Stereoselective Incorporation of Isoleucine into Cypridina Luciferin in Cypridina hilgendorfii (Vargula hilgendorfii)

The emission of light in the marine ostracod Cypridina hilgendorfii (presently Vargula hilgendorfii) is produced by the Cypridina luciferin-luciferase reaction in the presence of molecular oxygen. Cypridina luciferin has an asymmetric carbon derived from isoleucine, and the absolute configuration is identical to the C-3 position in L-isoleucine or D-alloisoleucine. To determine the stereoselective incorporation of the isoleucine isomers (L-isoleucine, D-isoleucine, L-alloisoleucine, and D-alloisoleucine), we synthesized four ²H-labeled isoleucine isomers and examined their incorporation into Cypridina luciferin by feeding experiments. Judging by these results, L-isoleucine is predominantly incorporated into Cypridina luciferin. This suggests that the isoleucine unit of Cypridina luciferin is derived from L-isoleucine, but not from D-alloisoleucine.     

Plankton cycles disguised by turbulent advection

Mathematical models used to represent plankton dynamics often display limit-cycle behavior in a range of realistic parameter values. However, experimental data do not show evidence of plankton oscillations besides externally driven seasonal blooms, casting doubts on the validity of the models themselves. In this work we show that spatial-temporal variability, coupled with advection by mesoscale turbulence, can disguise limit-cycle behavior to the point that it cannot be detected in fixed-point measurements of plankton abundance. The results presented here have more general implications as they indicate that the behavior of ecosystem models in the presence of advection can be very different from that occurring for homogeneous conditions. Care should thus be exercised in drawing general conclusions from the analysis of homogeneous ecosystem models.     

Stable isotope analysis as a tool to monitor dietary trends in little penguins Eudyptula minor

Long‐term dietary monitoring of seabirds can be used to relate population fluctuations to at‐sea events. Stomach flushing is a conventional dietary monitoring technique, but has a number of disadvantages. Stable isotope analysis (SIA) is a less invasive method that provides unbiased dietary information over a longer period. We evaluated stable isotope analysis as a potential tool for monitoring long‐term little penguin Eudyptula minor diet. We determined diet composition during the chick feeding stage using stomach flushing and SIA at three separate colonies, using spatial variation in diet as a surrogate for potential temporal variation. Bayesian isotopic mixing models were generated for blood and feathers to evaluate their ability to discriminate broad‐scale (fish, squid, crustaceans) and fine‐scale (individual prey species) diet composition. Differences in stable carbon and nitrogen isotope ratios were found between colonies: broad‐scale isotopic mixing models predicted different proportional contributions of broad taxa (fish, cephalopod, crustacean) to diet than was indicated by stomach samples, reflecting the bias incurred by one‐off stomach contents analysis. Fine‐scale isotopic mixing models predicted proportional contributions of prey items with less certainty. Blood isotopic mixing models had narrower confidence intervals than models for feathers, but trends in δ¹⁵N for feathers mirrored those for blood. Our results suggest that relying on stomach contents analysis to detect shifts in prey consumption in little penguins could be very misleading, resulting in a less‐than‐complete idea of total prey consumption. SIA of little penguin tissues could be used to monitor dietary shifts across dissimilar taxa that may affect population numbers, but would fail to detect shifts between fish species.     

Stable Isotope Evidence of Puma concolor (Felidae) Feeding Patterns in Agricultural Landscapes in Southeastern Brazil

We evaluated puma (Puma concolor) feeding patterns in southeastern Brazilian agricultural landscapes using carbon and nitrogen stable isotope analyses of hair collected from fecal samples (N = 64). We classified the samples into three groups: feeding patterns based on forest remnants, on the agricultural matrix or both. We observed a predominance of consumption of C₃ prey (~47% of individuals) in the area with the highest proportion of forest coverage. Conversely, C₄ prey were highly consumed (~40% of individuals) where the agricultural matrix was predominant. The δ¹³C values for pumas in both areas indicated that their food resources come from both forest remnants and the agricultural matrix and that some individuals preferentially consumed C₄ prey, indicating that food resources from the agricultural matrix make up most of their diet (~46% of prey individuals). The wide range of puma's δ¹⁵N values in both areas indicated a diet based on different types of prey. However, the C₄ group had higher values, indicating that both pumas and their prey feed on enriched resources from the agricultural matrix. The results confirm the high behavioral plasticity of pumas in using highly anthropogenic habitats. The stable isotope analyses conducted in this study yielded new information on large carnivore trophic ecology that might be useful in the development of new conservation strategies in disturbed areas.     

Biogeochemical properties of a tropical swamp forest ecosystem in southern Thailand

The distributions of organic matter in the tropical swamps in southern Thailand are reported. The concentrations of particulate and dissolved organic carbon (POC and DOC) in the Bang Nara River, which drains swamp forests and nearby paddy fields, were 2.9 ± 2.0 and 6.2 ± 1.3 mg C l⁻¹, respectively. Although the variation was large, DOC concentration in the Bang Nara River seemed to be higher than POC in November 1992 (DOC/POC ratio, 2.8 ± 2.2). River waters from the upland areas were characterized by low POC and DOC concentrations as compared with Bang Nara River water. The δ¹³C values of POC and river sediments were useful to distinguish between organic matter originating in upland and swamp areas. It is suggested that the distributions of organic matter and its isotopic composition reflect the difference in drainage characteristics between lowland swamp and upland areas. Isotopic analyses of plant leaves and soils revealed that the swamp forest ecosystems were characterized by low δ¹³C and low δ¹⁵N values, which suggested low efficiency of water use by plants and large contributions of atmospheric deposition of nitrogen, respectively. Although CO₂ recycling in the forest might be an important factor determining the δ¹³C values of understory plants, the main process in carbon metabolism of tropical swamp forests would be CO₂ exchange between the atmosphere and forest canopy. Received: May 1, 2001 / Accepted: September 28, 2001     

Comparative trophic impacts of two globally invasive cyprinid fishes reveal species‐specific invasion consequences for a threatened native fish

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An HPLC-CAD/fluorescence lipidomics platform using fluorescent fatty acids as metabolic tracers

Fluorescent lipids are important tools for live imaging in cell culture and animal models, yet their metabolism has not been well-characterized. Here we describe a novel combined HPLC and LC-MS/MS method developed to characterize both total lipid profiles and the products of fluorescently labeled lipids. Using this approach, we found that lipids labeled with the fluorescent tags, 4,4-difluoro-5,7-dimethyl-4-bora-3a,4a-diaza-s-indacene (BODIPY FL), 4,4-difluoro-5-(2-thienyl)-4-bora-3a,4a-diaza-s-indacene [BODIPY(558/568)], and dipyrrometheneboron difluoride undecanoic acid (TopFluor) are all metabolized into varying arrays of polar and nonpolar fluorescent lipid products when they are fed to larval zebrafish. Quantitative metabolic labeling experiments performed in this system revealed significant effects of total dietary lipid composition on fluorescent lipid partitioning. We provide evidence that cholesterol metabolism in the intestine is important in determining the metabolic fates of dietary FAs. Using this method, we found that inhibitors of dietary cholesterol absorption and esterification both decreased incorporation of dietary fluorescent FAs into cholesterol esters (CEs), suggesting that CE synthesis in enterocytes is primarily responsive to the availability of dietary cholesterol. These results are the first to comprehensively characterize fluorescent FA metabolism and to demonstrate their utility as metabolic labeling reagents, effectively coupling quantitative biochemistry with live imaging studies.     

TAILS Identifies Candidate Substrates and Biomarkers of ADAMTS7, a Therapeutic Protease Target in Coronary Artery Disease

Loss-of-function mutations in the secreted enzyme ADAMTS7 (a disintegrin and metalloproteinase with thrombospondin motifs 7) are associated with protection for coronary artery disease. ADAMTS7 catalytic inhibition has been proposed as a therapeutic strategy for treating coronary artery disease; however, the lack of an endogenous substrate has hindered the development of activity-based biomarkers. To identify ADAMTS7 extracellular substrates and their cleavage sites relevant to vascular disease, we used TAILS (terminal amine isotopic labeling of substrates), a method for identifying protease-generated neo–N termini. We compared the secreted proteome of vascular smooth muscle and endothelial cells expressing either full-length mouse ADAMTS7 WT, catalytic mutant ADAMTS7 E373Q, or a control luciferase adenovirus. Significantly enriched N-terminal cleavage sites in ADAMTS7 WT samples were compared to the negative control conditions and filtered for stringency, resulting in catalogs of high confidence candidate ADAMTS7 cleavage sites from our three independent TAILS experiments. Within the overlap of these discovery sets, we identified 24 unique cleavage sites from 16 protein substrates, including cleavage sites in EFEMP1 (EGF-containing fibulin-like extracellular matrix protein 1/Fibulin-3). The ADAMTS7 TAILS preference for EFEMP1 cleavage at the amino acids 123.124 over the adjacent 124.125 site was validated using both endogenous EFEMP1 and purified EFEMP1 in a binary in vitro cleavage assay. Collectively, our TAILS discovery experiments have uncovered hundreds of potential substrates and cleavage sites to explore disease-related biological substrates and facilitate activity-based ADAMTS7 biomarker development.     

同位素富集-稀释法研究食性转变对鱼类不同组织N同位素转化率的影响

稳定同位素技术广泛地用于描绘生态系统中食物网的食物来源和营养级关系,但是消费者不同组织转化率的研究相对较少。通过锦鲤摄食人工添加15N蓝藻的食性转化实验,研究不同组织N同位素转化率的差异,探讨组织生长和代谢对同位素转化的相对贡献,为不同时间尺度的稳定同位素研究取样奠定基础。结果表明,通过42d的加富蓝藻饲喂,各组织的N稳定同位素发生显著变化。肝的δ15N为(19.3±1.4)‰,显著高于其它组织,其次为鱼鳍((15.6±1.0)‰)和血液((12.6±0.4)‰),肌肉的δ15N‰最低,为(9.9±0.7)‰。在随后的同位素稀释实验中,锦鲤的体重增加,相对生长速率为0.011d-1,鳍肉的转化率最快,达到11.4%/d,半衰期仅为6.1d,其次是血液和肝,肌肉的转化率最低,仅有3.8%/d,半衰期最长,为18.4d。代谢衰减指数c和-1不存在显著差异,表明锦鲤各组织的N同位素转化主要由组织生长引起。结论显示,同位素富集-稀释法可以有效评价鱼类食性转变对不同组织同位素转化的差异,鳍肉和血液同位素分析可以作为锦鲤食性转变快速追踪的手段。     

Vertical distributions of stable isotopic compositions and bacteriochlorophyll homologues in suspended particulate matter in saline meromictic Lake Abashiri

We determined chloropigment composition as well as stable carbon and nitrogen isotopic compositions of dissolved and particulate species in saline meromictic Lake Abashiri. We observed a sharp peak of bacteriochlorophyll e in a narrow redox boundary zone and the upper monimolimnion, indicating a dense population of brown-colored strains of green sulfur bacteria around the chemocline. Nitrogen isotopic records of particulate nitrogen and dissolved ammonium suggested that the green sulfur bacteria in the redox boundary zone assimilated either ammonium or dinitrogen through the nitrogen fixation pathway. In the anoxic monimolimnion, several lines of evidence suggest that a major portion of particulate organic matter originated from the overlying mixolimnion and redox boundary zone.