A novel method for artificial lipid-bilayer formation

Many proposals have been made regarding the development of biosensors using single-channel recording with an artificial planar bilayer. The fragile nature of bilayer membranes is the major difficulty for the application of the artificial bilayer technique to the development of biosensors. We have developed an apparatus that promptly forms artificial bilayers. This technique is more efficient than other techniques for forming artificial bilayers. Bilayer membranes could be formed within 10s requiring 1 microl of analyte solution to record single-channel currents using our apparatus. A bilayer was formed by pressing the membrane on an agarose layer with hydraulic pressure. With this novel apparatus, we have recorded single-channel currents of various types of channels such as the BK-channel, the nicotinic receptor channel and the ryanodine receptor channel. The properties of the channels determined with this novel technique agreed well with those determined with conventional techniques.     

Sun Compass Orientation of Tylos europaeus (Crustacea, Isopoda) Under Artificial Light

To provide a first assessment of the parameters used by adult individuals of the supralittoral isopod Tylos europaeus to recognize the sun as a compass orienting reference, we used the apparatus designed and tested with the amphipod T. saltator. The apparatus reproduces a scenario similar to the natural one (with a false sun and sky illuminated artificially). The scenario produced inside the apparatus is sufficient to induce the isopods to exhibit solar orientation similar to that of conspecifics tested under the natural sun and sky. Nevertheless, this ability depends on some threshold values of illumination of the artificial sun and sky: to obtain a good orientation the irradiance of the artificial sun and sky should be more than 0.4 and 1.3 μW/cm² respectively. When the artificial sky is not illuminated, the individuals show only a photopositive tendency.     

Purification and concentration of albumin solutions by diafiltration using a parallel membrane dialyzer]

The last two years, artificial Kidneys have been used for purification and concentration of human serum albumin solutions coming from plasma cracking in two Blood Transfusion Centers. Results are easily reproducible and the apparatus is reliable and of low cost. The properties of dialysis and ultrafiltration of the A.N. 69 membrane are useful for eliminating ethanol and water. The artificial Kidneys are effective at low pressure. It is then possible to use peristaltic pumps and to have a closed circuit. The whole apparatus must be sterilized with chemical reagents.     

Microcapsule artificial kidney: treatment of patients with acute drug intoxication

The microcapsule artificial kidney was used in the treatment of three patients with acute drug intoxication. The apparatus contains 300 g. of microencapsulated activated charcoal with a total membrane area available for diffusion of more than 2m.² The membrane thickness is only 500 A. These properties make possible a compact artificial kidney whose efficiency for the removal of uremic metabolites and drugs is much higher than standard hemodialysis apparatus. The microcapsules are made blood-compatible by coating with human albumin. A roller pump was used to propel the blood through the microcapsule artificial kidney at a flow rate of 300 ml./min. for two to three hours. The clearance values for glutethimide, methyprylon and methaqualone were much higher than those achieved by standard hemodialysis. Hemoperfusion quickly lowered the drug level in the blood with resulting clinical improvement.     

Separation and determination of mass and radioactivity of fermentation gases.

A procedure is described for the separation of permanent gases on a gas chromatograph, the determination of each component by means of a thermal conductivity detector and the simultaneous measurement of radioactivity in each peak by means of a proportional counter.Procedures for calibration of the apparatus and for calculation of absolute radioactivities in samples are given.The capabilities of the apparatus are illustrated by some results of experiments with an artificial rumen using ¹⁴C- and ³H-labeled compounds.     

Moon orientation in adult and young sandhoppers under artificial light

Our experiments, carried out at night and during the day on adults and laboratory-born young of the sandhopper Talitrus saltator, deal with the identification and use of the moon as an orientating factor. Sandhoppers were released in an apparatus (a Plexiglas dome) that produced a scenario similar to the natural one (with artificial sky, moon or sun illuminated at different intensities).When tested at night, the adult and young sandhoppers used the artificial moon like the natural one, independently of the intensity of illumination of the artificial sky and moon. In other words, sandhoppers tested at night always identified the artificial moon as the moon and never as the sun. In daytime releases, the seaward orientation failed at low intensities of artificial sky and sun illumination (3.07 and 1.55 microW cm2, respectively), whereas the sun compass was used effectively at higher levels of artificial sun and sky illumination. The innate ability of moon compass orientation in inexpert young sandhoppers was demonstrated even under artificial light.     

光敏蛋白菌紫质固相支撑人工膜系统的界面电荷

以固相支撑的菌紫质人工膜系统,在分子电子器件研究中占有重要地位。本文对其两种类型,Ｎ固相型和Ｃ固相型的界面电荷进行了研究,并提出了相关的模型。     

黄土高原丘陵沟壑区抗侵蚀植物叶表皮的生态适应性

采用叶表皮临时装片法研究了黄土高原丘陵沟壑区不同土壤侵蚀环境沟间地、沟谷地和沟间人工刺槐林地6种抗侵蚀植物叶表皮组成细胞的形态特征.结果表明: 沟间地抗侵蚀植物叶的上表皮气孔开度、气孔密度、气孔指数、气孔器长宽比可塑性、气孔器面积可塑性、表皮毛〖JP2〗密度和表皮细胞密度分别比沟谷地提高93.8%、66.8%、17.9%、36.4%、42.3%、199.4%和46.5%,下表皮分别提高90.4%、76.6%、9.8%、47.1%、43.9%、98.2%和50.1%;沟间人工刺槐林地叶上表皮各形态指标分别比沟谷地提高66.7%、20.5%、11.9%、37.9%、19.8%、113.1%和10.8%,叶下表皮分别提高106.7%、45.8%、11.9%、41.3%、21.2%、52.2%和28.1%.沟间地植物叶上、下叶表皮毛长度分别比沟谷地缩短58.8%和29.7%,表皮细胞面积分别比沟谷地减少40.3%和37.0%.沟间人工刺槐林地叶上、下表皮毛长度分别比沟谷地缩短25.0%和23.6%,表皮细胞面积分别比沟谷地减少22.2%和19.2%.抗侵蚀植物通过增加叶表皮气孔开度、气孔密度、气孔指数、气孔器长宽比可塑性、气孔器面积可塑性、表皮毛密度、表皮细胞密度和减少表皮毛长度、表皮细胞面积来适应较强的土壤侵蚀环境.     

Retractile processes in T lymphocyte orientation on a stimulatory substrate: morphology and dynamics

T cells of the immune system target infected and tumor cells in crowded tissues with high precision by coming into direct contact with the intended target and orienting the intracellular Golgi apparatus and the associated organelles to the area of the cell-cell contact. The mechanism of this orientation remains largely unknown. To further elucidate it we used three-dimensional microscopy of living T cells presented with an artificial substrate mimicking the target cell surface. The data indicate that long, finger-like processes emanate from the T cell surface next to the intracellular Golgi apparatus. These processes come in contact with the substrate and retract. The retraction accompanies the reorientation of the T cell body which brings the Golgi apparatus closer to the stimulatory substrate. Numerical modeling indicates that considering the forces involved the retraction of a process attached with one end to the cell body near the Golgi apparatus and with the other end to the substrate can bring the Golgi apparatus to the substrate by moving the entire cell body. The dynamic scenarios that are predicted by the quantitative model explain features of the reorientation movements that we measured but could not explain previously. We propose that retraction of the surface processes is a force-generating mechanism contributing to the functional orientation of T lymphocytes.     

An artificial lipid bilayer formed on an agarose-coated glass for simultaneous electrical and optical measurement of single ion channels

The purpose of this study is to develop an apparatus for simultaneous measurement of electrical and spectroscopic parameters of single ion channels. We have combined the single channel recording apparatus with an artificial lipid bilayer and a fluorescence microscope designed to detect single fluorescent molecules. The artificial membranes were formed on an agarose-coated glass and observed with an objective-type total internal reflection fluorescence microscope (TIRFM). The lateral motion of a single lipid molecule (beta-BODIPY 530/550 HPC) was recorded. The lateral diffusion constant of the lipid molecule was calculated from the trajectories of single molecules as D = 8.5 +/- 4.9 x 10(-8) cm(2)/s. Ionic channels were incorporated into the membrane and current fluctuations were recorded at the single-channel level. After incorporation of Cy3-labeled alametithin molecules into the membrane, bright spots were observed moving rather slowly (D = 4.0 +/- 1.6 x 10(-8) cm(2)/s) in the membrane, simultaneously with the alametithin-channel current. These data show the possibility of the present technique for simultaneous measurement of electrical and spectroscopic parameters of single-channel activities.     

Method of isolating pathogenic enterobacteria from atmospheric air]

The authors present comparative results of determination of sedimentation and aspiration methods of air pollution by pathogenic enterobacteria during the artificial sewage "raining". Results (in per cent) of isolation of salmonellae from the air by both methods proved to be very close; however, a possibility of isolation of pathogenic bacteria from the air increased with the use of the apparatus for the examination of large air volumes.     

Protective cytokinin action switches to damaging during senescence of detached wheat leaves in continuous light

The effect of exogenous application of the cytokinin meta -topolin [mT; N⁶-( meta -hydroxybenzyl)adenine] on artificial senescence of detached wheat leaves ( Triticum aestivum L. cv. Hereward) was studied and compared in leaves senescing under continuous light (100 µmol photons m⁻² s⁻¹) and darkness. Senescence-induced deterioration in structure and function of the photosynthetic apparatus was characterized by reduction in chlorophyll content, maximal efficiency of photosystem (PS) II photochemistry ( F _v/ F _m) and the rate of CO₂ assimilation, by increase in the excitation pressure on PSII (1 −  q _P) and a level of lipid peroxidation and by modifications in chloroplast ultrastructure. While in darkened leaf segments mT effectively slowed senescence-induced changes in all measured parameters, in light-senescing segments the effect of mT changed into opposite a few days after detachment. We observed an overexcitation of photosynthetic apparatus, as indicated by pronounced increases in the excitation pressure on PSII and in a deepoxidation state of xanthophyll cycle pigments, marked starch grain accumulation in chloroplasts and stimulation of lipid peroxidation in light-senescing leaf segments in mT. Possible mechanisms of acceleration of senescence-accompanying decrease in photosynthetic function and increase in lipid peroxidation during mT influence are discussed. We propose that protective mT action in darkness becomes damaging during artificial senescence in continuous light due to overexcitation of photosynthetic apparatus resulting in oxidative damage.     

Microbial responses to salt-induced osmotic stress

Summary The construction and assembly of a model root region is described. The model was used to manipulate the soil matrix, soil microorganisms, and to simulate release of root exudates. The design of the apparatus facilitated long-term, direct microscopic observations of microbial activity in soil and on artificial roots. Preliminary studies indicate that microbial responses to osmotic stress and to changes in components of exudate solutions are easily monitored.     

The electron microscopic and classic Golgi apparatus

The relationship of the membrane structure, designated in electron microscopy as the Golgi apparatus, to the classic Golgi apparatus in the light microscope were studied withFagopyrum. Comparison of these structures in plant cells with the same or similar structures in animal cells led to the following conclusions: there exist two groups of formations, impregnable with osmium or silver, considered as the classic Golgi apparatus. The first group contains the active membrane structures. These are the dictyosomes and the anastomosing form of the electron microscopic Golgi apparatus. To this group belongs also the endoplasmatic reticulum, which in plant cells forms dense vacuoles, having the appearance of the classic Golgi apparatus, and in animal cells occasionally has a similar arrangement as the anastomosing form of the Golgi apparatus. The second group comprises formation containing reserve and secretion material, i.e. predominantly products of the activity of the electron microscopic Golgi apparatus and of the endoplasmic reticulum (matter of the dense vacuoles, lipochondria, secretory granula etc.). In the plant cells, especially ofFygopyrum, the dictyosomes contained in the structures of the first group are separated from the formations of a reserve character in the second group, formed in the lumen of the endoplasmic reticulum (dense vacuoles). The identity of the dictyosomes with the osmiophilic platelets, considered by some authors in the light microscope as the classic Golgi apparatus, has not been proved up to present, because of the one-sidedness of the methods used nowadays. WithFagopyrum no foundation has been observed for the assumed formation of net-form structures by grouping of the dictyosomes. Structures similar to the net-form of the classic Golgi apparatus in the animal cell form only dense vacuoles. On the basis of the differentiation of both types of formations in the plant cell, the foundations were laid for the characterization of the classic Golgi apparatus in the animal cell. The net-form of the classic Golgi apparatus in the animal cell is obviously not artificial, but reflects the ultrastructural arrangement of the electron microscopic Golgi apparatus or of the endoplasmic reticulum. The problem of the suitability and specification of the name Golgi apparatus in the animal and plant cell was also discussed. In contrast to the opinion of some authors, it does not appear useful to remove the name golgi apparatus, designating the dictyosomes and the anastomosing forms of the smooth membranes.     

The repellent scent-mark of the honeybeeApis mellifera tigustica and its role as communication cue during foraging

Summary Experimental evidence for flower-marking in honeybees (Apis mellifera ligustica), using pairs of workers from the same colony foraging on an artificial patch of flowers, is reported. Workers marked artificial flowers with scent and strongly rejected all flowers they had recently visited. The same rejection behavior, in a lower although significant proportion, was observed when bees visited flowers just abandoned by the other individual of the pair. The repellent nature of this scent-mark was demonstrated with the use of an air extractor connected to the patch of artificial flowers. When the apparatus was turned on, the rejection behavior disappeared and bees accepted both flowers just abandoned by themselves and flowers just abandoned by the other bee. Differences in the response level of bees to their own marks or to the partner's marks suggest that the repellent scent-mark applied by a bee during foraging would basically be a self-use signal, although it certainly has value in communicating with other workers.     

An "artificial tongue" for calibrating solution flow characteristics of taste stimulus delivery systems

An "artificial tongue" is described which can be used to calibratethe stimulus delivery characteristics of flow systems for liquidtaste stimuli. The calibrator, which functions on the basisof conductance measurements, is used to ascertain the behaviourof certain test solutions in the flow system. The test solutionsare so prepared that they mimic the hydraulic properties ofthe stimulus solutions to be used. The special apparatus requiredis neither complex nor costly, and the procedures involved arestraightforward.     

鮸鱼弧菌病病原菌(哈维氏弧菌)的分离与鉴定

【目的】2009年春季,浙江省舟山地区养殖鮸鱼暴发弧菌病。症状主要表现为体表病灶部位出血、肌肉溃烂、内脏器官有白斑等。【方法】从病鱼体表溃疡部位及内脏分离出优势菌株090212,经人工感染证实该菌即为致病菌。通过API系统和菌体常规形态特征、培养特性和生理生化反应指标测定以及16S rRNA测序分析等综合鉴定,【结果】确认090212为哈维氏弧菌(Vibrio harveyi),该菌为革兰氏阴性,菌体呈短杆状,极生单鞭毛。该菌对氟苯尼考、四环素等5种抗生素敏感。【结论】哈维氏弧菌是海水养殖鱼类的常见致病菌,但作为养殖鮸鱼的病原菌尚属首次报道,将对鮸鱼的病害防治和健康养殖具有重要的指导意义。     

Ultrastructural changes in the neurons and interneuronal synapses of the hypothalamus in the progeny of morphine-dependent rats

The electron microscopic method has revealed early maturation of the protein-synthesizing apparatus of some cells, on the one hand, and underdevelopment of neuroblast volumes, appearance of artificial contacts, axonal degeneration and destruction in large trunks, on the other hand. Compensatory-restorative processes in the cell on the 14th and 21st days of the postnatal development result in a complete restoration of morphological organization of neurons and interneuronal connections in the hypothalamus of offspring of the morphine-dependent rats.     

Microencapsulation of chloroplast particles

Chloroplast and photosystem I particles were encapsulated in small spheres (about 20 μm diameter) with an artificial membrane built up by cross-linking amino groups of protamine with toluenediisocyanate. The artificial membrane was permeable to small substrate and product molecules but not to soluble proteins. Photosystem I activity was retained by the encapsulated chloroplast particles. Washed photosystem I particles were encapsulated with the soluble proteins, ferredoxin, and ferredoxin-NADP oxidoreductase, and the microcapsules photoreduced NADP using ascorbate plus dichlorophenolindophenol as the electron donor. The photosystem I particles were also encapsulated with hydrogenase from Chromatium and a very low rate of photoevolution of hydrogen was obtained. The results show that chloroplast membrane fragments can be encapsulated with soluble proteins that couple transfer reactions to the primary photochemical apparatus.     

Use of magnetic particles isolated from magnetotactic bacteria for enzyme immobilization

Summary We report the novel use of magnetic particles isolated from magnetotactic bacteria. Magnetotactic bacteria were collected from enriched sludge by use of a magnetic harvesting apparatus. Magnetic particles separated from magnetotactic bacteria were shown to be pure magnetite. Glucose oxidase and uricase were immobilized on magnetic particles. The activity of glucose oxidase immobilized on biogenic magnetites was 40 times that immobilized on artificial magnetites or Zn-ferrite particles. Both glucose oxidase and uricase coupled with biogenic magnetic particles retained their activities when they were reused 5 times.