Diffusion and interaction mechanism of rejuvenating agent with virgin and recycled asphalt binder: a molecular dynamics study

The quality of asphalt binder recycling is largely dependent on molecular diffusion between virgin asphalt binder, aged asphalt binder, and rejuvenator. In this study, molecular dynamics (MD) simulations were used to study diffusion and interaction mechanism of rejuvenating agent in recycled asphalt binder. The diffusion process of rejuvenator into virgin and aged asphalt binder was studied using a three-layered model. A mixture model of virgin and aged asphalt binder was built to evaluate the effect of rejuvenator on the molecular structure of asphalt binder, such as nanoaggregate behaviour and translational mobility. The simulation results of the layered model suggest that rejuvenator may improve blending efficiency of virgin and aged asphalt binder depending on temperature. The calculated inter-diffusion coefficients indicate that the rejuvenator diffuses faster into virgin asphalt binder than aged asphalt binder. The radial distribution functions of asphaltene, resin, and aromatic pairs show that rejuvenator causes the molecular structures of virgin and aged asphalt binder more similar to that of virgin asphalt binder. The rejuvenator reduces the self-association trends of asphaltene molecules, but saturates from local aggregation inside the rejuvenated asphalt binder. On the other hand, rejuvenator increases translational mobility of saturate, aromatic, resin and asphaltene fractions.     

Modulation of calcium mobilization in aortic rings of pregnant rats: Contribution of extracellular calcium and of voltage-operated calcium channels

Pregnancy is associated with decreased vascular responsiveness to vasopressor stimuli. We have tested the involvement of Ca2+ mobilization in myotropic responses of aortic rings obtained from pregnant and virgin rats. Contractions of the rings to phenylephrine, in the absence of calcium in the bathing medium, were lower in tissues from virgin than from pregnant rats. Concentration-response curves to CaCl2 that were measured after stimulation by phenylephrine in the absence of Ca2+ were shifted to higher levels of contraction. This was not observed when KCl was used to prestimulate the aorta. D-600, a phenylalkylamine calcium channel blocker, similarly inhibited these responses to CaCl2 in tissues from both pregnant and virgin animals. D-600 exerted a concentration-dependent inhibition of responses to phenylephrine and KCl. However, the calcium antagonist was less effective in aortic rings of pregnant than of virgin rats. Basal 45Ca2+ uptake was lower in aortic rings from pregnant than from virgin rats, and Bay K 8644 was unable to reverse this difference. The time course of basal and stimulated (KCl) 45Ca2+ influx was lower in aorta of pregnant rats at all times studied. Moreover, when the intracellular calcium pools were emptied with phenylephrine, the refilling of these pools was delayed in aortic rings of pregnant rats. These results indicate an altered extracellular calcium mobilization of aortic rings from pregnant rats. These changes may be due to a functional alteration of the voltage-operated calcium channels during pregnancy.     

A stochastic model for CD8(+)T cell dynamics in human immunosenescence: implications for survival and longevity.

We propose here a stochastic model for the CD 8(+)T lymphocyte dynamics on the long time-scale of the human lifespan. Our purpose has been to test the hypothesis, recently proposed on the basis of our experimental data (Fagnoni et al., 2000), that the depletion of virgin CD8(+)T lymphocytes can be considered a reliable biomarker related to the risk of death. This hypothesis is embedded in a more general theory of immunosenescence according to which the accumulation of antigen experienced (AE) T cells and the concomitant exhaustion of antigen non-experienced (ANE) T cells with age, mostly due to the chronic lifelong exposure to antigens, is a major characteristic of the remodeling of the human immune system with age. In our model we considered a deterministic balance of ANE and AE T cell concentrations plus a stochastic forcing, which describes the chronic antigenic stress fluctuations, assuming a mean genetically determined capability of individuals to respond to antigens. The major results of our model is the validation of the above-mentioned hypothesis, since the model is capable of fitting the experimental data concerning the changes of ANE T cell concentration over age, and at the same time to reproduce survival curves similar to the demographic ones. Furthermore, the stochastic process results in being responsible for the peculiar shape of the survival curves.     

Influence of age and parity on the susceptibility of rat mammary gland epithelial cells in primary cultures to 7,12-dimethylbenz(a) anthracene

Summary Mammary gland epithelial cells from rats of different ages or with different reproductive histories vary in their proliferative properties and susceptibility to dimethylbenz(a)anthracene (DMBA) carcinogenesis in vivo. The present study was carried out to determine whether these differences are maintained under in vitro conditions. Primary cultures of mammary gland epithelial cells of young virgin, old virgin, and parous rats were treated with various doses of DMBA. Growth rates, DNA synthesis, and dose-response curves were determined; the toxicity of DMBA was measured by its effect on cell growth. Cell morphology was studied by transmission and scanning electron microscopy. Epithelial cells from the mammary gland of young virgin rats adapted rapidly to the culture conditions, behaving as if the cells were in the logarithmic phase of growth prior to plating. Mammary gland epithelial cells from old virgin and parous rats required a lag period prior to cell growth during which the proliferating cells adapted to the culture conditions. Cells from each group had comparable doubling times, and DNA synthesis peaked approximately 1 d after initiation of growth in culture. The numbers of proliferating cells decreased with increasing age and parity of the donor. Mammary gland epithelial cells of young virgin rats were more susceptible to both low and high doses of DMBA than those of old virgin and parous rats when the carcinogen was added either 24 h after plating or at the peak of DNA synthesis. These results indicate that age and parity influence the proliferative status of the cells and their susceptibility to DMBA in vitro, simulating in that way the in vivo situation. Supported by Public Health Service Grants CA-23539 and CA-27026 from the National Cancer Institute and by an Institutional grant from the United Foundation of Greater Detroit.     

The metabolism of acetone in the pregnant rat

The administration of a single dose of acetone (100 mg/kg bw) to virgin and 21-day pregnant rats resulted in the appearance of relatively high concentrations of 1,2-propanediol, acetol and methylglyoxal in plasma and liver. In the fetuses no methylglyoxal was detectable. The acetone metabolism curves tend to indicate that the capacity for acetone disposal may be enhanced in the 48 hr-fasted pregnant rat, thus enabling the animal to re-use acetone metabolically, possibly for accelerated gluconeogenesis.     

The effect of cleaving the reactive-site peptide bond Lys-15--Ala-16 on the conformation of bovine trypsin-kallikrein inhibitor (K unitz) as revealed by solvent-perturbation spectra, circular dichroism and fluorescence.

Spectroscopic measurements of virgin bovine trypsin-kallikrein inhibitor and its modified species (in which the reactive-site peptide bond Lys-15--Ala-16 is split) indicate a conformational difference between both proteins. The inhibitor contains four tyrosines but no tryptophan. In the modified inhibitor a tyrosyl blue shift is seen in the difference absorption spectrum of modified against virgin inhibitor. The solvent perturbation spectra show an increase of the fraction of exposed tyrosyls from 0.45 in the virgin inhibitor to 0.59 in the modified form. Comparison of the circular dichroism spectra of the modified and virgin inhibitors reveals a decrease of the mean residue ellipticity in the tyrosine and peptide bond region of the modified inhibitor. In the fluorescence spectra a 50% increase in the quantum yield of the tyrosine fluorescence is observed in the modified inhibitor. All these spectroscopic data support the idea, which is also evidenced by the X-ray crystallographic model, that in the modified inhibitor up to five residues from Ala-16 to Arg-20 gain rotational freedom.     

The evolutionary dynamics of adaptive virginity,sex‐allocation,and altruistic helping in haplodiploid animals

In haplodiploids, females can produce sons from unfertilized eggs without mating. However, virgin reproduction is usually considered to be a result of a failure to mate, rather than an adaptation. Here, we build an analytical model for evolution of virgin reproduction, sex‐allocation, and altruistic female helping in haplodiploid taxa. We show that when mating is costly (e.g., when mating increases predation risk), virginity can evolve as an adaptive female reproductive strategy. Furthermore, adaptive virginity results in strongly divergent sex‐ratios in mated and virgin queen nests (“split sex ratios”), which promotes the evolution of altruistic helping by daughters in mated queen nests. However, when helpers evolve to be efficient and increase nest production significantly, virgin reproduction is selected against. Our results suggest that adaptive virginity could have been an important stepping stone on the pathway to eusociality in haplodiploids. We further show that virginity can be an adaptive reproductive strategy also in primitively social haplodiploids if workers bias the sex ratio toward females. By remaining virgin, queens are free to produce sons, the more valuable sex in a female‐biased population. Our work brings a new dimension to the studies linking reproductive strategies with social evolution.     

Lymphokine-induction of memory B-cell differentiation: differential stimulation of large virgin and memory B-cell differentiation

In order to compare and contrast the requirements of virgin and memory B cells for B-cell differentiation factors, a model system was developed in which low-density rat B cells isolated from 4-week primed antigen-draining lymph nodes were cultured in vitro. This large low-density cell population contained B cells which were 90% surface IgM positive and 60% IgD positive and showed moderately elevated Ia staining. When the cell population was stimulated with antigen plus lymphokines or lymphokines alone, antigen-specific IgG antibody was secreted; this was used as a measure of memory cell differentiation. When the cell population was stimulated with mitogen (lipopolysaccharide plus dextran sulfate) plus lymphokines, polyclonal IgG and IgM secretion was seen and was used as a measure of virgin B-cell differentiation. Using this system, we found that lymphokines contained in a Con A-induced rat spleen cell supernatant (CSN) were sufficient to drive both memory and virgin B-cell differentiation. In contrast, lymphokines contained in the supernatant from the murine T-cell hybridoma B151K12 (B151CFS) were able to induce large amounts of polyclonal IgM and IgG secretion but did not support memory B-cell differentiation. When recombinant human IL-2 was added to these cultures, it acted synergistically to augment virgin B-cell differentiation, but this combination of lymphokines was still not able to support memory B-cell differentiation. Furthermore, recombinant rat interferon-gamma and a commercial source of human BCGF, with or without IL-2, were unable to promote significant virgin or memory B-cell differentiation. These data support the hypothesis that memory B cells and virgin B cells differ in their lymphokine requirements for differentiation into antibody-secreting cells.     

Induction of mammary gland differentiation in transgenic mice by the fatty acid-binding protein MRG

A mammary-derived growth inhibitor-related gene (MRG) was previously identified and characterized. MRG induces differentiation of mammary epithelial cells in vitro and its expression is associated with mammary differentiation. To further define the role of MRG on mammary gland differentiation, a MRG transgenic mice model under the control of mouse mammary tumor virus promoter was established and the effect of MRG on mammary gland differentiation was investigated at histological and molecular levels. Expression of endogenous mouse MRG gene was significantly increased from the non-differentiated gland of control virgin mice to the functionally differentiated gland of pregnant control mice. Whole mount analyses demonstrated that ductal development was not affected by MRG transgene expression. While there was no lobuloalveolar structure in control virgin mice, expression of MRG transgene in the mammary gland resulted in the development of lobuloalveolar-like structure, which mimics the gland from early pregnancy. Consistent with the morphological change, expression of MRG also increased milk protein beta-casein expression in the gland. To study the mechanism of MRG-induced mammary differentiation, we investigated the Stat5 activation in the glands from the transgenic mouse versus virgin control mouse. While activated Stat5 was expressed at the minimal level in the non-differentiated control virgin gland, a significant Stat5 phosphorylation was observed in the virgin transgenic gland. Our data indicate that MRG is a mediator of the differentiating effects of pregnancy on breast epithelium, and overexpression of MRG in young nulliparous mice can induce differentiation.     

Effect of insulin on glucose metabolism in adipocytes from virgin and late-pregnant rats.

Under basal conditions (zero insulin), paraovarian adipocytes from 19-day-pregnant rats exhibited the same rates of [U-14C]glucose conversion into CO2 and total lipids as did those from age-matched virgin rats. The dose-response curves for insulin stimulation of glucose metabolism were similar in both groups: maximal response (+100% over basal values) and high sensitivity (half-maximal effect at 0.05 nM-insulin). The present results suggest that the insulin resistance in vivo that occurs during late pregnancy may involve circulating factors lost in vitro.     

Comparison of structure,regeneration and dead wood in virgin forest remnant and managed forest on Grmeč Mountain in Western Bosnia

This paper compares the forest structure, regeneration and distribution of dead wood in a virgin forest remnant and a close-to-nature managed beech–conifer mixture situated on Grme? Mountain in Western Bosnia. The investigations were carried out in a 1 ha permanent sample plot and 35 circular plots (20 m radius) in the virgin forest and in 17 circular plots (25 m radius) in managed forests. The number of trees in the managed forest was significantly (p = 0.05) higher than that in virgin forest and the distribution of the number of trees per diameter classes had a decreasing trend, but with a different shape in the virgin forest compared to the managed stands. In the lower diameter classes, the stock volume recorded in virgin forest was half of that in the managed forest, whilst for higher diameter classes the cumulated volume of the growing stock was almost double in virgin forest. The young crops had a significantly lower presence in the virgin forest and a larger volume of dead wood was identified in the virgin forest than in managed stands. The study results are important in assessing the consequences of close-to-nature management on the forest structure and regeneration when compared to the condition in virgin forests.     

Coarse woody debris in virgin and managed forest

This study treats dead trees and their remnants in the managed and virgin forest of Rajhenavski Rog, Slovenia, at a location of Omphalodo-Fagetum omphalodetosum plant community. The study plots were selected in four forest cycle developmental phases (optimal, mixed, regeneration and juvenile phase) of both managed and virgin forest. The quantity (volume and dry matter) and the structure of coarse woody debris (CWD) were compared between the selected plots within the particular type of the forest, and between the virgin and the managed forest. Belowground CWD was quantified by modeling the tree's biomass and decaying processes of the trees. The results show significant differences in CWD between the virgin forest developmental phases. The highest concentration of CWD in the virgin forest was found in the regeneration phase (626.0 m³/ha and 179.3 t_dry matter/ha), while the juvenile phase (248.3 m³/ha and 40.2 t_dry matter/ha) has the smallest amount of CWD. Managed forest has very evenly distribution of CWD between developmental phases and it ranges from 41.0 m³/ha and 49.0 t_dry matter/ha in the mixed developmental phase to 67.0 m³/ha and 56.2 t_dry matter/ha in the juvenile phase. The main reasons for such a large differences are forest management measures (e.g. wood extraction, short rotation time, reduction of natural tree mortality), which decrease quantity, distribution and size of CWD. It was identified that forest management causes reduction and homogenization of CWD on our study plots, which can trigger degradation processes (e.g. soil erosion, reduction of site productivity, reduction of habitats). Tree heights curves show significant differences in maximum tree's height between the virgin and the managed forest. Maximum tree's height is lower in the managed forest which may indicate the reduction of forest productivity due to reduction of CWD. Study has shown some positive effects of forest management on accumulation of underground CWD in the managed forest (from 40.0 t_dry matter/ha to 48.2 t_dry matter/ha), which significantly exceeds underground CWD in the virgin forest (from 2.0 t_dry matter/ha to 22.8 t_dry matter/ha).     

Amino acid metabolism and protein synthesis in lactating rats fed on a liquid diet.

1. Amino acid metabolism was studied in control virgin rats, lactating rats and virgin rats protein-pair-fed with the lactating rats (high-protein virgin rats). 2. Urinary excretion of nitrogen and urea was higher in lactating than in control virgin rats, and in high-protein virgin rats it was higher than in lactating rats. 3. The activities of urea-cycle enzymes (units/g) were higher in high-protein virgin than in lactating rats, except for arginase. In lactating rats the activities of carbamoyl-phosphate synthase, ornithine carbamoyltransferase and argininosuccinate synthase were lower than in control virgin rats. When the liver size is considered, the activities in lactating rats were similar to those in high-protein virgin rats, except for arginase. 4. N-Acetylglutamate content was higher in high-protein virgin rats than in the other two groups. 5. The rate of urea synthesis from precursors by isolated hepatocytes was higher in high-protein virgin rats than in the other two groups. 6. The flooding-dose method (L-[4-3H]phenylalanine) for measuring protein synthesis was used. The absolute synthesis rates of mammary gland, liver and small-intestinal mucosa were higher in lactating rats than in the other two groups, and in high-protein virgin rats than in control virgin rats 7. These results show that the increased needs for amino acids during lactation are met by hyperphagia and by a nitrogen-sparing mechanism.     

Maternal condition reduces fear behaviors but not the endocrine response to an emotional threat in virgin female rats

Lactating dams and maternal virgin females are less fearful in behavioral tests compared with non-maternal animals, suggesting that maternal condition per se reduces the negative value of threatening stimuli. In addition, lactating females exhibit a diminished hypothalamic-pituitary-adrenal response to potential environmental threats. Can the maternal condition, independently of the endocrine profile of lactation, promote a reduction in the behavioral as well as in the endocrine response to an emotional stressor? To answer this question, anxiety-related and fear behaviors as well as the levels of corticosterone were evaluated in response to a bright-lit open field-loud noise model in maternal and non-maternal non-ovariectomized virgin females and lactating dams in the presence of the pups. Maternal animals, both lactating and virgin, presented an increased exploration of the bright-lit open field and a significant reduction of fear behaviors, indicated by the decreased flight and immobility responses to the subsequent activation of a loud noise, in comparison to non-maternal virgins. Interestingly, maternal virgin females, as non-maternal rats, showed high corticosterone plasma levels, in contrast to the lower endocrine response exhibited by lactating dams when confronted to this threat. Present results suggest that maternal condition allows females to take risks when caring for their young, a behavioral strategy that is independent of the reduced hypothalamic-pituitary-adrenal axis response characteristic of lactation. This evidence points towards a clear dissociation in the mechanisms regulating behavioral and endocrine responses to emotional stressors during motherhood.     

蜂群中不同亚家庭的工蜂寿命分析

在蜂群中,蜂王与多个雄蜂自然交尾,形成不同亚家庭.为了研究不同亚家庭中工蜂寿命是否有差异,我们以西方蜜蜂Apis mellifera L.为实验材料,随机从一群自然群中取270只刚孵化的工蜂,单独饲养于有蜜粉脾小蜂箱内,每天将自然死亡个体取出标记.并利用3对微卫星进行个体基因型分析,通过Matesoft软件划分亚家庭,然后分析了各亚家庭工蜂的自然寿命及生存曲线.结果表明:实验蜂群由12个亚家庭组成,其中2个亚家庭工蜂寿命与其他亚家庭存在显著差异(P＜0.05).     

Sustainable sourcing of strategic raw materials by integrating recycled materials

In this paper we investigate a manufacturer’s sustainable sourcing strategy that includes recycled materials. To produce a short life-cycle electronic good, strategic raw materials can be bought from virgin material suppliers in advance of the season and via emergency shipments, as well as from a recycler. Hence, we take into account virgin and recycled materials from different sources simultaneously. Recycling makes it possible to integrate raw materials out of steadily increasing waste streams back into production processes. Considering stochastic prices for recycled materials, stochastic supply quantities from the recycler and stochastic demand as well as their potential dependencies, we develop a single-period inventory model to derive the order quantities for virgin and recycled raw materials to determine the related costs and to evaluate the effectiveness of the sourcing strategy. We provide managerial insights into the benefits of such a green sourcing approach with recycling and compare this strategy to standard sourcing without recycling. We conduct a full factorial design and a detailed numerical sensitivity analysis on the key input parameters to evaluate the cost savings potential. Furthermore, we consider the effects of correlations between the stochastic parameters. Green sourcing is especially beneficial in terms of cost savings for high demand variability, high prices of virgin raw material and low expected recycling prices as well as for increasing standard deviation of the recycling price. Besides these advantages it also contributes to environmental sustainability as, compared to sourcing without recycling, it reduces the total quantity ordered and, hence, emissions are reduced.     

婚飞行为影响中华蜜蜂性成熟处女蜂王的基因表达

婚飞是性成熟处女蜂王与雄蜂交配过程中的一个重要前奏, 在该过程中蜂王体内伴随着一系列重要的生理变化。为了探究中华蜜蜂Apis cerana cerana处女蜂王婚飞过程中基因表达变化, 本研究利用数字基因表达谱(digital gene expression, DGE) 技术分析了中华蜜蜂性成熟处女蜂王飞行与未飞行之间的基因表达差异。经DGE测序, 分别从两个样品中获得5.98和6.01 百万条Clean标签。通过分析检测到250个基因有差异表达, 其中133个基因在飞行蜂王中上调表达, 117个基因在飞行蜂王中下调表达。这些差异基因可以归类到348个功能性类别和142个生化途径。结果表明中华蜜蜂性成熟处女蜂王在婚飞过程中大量基因的表达发生了变化。这些结果为进一步研究中华蜜蜂蜂王婚飞过程中生理变化的分子机制提供了重要的基因表达信息。     

基于修正的Gash模型模拟小兴安岭原始红松林降雨截留过程

原始红松林是我国小兴安岭地区的顶级植物群落,理解它的降雨截留过程,对区域生态、水文环境具有重要意义.采用定位研究法,于2010-2011年21场降雨数据对原始红松林的降雨截留和分配效应进行系统研究,并结合当地的气象和林分资料,利用修正的Gash模型对小兴安岭原始红松林进行林冠截留模拟.该模型对穿透雨、树干茎流、截留量的模拟值分别为:370.91、16.14、130.07 mm.穿透雨模拟值比实测值低2.28 mm,相对误差1.75％,树干茎流模拟值高于实测值8.12 mm,相对误差50.3％,林冠截留量模拟值比实测值低2.35 mm,相对误差为1.81％.穿透雨和林冠截留量的模拟值和实测值有较好的一致性,树干茎流的模拟值和实测值相差较大.综合结果表明:修正的Gash模型对小兴安岭地区原始红松林降雨截留拟合具有很好的适用性.     

棉铃虫蛾对寄主植物挥发油的嗅觉电生理及行为反应

本项研究以Tenax TA、Tenax GR为吸附剂,正己烷为洗脱剂,得到了玉米雄穗、玉米花丝、棉花花、棉花顶尖、棉蕾、芹菜、花生叶及花的挥发油。触角电位(EAG)测试结果表明,参试寄主植物及器官的挥发油均有一定的EAG活性,EAG的反应幅度为0.1～1.2mV,雌性棉铃虫Helicoverpa armigera对其产卵寄主或器官的EAG反应在1～10日龄内有随着日龄增加而增大的趋势,交配蛾对棉花花、棉花嫩尖、芹菜、棉蕾的挥发油的EAG反应显著高于处女蛾,雌蛾在经过花生、玉米花丝及棉花顶尖挥发油处理的滤纸上落卵量显著高于对照,表明这几种挥发油中含有可刺激棉铃虫选择其产卵的它感信息化合物。剂量反应曲线表明,交配可显著降低雌蛾对棉花嫩尖和棉花花挥发油的敏感阈值。四臂嗅觉仪测试棉铃虫对不同寄主植物挥发油的行为反应表明,棉蕾、棉花叶、玉米花丝、花生的挥发油均具有-定的诱蛾活性,选择系数分别为0.78、0.61、0.49、0.31。     

Pattern formation in one- and two-dimensional shape-space models of the immune system.

A large-scale model of the immune network is analyzed, using the shape-space formalism. In this formalism, it is assumed that the immunoglobulin receptors on B cells can be characterized by their unique portions, or idiotypes, that have shapes that can be represented in a space of a small finite dimension. Two receptors are assumed to interact to the extent that the shapes of their idiotypes are complementary. This is modeled by assuming that shapes interact maximally whenever their coordinates in the space-space are equal and opposite, and that the strength of interaction falls off for less complementary shapes in a manner described by a Gaussian function of the Euclidean "distance" between the pair of interacting shapes. The degree of stimulation of a cell when confronted with complementary idiotypes is modeled using a log bell-shaped interaction function. This leads to three possible equilibrium states for each clone: a virgin, an immune, and a suppressed state. The stability properties of the three possible homogeneous steady states of the network are examined. For the parameters chosen, the homogeneous virgin state is stable to both uniform and sinusoidal perturbations of small amplitude. A sufficiently large perturbation will, however, destabilize the virgin state and lead to an immune reaction. Thus, the virgin system is both stable and responsive to perturbations. The homogeneous immune state is unstable to both uniform and sinusoidal perturbations, whereas the homogeneous suppressed state is stable to uniform, but unstable to sinusoidal, perturbations. The non-uniform patterns that arise from perturbations of the homogeneous states are examined numerically. These patterns represent the actual immune repertoire of an animal, according to the present model. The effect of varying the standard deviation sigma of the Gaussian is numerically analyzed in a one-dimensional model. If sigma is large compared to the size of the shape-space, the system attains a fixed non-uniform equilibrium. Conversely if sigma is small, the system attains one out of many possible non-uniform equilibria, with the final pattern depending on the initial conditions. This demonstrates the plasticity of the immune repertoire in this shape-space model. We describe how the repertoire organizes itself into large clusters of clones having similar behavior. These results are extended by analyzing pattern formation in a two-dimensional (2-D) shape-space.(ABSTRACT TRUNCATED AT 400 WORDS)