Rapid measurement of deuterium-labeled long-chain fatty acids in plasma by HPLC-ESI-MS

Imbalanced fatty acid metabolism contributes significantly to the increased incidence of metabolic disorders. Isotope-labeled fatty acids (2H, 13C) provide efficient means to trace fatty acid metabolism in vivo. This study reports a new and rapid method for the quantification of deuterium-labeled fatty acids in plasma by HPLC-MS. The sample preparation protocol developed required only hydrolysis, neutralization, and quenching steps followed by high-performance liquid chromatography-electrospray ionization-mass spectrometry analysis in negative ion mode using single ion monitoring. Deuterium-labeled stearic acid (d7-C18:0) was synthesized to reduce matrix interference observed with d5 analog, which improved the limit of detection (LOD) significantly, depending on the products analyzed. Linearity > 0.999 between the LOD (100 nM) and 30 microM, accuracy > 90%, precision > 88%, and adequate recovery in the dynamic range were obtained for d7-C18:0 and d7-oleic acid (C18:1). Upon oral dosing of d7-C18:0 in rats, the parent compound and its desaturation and beta-oxidation products, d7-C18:1 and d7-C16:0, were circulating with a maximal concentration ranging from 0.6 to 2.2 microM, with significant levels of d7-fatty acids detected for up to 72 h.     

Levels of endemism are not necessarily biased by the co-presence of species with different range sizes: a case study of Vilenkin and Chikatunov's models

Aim  To illustrate problems in the methods proposed by B. Vilenkin and V. Chikatunov to study levels of endemism and species–area relationships.
Location  The study used data on the distribution of tenebrionid beetles (Coleoptera, Tenebrionidae) on the Aegean Islands (Greece).
Methods  A total of 32 islands and 170 taxa (species and subspecies) were included in this study. Levels of endemism were evaluated both as the proportion of endemic taxa, and according to the methods proposed by Vilenkin and Chikatunov, which are based on the number of non-endemic taxa and various relationships with area. A model of the species–area relationship proposed by these authors was also analysed.
Results  The number of endemic taxa was positively correlated with the number of taxa with different distribution types, but this positive correlation did not influence the estimation of the level of endemism. In fact, the commonly used estimate of endemicity as a percentage was strongly correlated with the endemism values calculated according to the method of Vilenkin and Chikatunov. The usual power function fitted the species–area relationship as well as the most complicated method of Vilenkin and Chikatunov.
Main conclusions  As hypothesized by Vilenkin and Chikatunov, the number of endemic taxa was influenced both by the number of taxa of other biogeographical ranks, and by an island's area. However, explanations for the positive relationship between the number of endemic taxa and taxa of different biogeographical ranks are equivocal. Importantly, this relationship did not necessarily influence the level of endemism, which could be expressed adequately by percentages. The method proposed by Vilenkin and Chikatunov to estimate the species–area relationship cannot be clearly justified on theoretical grounds and is of questionable practical utility.     

鸡粪有机肥对设施菜地土壤重金属和微生物群落结构的影响

通过田间试验,研究在设施菜地上施用不同剂量的鸡粪有机肥对土壤-植物系统中重金属的累积、重金属有效性和土壤微生物群落结构的影响,进一步探讨土壤微生物群落结构与土壤重金属之间的相关关系。结果表明,与对照相比,施用有机肥可提高小白菜地上部生物量,其中施肥量为60 t/hm²时值最大,增幅为59.92%;小白菜地上部Cd、Cr、Cu、Zn和As含量均大幅增加,但Pb含量无明显变化。土壤重金属Cd、Cr、Cu、Zn和As的全量均随鸡粪有机肥施加量的增加而增大,最高增幅分别为21.30%、21.58%、17.40%、19.40%和17.43%,出现明显的累积现象;施用有机肥均增加了土壤Cd、Cr、Cu、Zn和As的有效态含量,而Pb的全量和有效态含量无显著变化;除重金属Pb外,不同重金属元素全量与有效态含量均显著正相关,其中元素Zn的全量与有效态含量相关性最强。磷脂脂肪酸（PLFA）分析结果表明,土壤中含量较高的PLFA为16：0、18：1ω7c、10Me16：0和18：1ω9c,土壤微生物总PLFA和各类群PLFA含量均呈现M0.5 > M1 > CK > M2 > M4;相关性分析结果表明,土壤Cu、As全量和Cd、Cr、Cu、As有效态含量与微生物总PLFA和各类群PLFA含量均呈现显著负相关关系,其中有效态Cr和Cu含量对微生物群落结构的影响最为显著。     

Effect of visible light on superoxide dismutase (SOD) activity in the red alga Gracilariopsis tenuifrons (Gracilariales, Rhodophyta)

Superoxide dismutase (SOD) was studied in the agarophyte Gracilariopsis tenuifrons. Similar SOD activity (130 ± 9U mg^-1) was observed in material from different regions of SouthAmerica, from different phases of the life cycle (gametophytes andtetrasporophytes), and from apical and basal sections of the thallus.In alga grown under a light-dark cycle, SOD activity in samples takenat different times exhibited a diurnal rhythm. The activity measured duringthe day phase was twice as much as during the night phase. This rhythm didnot persist under constant light, indicating light regulation of SOD activity.SOD activity was tested in algae submitted to different light intensities anddifferent wavelengths. It increased with the light intensity. The blue lightwavelength exerted a greater induction of SOD activity than other specificwavelengths.     

Factors influencing the seasonal life history of the pitcher-plant mosquito, Wyeomyia smithii

Abstract.  1. The effects of resource levels, thermal microclimate, and seasonal oviposition patterns on fecundity and survivorship in the pitcher-plant mosquito, Wyeomyia smithii (Coq.), were examined at a northern Wisconsin bog over the course of 2 years. Wyeomyia smithii are bivoltine at this locality, thereby enabling the study of summer and overwintering generations separately.
2. Nutrient resources of W. smithii were not limiting and there was no indication of density-dependent survivorship or fecundity.
3. Oviposition rates were highest in young, large pitchers and individual mosquitoes appeared to allocate only a few eggs to any one leaf.
4. Winter was the harsh season, and the principal manifestation of seasonal harshness was reduced survivorship.
5. Overwintering W. smithii that had been oviposited later in the summer had a higher odds of survival than those oviposited earlier in the summer.
6. It was concluded that dispersal of eggs among many pitchers serves to spread the risk of encountering lethal winter temperatures among spatially unpredictable patches.     

Quantitative genetics of natural variation of behavior in Drosophila melanogaster: the possible role of the social environment on creating persistent patterns of group activity

Using a set of nine effectively isogenic lines collected from nature in 1998, we observed unperturbed behaviors of mixed-sex groups of Drosophila melanogaster. We repeatedly scanned replicated groups of genetically identical individuals, five females and five males, and recorded the behavior of each individual (i.e., walking, feeding, grooming, flying, courting, mating, fighting, or resting). From these behaviors, we made a composite variable of activity for our quantitative genetic analysis. Genotypes differed in activity, explaining 14.41% of the variation in activity; 8.60% of the variation was explained by a significant genotype x sex interaction, which signifies genetic variation for sexual dimorphism in behavior. Phenotypic plasticity explained 11.13% of the variation in activity. Different genotypes and sexes within genotypes had different rank orders of the component behaviors that contribute to activity. We found no effect of common rearing environment. Instead, differences between replicate groups within genotype accounted for 19.47% variation in activity, and activity was significantly repeatable across scans. This emergent group behavior is likely caused by differences between groups of interacting individuals, even though individuals were genetically identical across groups. Thus, emergent group behavior explained almost as much variation in activity as the combined sources of genetic variation (23.01%), and this is an additional level on which selection could operate: individuals and groups. We discuss how differences among groups could change patterns of additive genetic variation available for evolution. Furthermore, because the behavior of an individual is influenced by conspecifics, genotype interactions between individuals could contribute to indirect selection. Finally, if we consider activity as a syndrome governing all component behaviors with strong genetic correlations among behaviors within an individual, then these component behaviors cannot evolve independently. These results suggest that reductionist approaches of molecular behavior genetics may be incomplete and/or misleading when considering similar phenotypes at the population level or when trying to understand how behaviors evolve.     

Metabolic preconditioning of cells with AICAR-riboside: Improved cryopreservation and cell-type specific impacts on energetics and proliferation

In species whose evolutionary history has provided natural tolerance to dehydration and freezing, metabolic depression is often a pre-requisite for survival. We tested the hypothesis that preconditioning of mammalian cells with 5-aminoimidazole-4-carboxamide-1-b-d-ribofuranoside (AICAR) to achieve metabolic depression will promote greater survivorship during cryopreservation. AICAR is used extensively to stimulate AMP-activated protein kinase (AMPK), which can result in downregulation of biosynthetic processes. We showed that the metabolic interconversion of AICAR was cell-type dependent. Accumulation of 5-aminoimidazole-4-carboxamide-1b-d-ribofuranosyl-5′-monophosphate (ZMP), as well as other metabolites that possess multiple phosphates (i.e., ZDP, ZTP), varied approximately 3.5-fold across the cell lines tested. AICAR treatment also significantly influenced the concentrations of cellular adenylates (ATP, ADP, and AMP). Depression of cell metabolism and proliferation with AICAR treatment differed among cell lines. Proliferation for a given cell line was negatively correlated with the fold-increase achieved in the ‘effective adenylate ratio’ ([AMP] + [ZMP])/[ATP]) after AICAR treatment. Metabolic preconditioning with AICAR promoted a significant increase in viability post-freezing in J774.A1 macrophages, HepG2/C3A cells and primary hepatocytes but not in NIH/3T3 fibroblasts or OMK cells. The effect of AICAR on viability after freezing was positively correlated (r² = 0.94) with the fold-increase in the ‘effective adenylate ratio’. Thus for each cell line, the greater the depression of metabolism and proliferation due to preconditioning with AICAR, the greater was the survivorship post-freezing.     

Numerical exploration of the influence of neural noise on the psychometric function at low stimulation intensity levels

The relationship between stimulus intensity and the probability of detecting the presence of the stimulus is described by the psychometrical function. The probabilistic nature of this relationship is based on the stochastic behaviour of sensory neural channels and sensory networks involved in perceptual processing (Kiang 1968). This study tries to establish a continuum of variability across different levels of integration in the central nervous system. Once the opening and closing times of ionic channels was simulated, a threshold to the collective behaviour of voltage-gated ionic channels was imposed in order to generate the spike train of a single neuron. Afterwards, the trains of spikes of different neurons were added up, simulating the activity of a sensory nerve. By adding the activity due to the stimulus to the spontaneous neural behaviour, the psychometric function was simulated using a thresholding approach. The results can replicate the stochastic resonance phenomenon, but also open up the possibility that attentional phenomena can be mediated not only by increasing neural activity (bursting or oscillatory), but also by increasing noise at the neural level.     

Behaviour of interleukin-2 serum levels in advanced non-small-cell lung cancer patients: relationship with response to therapy and survival

 Interleukin(IL)-2 is a T helper (Th) 1 type cytokine that has been shown to play an important role in antitumour immune responses. In this study, the prognostic significance of serum IL-2 levels was investigated in 60 advanced non-small-cell lung cancer (NSCLC) patients. IL-2 serum levels were determined before chemotherapy, at the end of chemotherapy and during follow-up, using a commercially available enzyme-linked immunoadsorbent assay kit. The results were analysed according to the response to therapy and were used to generate a model predicting overall survival and time to treatment failure. All 60 patients were shown to have higher IL-2 serum levels than controls (P < 0.0001). Stage IV patients had significantly lower IL-2 levels than stage III patients (P < 0.0001), although they were still significantly higher than controls (P < 0.0001). It is interesting that, when patients were divided into responders and non-responders according to the response to therapy, the former were shown to have significantly higher pre-chemotherapy levels than the latter (P < 0.0001). Moreover, a further significant increase in IL-2 serum levels (P=0.004) and a significant decrease (P < 0.0001) were shown in responders and non-responders, respectively at the end of the therapy. Using univariate and multivariate analyses, both overall survival and time to treatment failure were shown to be affected by the mean pathological levels of IL-2. Furthermore, the prognostic significance of the serum level of IL-2 was confirmed by the stepwise regression analysis. In conclusion, determination of pre-treatment IL-2 serum levels was shown to be of independent prognostic utility in patients with advanced NSCLC; therefore, its possible use for prediction of outcome is proposed. Received: 16 March 2000 / Accepted: 27 July 2000