Chemical constituents of marine mangrove-derived endophytic fungus Alternaria tenuissima EN-192

A chemical investigation of the ethyl acetate extract of the fermentation broth of Alternaria tenuissima EN-192,an endophytic fungus obtained from the stems of the marine mangrove plant Rhizophora stylosa,resulted in the isolation of nine known secondary metabolites,including four indole-diterpenoids:penijanthine A(1),paspaline(2),paspalinine(3),and penitrem A(4);three tricycloalternarene derivatives:tricycloalternarene 3a(5),tricycloalternarene 1b(6),and tricycloalternarene 2b(7);and two alternariol congeners:djalonensone(8) and alternariol(9).The chemical structures of these metabolites were characterized through a combination of detailed spectroscopic analyses and their comparison with reports from the literature.The inhibitory activities of each isolated compound against four bacteria were evaluated and compounds 5 and 8 displayed moderate activity against the aquaculture pathogenic bacterium Vibrio anguillarum,with inhibition zone diameters of 8 and 9 mm,respectively,at 100 μg/disk.To the best of our knowledge,this is the first report on the secondary metabolites of mangrove-derived Alternaria tenuissima and also the first report of the isolation of indole-diterpenoids from fungal genus Alternaria.     

Secondary metabolites of a deep sea derived fungus Aspergillus versicolor CXCTD-06-6a and their bioactivity

In order to obtain novel secondary metabolites, a deep sea inhabiting fungus Aspergillus versicolor CXCTD-06-6a was investigated. One new diketopiperazine brevianamide W(1a), as well as five known diketopiperazine alkaloids, diketopiperazine V(1b), brevianamide Q(2), brevianamide R(3), brevianamide K(4), and brevianamide E(5), were isolated from the Et OAc extract of the fermentation broth. Their structures were elucidated by spectroscopy techniques(NMR, MS). The six compounds exhibited moderate radical scavenging activity against DPPH with clearance ratio of 55.0%(1a and 1b), 53.7%(2), 46.2%(3), 61.4%(4) and 19.3%(5) at a concentration of 13.9 μmol L-1, respectively; while the positive control ascorbic acid showed a ratio of 70.3% at the concentration of 28.4 μmol L-1.     

Chemical profile of the secondary metabolites produced by a deep-sea sediment-derived fungus Penicillium commune SD-118

Bioassay-guided fractionation of the crude extract from Penicillium commune SD-118, a fungus obtained from a deep-sea sediment sample, resulted in the isolation of a known antibacterial compound, xanthocillin X (1), and 14 other known compounds comprising three steroids (2-4), two ceramides (5 and 6), six aromatic compounds (7-12), and three alkaloids (13-15). Xanthocillin X (1) was isolated for the first time from a marine fungus. In the bioassay, xanthocillin X (1) displayed remarkable antimicrobial activity against Staphylococcus aureus and Escherichia coli, and significant cytotoxicity against MCF-7, HepG2, H460, Hela, Du145, and MDA-MB-231 cell lines. Meleagrin (15) exhibited cytotoxicity against HepG2, Hela, Du145, and MDA-MB-231 cell lines. This is the first report of the cytotoxicity of xanthocillin X (1).     

Study on the Preparation and Antileukemic Activity of New Lipophilic 1-β-D-arabinofuranosylcytosine Derivatives

Cytarabine(1-β-D-arabinofuranosylcytosine, Ara-C), isolated from a Caribbean sponge species Tethyacrypta, is the first antitumor drug from a marine resource. In 1980, the US Food and Drug Administration approved this drug for the treatment of different types of leukemia. This drug has a short plasma half-life, low stability, limited bioavailability, and severe side effects. To improve stability and bioavailability, we synthesized nine novel derivatives by blocking the cytarabine metabolic sites and improving lipophilicity. The c Log P values of the newly synthesized compounds were calculated. All the synthesized compounds were more lipophilic than cytarabine, resulting in membrane permeability and bioavailability improvement. The antitumor activities against leukemia cell line HL-60 were evaluated by using the MTT assay. The bioassay results revealed that the IC_(50) values of compounds 5, 8 and 9 were 0.080, 0.090 and 0.057 μmol L~(-1), respectively, which was similar with that of cytarabine(0.056 μmol L~(-1),). In comparison, compound 4 with a phosphate group at O5' was inactive. Because phosphoester bonds are easily hydrolyzed by alkaline phosphatase and are commonly used in producing prodrug strategies, compound 4 might also be metabolized in vivo and generate compound 3 or even cytarabine through a multi-step reaction. Thus, compound 4 might be a promising compound to be developed as a prodrug.     

Isolation,identification, and cytotoxicity of a new isobenzofuran derivative from marine <Emphasis Type="Italic">Streptomyces</Emphasis> sp. W007

A new isobenzofuran derivative (1) was isolated from the marine Streptomyces sp. W007 and its structure was determined through extensive spectroscopic analyses, including 1D-NMR, 2D-NMR, and ESI-MS. The absolute configuration of compound 1 was determined by a combination of experimental analyses and comparison with reported data, including biogenetic reasoning, J-coupling analysis, NOESY, and ¹H-¹HCOSY. Compound 1 exhibited no cytotoxicity against human cells of gastric cancer BGC-823, lung cancer A549, and breast cancer MCF7.     

Alkaloids from an algicolous strain of <Emphasis Type="Italic">Talaromyces</Emphasis> sp.

Compounds isolated and identified in a culture of the alga-endophytic fungus Talaromyces sp. cf-16 included two naturally occurring alkaloids, 2-[(S)-hydroxy(phenyl)methyl]-3-methylquinazolin-4(3H)-one (1a) and 2-[(R)-hydroxy(phenyl)methyl]-3-methylquinazolin-4(3H)-one (1b), that were identified for the first time. In addition, seven known compounds (2–8) were obtained from the culture. Following chiral column chromatography, compounds 1a and 1b were identified as enantiomers by spectroscopic analyses and quantum chemical calculations. Bioassay results showed that 5 was more toxic to brine shrimp than the other compounds, and that 3–6 could inhibit Staphylococcus aureus.     

Phenylquinolinones with antitumor activity from the Indian Ocean-derived fungus <Emphasis Type="Italic">Aspergillus versicolor</Emphasis> Y31-2

Two phenylquinolinones, including one new compound (1) and a previously isolated compound (2), were isolated from the ethyl acetate extracts of the fungus Aspergillus versicolor Y31-2, which was obtained from seawater samples collected from the Indian Ocean. The structures of these compounds were established by spectroscopic analyses. 4-(3-Hydroxyphenyl)-3-methoxyquinolin-2(1H)-one (1) exhibited moderate cytotoxicity against MCF-7 (human breast carcinoma cell line) and SMMC-7721 (human liver cancer cell line) cells with IC₅₀ values of 16.6 and 18.2 μmol/L, respectively. To the best of our knowledge, this study represents the first reported account of the isolation of compounds 1 and 2 as the secondary metabolites of the seawater derived fungus Aspergillus versicolor from the Indian Ocean.     

Geochemistry of re and trace elements in ferromanganese nodules and their genesis

Trace and RE element geochemistry and genesis have been studied with respect to ferromanganese nodules from the sediments of the Pacific, Atlantic and Indian oceans.     

Immunogenicity and protective role of antigenic regions from five outer membrane proteins of <Emphasis Type="Italic">Flavobacterium columnare</Emphasis> in grass carp <Emphasis Type="Italic">Ctenopharyngodon idella</Emphasis>

Flavobacterium columnare causes columnaris disease in freshwater fish. In the present study, the antigenic regions of five outer membrane proteins (OMPs), including zinc metalloprotease, prolyl oligopeptidase, thermolysin, collagenase and chondroitin AC lyase, were bioinformatically analyzed, fused together, and then expressed as a recombinant fusion protein in Escherichia coli. The expressed protein of 95.6 kDa, as estimated by 10% sodium dodecyl sulfate-polyacrylamide gel electrophoresis, was consistent with the molecular weight deduced from the amino acid sequence. The purified recombinant protein was used to vaccinate the grass carp, Ctenopharyngodon idella. Following vaccination of the fish their IgM antibody levels were examined, as was the expression of IgM, IgD and IgZ immunoglobulin genes and other genes such as MHC Iα and MHC IIβ, which are also involved in adaptive immunity. Interleukin genes (IL), including IL-1β, IL-8 and IL-10, and type I and type II interferon (IFN) genes were also examined. At 3 and 4 weeks post-vaccination (wpv), significant increases in IgM antibody levels were observed in the fish vaccinated with the recombinant fusion protein, and an increase in the expression levels of IgM, IgD and IgZ genes was also detected following the vaccinations, thus indicating that an adaptive immune response was induced by the vaccinations. Early increases in the expression levels of IL and IFN genes were also observed in the vaccinated fish. At four wpv, the fish were challenged with F. columnare, and the vaccinated fish showed a good level of protection against this pathogen, with 39% relative percent survival (RPS) compared with the control group. It can be concluded, therefore, that the five OMPs, in the form of a recombinant fusion protein vaccine, induced an immune response in fish and protection against F. columnare.     

Palynological assemblages from the holocene sediments of the Changjiang (Yangtze) River Delta and its implications

The palynolpgical study on the Quaternary sediments of 42 cores in the region of the Yangtze River Delta and the sections of 4 relics had been done since 1960. Palynological assemblages from the region of the Yangtze River Delta, showing obviously .common characteristics but with slight differences because of the vast areas covered, can be divided into five spores and pollen zones and three subzones in ascendant order:I.Quercus alienc-Pinus-Cupressaceae-Chenopodiaceae spores and pollen zone reflecting the cool, then cold and dry climate--Preboreal.II.Quercus dentata-Quefcus acutissima-Pinus-Castanea-Artemisia spores and pollen zone reflecting the temperate and cool, but a little dry climate--Boreal.III.Quercus glauca-Caslanopsis-Myrica-Caslanea spores and pollen zone reflecting the hot and moist climate--Atlantic.IV.Quercus acutissima-Pinus-Castanea-Quercus glauca- Chenopodiaceae spores and pollen zone reflecting the warm, and a little dry climate--Subboreal.V.Quercus acutissima-Costarica-Quercus glauca     

Combined effects of light intensity and NH^＋4-enrichment on growth,pigmentation, and photosynthetic performance of Ulva prolifera （Chlorophyta）

The aim of this study was to investigate the effects of light intensity and enhanced nitrogen supply on the growth and photosynthesis of the green-tide macroalga, Ulva prolifera. Thalli of U. prolifera were grown in natural or NH 4 +-enriched seawater under two different light intensities for 7 days, and then the growth rate, pigmentation, and photosynthetic performance of the thalli were evaluated. The results show that the relative growth rate(RGR) was markedly higher under the high light level than under the low light level. Enrichment with NH 4 + enhanced the RGR under high light intensity, but did not affect RGR under low light intensity. In low light conditions, NH 4 +-enrichment resulted in a marked decrease in the maximal photosynthetic rate( P m) and the maximum carbon fixation rate( V max), but it did not affect the half saturation constant for carbon( K 0.5) or the ratio of V max to K 0.5, which reflects the carbon acquisition efficiency. In high light conditions, P m, K 0.5, and the dark respiration rate( R d) increased under NH 4 + enrichment, but V max and the V max / K 0.5 ratio decreased. Regardless of the light intensity, NH 4 +-enrichment did not affect the apparent photosynthetic efficiency( α), which refl ects the ability of the alga to use light energy at low light levels. Under both low and high light intensities, the chlorophyll a(Chl a), chlorophyll b(Chl b), and carotenoids(Car) contents in thalli were higher in NH 4 +-enriched than in natural seawater, except that there was a decrease in the Chl b content of thalli in NH 4 +-enriched seawater under low light intensity. Therefore, NH 4 + enrichment improved the growth and photosynthetic performance of U. prolifera under high light intensity, but not under low light intensity. We discuss the possible mechanisms underlying these physiological responses.     

Microanalysis and preliminary pharmacokinetic studies of a sulfated polysaccharide from <Emphasis Type="Italic">Laminaria japonica</Emphasis>

A rapid, sensitive and reproducible high performance liquid chromatography (HPLC) method with post-column fluorescence derivatization has been developed to determine the amount of low-molecular-weight sulfated polysaccharide (GFS) in vivo. The metabolism of GFS has been shown to fit a two component model following its administration by intravenous injection, and its pharmacokinetic parameters were determined to be as follows: half-time of distribution phase (t _1/2α)=11.24±2.93 min, half-time of elimination phase (t _1/2β)=98.20±25.78 min, maximum concentration (C _max)=110.53 μg/mL and peak time (T _max)=5 min. The pharmacokinetic behavior of GFS was also investigated following intragastric administration. However, the concentration of GFS found in serum was too low for detection, and GFS could only be detected for up to 2 h after intragastric administration (200 mg/kg body weight). Thus, the bioavailability of GFS was low following intragastric administration because of the metabolism of GFS. In conclusion, HPLC with postcolumn derivatization could be used for quantitative microanalysis and pharmacokinetic studies to determine the presence of polysaccharides in the serum following intravenous injection.     

Bioreactor technology for production of valuable algal products

INTRODUCTIONManystudiesonvascularplantSduringthenontwperiodhavedrireP0redinChinaandabroad.TherearefeWreportSonwhethertherearesomevasctilarplantS,andhowmanyspedessurvive,underice,andontheirbiomassesandphotosyntheticproduCtivitiesduringwinter,espedal1yduringbeAfpinchLillyamepe0ylenctal.,l976,Liuetal.,l994).WeimtlycarriedoutaseadriontheunderforvascularplantsdurinwinderinsomewaterbodieswherevascularplantSgrowluxdrianhydurinInildareonsinnorthemareas,soastoimproveourknowedgeofvascularplaflt…     

Effects of nitrogen specification and culture method on growth of Enteromorpha prolifera

Eutrophication, which is the enrichment of a water mass with inorganic and organic nutrients that support plant growth, is a key factor in stimulating phytoplankton growth. In this study, we determined the effects of various nitrogen sources, different nitrogen concentrations in the culture medium, and two culture methods on the growth of the green alga, Enteromorpha prolifera. The relationship between the specific growth rate of E. prolifera and NO3--N concentration was consistent with that estimated using the Monod equation (R2 = 0.9713, P < 0.01). In the NO3--N medium, the maximum specific growth rate was calculated to be 0.1634/d and the semi-saturation constant was calculated to be 16.86 μmol/L. Our results show that E. prolifera can effectively utilize NH4+-N, NO3--N, and NO2--N and urea-N in the range of 5 to 50 μmol/L. NH4+-N was preferentially assimilated by E. prolifera, and urea-N was favorable for long-term growth.     

Distribution and migration of the common squidTodarodes pacificus in the Yellow Sea

Study of the distribution and migration of the common squid,Todarodes pacificus Steenstrup,basedon the index of important fishing ground(P) and fisheries statistics on the Yellow Sea and northern EastChina Sea during 1980—1991 showed that:1.Its catch in the fishing period(June to November) is 91.77% of the annual yield.The fishingground distributes over the northem and middle Yel1ow Sea and adjacent area of the Changjiang Estuary.2. It over-winters in the northem East China Sea and waters adjacent to Goto Island from De-cember to February and spawns in waters near Haijiao Is1and and west of Kyushu. The main stock mi-grates along 123°30′E to the ChangJiang Estuary, Haizhou Bay. offsea from Shidao to Qingdao,mideastern Yellow Sea, and offsea Weihai and Haiyang Island succesively for feeding after April. The sur-plus stock migrates again to the wintering ground in December.3.The favorable feeding temperature is 6-23℃(optimum of l3-20℃ in the Changjiang Estua-ry and 7-13℃ in the northern and middle Yel     

Influence of principal stress rotation of unequal tensile and compressive stress amplitudes on characteristics of soft clay

Soil behavior can reflect the characteristics of principal stress rotation under dynamic wave and traffic loads. Unequal amplitudes of tensile and compressive stresses applied to soils have complex effects on foundation soils in comparison with the pure principal stress rotation path. A series of undrained cyclic hollow torsional shear tests were performed on typical remolded soft clay from the Hexi area of Nanjing, China. The main control parameters were the tensile and compressive stress amplitude ratio(α) and the cyclic dynamic stress ratio(η). It was found that the critical η tended to remain constant at 0.13, when the value of the compressive stress amplitude was higher than the tensile stress amplitude. However, the influence of the tensile stress was limited by the dynamic stress level when α= 1.For obvious structural change in the soil, the corresponding numbers of cyclic vibration cycles were found to be independent of α at low stress levels and were only related to η. Finally, a new method for evaluating the failure of remolded soft clay was presented. It considers the influence of the tensile and compressive stresses which caused by complex stress paths of the principal stress rotation. This criterion can distinguish stable, critical, and destructive states based on the pore-water-pressure-strain coupling curve while also providing a range of failure strain and vibration cycles. These results provide the theoretical support for systematic studies of principal stress rotation using constitutive models.     

Equality testing for soil grid unit resolutions to polygon unit scales with DNDC modeling of regional SOC pools

Matching soil grid unit resolutions with polygon unit map scales is important to minimize the uncertainty of regional soil organic carbon(SOC) pool simulation due to their strong influences on the modeling.A series of soil grid units at varying cell sizes was derived from soil polygon units at six map scales,namely,1:50 000(C5),1:200 000(D2),1:500 000(P5),1:1 000 000(N1),1:4 000 000(N4) and 1:14 000 000(N14),in the Taihu Region of China.Both soil unit formats were used for regional SOC pool simulation with a De Nitrification-DeC omposition(DNDC) process-based model,which spans the time period from 1982 to 2000 at the six map scales.Four indices,namely,soil type number(STN),area(AREA),average SOC density(ASOCD) and total SOC stocks(SOCS) of surface paddy soils that were simulated by the DNDC,were distinguished from all these soil polygon and grid units.Subjecting to the four index values(IV) from the parent polygon units,the variations in an index value(VIV,%) from the grid units were used to assess its dataset accuracy and redundancy,which reflects the uncertainty in the simulation of SOC pools.Optimal soil grid unit resolutions were generated and suggested for the DNDC simulation of regional SOC pools,matching their respective soil polygon unit map scales.With these optimal raster resolutions,the soil grid units datasets can have the same accuracy as their parent polygon units datasets without any redundancy,when VIV 1% was assumed to be a criterion for all four indices.A quadratic curve regression model,namely,y = – 0.80 × 10~(–6)x~2 + 0.0228 x + 0.0211(R~2 = 0.9994,P 0.05),and a power function model R? = 10.394?~(0.2153)(R~2 = 0.9759,P 0.05) were revealed,which describe the relationship between the optimal soil grid unit resolution(y,km) and soil polygon unit map scale(1:10 000x),the ratio(R?,%) of the optimal soil grid size to average polygon patch size(?,km~2) and the ?,with the highest R~2 among different mathematical regressions,respectively.This knowledge may facilitate the grid partitioning of regions during the investigation and simulation of SOC pool dynamics at a certain map scale,and be referenced to other landscape polygon patches' mesh partition.     

Experimental study on large wood filtration performance by herringbone water-sediment separation structure

A herringbone water-sediment separation structure(hereinafter referred to as "herringbone structure") has been shown to be effective in separating coarse inorganic debris; however, less is known regarding the large wood(LW) filtration effect in this structure. This paper presents preliminary research on the wood filtration effect of the herringbone structure based on physical model tests.The results show that the herringbone structure exhibited effective performance in large wood size segregation, with a 100% component filtration rate for LW that diameter(D) larger than ribbed beam opening width(a). The total filtration rate also exceeded 80% when the Fraud number(Fr) is larger than 2.64 and increased with the increase of Fr. After exceeding Frmax, total filtration rate would be decreased due to overflow. Beside flow condition,structure parameters influence significantly on LW filtration rate. We attempt to explain the filtration process via particle contact trajectory and particle movement trajectory. The inclined angle of ribbed beam(γ) contributed the most variation to the filtration rate via influencing the coincidence with particle contact trajectory and particle movement trajectory. The high sensitivity coefficient of ribbed beam(θ) under relatively low Fr conditions implies remarkable influences on LW filtration effects by causing clogging problem. The ribbed beam opening width(a) together with LW diameter(D) influenced the size segregation performance.