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).     

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.     

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.     

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.     

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.     

Structures and antiviral activities of butyrolactone derivatives isolated from Aspergillus terreus MXH-23

A new butyrolactone derivative, namely butyrolactone VIII(1), and six known butyrolactones(2-7) were separated from the ethyl acetate(Et OAc) extract of the fermentation broth of a fungus, Aspergillus terreus MXH-23. The chemical structures of these metabolites were identified by analyzing their nuclear magnetic resonance(NMR) and mass spectrometry(MS). Known butyrolactone derivatives contain an α,β-unsaturated γ-lactone ring with α-hydroxyl and γ-benzyl, and butyrolactone VIII(1) was the first butyrolactones contains α-benzyl and γ-hydroxyl on α,β-unsaturated lactone ring. All of the butyrolactone derivatives were tested for their anti-influenza(H1N1) effects. Derivatives 4 and 7 showed moderate antiviral activities while the newly-identified, derivative 1, did not.     

Effects of nitrogen and phosphorus on the growth of <Emphasis Type="Italic">Levanderina fissa</Emphasis>: How it blooms in Pearl River Estuary

Effects of nitrogen (N) and phosphorus (P) from different sources and at different concentrations on the growth of Levanderina fissa (= Gyrodinium instriatum) were studied in laboratory conditions. The findings might explain the recurrent blooms of this species in Pearl River Estuary, China. Results showed that nutrient limitation significantly inhibited the growth of L. fissa. The values of specific growth rate (μ _max) and half-saturation nutrient concentration (K _S) were 0.37 divisions/d and 8.49 μmol L^?1 for N, and 0.39 divisions/d and 1.99 μmol L^?1 for P, respectively. Based on K _S values, dissolved inorganic N level in PRE was sufficient to support the high proliferation of L. fissa, while dissolved inorganic P concentration was far lower than the minimum requirement for its effective growth. L. fissa was not able to utilize dissolved organic N (DON) compounds such as urea, amino acids, and uric acid. However, it grew well by using a wide variety of dissolved organic P (DOP) sources like nucleotides, glycerophosphate, and 4-nitrophenylphosphate. The results from this study suggested that the ability in DOP utilization of L. fissa offers this species a competitive advantage in phytoplankton communities. The high level and continuous supply of DIN, enrichment of DOP, together with warm climate and low salinity in the Pearl River Estuary provided a suitable nutrient niche for the growth of L. fissa, and resulted in the recurrent blooms in the estuary.     

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.     

Effect of benzo[a]pyrene on detoxification and the activity of antioxidant enzymes of marine microalgae

The objective of this study was to examine the effect of benzo[a]pyrene(Ba P) on the detoxification and antioxidant systems of two microalgae,Isochrysis zhanjiangensis and Platymonas subcordiformis.In our study,these two algae were exposed to Ba P for 4 days at three different concentrations including 0.5 μg~(L-1)(low),3 μg~(L-1)(mid) and 18 μg~(L-1)(high).The activity of detoxification enzymes,ethoxyresorufin O-deethylase(EROD) and glutathione S-transferase(GST) increased in P.subcordiformis in all Ba P-treated groups.In I.zhanjiangensis,the activity of these two enzymes increased at the beginning of exposure,and then decreased in the groups treated with mid-and high Ba P.The activity of antioxidant enzyme superoxide dismutase(SOD) increased in I.zhanjiangensis in all Ba P-treated groups,and then decreased in high Ba P-treated group,while no significant change was observed in P.subcordiformis.The activity of antioxidant enzyme catalase(CAT) increased in I.zhanjiangensis and P.subcordiformis in all Ba Ptreated groups.The content of malondialdehyde(MDA) in Isochrysis zhanjiangensis increased first,and then decreased in high Ba P-treated group,while no change occurred in P.subcordiformis.These results demonstrated that Ba P significantly influenced the activity of detoxifying and antioxidant enzymes in microalgae.The metabolic related enzymes(EROD,GST and CAT) may serve as sensitive biomarkers of measuring the contamination level of Ba P in marine water.     

The effect of chemical cues on the settlement of sea cucumber (Apostichopus japonicus) larvae

The effects of four ions and eight neuroactive compounds on inducing larval settlement of A. japonicus were assessed in the present study. All bioassays were conducted in 60 × 9 mm Petri dishes, each contained 10 mL of the test solution and 10 doliolaria larvae. There were significant inductive effects of K+(10- mmol L-1), NH+4(0.1 mmol L1), GABA(10-3 mol L-1), acetylcholine(10-5 mol L-1), L-DOPA(10-5 mol L-1), norepinephrine(10-5 mol L-1) and dopamine(10-7 mol L-1 and 10-5 mol L-1) on the settlement of sea cucumber larvae. L-DOPA and dopamine are the most efficient chemical cues to induce A. japonicus larvae to settle. The highest percentage of larval settlement was induced by 10-5 mol L-1 L-DOPA and dopamine(33% and 40%) compared to the control(7%). However, Ca2+, Mg2+, choline, serotonin, and epinephrine were less effective on larval settlement at all tested concentrations. This study evaluated the stability and feasibility of chemical cues for larval settlement in different culture systems, which can be applied to improve the hatchery production of this valuable species.     

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.     

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.     

Distribution of sediment chloroplastic pigments in the Southern Yellow Sea,China

The distribution of sediment chloroplastic pigments (Chl-a, i.e. chlorophyll a and Pha-a, i.e. phaeophorbide a) in the Southern Yellow Sea of China was studied. Samples were collected from four cruises in January and June 2003, and January and June 2004. The results show that the vertical distribution of Chl-a and Pha-a in the sediment layers 0-2cm, 2-5cm and 5-8cm, follows a stable ratio, 5:3:2. The average ratio of Pha-a to Chl-a in sediment is 2.83. Spearman 2-tailed rank correlation analysis shows that Chl-a and Pha-a contents in each sediment layer have a highly significant correlation. The average contents of Chl-a and Pha-a in the sediment of the 0-8cm layer in the investigated area are 0.31 -0.47μgg-1 and 1.28-1.40 μgg-1 sediment (dry weight), respectively. The average Chl-a and Pha-a contents in sediment are higher in summer than in winter. ANOVA analysis shows that there is a highly significant variation among the Chl-a contents (P = 0.002 <0.01) of the four cruies, but this is not true for the case of Pha-a content (P = 0.766>0.05). The average Chl-a and Pha-a contents in the 2 sediment layers (0-2cm and 2-5cm) have significant or highly significant correlations with organic matter (OM), median diameter (Mdφ), silt plus clay percentage in the January 2003 cruise. In the June 2003 cruise, the average Chl-a content in the 3 sediment layers (0-2cm, 2-5cm, and 5-8cm) has a significant correlation with meiofauna biomass, and Pha-a content has highly significant correlations with water depth, bottom water temperature, OM and Mdφ The contents of Chl-a and Pha-a are lower than those in estuaries and intertidal areas, but close to those in the same area studied previously.     

De-eutrophication of effluent wastewater from fish aquaculture by using marine green alga Ulva pertusa

The de-eutrophication abilities and characteristics of Ulva pertusa, a marine green alga, were investigated in Qingdao Yihai Hatchery Center from spring to summer in 2005 by analyzing the dynamic changes in NH ₄ ⁺ , NO ₃ ^? , NO ₂ ^? as well as the total dissolved inorganic nitrogen (DIN). The results show that the effluent wastewater produced by fish aquaculture had typical eutrophication levels with an average of 34.3 μmol L^?1 DIN. This level far exceeded the level IV quality of the national seawater standard and could easily lead to phytoplankton blooms in nature if discarded with no treatment. The de-eutrophication abilities of U. pertusa varied greatly and depended mainly on the original eutrophic level the U. pertusa material was derived from. U. pertusa used to living in low DIN conditions had poor DIN removal abilities, while materials cultured in DIN-enriched seawater showed strong de-eutrophication abilities. In other words, the de-eutrophication ability of U. pertusa was evidently induced by high DIN levels. The de-eutrophication capacity of U. pertusa seemed to also be light dependent, because it was weaker in darkness than under illumination. However, no further improvement in the de-eutrophication capacity of U. pertusa was observed once the light intensity exceeded 300 μmol M² S^?1. Results of semi-continuous wastewater replacement experiments showed that U. pertusa permanently absorbed nutrients from eutrophicated wastewater at a mean rate of 299 mg/kg fresh weight per day (126 mg/kg DIN during the night, 173 mg/kg in daytime). Based on the above results, engineered de-eutrophication of wastewater by using a U. pertusa filter system seems feasible. The algal quantity required to purify all the eutrophicated outflow wastewater from the Qingdao Yihai Hatchery Center into oligotrophic level I clean seawater was also estimated using the daily discharged wastewater, the average DIN concentration released and the de-eutrophication capacity of U. pertusa.     

Assessment and comparison of <Emphasis Type="Italic">in vitro</Emphasis> immunoregulatory activity of three astaxanthin stereoisomers

In recent years, the immune-modulatory role of all-trans astaxanthin from different pigment sources has been studied. It was reported that all-trans astaxanthin might exist as three stereoisomers, and the composition of all-trans stereoisomers in natural materials differs from that of synthetic products. However, the different biological effects of various all-trans stereoisomers still remain unclear. In the present study, we evaluated the bioactivity of three astaxanthin stereoisomers, (3S,3’S)-trans-, (3R,3’R)-trans-and meso-trans-astaxanthin, in regulating cell-mediated immune response using mice lymphocytes and peritoneal exudates cells (PECs) systems. After the treatment with three astaxanthin stereoisomers (20 μmol L^?1), the lymphocyte proliferation capacity, neutral red phagocytosis of PECs and natural killer (NK) cell cytotoxic activity were comparatively assessed. The results showed that all three astaxanthin stereoisomers significantly promoted lymphocyte proliferation, phagocytic capacity of PECs, and cytotoxic activity of NK cells. Moreover, the (3S,3’S)-trans-astaxanthin exhibited a much higher response than others.     

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.     

Distribution patterns of chlorophyll <Emphasis Type="Italic">a</Emphasis> in spring and autumn in association with hydrological features in the southern Yellow Sea and northern East China Sea

Two field studies were conducted to measure pigments in the Southern Yellow Sea (SYS) and the northern East China Sea (NECS) in April (spring) and September (autumn) to evaluate the distribution pattern of phytoplankton stock (Chl a concentration) and the impact of hydrological features such as water mass, mixing and tidal front on these patterns. The results indicated that the Chl a concentration was 2.43±2.64 (Mean ± SD) mg m^?3 in April (range, 0.35 to 17.02 mg m^?3) and 1.75±3.10 mg m^?3 in September (from 0.07 to 36.54 mg m^?3) in 2003. Additionally, four areas with higher Chl a concentrations were observed in the surface water in April, while two were observed in September, and these areas were located within or near the point at which different water masses converged (temperature front area). The distribution pattern of Chl a was generally consistent between onshore and offshore stations at different depths in April and September. Specifically, higher Chl a concentrations were observed along the coastal line in September, which consisted of a mixing area and a tidal front area, although the distributional pattern of Chl a concentrations varied along transects in April. The maximum Chl a concentration at each station was observed in the surface and subsurface layer (0–10 m) for onshore stations and the thermocline layer (10–30 m) for offshore stations in September, while the greatest concentrations were generally observed in surface and subsurface water (0–10 m) in April. The formation of the Chl a distributional pattern in the SYS and NECS and its relationship with possible influencing factors is also discussed. Although physical forces had a close relationship with Chl a distribution, more data are required to clearly and comprehensively elucidate the spatial pattern dynamics of Chl a in the SYS and NECS.     

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.     

Speciation analysis of arsenic compounds in seafood by ion chromatography-atomic fluorescence spectrometry

Ion chromatography-ultra violet-hydride generation-Atomic Florescence Spectrometry was applied to detect 5 arsenic species in seafoods. The arsenic species studied include arsenobetaine(As B), arsenite(As(III)), dimethylarsinic acid(DMA), monomethylarsonic acid(MMA), and arsenate(As(V)), which were extracted from samples using 2% formic acid. Gradient elution using 33 mmol L~(-1) CH_3COONH_4 and 15 mmol L~(-1) Na_2CO_3 with 10 mL CH_3CH_2OH at pH 8.4 allowed the chromatographic separation of all the species on a Hamilton PRP-X100 anion-exchange column in less than 8 min. In this study, an ultrasound extraction method was used to extract arsenic species from seafood. The extraction efficiency was good and the recoveries from spiked samples were in the range of 72.6%–109%; the precision between sample replicates was higher than 3.6% for all determinations. The detection limits were 3.543 μg L~(-1) for As B, 0.4261 μg L~(-1) for As(III), 0.216 μg L~(-1) for DMA, 0.211 μg L~(-1) for MMA, and 0.709 μg L~(-1) for As(V), and the linear coefficients were greater than 0.999. We also developed an application of this method for the determination of arsenic species in bonito, Euphausia superba, and Enteromorpha with satisfactory results. Therefore, it was confirmed that this method was appropriate for the detection of arsenic species in seafood.     

Coral Ba/Ca and Mn/Ca ratios as proxies of precipitation and terrestrial input at the eastern offshore area of Hainan Island

Geochemical ratios in coral reef skeletons could be used as proxies to reconstruct past climatological and environmental records in data-poor regions. Using a 103-year data set(1902 to 2005), the annual variations in Ba/Ca and Mn/Ca ratios of Porites lutea skeletons at an eastern offshore area of Hainan Island(19?12'28.4'N, 110?37'38.8'E) were analyzed using inductively coupled plasma-optic emission spectrometry(ICP-OES). The analysis results showed that Ba/Ca ratios varied from a minimum of 3.120 μmol mol~(-1) in 1903 to a maximum of 10.064 μmol mol~(-1) in 1944, with an average of 5.256 μmol mol~(-1). Mn/Ca ratios varied from 0.206 to 5.708 μmol mol~(-1) with an annual average of 1.234 μmol mol~(-1), with peak values in 2001, 1964 and 1932, that correlated with strong rainfall events caused by typhoons. Variation in Ba/Ca and Mn/Ca ratios were compared with available river discharge and precipitation records, providing insight into past climatological events. Human activities and their indirect effects could impact the strength of the relationship between Ba/Ca and Mn/Ca ratios and observed precipitation and terrestrial input in the future.