Die Kontrolle der Wanderung und Differenzierung von Neuralleistezellen durch die Extrazelluläre Matrix

Neural crest cells give rise to various essential tissues in vertebrates. Among these are the teeth, the ganglia of the peripheral nervous system, endocrine cells and pigment cells. Neural crest cells are actively migrating cells, but their migration needs to be stimulated by an appropriate extracellular matrix (ECM). In the present article the role of the epidermis and subepidermal ECM during the migration of neural crest cells and their differentiation into pigment cells in amphibian embryos will be considered. Furthermore, a new microcarrier technique will be presented with which the influence of natural ECM and various ECM components on the behavior and differentiation of neural crest derivatives can be investigated in amphibian embryos.     

剪切应力对好氧颗粒污泥形态结构和微生物活性的影响机制研究

对不同剪切应力(0.189、0.267、0.327和0.377 N/m2)下4个序批式反应器(SBR)中好氧颗粒污泥的形态结构、比耗氧速率(SOUR)以及胞外聚合物进行了对比分析.结果表明,好氧颗粒污泥具有稳定的基本形态特征,其微生物主要由杆菌、球菌和丝状菌组成;其中杆菌能承受高剪切作用.是剪切应力为0.377 N/m2时的优势菌群.4个反应器中污泥粒径分布范围分别为0.2-0.5、0.5-1.5、0.5-1.5和0.3-0.5 mm;SOUR分别为34.54、40.08、46.26和46.42 mg/(g·h),胞外多聚糖分别为59.71、66-81、80.88和109.99 mg/g,胞外蛋白质分别为9.29、9.80、12.35和17.02 mg/g.好氧颗粒污泥比耗氧速率SOUR和胞外聚合物与剪切应力有很好的正相关性.确定了好氧颗粒污泥微生物活性与剪切应力的响应关系.     

Humic acid-enhanced electron transfer of in vivo cytochrome c as revealed by electrochemical and spectroscopic approaches

Out-membrane cytochrome c （Cyt c） plays an important role carrying electrons from the inside of microbes to outside electron acceptors. However, the active sites of Cyt c are wrapped by non- conductive peptide chains, hindering direct extracellular electron transfer （EET）. Humic acids （HA） have been previously proven to efficiently facilitate EET. However, the inherent mechanism of HA- stimulated EET has not been well interpreted. Here, to probe the mechanism behind HA-stimulated EET, we studied the interaction between Cyt c and HA. The attachment of active in vivo Cyt c on a graphite electrode was achieved when MR-1 cells were self-assembled on the electrode surface. Pure horse-heart Cyt c was covalently immobilized on an electrode via 4-aminobenzoic acid to create an active in vitro Cyt c-enriched surface. Cyclic voltammetric measurements and scanning electron microscopy confirmed the immobilization of bacterial cells and pure Cyt c protein. Electrochemical methods revealed that HA could enhance the electrocatalytic current of both in vitro and in vivo Cyt c towards oxygen and thiosulfate, suggesting enhanced EET. The blue-shifted soret band in the UV-Vis spectra and changes in the excitation/emission matrix fluorescence spectra demonstrated that Cyt c interacted with HA to form organic complexes via electrostatic or hydrogen-bonding interactions. The results will help understand electron shuttle-stimulated EET and develop bacteria- based bioremediation and bioenergy technologies.     

Influence of extracellular matrix on the proliferation of human amniotic fluid cells in vitro

Factors which influence the proliferation of human amniotic fluid cells in vitro have potential importance in reducing the time for prenatal diagnosis. Fibroblast growth factor (FGF) has been shown to be mitogenic for human amniotic fluid cells. The observation that cells which respond to FGF in vitro produce their own extracellular matrix (ECM), led to the use of an ECM as a substrate to assess proliferation. Pooled amniotic fluid cells maintained on an ECM prepared from bovine corneal endothelial cells demonstrated a significant increase in proliferation when compared with cells maintained on plastic substrate in the presence or absence of FGF. If FGF was added to cultures of amniotic fluid cells maintained on ECM, further increases in proliferation were noted compared with cells maintained on ECM in the absence of FGF. These results indicate that the substrate upon which amniotic fluid cells are maintained can have a profound influence on their proliferation.     

Glial and neuronal cells in amniotic fluid of anencephalic pregnancies

Cultured amniotic fluid cells from four anencephalic pregnancies were characterized in indirect immunofluorescence (IIF) microscopy using specific antibodies against different types of cytoskeletal intermediate filaments. Most of the cells showed a fine fibrillar cytoplasmic fluorescence with antibodies against glial fibrillary acidic protein (GFA), indicating that amniotic fluid cells in anencephalic pregnancies are of glial origin. The GFA-positive cells were rapidly adhering and proliferating. They remained as the major cell type also in long term cultures, and could easily be recovered from liquid nitrogen without losing their GFA positivity. GFA-positive cells were pleomorphic in appearance, and occurred in several morphologically different shapes. Amniotic fluid from one of the anencephalic cases contained typical neuronal cells, which in IIF were GFA-negative but could specifically be stained with anti-neurofilament antibodies. Most of the GFA-negative cells in all the cases were fibroblasts, identified by their fluorescence only with antibodies against vimentin. Epithelial cells showing positive keratin-fluorescence in IIF, were seen only occasionally.     

Forming a tough shell via an intracellular matrix and cellular junctions in the tail epidermis of <Emphasis Type="Italic">Oikopleura dioica</Emphasis> (Chordata: Tunicata: Appendicularia)

A postanal tail is a major synapomorphy of the phylum Chordata, which is composed of three subphyla: Vertebrata, Cephalochordata, and Tunicata (Urochordata). Among tunicates, appendicularians are the only group that retains the tail in the adult, and the adult tail functions in locomotion and feeding in combination with a cellulose-based house structure. Given the phylogenetic position of tunicates, the appendicularian adult tail may possess ancestral features of the chordate tail. We assess the ultrastructural development of the tail epidermis of the appendicularian Oikopleura dioica. The epidermis of the larval tail is enclosed by the larval envelope, which is a thin sheet similar to the outer tunic layer of ascidian larvae. The epidermis of the adult tail seems to bear no tunic-like cellulosic integuments, and the tail fin is a simple folding of the epidermis. Every epidermal cell, except for the triangular cells at the edge of the tail fin, has a conspicuous matrix layer of fibrous content in the apical cytoplasm without enclosing membranes. The epidermis of the larval tail does not have a fibrous matrix layer, suggesting the production of the layer during larval development and metamorphosis. Zonulae adhaerentes firmly bind the epidermal cells of the adult tail to one another, and the dense microfilaments lining the cell borders constitute a mechanical support for the cell membranes. The intracellular matrix, cell junctions, and cytoskeletons probably make the tail epidermis a tough, flexible shell supporting the active beating of the oikopleuran adult tail.     

微生物燃料电池恒电流预培养阳极生物膜分析表征

阳极性能是影响微生物燃料电池(Microbial Fuel Cells,MFCs)性能的关键因素之一,同时阳极的接种挂膜过程是影响微生物燃料电池启动效率的关键因素.因此,本课题组提出了预培养阳极作为微生物燃料电池的一种新型阳极的概念,在三电极体系下,通过外加恒定电流预培养阳极,在不同条件下对阳极表面进行电化学选择和生物膜驯化以丰富生物膜结构和厚度.结果表明,阳极的性能与预培养电流大小密切相关,预培养阳极CF-4i(外加4 A·m-2电流密度)通过循环伏安法、塔菲尔曲线、电化学阻抗谱测试,性能好于其他测试组及对照组,装配阳极CF-4i的微生物燃料电池能实现最大功率密度968.20 m W·m-2,是对照组的2.53倍.同时,通过共聚焦显微镜观察发现,生物膜大体分两层,外层的活细胞及内层的死细胞,外层活细胞长在内层死细胞之上.这种结构分布表明,阳极生物膜中的活细胞部分绝大多数都分布于生物膜的外侧,而不是均布于整个阳极生物膜中;同时这也表明不是整个阳极生物膜都具有新陈代谢活性,但死亡的细胞可以继续积累在电极表面附近,活的外层膜负责电流的产生,而内层的死细胞作为一种导电基质.     

The brain as a photoreceptor: intracerebral ocelli in the firefly

 This paper deals with the structure and function of the intracerebral ocelli in the caudal area of the brain of the Japanese firefly. A pair of epilaterally placed specialized pigmented organs was found at the caudal ends of the brains of the fireflies Luciola cruciata and L. lateralis. On the basis of light and transmission electron micrographs of both male and female individuals these organs seemed photoreceptive in nature. Intracellular and extracellular recordings were obtained from the intracerebral ocelli of the fireflies with microelectrodes. The physiological evidence revealed that the cells found in the brain were, indeed, photoreceptors. Received: 7 April 2000 / Accepted in revised form: 16 June 2000     

Detection and isolation of fetal cells from maternal blood using the flourescence-activated cell sorter (FACS)

The presence of fetal cells in the maternal circulation during pregnancy has been suggested by repeated observations of small numbers of cells containing Y chromatin or a Y chromosome in the blood of pregnant women. With the fluorescence-activitated cell sorter (FACS), we have used antibodies to a paternal cell surface (HLA) antigen, not present in the mother, to select fetal cells from the lymphocyte fractions of a series of maternal blood samples, collected as early as 15 weeks of gestation. These sorted cells have been examined for a second paternal genetic marker, Y chromatin. Y chromatin-containing cells were found among the sorted cells from prenatal maternal blood specimens in 8 pregnancies subsequently producing male infants whose lymphocytes reacted with the same antibodies to paternal antigen used for sorting with the FACS. In each of 17 pregnancies resulting in male infants who failed to inherit the antigen detected by the antibodies used for cell sorting, Y chromatin-containing cells were not found prenatally. The use of two paternal genetic markers, a cell surface antigen and nuclear Y chromatin, to identify fetal cells in maternal blood permits us to conclude that these cells are present in the mother's circulation, as early as 15 weeks gestation. Further development of the techniques reported here could lead to widespread screening of maternal blood samples during pregnancy for detection of fetal genetic abnormalities.     

自养菌和异养菌胞外聚合物对活性污泥絮凝特性的影响

为探讨自养菌和异养菌胞外聚合物(Extracellular polymeric substances,EPS)的差异及其对活性污泥表面及絮凝特性的影响,研究了有机负荷为0、0.30和0.74 kg·kg-1·d-1时,污泥松散附着胞外聚合物(loosely bound EPS,LB-EPS)和紧密粘附胞外聚合物(tightly bound EPS,TBEPS)的组分含量、形态、表面特性以及絮凝性能的差异,并就LB-EPS与TB-EPS对污泥絮凝特性影响的作用机理进行了探讨.结果表明,有机负荷越高,多糖含量越高,EPS总量及蛋白质含量越低.由三维荧光光谱测定结果可知,类富里酸物质为异养菌而非自养菌的代谢产物,且有机负荷越高,TB-EPS中的类富里酸荧光峰强度越大.自养和异养污泥的絮凝机理不同,自养活性污泥通过自养菌菌体自身生物絮凝作用形成密实的污泥絮体,LB-EPS与TB-EPS对其絮体形态、表面及絮凝特性影响较小;异养活性污泥絮体形态在TB-EPS提取后发生明显改变,TB-EPS在异养活性污泥絮体聚集时发挥着主要作用.包裹着LB-EPS的污泥表面负电荷多且疏水性差,不利于异养污泥生物絮凝.有机负荷越高,污泥表面Zeta电位负电性越强,相对疏水性(relative hydrophobity,RH)越小,污泥稳定性越差.     

用于生物降解酚类毒物的固定化细胞性能改进的研究

在获得了一种耐酚能力达915mg/L的菌种的基础上,进一步研究了以海藻酸钠包埋法制得的该菌种固定化细胞的性能。同时,采用了2种方法,即添加硅藻土和用己二胺-戊二醛对固定化细胞表面进行化学处理,使固定化细胞的机械强度,降酚活性,稳定性均得到了提高。实验证明这2种方法都有令人满意的效果。      

Monitoring biofouling based on aerobic respiration in reverse osmosis system

A monitoring method of biofouling in reverse osmosis (RO) system was proposed based on the fluorescent signal of resorufin, which is reduced by nicotinamide adenine dinucleotide released from viable cells during aerobic respiration. The fluorescent signal of resorufin reduced by planktonic cells and microorganisms of biofilm showed linearity, indicating its feasibility to monitor biofouling in a RO system. For the application of the method to the lab-scale RO system, the injection concentration of resazurin and the injection flow rate were optimized. Biofilm on RO membranes continuously operated in a lab-scale RO system was estimated by resorufin fluorescence under optimized detection condition. As a result, resorufin fluorescence on RO membrane showed a significant increase in which the permeability of RO system decreased by 30.48%. Moreover, it represented the development of biofilm as much as conventional biofilm parameters such as adenosine triphosphate, extracellular polymeric substances, and biofilm thickness. The proposed method could be used as a sensitive and low-cost technology to monitor biofouling without autopsy of membranes.     

Nutrients regeneration pathway, release potential, transformation pattern and algal utilization strategies jointly drove cyanobacterial growth and their succession

In order to better understand the contribution of nutrients regeneration pathway, release potential and transformation pattern to cyanobacterial growth and succession, 7 sampling sites in Lake Chaohu with different bloom degree were studied every two months from February to November 2018. The carbon, nitrogen (N) and phosphorus (P) forms or fractions in surface, interstitial water and sediments as well as extracellular enzymatic activities, P sorption, specific microbial abundance and community composition in sediments were analyzed. P regeneration pathway was dominated by iron-bound P desorption and phosphorus-solubilizing bacteria solubilization in severe-bloom and slight-bloom area respectively, which both resulted in high soluble reactive phosphorus (SRP) accumulation in interstitial water. However, in severe-bloom area, higher P release potential caused the strong P release and algal growth, compared to slight-bloom area. In spring, P limitation and N selective assimilation of Dolichospermum facilitated nitrate accumulation in surface water, which provided enough N source for the initiation of Microcystis bloom. In summer, the accumulated organic N in Dolichospermum cells during its bloom was re-mineralized as ammonium to replenish N source for the sustainable development of Microcystis bloom. Furthermore, SRP continuous release led to the replacement of Dolichospermum by Microcystis with the advantage of P quick utilization, transport and storage. Taken together, the succession from Dolichospermum to Microcystis was due to both the different forms of N and P in water column mediated by different regeneration and transformation pathways as well as release potential, and algal N and P utilization strategies.     

Stimulation of human amniotic fluid cell proliferation and colony formation by cell plating on a naturally produced extracellular matrix

Human amniotic fluid cells exhibit a higher cloning efficiency and rate of cell proliferation when maintained on dishes coated with a naturally produced extracellular matrix (ECM) in comparison with the regular tissue culture plastic. In 22 out of 31 amniotic fluid samples there was by plating the cells on ECM a 2–6 fold increase in number and size of colonies and in the cell density per colony as detected by actual staining and viewing of each colony. These effects yielded, in 21 of 41 additional samples, a reduction ranging from 2–8 days, in the culture time elapsing between amniocentesis and the first harvesting of cells for chromosomal analysis. An even greater effect was obtained with primary cells that failed to attach to plastic surfaces and stayed floating in the medium but did attach and proliferate when seeded on ECM. Cells that were left firmly attached to ECM after the first trypsinization and harvesting of cells for chromosomal analysis yielded colonies ready for second karyotyping in less than half the time required for cells maintained on plastic. Studies with secondary cultures of human amniotic fluid cells have demonstrated a 5–10 fold decrease in serum requirement of cells cultured on ECM as compared with plastic. Addition of fibroblast growth factor (FGF) to the cultures further potentiated the effects of ECM. The ECM induced stimulation of cell attachment and proliferation was not associated with any chromosomal anomalies, nor did it interfere with the handling procedure. ECM coated dishes may be useful to reduce the time interval between amniocentesis and diagnosis, in particular when the amniotic cells exhibit an exceedingly slow rate of proliferation on plastic or when large quantities of cells are required for enzymatic studies.     

Adopting and applying eco-design techniques: a practitioners perspective

This paper discusses the findings of a small scale research project which explored the possibility of adopting eco-design techniques. The paper focuses on identifying how eco-design techniques can be determined as being compatible with new product development processes. Via the development of a five stage ‘applicability framework’, this study demonstrates how a compatible suite of tools can be identified for application to product development processes. Testing and validation of this ‘applicability framework’, which was used to identify three key eco-design techniques; namely checklists, guidelines, and a material, energy and toxicity (MET) matrix, is shown to have taken place in relation to the development of a lightweight chemical detector product. It is established that checklists, guidelines and the MET matrix can be used both on a specific product, and also more generally in the design process. In particular, the MET matrix is shown as being used to successfully identify key environmental aspects of the product during its lifetime. The paper concludes by arguing that eco-design techniques may not have been more widely adopted by businesses because such methods are not necessarily generic and immediately applicable, but instead require some form of process-specific customisation prior to use, which can in turn act as a barrier to adoption. It is also highlighted that the shear diversity of pressures that come to bear during the product development process can also act as a barrier to adoption, and that the full integration of eco-design techniques will have to encompass approaches which overcome such pressures.     

生物电耦合对活性炭-超滤工艺处理地表微污染水的影响机理研究

利用生物电化学系统处理地表水时，弱电压不但会刺激电活性菌的呼吸作用，而且会引起氧化胁迫导致胞外聚合物（Extracellular Polymeric Substance，EPS）过量分泌.为确认在地表水处理中弱电压对膜污染和净水效率的影响，本研究设置了外加1.0 V直流电压（记为"BES系统"）和无外加电压（记为"CK系统"）的两组平行对照生物活性炭-超滤系统.经过50 d的运行，BES系统（36.1 kPa）相比CK系统（19.1 kPa）跨膜压差（Trans Membrane Pressure，TMP）增加更为显著，膜污染更严重.电路电流、电极电势和循环伏安（Cyclic Voltammetry，CV）曲线显示，随着装置运行，两系统生物膜逐渐稳定，并且产生具有氧化还原活性的EPS导致阳极电容增加.相比之下，BES系统产生了更多EPS，具有更高的阳极电容.水质指标显示，BES系统中比紫外吸光度（Specific Ultraviolet Absorbance，SUVA）、NH₄⁺-N和PO₄^3--P的去除增强但总有机碳（Total Organic Carbon，TOC）去除减弱；三维荧光光谱（Excitation-Emission-Matrix，EEM）和尺寸排阻色谱（Size Exclusion Chromatography，SEC）分析结果表明，BES系统膜池中产生了更多的生物聚合物，但腐殖质类有机物明显减少.活性炭表面EPS含量和腺嘌呤核苷三磷酸（Adenosine Triphosphate，ATP）含量的测试结果证实，弱电压刺激了生物膜的生长，同时增加了氧化还原活性EPS的含量.对膜表面累积多糖、蛋白质含量的测试分析进一步揭示了多糖类大分子生物聚合物在膜表面的累积是导致严重膜污染的直接原因.研究可为生物电化学系统在微污染水处理的研究和应用提供新的见解.     

2013年秋末黄渤海海水中胞外酶活性水平和垂直变化

于2013年11月6~23日对黄渤海进行了现场调查研究,对42个大面站以及A7、E7两个垂直站位进行了采样及环境参数的测定,研究了该海域海水中的9种胞外酶活性的水平及垂直变化.结果表明:秋末黄渤海表层海水中,9种胞外酶的平均活性由高到低排列为碱性磷酸酶[77.31nmol/(L·h)]>脂肪酶[23.59nmol/(L·h)]>β-D-葡萄糖苷酶[3.87nmol/(L·h)]>木糖苷酶[2.63nmol/(L·h)]>α-D-葡萄糖苷酶[1.58nmol/(L·h)]≈纤维素酶[1.47nmol/(L·h)]≈几丁质酶[1.29nmol/(L·h)]>肽酶[1.09nmol/(L·h)]>β-D-半乳糖苷酶[0.74nmol/(L·h)];表层海水9种胞外酶的活性大都呈现出黄海大于渤海,即南高北低的变化规律;表层海水中的β-D-葡萄糖苷酶活性和纤维素酶活性与温度呈现弱的正相关(P0.05);A7和E7两个站位的胞外酶活性的垂直分布显示不同胞外酶的活性在不同水深有明显的差异,基本表现为表层、10m和30m的活性高于50m、75m和底层的活性.     

Antenatal detection of neural tube defects: Comparison of biochemical and immunofluorescence methods

The aim of this study was to determine whether identification of glial cells in amniotic fluid samples could form a useful supplementary test in the antenatal diagnosis of neural tube defects (NTDs). In a 5-year study, 1452 samples of middle trimester amniotic fluid were examined blind to the results of other antenatal diagnostic tests and to the outcome of pregnancy. Reasons for amniocentesis included raised serum alpha-fetoprotein (329), previous NTD (73), and a family history of NTDs (71). Duplicate cytospin preparations were stained with Giemsa and an antibody to glial fibrillary acidic protein (GFAP), and on this basis a prediction of fetal NTD status was made which was not communicated to clinicians. Subsequent management of pregnancies was influenced only by the results of routine antenatal testing for NTDs. Twenty cases of NTDs occurred among the 1406 cases in which the outcome was subsequently known. Of these 20 cases, only five (four anencephalic, one spina bifida) were correctly predicted by immunofluorescent identification of GFAP-positive cells in the amniotic fluid. The remaining 15 cases (two anencephalic, 13 spina bifida) were not so identified. In a further 18 cases, apparently GFAP-positive cells were identified in the absence of NTDs. We conclude that GFAP immunofluorescence examination of routine amniocentesis samples of amniotic fluid is not a useful predictive test for NTDs.     

污水氯和二氧化氯消毒过程中溶解性有机物变化的三维荧光光谱解析

采用三维荧光光谱（3DEEM）技术对污水在氯和二氧化氯消毒过程中溶解性有机物（DOM）的变化进行了初步解析.结果表明,与饮用水、地表水等不同,生物处理出水含有较多的芳香族蛋白质和溶解性微生物代谢产物,且其中的腐殖质主要为生物源,芳香性较弱.在氯和二氧化氯消毒后,污水DOM中的芳香族蛋白质和微生物代谢产物类物质的荧光峰发生蓝移,即峰值的激发或发射波长减少了几个nm,这可能由芳香环结构的破坏引起;而腐殖质类物质的荧光峰发生红移,即峰值的激发或发射波长增加了几个到二十几个nm,此现象与地表水相反,可能与腐殖质的来源不同有关.与遗传毒性的变化类似,氨氮对水样氯消毒后的荧光变化有明显影响,但对二氧化氯消毒后的荧光变化无明显影响.     

Cells of the connective tissue differentiate and migrate into pollen sacs

In angiosperms, archesporial cells in the anther primordium undergo meiosis to form haploid pollen, the sole occupants of anther sacs. Anther sacs are held together by a matrix of parenchyma cells, the connective tissue. Cells of the connective tissue are not known to differentiate. We report the differentiation of parenchyma cells in the connective tissue of two Gordonia species into pollen-like structures (described as pseudopollen), which migrate into the anther sacs before dehiscence. Pollen and pseudopollen were distinguishable by morphology and staining. Pollen were tricolpate to spherical while pseudopollen were less rigid and transparent with a ribbed surface. Both types were different in size, shape, staining and surface architecture. The ratio of the number of pseudopollen to pollen was 1:3. During ontogeny in the connective tissue, neither cell division nor tetrad formation was observed and hence pseudopollen were presumed to be diploid. Only normal pollen germinated on a germination medium. Fixed preparations in time seemed to indicate that pseudopollen migrate from the connective tissue into the anther sac.