Densitometric measurement of two cellular parameters related to the secretory activity of the subcommissural organ

The subcommissural organ (SCO) of vertebrates is located in the roof of the third brain ventricle, and secretes into this ventricle a glycoproteinaceous, fibre-like structure, the liquor fibre or Reissner's fibre. A method is described for the densitometric measurement by means of computer-controlled scanning cytophotometry of two cellular parameters directly related to the secretory activity of the subcommissural organ in European green frogs. These parameters are: i) the amount of stained secretory material in the SCO, and ii) the amount of secretory material in the SCO labelled by a radioactive precursor. It appears that scanning cytophotometry offers a fairly rapid, accurate, objective and reproducible method to measure these parameters in stained sections and in autoradiographs of the SCO if this histological material fulfils certain conditions.     

Searching for specific binding sites of the secretory glycoproteins of the subcommissural organ

The molecular organization of Reissner's fiber (RF), the structure of its proteins, and the permanent turnover of these proteins are all facts supporting the possibility that RF may perform multiple functions. There is evidence that CSF-soluble RF-glycoproteins may occur under physiological conditions. The present investigation was designed to investigate the probable existence within the CNS of specific binding sites for RF-glycoproteins. Three experimental protocols were used: (1) immunocytochemistry of the CNS of bovine fetuses using anti-idiotypic antibodies, raised against monoclonal antibodies developed against bovine RF-glycoproteins; (2) in vivo binding of the RF glycoproteins, perfusing into the rat CSF 125I-labeled RF-glycoproteins, or grafting SCO into a lateral ventricle of the rat; (3) in vitro binding of unlabeled RF-glycoproteins to rat and bovine choroid plexuses maintained in culture. One of the anti-idiotypic antibody generated by a Mab raised against RF-glycoproteins binds to choroidal cells. Furthermore, binding of RF-glycoproteins to the rat choroid plexus was obtained when: (1) the choroid plexus was cultured in the presence of unlabeled RF-glycoproteins; (2) the concentration of soluble RF-glycoproteins in the CSF was increased by isografting SCOs into a lateral ventricle; (3) radiolabeled glycoproteins were perfused into the ventricular CSF. This evidence suggests that the apical plasma membrane of the ependymal cells of the choroid plexus has specific binding sites for RF-glycoproteins, of unknown functional significance. The radiolabeled RF-glycoproteins perfused into the rat CSF also bound to the paraventricular thalamic nucleus, the floor of the Sylvian aqueduct and of the rostral half of the fourth ventricle, and the meninges of the brain and spinal cord. The labeling of the paraventricular thalamic nucleus points to a functional relationship between this nucleus and the SCO. The possibility that the SCO may be a component of the circadian timing system is discussed.     

Cell lineage of the subcommissural organ secretory ependymocytes: differentiating role of the environment

SCO-ependymocytes have a secretory activity and a neural innervation relating them to neurosecretory nerve cells. To elucidate the cell lineage of the SCO-ependymocytes and emphasize the role of the neural innervation in their differentiation, in particular 5-HT innervation, we analyzed the developmental pattern of expression of several glial and neuronal markers: (1) in the SCO of mammals possessing (rat, cat) or devoid (mouse, rabbit) of 5-HT innervation, (2) in rat 5-HT deafferented SCO, and (3) in rat SCO transplanted in a foreign environment, the fourth ventricle. The ability of SCO-ependymocytes to transiently express GFAP during development and express the glial alpha alpha-enolase confirms the glial lineage of the SCO-ependymocytes. Synthesis of vimentin by SCO-ependymocytes relates them to the classical ependymocytes. The ability of mature SCO-ependymocytes to take up GABA only when they are innervated by 5-HT terminal underlines the role of the neural environment on the differentiation of these ependymocytes and suggests that differential maturation of the SCO according to its innervation, may lead to specific functional specialization of this organ in different species.     

Analysis and quantification of the secretory products of the subcommissural organ by use of monoclonal antibodies

Bovine Reissner's fiber (RF) glycoproteins were used as antigen for the production of polyclonal and monoclonal antibodies (Mabs). We also produced Mabs against intracellular secretory glycoproteins of the bovine subcommissural organ (SCO). These Mabs were used for immunodetection of secretory proteins in situ (structural and ultrastructural immunocytochemistry), in blots, and in solutions. Three different antigen-mediated ELISA were designed to evaluate the affinity of the Mabs, to study the nature of the epitopes, and for competition test among Mabs. Two double antibody sandwich ELISA were designed to detect and quantify soluble secretory materials in different samples, to study coexistence of epitopes, and to elucidate whether epitopes for Mabs are repeated or not in the RF-glycoproteins. Twenty-three Mabs recognizing the bovine RF- and SCO-glycoproteins in solutions (ELISA) as well as in tissue sections, were obtained. Nineteen of these Mabs also recognized the pig SCO, 11 the rabbit SCO, 6 the dog SCO, and 5 the rat SCO. None of the Mabs recognized the SCO of non-mammalian species. The different types of ELISA demonstrated that: (1) the epitopes reside in the proteinaceous moiety of the secretion, (2) they coexist in the same molecular forms and, with few exceptions, they did not overlap, (3) they were not repeated in the secretory molecule(s). Three Mabs were used for immunoblotting of RF; one of them revealed the same band pattern as that shown by an anti-RF serum. It is concluded that all Mabs raised in our laboratory are directed against non-repeated sequences of RF-glycoproteins that have not been conserved in vertebrate phylogeny.     

Organ culture of the bovine subcommissural organ: evidence for synthesis and release of the secretory material

The subcommissural organ (SCO) is a brain circumventricular organ formed by ependymal and hypendymal secretory cells. It secretes glycoproteins into the cerebrospinal fluid of the third ventricle where they condense into a thread-like structure known as Reissner's fiber (RF). The present study was designed to investigate whether or not the bovine SCO continues to synthesize and release glycoproteins after a long-term culture. Cultured explants of SCO survive for several months. The content of the secretory granules present in the cultured ependymocytes displayed immunoreactive and lectin-binding properties similar to those of the core glycosylated glycoproteins found in the bovine SCO. The explants actively incorporated (35)S-cysteine. In the cultured ependymocytes, the pattern of distribution of the radioactive label and that of the immunoreactive secretory material was similar, thus indicating that this material has been synthesized during culture. At the ultrastructural level, the cultured tissue exhibited a high degree of differentiation comparable to that of the bovine SCO in situ. A striking finding was the observation of similar results when cerebrospinal fluid was used as a culture medium. The addition of antibodies against RF-glycoproteins into the culture medium allowed visualization, by means of different immunocytochemistry protocols, deposits of extracellular immunoreactive secretory material on the free surface of the cultured ependymocytes, indicating that release of secretory glycoproteins into the culture medium does occur. Primary culture of dispersed SCO ependymocytes, obtained either from fresh or organ cultured bovine SCO, showed that these cells release RF-glycoproteins that aggregate in the vicinity of each cell. The present investigation has shown that: (1) two types of secretory ependymocytes become evident in the cultured SCO; (2) under culture conditions, the SCO cells increase their secretory activity; (3) explants of bovine SCO synthesize RF-glycoproteins and release them to the culture medium; (4) after release these proteins aggregate but do not form a RF; (5) a pulse of anti-RF antibodies into the culture medium blocks the secretion of RF-glycoproteins for several days.     

Functional aspects of the subcommissural organ-Reissner's fiber complex with emphasis in the clearance of brain monoamines

Reissner's fiber (RF) extends along the cerebral aqueduct, fourth ventricle, and the entire length of the central canal of the spinal cord. It grows continuously in the caudal direction by addition of newly released glycoproteins by the subcommissural organ (SCO) to its proximal end. Several hypotheses about RF function have been advanced. One of them postulates that RF binds biogenic amines present in the CSF and clears them away. In recent years, this hypothesis has been tested in our laboratory by using several experimental protocols. Firstly, the CSF concentration of monoamines was investigated in RF-deprived rats subjected to immunological neutralization of the SCO-RF complex. Secondly, the capacity of RF to bind monoamines in vivo was studied by injecting radiolabeled serotonin or noradrenaline into the rat CSF, and by perfusing them into the CSF, during one week, using an Alzet's osmotic pump. In vitro binding studies were performed using isolated bovine RF. All the findings obtained indicate that RF binds monoamines present in the ventricular CSF and then transports them along the central canal. In the absence of RF, the CSF concentration of monoamines increases sharply.     

SCO-spondin, a glycoprotein of the subcommissural organ/Reissner's fiber complex: evidence of a potent activity on neuronal development in primary cell cultures

In the cattle, SCO-spondin was shown to be a brain-secreted glycoprotein specifically expressed in the subcommissural organ (SCO), an ependymal differentiation located in the roof of the Sylvian aqueduct. Furthermore, SCO-spondin makes part of Reissner's fiber (RF), a structure present in the central canal of the spinal cord. Sequencing of overlaping cDNA inserts after successive screening of a cattle SCO cDNA expression library allowed characterization of the complete sequence of this novel protein. Conserved domains were identified including twenty-six thrombospondin type 1 repeats (TSRs), nine low-density lipoprotein receptor LDLr type A domains (LDLRA), two epidermal growth factor EGF-like domains, and homologies to mucins and the von Willebrand factor were found in the amino- and carboxy- termini. In addition, SCO-spondin shows a unique arrangement "in mosaic" of these domains. The putative function of SCO-spondin in neuronal differentiation is discussed regarding these features and homologies with other developmental molecules of the central nervous system exhibiting TSR domains, and involved in axonal guidance.To correlate molecular and functional features of SCO-spondin, we tested the effect of oligopeptides whose sequences include highly conserved regions of the TSRs, LDLRA repeats, and a potent site of attachment to glycosaminoglycan, on cortical and spinal cord neurons in primary cell cultures. Peptides corresponding to SCO-spondin TSRs markedly increased adhesivity and neuritic outgrowth of cortical neurons and induced disaggregation of spinal cord neurons. Thus, SCO-spondin is a candidate to interfere with neuronal development and/or axonal guidance during ontogenesis of the central nervous system in modulating side-to-side and side-to-substratum interactions, and in promoting neuritic outgrowth. RF proper has a wide range of activity on neuronal differentiation, including survival, aggregation, and disaggregation effects and neurite extension of cortical and spinal cord neurones "in vitro." Thus, the SCO/RF complex may interact with developmental processes of the central nervous system including the posterior commissure and spinal cord differentiation.     

Peeling of the built-up edge of a cutting tool during the machining of a free cutting steel and its effect on tool life

A built-up edge repeatedly forms on the rake face of a cutting tool and then peels away during the machining of a carbon steel. The mode of peeling of the built-up edge from the rake face and its effect on tool life during the machining of free cutting steels was investigated. It was found that the peeling mode changes with the composition of free cutting steels and greatly affects tool life.     

浅谈汽车零部件磨损对其寿命的影响

分析零部件磨损的变化特性，及其对汽车技术状况的影响，以便采取有效措施，提高汽车利用率。     

Modeling of aluminum particle combustion with emphasis on the oxide effects and variable transport properties

A simplified analytical modeling of single aluminum particle combustion was conducted. Ignition and quasi-steady combustion (QSC) were separately formulated and integrated. Both the heat transfer from the hot ambient gas and the enthalpy of heterogeneous surface reaction (HSR) served to cause the particle ignition. Conservation equations were solved for QSC parameters in conjunction with conserved scalar formulation and Shvab-Zeldovich function. Limit temperature postulate was formulated by a sink term pertinent to the dissociation of the aluminum oxide near the flame zone. Effective latent heat of vaporization was modified for the thermal radiation. Ignition and QSC of the aluminum particle were predicted and discussed with emphasis on the effect of the aluminum oxide and variable properties. The model was validated with the experiments regarding ignition delay time, burning rate, residue particle size, flame temperature, QSC duration, and stand-off distance of the envelop flame. Agreement was satisfactory and the prediction errors were limited within 10%.     

An economical, semi-automatic system for measuring cellular tree structures in three dimensions, with special emphasis on Golgi-impregnated neurons.

The hardware and software of a semi-automatic system are described for the measurement of cellular tree structures in thick microscopical sections such as the process of Golgi-impregnated neurons. With this system, the morphologist--viewing the neuron directly through the microscope--measures the tree structures using three cartesian coordinates of characteristic points, e.g. centre of cell, origin of processes, bifurcation points, end points, deflection points, etc. The system requires an on-line computer, but does not demand a dedicated computer since a joy-stick generator usually drives the microscopical stage during cell tracking. A digital computer is utilized for acquisition of data, and, when needed, to return the stage to a required position. The microscope stage is coupled to three stepping motors with 0.5 micron as the smallest step. The stepping motor for movement of the stage parallel to the optical axis (z-direction) is coupled directly to a spindle mechanism, replacing the original cog wheel mechanism. This construction shows no measurable backlash. The practical advantages of this new measuring system in relation to the existing digital systems currently in use, its accuracy and its costs, are discussed.     

Morphological observations of small granule-containing (Chromaffin) cells in the celiac ganglion of the guinea pig,with emphasis on cell contacts

Utilizing electron microscopic observation, several contacts between small, granule-containing cells (SGC) and postganglionic neurons (PGN) in the celiac ganglion of the guinea pig have been observed. A SGC in very close association with a PGN was seen to receive a distinct synaptic contact that contained many vesicles with dense cores. This contact was morphologically unlike cholinergic synapses previously reported on chromaffin cells. Because the SGC and PGN were clearly separated by a thin rim of satellite cell cytoplasm mutual to both cells, it is not known how or if the SGC would possibly exert a synaptic or paracrine effect on the PGN. Also, intraganglion SGC existed as large well-vascularized islands within the celiac ganglion. These intraganlion clusters sometimes contained more than 50 cells and perhaps could be considered to function as localized neuroendocrine components within the ganglion by secreting granule products into the nearby blood vessels for local or distant effects, although this certainly is not known. This work reports a unique synaptic ending upon a single-occurring SGC, which, in turn, closely approximates a ganglion neuron in a soma-somatic relationship. In addition, a very close association (but no actual contact) was observed between granule-containing processes, presumably emanating from the intraganglion clusters, and PGN. Whatever the function of ganglionic SGC may be, the exact relationship between SGC and PGN presumably would be of great interest and potential importance. © 1994 Wiley-Liss, Inc.     

A study of the peeling of the built-up edge from a cutting tool during the discontinuous machining of a carbon steel and its effect on tool life

A built-up edge repeatedly forms and peels from the rake face of a cutting tool during the machining of a carbon steel. The mode of peeling and its effect on the tool life during the discontinuous machining of a carbon steel was investigated. The built-up edge peeled from the tool rake face when the tool entered a groove in the surface of the carbon steel. This significantly affected the tool life.     

人体血液及其润滑性能研究的现状与展望

血液的润滑性能研究涉及到血液流变学、生物力学、流体力学、材料科学、摩擦学等多学科领域。目前国内外对血液作了大量的研究工作,但在血液润滑机制的研究上仍处于探索阶段。总结了血液及其摩擦润滑性能研究方面所取得的成果,指出目前血液及其摩擦润滑性能的研究主要是从宏观的角度进行探讨,没有从微观尺度研究血液摩擦润滑的机制。提出了今后血液润滑性能研究的主要内容,包括基于细胞生物力学的血液润滑性能基础理论研究以及物理环境对血细胞生物行为过程及润滑性能影响等方面的研究。     

Layer formation on the rake face of a cutting tool and its effect on tool life

The relation between the formation of a deposited layer on the rake face and the life of the cutting tool during the machining of carbon steels was examined. The chemical composition of the layer depended upon the types of non-metallic inclusions in the steels, and the life of the tool was considerably affected by them.     

创造性思维方式在工业产品全生命周期中的应用探讨

分析了入世后中国工业产品设计的状况，从创造性思维的概念出发，阐述了创造性思维在产品设计中的意义以及在产品全生命周期中的应用。     

Morphological differentiation of seven parthenogenetic Artemia (Crustacea: Branchiopoda) populations from China,with special emphasis on ploidy degrees

Parthenogenetic Artemia from seven Chinese locations with different elevations and various ploidies are characterized by phenotypic and morphometric analyses. Our findings show that the studied populations exhibit dissimilar patterns of ovisac. Four phenotypic patterns of furca are qualified and one of them is shared among di‐, tetra‐ and pentaploid Artemia. Results of discriminant analysis based on morphometric data reveal that tetra‐ and pentaploid populations are grouped together, but the Aqqikkol Lake population is clearly differentiated. Previous hypothesis/conclusion that polyploid Artemia are larger than diploids is only partly supported by the present results, which show that pentaploid and tetraploid populations are larger than the mostly diploid populations in terms of the total length, but the body size of the Aibi Lake triploids has not significant difference with the sympatric diploids and the mostly diploid Aqqikkol population that inhabit in very high altitude has the largest body size among all parthenogenetic populations. The founding confirms that body size of Artemia is following with Bergmann's rule. Microsc. Res. Tech. 79:258–266, 2016. © 2016 Wiley Periodicals, Inc.     

Subpixel position measurement using 1D, 2D and 3D centroid algorithms with emphasis on applications in confocal microscopy

The accuracy with which centroid algorithms in 1D, 2D and 3D can estimate an object's position has been investigated. Three factors that can influence the method's accuracy have been investigated: systematic error of the algorithm, influence of photon noise and the influence of perturbations such as scanning nonlinearity. The variation of the accuracy with parameters that are relevant for confocal microscopy, such as object diameter and photon noise/pixel, has been considered. Theory and simulations presented show that the variation of the accuracy with respect to such parameters can differ drastically between the 1D, 2D and 3D cases. Experiments performed using microspheres show that the magnitudes of the three types of error can be approximately the same under normal operating conditions and that it is therefore necessary to take all three into account when assessing the total error.