Hybrid wastewater treatment system using anaerobic microorganisms and reed (Phragmites communis)

A small hybrid wastewater treatment system consisting of a settling tank in series with an anaerobic filter-reed (Phragmites communis) treatment cell was evaluated and compared with a similar plant-free system. Data demonstrated that by combining anaerobic filters, also referred to as attached film filters, and vascular aquatic plants a synergistic effect is produced which increases the treatment efficiency of each individual system. The plant-free system reduced the BOD₅ from 114 to 31 mg/l in 6 h as compared to a reduction of 110 to 9 mg/l in the anaerobic filter-reed system in the same length of time. The BOD₅ and TSS after 24 h in each component of plant-free system were reduced from 114 mg/l to 14 mg/l and 51 to 15 mg/l, respectively. Under the same conditions, the hybrid system reduced the BOD₅ from 110 to 3 mg/l and the TSS from 68 to 6 mg/l. The hybrid system also reduced the total Kjeldahl nitrogen (TKN) from 16.1 to 2.9 mg/l, total phosphorus (TP)from 4.4 to 2.0 mg/l, and the ammonia (NH₃-N)from 12.4 to 0.6 mg/l after 24 h of exposure while the plant-free system demonstrated insignificant reduction of these components.     

A Model for Single-Substrate Trimolecular Enzymatic Kinetics

We developed a kinetic model for a single-substrate trimolecular enzymatic system, where a receptor binds and stretches a substrate to expose its cleavage site, allowing an enzyme to bind and cleave it into product. We demonstrated that the general kinetics of the trimolecular enzymatic system is more complex than the Michaelis-Menten kinetics. Under a limiting condition when the enzyme-substrate binding is in fast equilibrium, the enzymatic kinetics of the trimolecular system reduces to the Michaelis-Menten kinetics. In another limiting case when the receptor dissociates negligibly slowly from the substrate, the trimolecular system is simplified to a bimolecular system, which follows the Michaelis-Menten equation if and only if there is no enzyme-substrate complex initially. We applied this model to a particular trimolecular system important to hemostasis and thrombosis, consisting of von Willebrand factor (substrate), platelet glycoprotein Ibα (receptor), and ADAMTS13 (enzyme). Using parameters from independent experiments, our model successfully predicted published data from two single-molecule experiments and fitted/predicted published data from an ensemble experiment.     

Glutamine transport in C6 glioma cells

Glutamine transport across the cell membranes of a variety of mammalian tissues is mediated by at least four transport systems: a sodium-independent system L, and sodium-dependent systems A, ASC and N, the latter occurring in different tissue-specific variants. In this study we assessed the contribution of these systems to the uptake of [(3)H]glutamine in C6 rat glioma cells. The sodium-dependent uptake, which accounted for more than 80% of the total uptake, was not inhibited by 2-methylaminoisobutyric acid (MeAIB), indicating that system A was inactive, possibly being depressed by glutamine present in the culture medium. About 80% of the sodium-dependent uptake was mediated by system ASC, which differed from system ASC common to other CNS- and non-CNS tissues by its pH-dependence and partial lithium tolerance. The residual 20% of sodium-dependent uptake appeared to be mediated by system N, which was identified as a component resistant to inhibition by MeAIB+threonine. The system N in C6 cells appeared to be neither fully compatible with the neuronal system Nb, nor with the N system described in astrocytes: it differed from the former in being strongly inhibited by histidine and showing fair tolerance for lithium, and from the latter in its pH-insensitivity and strong inhibition by glutamate. The sodium-independent glutamine uptake differed from the astrocytic or neuronal uptake in its relatively weak inhibition by system L substrates and a strong inhibition by system ASC substrates, indicating a possible contribution of a variant of the ASC system.     

Secretome analysis using a hollow fiber culture system for cancer biomarker discovery

Secreted proteins, collectively referred to as the secretome, were suggested as valuable biomarkers in disease diagnosis and prognosis. However, some secreted proteins from cell cultures are difficult to detect because of their intrinsically low abundance; they are frequently masked by the released proteins from lysed cells and the substantial amounts of serum proteins used in culture medium. The hollow fiber culture (HFC) system is a commercially available system composed of small fibers sealed in a cartridge shell; cells grow on the outside of the fiber. Recently, because this system can help cells grow at a high density, it has been developed and applied in a novel analytical platform for cell secretome collection in cancer biomarker discovery. This article focuses on the advantages of the HFC system, including the effectiveness of the system for collection of secretomes, and reviews the process of cell secretome collection by the HFC system and proteomic approaches to discover cancer biomarkers. The HFC system not only provides a high-density three-dimensional (3D) cell culture system to mimic tumor growth conditions in vivo but can also accommodate numerous cells in a small volume, allowing secreted proteins to be accumulated and concentrated. In addition, cell lysis rates can be greatly reduced, decreasing the amount of contamination by abundant cytosolic proteins from lysed cells. Therefore, the HFC system is useful for preparing a wide range of proteins from cell secretomes and provides an effective method for collecting higher amounts of secreted proteins from cancer cells. This article is part of a Special Issue entitled: An Updated Secretome.     

制度供给视角下的基本医疗体制改革之决策

基本医疗改革的财政投入模式应从单一关注医疗服务供给转向协同关注医疗服务产出，无过错医疗损害救济应纳入国家财政投入的预算体系。面对医疗损害的高发性，以救济为心的医疗侵权责任制度应向以预防中心的制度进行改革。医疗过失诉讼制度对医疗体系有着深刻的负面制约作用。基本医疗改革的决策应从更宏观的视角进行制度供给设计。     

Sedimentology of geomorphologically controlled Quaternary tufas in a valley in southern Spain

Four vertical facies sequences characterizing different environments have been identified in relation to two phases of tufa deposition (probably Pleistocene and Holocene) within a valley located in the central sector of the Betic Cordillera (southern Spain): (1) alluvial-fluvial (non-tufa) deposits, (2) fluvial tufa, (3) fluvio-lacustrine facies, and (4) spring with waterfall tufa. The first three created a stepped axial depositional system and the spring with waterfall tufa formed a transverse system. During the first phase of tufa construction, the vertical evolution in the axial system from alluvial-fluvial (non-tufa) to fluvio-lacustrine tufa deposits is interpreted as the consequence of geomorphological control. The evolution through time from a high-gradient and stepped fluvial system to a low-gradient fluvio-lacustrine system occurred upstream from where the progradation of the transverse, perched spring tufa system became narrower and finally dammed the valley. The tufa barrage caused an increase in the vertical accumulation within the axial system and a consequent upstream decrease of accommodation space and slope of the longitudinal profile. After a period of fluvial incision, tufa formed again during the Holocene in a high-gradient and stepped fluvial system.     

一种新的基于STR的染色体三体性疾病诊断策略中遗传标记的评估

背景:在此前发表的文章中,我们提出了一种新的基于短串联重复序列(Short Tandem Repeats,STR)的染色体三体性疾病的诊断策略.当应用这种策略来检测特定染色体拷贝数时,需要从人类基因组众多的STR中选择适宜的染色体特异性STR基因座构建一个诊断系统,根据系统中的STR基因座是否检测到三种不同的等位基因产物来判断个体是否为三体患者.目的:本研究拟进一步提出并验证对单个STR基因座及由多个STR基因座构成的诊断系统的评估方法,旨在帮助选择适宜的STR基因座构建一个高效能的诊断系统.方法:我们提出一个新的参数--三等位基因栓出率,并推导出该参数的计算公式,用于定量评估一个STR基因座在这种诊断策略中的效能.在此基础上,推导出另一个数学公式,计算一个非整倍体诊断系统能够在一个三体性患者?哌 检测到三个不同等位基因的概率,根据这个概率的大小来衡量系统诊断效能的高低.最后,我们将所提出的两个公式用于评估我们在先前研究中构建的一个21三体的诊断系统.结果:这个21三体诊断系统由9个21号染色体特异性STR基因座构成.根据我们所提出的两个公式,这些STR基因座的三等位基因检出率在0.203-0.638之间,该系统在21三体患者能够检测到三个不同等位基因的概率大于0.95.结论:我们所提出并验证的公式可以对单个STR基因座和系统的诊断效能进行定量评估,帮助选择适宜的STR遗传标记,并确定一个高效能诊断系统所需的STR基因座的数量,从而为这种诊断策略的广泛应用提供基础.     

Evaluation of Radiometric System for Detecting Bacteremia

An automated radiometric system (BACTEC, Johnston Laboratories) for detection of bacteremia was evaluated in parallel with a standard blood culture system in use in our laboratory. Of 1,445 blood cultures from 484 patients with possible bacteremia, 106 sets of cultures (excluding 39 presumed contaminated), representing 56 patients, were positive by both methods. The conventional system yielded 85 positive cultures from 48 patients, whereas the BACTEC system yielded 84 positive cultures from 43 patients. The BACTEC system failed to detect 22 cultures that were positive in the conventional system, and the conventional system failed to detect 21 cultures that were positive in the BACTEC system. The detection efficiency was generally equivalent in the two systems except for the lower detection rates of anaerobes and Enterobacter aerogenes by the BACTEC system and the lower detection rates of Torulopsis glabrata and, possibly, Pseudomonas sp. (group IVD) in the conventional system. The BACTEC system had a slight advantage over the conventional system in the time interval to detection of positivity. Approximately 20% of the positive cultures detected by the BACTEC system were detected on the first day of incubation compared with 7% by the conventional system. The recovery rates and detection times of anaerobes were less efficient by the BACTEC system than by the conventional system. It does not appear that the radiometric method has much advantage over available conventional methods.     

Reconstructing the dynamics of unobserved variables in spatially extended systems

Attractor reconstruction using embedding techniques is a widely used tool when analysing data from real systems. It allows reconstruction of the system dynamics from only one observable and is thus extremely powerful. We show here that this reconstruction is also possible from spatially coupled systems. We use a common host–parasitoid model as an example as ecological systems are virtually always spatially extended. Additionally, data from ecological systems has often only one observable, e.g. population density, from a potentially much higher-dimensional system. Singular value decomposition is used to show the existence of a functional relationship mapping the time delayed coordinates of one variable to the full spatially coupled system. We investigate the effects of noise and indicate two important spatial scales. Finally, we illustrate that a reconstruction can be obtained from a system that is only partially sampled.     

Dopamine Enhances Model-Based over Model-Free Choice Behavior

Decision making is often considered to arise out of contributions from a model-free habitual system and a model-based goal-directed system. Here, we investigated the effect of a dopamine manipulation on the degree to which either system contributes to instrumental behavior in a two-stage Markov decision task, which has been shown to discriminate model-free from model-based control. We found increased dopamine levels promote model-based over model-free choice.     

基于景观格局和网络分析法的崇明绿地系统现状和规划的评价

以崇明2005年卫星影像图、土地利用现状图为基础资料,结合崇明绿地系统规划,在地理信息系统支持下,选择崇明本岛面积大于10hm²的绿地作为节点,依照网络分析法,构建了不同的生态网络方案,通过α指数、β指数、γ指数以及成本比等网络结构指数进行不同网络方案的比较,从中选出最优的规划方案。然后在景观分析软件Fragstats支持下,运用景观格局分析方法,选用多种景观分析指数,在斑块类型层次上和景观层次上对崇明绿地系统现状、绿地系统规划以及优化后的生态网络规划方案从景观生态学角度进行了定量分析与评价,并针对现有的绿地系统规划提出了进一步完善的建议。研究结果表明崇明绿地系统规划有效地改善了绿地景观破碎化的问题,绿地斑块形状的复杂性程度增大,景观连接度得到提高,景观多样性和均匀性指数均得到较大幅度提升。而在现有绿地系统规划基础上做出的优化生态网络方案可进一步改善崇明本岛现有绿地系统的数量和质量。同时表明,生态网络分析结合景观格局分析是评价和改善绿地系统规划有效的途径。     

Cell-free translation of mammalian myosin heavy-chain messenger ribonucleic acid from growing and fused-L6E9 myoblasts.

An mRNA-dependent reticulocyte cell-free protein synthesizing system very efficient in the translation of myosin heavy-chain mRNA from a rat myogenic cell line is described. This system exhibits a high degree of fidelity with regard to the spectrum and relative proportion of the different proteins synthesized from a sample of cytoplasmic RNA as compared to the proteins synthesized in vivo by the cells from which the RNA is prepared. The main feature of this system is the use of a K+ and Cl- concentration similar to those of the reticulocyte cytoplasm. Using this system, myosin heavy chain, identified by low-salt precipitation, electrophoretic mobility, and partial peptide analysis, represents 17% of the total protein synthesis when cytoplasmic RNA from well-fused L6E9 cells is used. Furthermore, when RNA preparations from growing myoblasts, that when analyzed in other cell-free translational systems seem not to contain any myosin heavy-chain mRNA, are tested in the system reported here, they are proven to contain high amounts of translatable myosin heavy-chain mRNA.     

A Comparative Study and Partial Characterization of Multi-Uptake Systems for γ-Aminobutyric Acid

Previous work by the authors had indicated that synaptosome-enriched preparations from the cerebral cortex of the rat contained a high-, a medium-, and a low-affinity uptake system for gamma-aminobutyric acid (GABA). The present study demonstrated that this phenomenon also prevailed in synaptosomes from rat diencephalon, mesencephalon, and cerebellum, although the Vmax values for the high- and medium-affinity systems in the cerebellum were very low relative to those of the other regions. When a different type of preparation containing nerve endings (glomeruli) was obtained from the cerebellum, it possessed a Vmax value for the high-affinity system that was more similar to that for the corresponding system in synaptosomes from the other brain regions. In contrast to the above situation, synaptosomes from rat olfactory bulb lacked the low-affinity uptake system, as did synaptosomes from dog olfactory bulb. The aspartate/glutamate uptake systems, as measured with D-aspartate, provided a regional pattern quite different from those of GABA uptake. Only two uptake systems, a high- and low-affinity system, were observed in all regions tested. All three GABA uptake systems were present in cortical synaptosomes from the mouse, hamster, and guinea pig, and all three systems were sodium dependent, energy dependent, temperature sensitive, and totally inhibited by nipecotic acid.     

Concentration of bovine plasmalogen using phospholipase A1 in two-phase system

Plasmalogen that contains choline was concentrated from bovine heart to 95% purity from an initial value of 45%, and ethanolamine-containing plasmalogen from bovine brain to 99% purity from initial 64% using phospholipase A₁ from Serratia sp. MK1 in a two-phase system. In the two-phase system, Ca2⁺ was not required and Tris/HCl buffer and butyl acetate were used as an aqueous phase and a solvent phase of the two-phase system, respectively.     

Renal transport of monocarboxylic acids. Heterogeneity of lactate-transport systems along the proximal tubule.

The characteristics of D- and L-lactate transport in luminal-membrane vesicles derived from whole cortex, from the pars convoluta and from the pars recta of rabbit kidney proximal tubule were studied. It was found that uptake of both isomers in vesicles from whole cortex occurred by means of dual electrogenic transport systems, namely a low-affinity system and a high-affinity system. Uptake of both isomers in vesicles from the pars recta was strictly Na+-dependent and is mediated via a single high-affinity common transport system. Vesicles from the pars convoluta contained a cation-dependent but Na+-unspecific low-affinity common transport system for these compounds. The physiological importance of this system is briefly discussed.     

Regulatory dynamics of synthetic gene networks with positive feedback

Biological processes are governed by complex networks ranging from gene regulation to signal transduction. Positive feedback is a key element in such networks. The regulation enables cells to adopt multiple internal expression states in response to a single external input signal. However, past works lacked a dynamical aspect of this system. To address the dynamical property of the positive feedback system, we employ synthetic gene circuits in Escherichia coli to measure the rise-time of both the no-feedback system and the positive feedback system. We show that the kinetics of gene expression is slowed down if the gene regulatory system includes positive feedback. We also report that the transition of gene switching behaviors from the hysteretic one to the graded one occurs. A mathematical model based on the chemical reactions shows that the response delay is an inherited property of the positive feedback system. Furthermore, with the aid of the phase diagram, we demonstrate the decline of the feedback activation causes the transition of switching behaviors. Our findings provide a further understanding of a positive feedback system in a living cell from a dynamical point of view.     

Organizational levels analysis: a key to understanding processes in natural systems

Recent research conducted on a hypothetical four-parameter system generated by two stochastic processes has shown that errors of interpretation are likely to result when the relative importance of an underlying generating process is inferred from the analysis of an emergent pattern of a natural system. This paper presents an approach which substantially reduces that error for the hypothetical system, taken as an analog for a natural system. This approach is based on recognition of the organizational levels composing the system and utilizes observations on structure at a lower organizational level to resolve the properties of the generating processes at the lower level. The resulting knowledge of those underlying structures and the processes which created them are then used to determine the relative importance of those processes in affecting the emergent pattern of the system at a higher organizational level. It is concluded that in order to understand the role of a process in a natural system, measurements of system structure must be performed at a frequency which isolates the process from the other processes in the system.     

Solutions for determining equibiaxial substrate strain for dynamic cell culture

In this work, empirical and analytical solutions of equibiaxial strain on a flexible substrate are derived for a dynamic cell culture system. The empirical formula, which fulfills the mechanistic conditions of the culture system, is based on a regression analysis from finite element analyses for a substrate undergoing large strains (<15%). The analytical (closed-form) solution is derived from the superposition of two elastic responses induced in the equibiaxial strain culture system after applying pressure to a substrate undergoing small strains (microstrains). There is good agreement between the strain predicted from the solutions and from the direct measurement. Using material and geometric properties of the culture system, the solutions developed here are straightforward and can be used to circumvent experimental measurements or finite element analysis to establish substrate pressure–strain relationships.     

Self-sustaining populations, population sinks or aggregates of strays: chum (Oncorhynchus keta) and Chinook salmon (Oncorhynchus tshawytscha) in the Wood River system, Alaska

Small populations can provide insights into ecological and evolutionary aspects of species distributions over space and time. In the Wood River system in Alaska, USA, small aggregates of Chinook (Oncorhynchus tshawytscha) and chum salmon (O. keta) spawn in an area dominated by sockeye salmon (O. nerka). Our objective was to determine whether these Chinook and chum salmon are reproductively isolated, self-sustaining populations, population sinks that produce returning adults but receive immigration, or strays from other systems that do not produce returning adults. DNA samples collected from adult chum salmon from 16 streams and Chinook salmon from four streams in the Wood River system over 3 years were compared to samples from large populations in the nearby Nushagak River system, a likely source of strays. For both species, microsatellite markers indicated no significant genetic differentiation between the two systems. Simulations of microsatellite data in a large source and a smaller sink population suggested that considerable immigration would be required to counteract the diverging effects of genetic drift and produce genetic distances as small as those observed, considering the small census sizes of the two species in the Wood River system. Thus, the Wood River system likely receives substantial immigration from neighbouring watersheds, such as the Nushagak River system, which supports highly productive runs. Although no data on population productivity in the Wood River system exist, our results suggest source-sink dynamics for the two species, a finding relevant to other systems where salmonid population sizes are limited by habitat factors.     

On the fidelity of DNA replication: use of synthetic oligonucleotide-initiated reactions

The phi X174 fidelity system provides a biological assay for quantitating the accuracy of DNA polymerases. Expansion of this system to cell extracts and DNA replication complexes from eucaryotes has been limited by the presence of nucleases in these preparations. We have overcome these limitations by priming the phi X template with a synthetic oligodeoxynucleotide, with its free 3'-hydroxyl terminus only a short distance from the amber locus that is the site for determining the frequency of misincorporation. In this paper, this modified phi X system is characterized and compared to that using defined natural DNA restriction fragments as primers. The modified system has been applied to studies on the fidelity of DNA synthesis using different forms of purified DNA polymerase-alpha from calf thymus, as well as crude extracts from human lymphocytes.