Müller glial cells in anuran retina

Whereas in the brain, the activity of the neurons is supported by several types of glial cells such as astrocytes, oligodendrocytes, and ependymal cells, the retina (evolving from the brain during ontogenesis) contains only one type of macroglial cell, the Müller (radial glial) cells, in most vertebrates including the anurans. These cells span the entire thickness of the tissue, and thereby contact and ensheath virtually every type of neuronal cell body and process. This intimate topographical relationship is reflected by a multitude of functional interactions between retinal neurons and Müller glial cells. Müller cells are the principal stores of retinal glycogen, and are thought to fuel retinal neurons with substrate (lactate/pyruvate) for their oxidative metabolism. Furthermore, Müller cells are involved in the control and homeostasis of many constituents of the extracellular space, such as potassium and perhaps other ions, signaling molecules, and of the extracellular pH. They also seem to play important roles in recycling mechanisms of photopigment molecules and neurotransmitter molecules such as glutamate and GABA. By containing the main retinal stores of glutathione, Müller cells may protect retinal neurons against free radicals. Moreover, Müller cells express receptors for many neuroactive substances, and may also release such substances to their neighbouring neurons. Thus, Müller cells exert many functions crucial for signal processing in the normal retina. Moreover, Müller cells change their properties in cases of retinal disease and injury, and may either support the survival of neuronal cells or accelerate the progress of neuronal degeneration.     

Cellular and subcellular distribution of metals in molluscs

The cellular processes involved in metal metabolism in molluscs are reviewed, with emphasis on the contribution of microscopy (AMG, ARG, EPMA, and SIMS) to both basic research of metal cell biology and applied environmental research. In molluscs, metal uptake may occur by facilitated diffusion, active transport, or endocytosis, and can be enhanced by MT synthesis or formation of mineralized granules. In aquatic molluscs, gills constitute a key interface for dissolved metal uptake, where metals are bound to MT, incorporated into lysosomes, and released basally towards the blood plasma and circulating hemocytes. However, particulate metal uptake is mainly achieved via the digestive tract by endocytosis; further metals are transferred first to lysosomes and then to residual bodies, especially in the digestive cells of the digestive gland. Additionally, metals can be accumulated selectively in specific cell types. As ligands pools differ from cell to cell, different metals may be retained in different cell types. Class "a" metals are localized in cells with granules composed of carbonate, oxalate, phosphate, and sulfate (oxygen donors), whereas "b" metals are associated with those cell types rich in sulfur and nitrogen ligands (sulfur donors). In molluscs, oxygen donors occur in connective tissue calcium cells and basophilic cells, whereas sulfur donors are present in digestive cells, podocytes, nephrocytes, and rhogocytes. Hemocytes, which constitute the most relevant system for metal transport between tissues, move around the body and may penetrate tissues and remove metals from the inner medium to be accumulated in lysosomes as nondigested products. Rhogocytes also participate in metal mobilization, accumulation, and release. The assessment of metal levels in target cells of sentinel molluscs by microscopic techniques provides an early-warning measure, with promising applications as an exposure biomarker for environmental monitoring programs.     

The clinical significance of metal ion release from cobalt-chromium metal-on-metal hip joint arthroplasty

Metal-on-metal (MOM) bearings offer extremely low wear and the avoidance of polyethylene but generate metallic wear particles. Although their total volume is dramatically smaller than polyethylene debris, these particles are in the nanometre size range and are many times more numerous. Metallic particles are ingested by macrophages or may be disseminated via lymphatics to the reticuloendothelial system. They corrode, and metal ions are present in the circulation and concentrated in erythrocytes. Excretion of metal ions via the kidneys seems to balance their generation in patients with MOM implants. However, highly sensitive detection methods can be used to show that levels of circulating cobalt and chromium ions are several times the normal level. These concentrations are well within the limits identified as dangerous to health in workers exposed to industrial chemicals, and also considerably lower than the levels found to cause cell toxicity in vitro. The local concentrations of particles and metal ions in the synovial tissue may occasionally exceed these limits and cause tissue necrosis. Clinical experience of lysis is rare in association with MOM bearings, as are hypersensitivity reactions and MOM bearings have had an excellent record over four decades and have a favourable benefit to risk ratio. Further reduction in risk will be achieved by improvement of materials, engineering, and accuracy of insertion.     

Infrared consequence spectroscopy of gaseous protonated and metal ion cationized complexes

In this article, the new and exciting techniques of infrared consequence spectroscopy (sometimes called action spectroscopy) of gaseous ions are reviewed. These techniques include vibrational predissociation spectroscopy and infrared multiple photon dissociation spectroscopy and they typically complement one another in the systems studied and the information gained. In recent years infrared consequence spectroscopy has provided long‐awaited direct evidence into the structures of gaseous ions from organometallic species to strong ionic hydrogen bonded structures to large biomolecules. Much is being learned with respect to the structures of ions without their stabilizing solvent which can be used to better understand the effect of solvent on their structures. This review mainly covers the topics with which the author has been directly involved in research: structures of proton‐bound dimers, protonated amino acids and DNA bases, amino acid and DNA bases bound to metal ions and, more recently, solvated ionic complexes. It is hoped that this review reveals the impact that infrared consequence spectroscopy has had on the field of gaseous ion chemistry. © 2009 Wiley Periodicals, Inc., Mass Spec Rev 28:586–607, 2009     

Influence of calcium and amino acids on the osmium impregnation of the endoplasmic reticulum

The aim of this study was to define further the interaction between osmium and organelle content in cells prefixed with glutaraldehyde. We have studied the reaction of osmium with divalent or trivalent cations (calcium, barium, zinc, aluminum, and iron) and various amino acids in the same conditions prevalent in histological techniques, in particular with Thiéry's technique of metal impregnation. Experiments were carried out in vitro in test tubes, on cellulose acetate discs, or with an immunodiffusion apparatus. Some experiments were also carried out with tissue extracts (kidney and intestine). Our studies suggest that calcium is in general essential for the formation of osmium black, but also that lysine is reactive even in the absence of calcium and that a few amino acids–such as tryptophan, ornithine, cysteine, and aspartic acid–are only slightly reactive in the absence of calcium. Other amino acids do not seem to participate in the endoplasmic reticulum osmium impregnation even in the presence of calcium ions. Our studies also suggest that osmium reactivity reflects calcium binding sites and not only calcium content.     

Structural analysis of organometallic compounds with soft ionization mass spectrometry

The analysis of organometallic compounds with mass spectrometry has some special features in comparison with organic and bioorganic compounds. The first step is the choice of a suitable ionization technique, where the electrospray ionization is certainly the best possibility for most classes of organometallic compounds and metal complexes. Some ionization mechanisms of organometallic compounds are comparable to organic molecules, such as protonation/deprotonation, and adduct formation with sodium or potassium ions; however, in many cases, different mechanisms and their combinations complicate the spectra interpretation. Organometallics frequently undergo various types of adduct and polymerization reactions that result in significantly higher masses observed in the spectra in comparison to molecular weights of studied compounds. Metal elements typically have more natural isotopes than common organic elements, which cause characteristic wide distributions of isotopic peaks; for example, tin has ten natural isotopes. The isotopic pattern can be used for the identification of the type and number of metal elements in particular ions. The ionization and fragmentation behavior also depend on the type of metal atom; therefore, our discussion of mass spectra interpretation is divided according to the different type of organometallic compounds. Among various types of mass spectrometers available on the market, trap-based analyzers (linear or spherical ion-traps, Orbitrap) are suitable to study complex fragmentation pathways of organometallic ions and their adducts, whereas high-resolution and high-mass accuracy analyzers (time-of-flight-based analyzers, or Fourier transform-based analyzers-Orbitrap or ion cyclotron resonance mass spectrometers) provide accurate masses applicable for the determination of the elemental composition of individual ions.     

Localization of chemical elements and isotopes in the leaf of soybean (Glycine max) by secondary ion mass spectrometry microscopy: critical choice of sample preparation procedure

Secondary ion mass spectrometry (SIMS) is one of the few microscopical methods that potentially can detect and in situ localize the various isotopes of virtually all elements. Recent work with SIMS has demonstrated the possibility of imaging the distribution of various elements in plant cell and tissues. However, in these studies, the elements were incorporated in cell macromolecules or associated with structural polymers, precipitated or immobilized in dry seeds. The localization of mineral ions is of particular significance for the physiology of higher plants owing to their quantitative importance and the impact of their cellular distribution on metabolic regulation. Here we analyse the possibility of mapping different elements (K, Ca, Mg, P, S, ¹⁵N and ¹⁴N) present as soluble and/or bound forms in highly vacuolated leaf cells. Cryoprocedures to prepare samples for SIMS detection are described and discussed. The quality of the results is assessed at each step of the sample preparation and analysis. Various methodologies are used, including photonic and electronic microscopies, and the agreement of the observed ion distribution with current knowledge of ion compartmentalization in plant cells. The K/Ca emission ratio is proposed as an index of the degree of preservation of the natural ion distribution to critically evaluate the results and identify where artefacts are likely to occur.     

Ultrastructural neurobiology of the olfactory mucosa of the brown trout, Salmo trutta.

This paper describes four investigations of the olfactory mucosa of the brown trout: 1) the ultrastructure of the olfactory mucosa as revealed by scanning (SEM), conventional transmission (TEM), and high voltage (HVEM) electron microscopy; 2) light and electron-microscopic investigations of retrograde transport of the tracer macromolecule horseradish peroxidase (HRP) when applied to the cut olfactory nerve; 3) SEM and TEM investigations of the effects of olfactory nerve transection on cell populations within the olfactory epithelium; and 4) ultrastructural investigations of reversible degeneration of olfactory receptors caused by elevated copper concentrations. The trout olfactory epithelium contains five cell types: ciliated epithelial cells, ciliated olfactory receptor cells, microvillar olfactory receptor cells, supporting cells, and basal cells. The ciliated and microvillar olfactory receptor cells and a small number of basal cells are backfilled by HRP when the tracer is applied to the cut olfactory nerve. When the olfactory nerve is cut, both ciliated and microvillar olfactory receptor cells degenerate within 2 days and are morphologically intact again within 8 days. When wild trout are taken from their native stream and placed in tanks with elevated copper concentrations, ciliated and microvillar cells degenerate. Replacement of these trout into their stream of origin is followed by morphologic restoration of both types of olfactory receptor cells. Ciliated and microvillar receptor cells are primary sensory bipolar neurons whose dendrites make contact with the environment; their axons travel directly to the brain. Consequently, substances can be transported directly from the environment into the brain via these "naked neurons." Since fish cannot escape from the water in which they swim, and since that water may occasionally contain brain-toxic substances, the ability to close off--and later reopen--this anatomic gateway to the brain would confer a tremendous selective advantage upon animals that evolved the "brain-sparing" capacity to do so. Consequently, the unique regenerative powers of vertebrate olfactory receptor neurons may have their evolutionary origin in fishes.     

Tribochemistry and the development of AW and EP oil additives — a review

The antiwear (AW) and extreme pressure (EP) properties of chlorine, sulphur, phosphorus-containing organic compounds and zinc dialkyldithiophosphate (ZDDP) oil additives are affected greatly by their reactive ability to the metal surface, as well as by the chemical composition, chemical state, physical and mechanical properties of the protective films formed. Over four decades, research has taken place on boundary lubrication, and a much better understanding of the AW and EP action mechanism of one additive by itself in base oil has been obtained, and much more knowledge on the relationship between wear and decomposition of additives, adsorption and reaction of the additive or its decomposed products with metal, has been gained. A series of analytical methods and a great number of modern surface analytical tools have been set up and used for research in this area. The problems and some suggestions for the future study on boundary lubrication of oil additive are proposed.     

Nanometer Scale Chemomechanical Characterization of Antiwear Films

We report the first nanometer scale chemical and mechanical (chemomechanical) characterization of selected features of a tribologically derived zinc dialkyl-dithiophosphate (ZDDP) antiwear film. AFM permits identification of the features responsible for preventing wear. These features are identified by nearby microscale fiducial marks, and their mechanical properties are determined by imaging nanoindentation. The same features are then studied by X-ray photoelectron emission microscopy (X-PEEM), which provides both elemental and chemical information at 200 nm spatial resolution. The mechanical properties are then determined for the same features, which are formed of a polyphosphate glass. This information provides new insights into the mechanisms by which ZDDP antiwears films are effective at inhibiting asperity contact between two metal surfaces     

One hundred years of interstitial cells of Cajal

This review is a portrayal of the evolution of ideas involving the interstitial cells of Cajal in changing disguises as dull fibroblasts, not very exciting Schwann cells, or perhaps quite important, though primitive neurons. However, today unmasked (we believe), they reveal themselves as myoid cells, a role that, judging by current interest, is far more exciting than former ones. Close to 500 publications from 1860-1999 have contributed to the discussion in one way or the other. This literature contains a wealth of correct observations but obviously also wrong interpretations, which are seen as a result of too blind a belief in specificities of visualization methods, combined with a desire to interpret even the hidden detail. It has been my objective to attempt to trace the origins of viable ideas, and I have therefore focused on relatively few authors. The most recent development from 1980 until today is so well covered by easily accessible reviews that I have resorted to a mere, but hopefully complete, list of them. Modern ICC'ists have so far been caught in the external muscle of the gut and kept their hands off its internal affairs. However, while working my way through the literature it struck me that a number of recent studies may provide the elements of a plausible model for the villous contraction mechanism. In the present context, an important point is that the very first published interstitial "neurons" from Cajal's hand-of the intestinal villus, 1889-may achieve new significance as a possible correlate to the regulatory ICC of the intestinal muscularis. Partly to make this point, I have taken the liberty of giving a short account of recent results from our lab.     

Active dendritic properties constrain input-output relationships in neurons of the central olfactory pathway in the crayfish forebrain

Parasol cells are multimodal sensory interneurons of the hemi-ellipsoid body in the decapod forebrain. In reptant crustaceans, the hemi-ellipsoid body resides in the base of the eyecup, as an appendage to the terminal medulla. Parasol cells exhibit periodic depolarizations at a frequency of 0.5-1.0 Hz. I have investigated the role of these periodic depolarizations and their superimposed impulse bursts in affecting the input/output properties of these neurons. Parasol cells receive input from photic, olfactory, and mechanosensory pathways. Strong stimulation over any one of these pathways can lead to the generation of one or more impulse bursts in a subset of parasol cells, timed to occur at the peak of successive periodic depolarizations. A role for the periodic depolarizations in the function of the parasol cells has yet to be established. I suggest the possibility that they may act as a nonlinear amplifier that boosts spatially-summated excitatory synaptic potentials from strong or appropriate stimuli above threshold for burst generation. Another possibility includes modification of voltage-sensitive ion channels in the dendritic membrane, permitting a more effective spread of excitatory synaptic currents to impulse or burst initiating zones. Impulse bursts may be a highly effective mode of output for these neurons, especially so as they occur synchronously in a subset of cells in response to strong sensory input. Furthermore, backfiring of bursts into the dendritic tree has a brief (2-3 second) but effective suppressive action upon weak sensory input, which can thereby be masked by stronger, burst-generating input. This masking phenomenon is seen in other arthropod sensory interneurons, where its physiological basis appears to be a transient accumulation of intracellular Ca(++) ions that open calcium-sensitive potassium channels.     

Limit analysis on surface roughness and dimensional accuracy of spray metallic crust for rapid tooling production by metal arc spraying process

Repetitively prototyping several prototypes of the same component using currently available rapid-prototyping equipment can be very costly. Substantial savings can be attained with the use of a metal spray process to produce a crust shell on a rapid-prototyping pattern coated with mould release agent (PVA), and then backing the crust with aluminium granules so as to form rapid mould/tooling (RT). The smoothness of the innermost spray layer and its dimensional accuracy in relation to the RP prototype are factors that ensure good quality of parts replicated by RT. This paper establishes a preliminary model for predicting the upper bound of the former and the lower bound of the latter, which have been experimentally verified by arc spraying zinc onto PVA. The theoretical model permits the prediction of spraying parameters for controlling the achievable surface finishing and dimensional accuracy of a spray in RT production.     

液体样本重金属离子检测微流控传感器与系统

重金属离子是水污染的主要污染物之一,即使微量的重金属离子也会对生态环境和生物造成危害。 水体中的重金属离 子通过食物链在人体中积累,汗液、血清等体液中的重金属离子可作为特定疾病的标志物。 实现液体样本中重金属离子的快 速、灵敏检测,对生态环境监管和精准医疗具有重要意义。 首先介绍了基于微机电系统(MEMS)技术的重金属离子传感器,包 括紫外-可见分光光度法、比色法、荧光检测法、发光化学法、电化学法和悬臂梁式检测方法。 其次论述了与上述检测方法匹配 的便携式仪器,最后对未来重金属离子检测传感器与系统的发展趋势和应用前景进行了展望。     

The terminal nerve and its relation with extrabulbar "olfactory" projections: lessons from lampreys and lungfishes

The definition of the terminal nerve has led to considerable confusion and controversy. This review analyzes the current state of knowledge as well as diverging opinions about the existence, components, and definition of terminal nerves or their components, with emphasis on lampreys and lungfishes. I will argue that the historical terminology regarding this cranial nerve embraces a definition of a terminal nerve that is compatible with its existence in all vertebrate species. This review further summarizes classical and more recent anatomical, developmental, neurochemical, and molecular evidence suggesting that a multitude of terminalis cell types, not only those expressing gonadotropin-releasing hormone, migrate various distances into the forebrain. This results in numerous morphological and neurochemically distinct phenotypes of neurons, with a continuum spanning from olfactory receptor-like neurons in the olfactory epithelium to typical large ganglion cells that accompany the classical olfactory projections. These cell bodies may lose their peripheral connections with the olfactory epithelium, and their central projections or cell bodies may enter the forebrain at several locations. Since "olfactory" marker proteins can be expressed in bona fide nervus terminalis cells, so-called extrabulbar "olfactory" projections may be a collection of disguised nervus terminalis components. If we do not allow this pleiomorphic collection of nerves to be considered within a terminal nerve framework, then the only alternative is to invent a highly species- and stage-specific, and, ultimately, thoroughly confusing nomenclature for neurons and nerve fibers that associate with the olfactory nerve and forebrain.     

Recent developments in the ion/ion chemistry of high-mass multiply charged ions

The ability to form multiply charged high-mass ions in the gas-phase, most notably via electrospray ionization (ESI), has allowed the study of many different combinations of positively and negatively charged ions. The charged products are directly amenable to study with mass spectrometry. Ion/ion reactions have proved to be "universal" in the sense that the high exothermicities and large rate constants associated with essentially any combination of oppositely charged ions lead to reaction regardless of the chemical functionalities associated with the ions. These characteristics make ion/ion reactions potentially analytically useful provided reagent ion densities and spatial overlap of the oppositely charged ions are high. These conditions can be readily met by several instrumental configurations. The focus of this review is to highlight developments in this field since 1998. Novel instrumentation has been developed to study ion/ion reactions, such as atmospheric pressure ion/ion reactors followed by mass analysis, or electrodynamic ion trap mass spectrometers, which are used as reaction vessels at sub-atmospheric pressures. A wide variety of reaction phenomenologies have been observed in various ion/ion reactions, with proton transfer being the most common. New phenomenologies have been observed in the reactions of multiply charged positive ions with singly charged negative ions, including cation transfer and cation exchange. A new series of reactions between multiply charged positive ions and multiply charged negative ions have been made possible by recent instrumentation developments. These reactions have led to the observation of proton transfer and complex formation. These observations have provided new insights into ion/ion reaction dynamics and a bound orbit model appears to best account for experimental results. New applications are also discussed for a several ion/ion reaction.     

Stereological methods in the analysis of neuronal parameters in the central nervous system*

The structure of the central nervous system (CNS) is briefly explained. It is compared with the electronic computer and the differences between the two are stated. The methods of measuring this structure and its function are reported. The surface of the cortex as well as the volume of different grey and white matters making up the brain can macroscopically be evaluated on serial slices with the aid of stereological procedures. There is an exponential correlation between volume of the cortex and volume of brain and body in mammals (Fig. 3). The density of neurons and the lengths of their processes can be examined by light microscopy. An improved procedure of counting neurons in the cortex is described. With regard to mammals it is interesting that the densities of neurons in larger brains are lower than in smaller ones. The correlation is negatively exponential. Man and primates have a higher density of cells than the rest of the mammals; their neurons, however, are small. This corresponds to a specialization that can be considered to represent a miniaturization of the neurons (Fig. 4). The topological analysis of dendritic trees is mentioned. The lengths, surfaces and volumes of processes of cells and the amount of synapses can be measured on electron micrographs. The principles and problems of stereology are described by an example of how to measure the myelinated fibres. Some results of the development of these fibres from birth on are reported upon (Fig. 6). In conclusion, some data of the probable dimensions of the human brain are given.     

Immunohistochemical characterization of TH13‐L2 spinal ganglia neurons in sheep (Ovis aries)

Spinal ganglia (SG) neurons are commonly classified according to various specific features. The most widespread classification based on morphological and ultrastructural features subdivides SG neurons into light and small dark neurons. Using immunohistochemical, histochemical and lectin methods, it is possible to further subdivide the small dark neurons into two subpopulations: peptidergic and nonpeptidergic neurons. The majority of studies on SG neurons were carried out on mice and rats; there are few or no studies on large mammals. In this study, some of the widely used neuronal markers, neurofilament 200 kDa (NF200), substance P (SP), calcitonin gene‐related peptide (CGRP) and isolectin B4 (IB4), were employed to characterize neuronal nitric oxide synthase (nNOS)‐immunoreactivity (‐IR) in sheep (Ovis aries) SG (Th13‐L2) neurons. The majority of the SG neurons were IB4‐labeled (79 ± 10%), followed by NF200‐ (45 ± 4%), NOS‐ (44 ± 10%), SP‐ (42 ± 5%) and CGRP‐IR (35 ± 7%) neurons. The triple staining experiments showed that a higher percentage (75 ± 16%) of NOS‐IR neurons bound both IB4 and CGRP, or both IB4 and SP (49 ± 6%). The IB4 marker showed an unexpected staining pattern; in fact, IB4‐labeled neurons largely colocalized with NF200, usually considered a marker of light SG neurons, and with CGRP and SP. For this reason, IB4 cannot be employed in sheep to differentiate between light and dark neurons, or between peptidergic and nonpeptidergic neurons. These results suggest the importance of being cautious when comparing data among different species. Microsc. Res. Tech., 2010. © 2009 Wiley‐Liss, Inc.     

Cam and Tappet Lubrication. IV–Radioactive Study of Sulfur in the EP Film

A radioactive tracer, sulfur-35, synthesized into zinc dialkyl dithiophosphate molecules, was used to study the EP film formed on cast iron cams and tappets run in motor oils containing this additive. The sulfur content of static films increased with immersion time and temperature, and the presence of phosphate-coated metal surfaces. The bound sulfur of films formed during dynamic tests increased with running time, load, and with the use of phosphate-coated surfaces. These conditions also influenced the ratio of Zn:P:S contained in both static and dynamic films. Zinc, and particularly phosphorus, in the films increased more rapidly than the sulfur with increased temperature and/or pressure. The dynamic films are not easily worn off by running in nonadditive oil. The mechanism of action of zinc dithiophosphates appears to be related to chemical reactions of additive decomposition products with the metal surfaces to form tightly-bound solid films which reduce damage under extreme pressure conditions. Radioactive counting and X-ray spectroscopy were used to obtain the amount of sulfur and zinc on tappets. Densitometer traces of the autoradiographs were utilized to determine the distribution of the sulfur on cam and tappet surfaces.