RACE, respectively. The full length of carp TCR gamma chain is 1368 bp and encodes 326 amino acids including a signal peptide region of 19 amino acids and a transmembrane region of 23 amino acids at the C-terminal region from aa 291 to 313. The V region of carp TCR gamma contains 109 amino acids, the core motif FGXG in J segment was also found in carp TCR gamma. The C region of carp TCR gamma contains the characteristic CX6PX6WX45C motif. The CP region of carp TCR C gamma contains 37 amino acids. The full length of carp CD3 gamma/delta is 790 bp and encodes 175 amino acids including a signal peptide region of 17 amino acids and a transmembrane region of 23 amino acids from aa 93 to 115. Similar to other known CD3 gamma/delta s, four cysteine residues in the extracellular domain and an immunoreceptor tyrosine-based activation motif ITAM (YxxL/Ix6-8YxxL/I) in the intracellular domain are also included in carp CD3 gamma/delta. Differing from other known CD3 gamma/delta s, carp CD3 gamma/delta tacks the CXXCXE motif in the extracellular domain. RTPCR analysis demonstrated that the expression of TCR gamma gene was mainly in the thymus and gill of 6-month carp, but in 18-month carp, TCR gamma gene was detected in all the examined tissues. The expression of CD3 gamma/delta gene was detected in all examined tissues of 6 and 18-month carp; among them, the highest expression level was in the thymus of 6-month carp. In situ hybridization showed that CD3 gamma/delta-expressing cells were widely distributed in the head kidney, spleen and kidney of carp, whereas in the thymus, they were densely distributed in the lymphoid outer zone and scattered in the epithelioid inner zone. (c) 2007 Published by Etsevier Ltd.

Sediment sources and the flood record from Wanghu lake, in the middle reaches of the Yangtze River

A sediment core was collected from the centre of Wanghu Lake, in the Middle Reaches of the Yangtze River. The recent part of the core was dated using a combination of Pb-210 and spheroidal carbonaceous particle (SCP) techniques. Extrapolating this chronology dated the laminated section of the core, between 723 and 881 mm, to the first half of the 18th century and this section was selected for detailed study. The thicknesses of the laminae were measured using reflecting and polarizing microscopes whilst geochemistry was determined by an electron probe. The thickness of the dark layers was found to be positively correlated with titanium concentrations, and negatively correlated with aluminium and potassium concentrations. The thickness of the light layers was found to be negatively correlated with the concentrations of titanium. It is concluded that the dark layers were deposited from the Fushui River, a tributary of the Yangtze River, under periods of normal flow whilst the light Layers were mainly deposited from the Yangtze River itself during flood periods. Documentary evidence for floods occurring in the take catchment corresponded with thick laminations of high titanium concentration. Further, two of the three thickest, light laminations with low titanium concentrations were found to be synchronous with recorded flood dates of the main Yangtze River in its Middle Reaches, but one was synchronous with a local drought. These data suggest that the Lake sediment provides an archive of the relative water levels of the Yangtze and Wanghu including floods of both the main Yangtze River and the local hydrological regime. (c) 2006 Elsevier B.V. All rights reserved.

Effect of dietary carbohydrate sources on growth performance and utilization for gibel carp (Carassius auratus gibelio) and Chinese longsnout catfish (Leiocassis longirostris Gunther)

The nutritional function of monosaccharides, disaccharides and polysaccharides for omnivorous gibel carp and carnivorous Chinese longsnout catfish were investigated and the ability of these two species to utilize carbohydrates was compared. For each species, triplicate groups of fish were assigned to each of five groups of isoenergetic and isonitrogenous experimental diets with different carbohydrate sources: glucose, sucrose, dextrin, soluble starch (acid-modified starch) and alpha-cellulose. The carbohydrates were included at 60 g kg(-1) in Chinese longsnout catfish diets and at 200 g kg(-1) in gibel carp diets. A growth trial was carried out in a recirculation system at 27.8 +/- 1.9 degrees C for 8 weeks. The results showed that fish with different food habits showed difference in the utilization of carbohydrate sources. For gibel carp, better specific growth rate (SGR) and feed efficiency (FE) were observed in fish fed diets containing soluble starch and cellulose, but for Chinese longsnout catfish, better SGR and FE were observed in fish fed diets containing dextrin and sucrose. Apparent digestibility coefficient of dry matter (ADC(d)) and apparent digestibility coefficient of energy (ADC(e)) were significantly affected by dietary carbohydrate sources in gibel carp. ADC(d) and ADC(e) significantly decreased as dietary carbohydrate complexity increased in Chinese longsnout catfish except that glucose diet had medium ADC(d) and ADC(e). In both species, no significant difference of apparent digestibility coefficient of protein was observed between different carbohydrate sources. Dietary carbohydrate sources significantly affected body composition, and liver phosphoenolpyruvate carboxykinase (PEPCK), pyruvate kinase (PK), glucose 6-phosphate dehydrogenase (G6PD) and malic enzyme (ME) activities also varied according to dietary carbohydrate complexity. Fish with different food habits showed different abilities to synthesize liver glycogen, and the liver glycogen content in gibel carp was significantly higher than in Chinese longsnout catfish. The influence of carbohydrate source on gluconeogenesis and lipogenesis was also different in the two fish species.

Discharges and sediments as sources of extracellular alkaline phosphatase in a shallow Chinese eutrophic lake

Total alkaline phosphatase activity (APA) and soluble reactive phosphorus (SRP) concentrations were measured in municipal wastewater, and a shallow Chinese freshwater lake receiving it. Activities of Dissolved alkaline phosphatase ( ADAP) in overlying and interstitial water were also analyzed monthly at three sites for several years. The lake was enriched with SRP and alkaline phosphatase by discharge of the wastewater, indicating that the inclusion of APA for estimating water pollution was reasonable. Annual data showed that APA in coarser fraction was significantly higher at the site receiving more wastewaters, both in surface and overlying water, suggesting that resuspension of enzyme most likely occurred in the basin heavily discharged. ADAP was an order of magnitude higher in the wastewater than those in lake waters, and was generally higher in interstitial water, a feature more striking at the site receiving more discharges. Besides, it was irrespectively inhibited by Na2WO4, L-cysteine and EDTA-Na, but stimulated by Cu2+, Zn2+, CTAB and Triton X-100 in interstitial, overlying and surface waters. This similarity of responding patterns to the stressors indicated an analogy between dissolved alkaline phosphatase in water column and that in interstitial water, supporting the hypothesis that the polluted sediments act as source of dissolved alkaline phosphatase in eutrophic lakes.

Optimization of growth and fatty acid composition of a unicellular marine picoplankton, Nannochloropsis sp., with enriched carbon sources

A unicellular marine picoplankton, Nannochloropsis sp., was grown under CO2-enriched photoautotrophic or/and acetate-added mixotrophic conditions. Photoautotrophic conditions with enriched CO2 of 2800 mul CO2 l(-1) and aeration gave the highest biomass yield (634 mg dry wt l(-1)), the highest total lipid content (9% of dry wt), total fatty acids (64 mg g(-1) dry wt), polyunsaturated fatty acids (35% total fatty acids) and eicosapentaenoic acid (EPA, 20:5omega3) (16 mg g(-1) dry wt or 25% of total fatty acids). Mixotrophic cultures gave a greater protein content but less carbohydrates. Adding sodium acetate (2 mM) decreased the amounts of the total fatty acids and EPA. Elevation of CO2 in photoautotrophic culture thus enhances growth and raises the production of EPA in Nannochloropsis sp.