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PubMed1998

Behavioral constraints in the development of neuronal properties: a cortical model embedded in a real-world device.

The ability of organisms to categorize diverse and often novel stimuli depends on ongoing interactions with their environment. In a modality such as vision, categorization requires the generation of both selective and invariant responses of cortical neurons to complex visual stimuli. How does behavior contribute to shaping the responses of these neurons? Analysis of this question is made difficult by the complex multilevel interactions between many neural and behavioral variables. To mitigate this difficulty, we studied the development and ongoing plasticity of pattern-selective neuronal responses by means of synthetic neural modeling. For this purpose, we constructed Darwin V, which consists of a simulated neuronal model embedded in a real-world device that is capable of motion and autonomous behavior. The neuronal model consists of four major components: a visual system (containing cortical and subcortical networks); a taste system based on conductance; sets of motor neurons capable of triggering behavior; and a diffuse ascending (value) system. The modeled visual cortex consists of two areas: a topographic map responsive to elementary features connected to a higher-order map composed of initially non-selective neuronal units. During behavior over time in its environment, Darwin V encounters numerous objects consisting of black metal cubes displaying different patterns of white blobs and stripes. Initially, the lack of specific higher-order visual responses does not allow visual pattern discrimination, and appetitive and aversive behaviors are triggered by the 'taste' (surface conductivity of objects) alone. In the course of sensory experience, however, changes occur in visual and sensorimotor connection strengths, with two major consequences. First, units within the higher visual area acquire responses that are both pattern selective and translation invariant. Second, as a result of the operation of the value system, these responses become linked to appropriate behaviors. Analysis of Darwin V after such changes indicates that the continuity of self-generated movements is essential for the development of pattern-selective and translation-invariant responses. The concomitant development of a preference for foveal over parafoveal objects was found to be due to increased behavioral interactions with object cubes gripped by the centrally mounted effector (snout) of Darwin V. Finally, even after development of higher-order visual responses, visual responses to more frequently encountered objects continued to be enhanced, while other responses were diminished. Overall, the detailed study of Darwin V over multiple levels of organization provides a heuristically revealing example of the crucial role played by behavioral and environmental interactions in the development of complex responses by specialized neurons.

Behavior↗
PubMed1984

Basic and 'special' stains for plastic sections in bone marrow histopathology, with special reference to May-Grünwald Giemsa and enzyme histochemistry.

A battery of morphological, histochemical, and enzyme histochemical stains have been experimented on semithin sections of glycol-methacrylate-embedded bone marrow biopsies. We have been able to reproduce on sections the typical 'Romanowsky effect' which characterizes May-Grünwald Giemsa-stained smears of bone marrow or peripheral blood. This appears to be of critical importance for proper routine morphological evaluation of bone marrow biopsies. Conventional histochemical stains, and the enzyme histochemistry reactions that are most useful and widely used in the study of marrow aspiration smears have been successfully applied to plastic sections: in this way the evaluation of the cytochemical profiles of marrow diseases, especially leukemias, may be included in the histopathologist's diagnostic approach, with the additional advantage of preserving the architecture of the tissue and the relationship between haemapoietic cells and stromal components.

Biopsy, Needle↗
PubMed1993

Environmental scanning electron microscope imaging examples related to particle analysis.

This work provides examples of some of the imaging capabilities of environmental scanning electron microscopy applied to easily charged samples relevant to particle analysis. Environmental SEM (also referred to as high pressure or low vacuum SEM) can address uncoated samples that are known to be difficult to image. Most of these specimens are difficult to image by conventional SEM even when coated with a conductive layer. Another area where environmental SEM is particularly applicable is for specimens not compatible with high vacuum, such as volatile specimens. Samples from which images were obtained that otherwise may not have been possible by conventional methods included fly ash particles on an oiled plastic membrane impactor substrate, a one micrometer diameter fiber mounted on the end of a wire, uranium oxide particles embedded in oil-bearing cellulose nitrate, teflon and polycarbonate filter materials with collected air particulate matter, polystyrene latex spheres on cellulosic filter paper, polystyrene latex spheres "loosely" sitting on a glass slide, and subsurface tracks in an etched nuclear track-etch detector. Surface charging problems experienced in high vacuum SEMs are virtually eliminated in the low vacuum SEM, extending imaging capabilities to samples previously difficult to use or incompatible with conventional methods.

Membranes, Artificial↗
PubMed1999

Muscle structure and innervation are affected by loss of Dorsal in the fruit fly, Drosophila melanogaster.

In Drosophila, the Rel-protein Dorsal and its inhibitor, Cactus, act in signal transduction pathways that control the establishment of dorsoventral polarity during embryogenesis and the immune response during postembryonic life. Here we present data indicating that Dorsal is also involved in the control of development and maintenance of innervation in somatic muscles. Dorsal and Cactus are colocalized in all somatic muscles during postembryonic development. In larvae and adults, these proteins are distributed at low levels in the cytoplasm and nuclei and at much higher levels in the postsynaptic component of glutamatergic neuromuscular junctions. Absence of Dorsal, in homozygous dorsal mutant larvae results in muscle misinsertions, duplications, nuclear hypotrophy, disorganization of actin bundles, and altered subcellular distribution of Cactus. Some muscles show very abnormal neuromuscular junctions, and some motor axon terminals are transformed into growth cone-like structures embedded in synaptotagmin-enriched vesicles. The detailed phenotype suggests a role of Dorsal signalling in the maintenance and plasticity of the NMJ.

Animals↗
PubMed1977

Autoradiographic detection of mucopolysaccharide accumulation in single fibroblasts.

A method is described for localizing acid mucopolysaccharides autoradiographically in cultured cells. Normal fibroblasts and fibroblasts, from patients suffering from Mucopolysaccharidosis II disease (MPS II), were cultured for six days in the presence of 35SO4 and one day in unlabelled medium. The cultured cells were transferred to plastic film dish and, after settling, they were rapidly quenched, freeze-dried, fixed in osmium tetroxide vapour and embedded in Epon. Grain counting after autoradiography in 2 mum sections revealed a significant difference (P greater than 0.001) in 35SO4 incorporation in the perinuclear cytoplasm of MPS II cells and control cells grown under the same conditions. Autoradiography was also performed after mixing MPS II cells and control fibroblasts in a ratio 1:1-8 prior to freezing and the same ratio was found between labelled and unlabelled fibroblasts. These results demonstrate the feasibility of the present autoradiographic technique for the detection of the acid mucopolysaccharide storage at the single cell level.

Autoradiography↗
PubMed1975

Studies of intestinal lymphoid tissue. I. Electron microscopic evidence of 'blast transformation' in epithelial lymphocytes of mouse small intestinal mucosa.

The morphology of epithelial lymphocytes in osmium-fixed, Epon-embedded jejunum of adult mice was studied by light and electron microscopy. Toluidine blue-stained 1 mum 'thick' plastic sections were compared with adjacent thin sections, thereby permitting precise ultrastructural identification and description of selected epithelial lymphocytes. Their size and appearances varied considerably, ranging from typical small lymphocytes through medium-sized lymphocytes to large immunoblasts. A high proportion of medium-sized epithelial lymphocytes (mean diameter 6-9 +/- 1-1 mum) contained several lysosomes, extensive Golgi complexes, prominent centrioles and abundant ribosomes. Their appearances, therefore, corresponded directly to mitogen-stimulated lymphocytes. In contrast, immunoblasts were big cells (mean diameter 11-0 +/- 0-8 mum) with large, euchromatic nuclei and prominent nucleoli. The majority had pale-staining, ribosome-studded cytoplasm and thus resembled type I, or T blasts. Very rarely, densely staining blasts containing ribosomes and well developed rough endoplasmic reticulum were observed; these corresponded to type II or B blasts. These observations indicate that transformation of lymphocytes occurs within the interepithelial cell spaces of the small intestinal mucosa, suggesting that epithelial lymphocytes are immunocompetent cells which may be responsive to local antigenic stimulation.

Animals↗
PubMed1991

[Massive incrustation of a plastic urethral prosthesis after implantation for 5 months].

A male aged 73 years in poor cardio-pulmonary condition and with urinary retention due to prostatic infravesical obstruction was treated with implantation of a new type of plastic urethral catheter. Prostatectomy was planned after improvement in the cardio-pulmonary condition. This period was prolonged and, five months after implantation of the urethral catheter, massive incrustation in the proximal intravesical part had formed. The catheter was found to be firmly embedded in the prostatic part of the urethra and cystotomy proved necessary to remove it.

Aged↗
PubMed1980

A method for the demonstration of pores in calcified eggs of vertebrates and invertebrates.

Determination of pore number, pore size as well as pore canal distribution is important in understanding the biology of calcified eggs among vertebrates as well as invertebrates. Various techniques have been applied to elucidate pore structure of avian egg shells, including fracturing coupled with scanning electron microscopy and differential histochemical staining of the matrix. The best results reported have thus far been from plastic impregnated, etched specimens (Board & Tullett, 1974). The present report is a modification of this latter technique, utilizing the very low viscosity, highly penetrating electron microscopic embedding medium developed by Spurr (1969). This modification is used in this report of the previously undescribed pores from the egg shells of the giant South American land snail, Strophocheilus oblongus. These eggs, which can be up to 50 mm in length, have recently been investigated with respect to the calcium carbonate mineralogy (Tompa, 1976) as well as with respect to the matrix biochemistry Tompa et al., 1977). (Similar resin has been used successfully for the plastic impregnation and ultrathin sectioning of that region of the snail shell where the body muscles insert into the shell comumella (Tompa & Watabe, 1976).) This technique, which can be utilized on avian eggs or on any other calcified material in probing for pores, relies on the high penetrating ability of the resin as well as on its easy commercial availability around the world.

Animals↗
PubMed1993

Application of Levanol fast cyanine 5RN stain for demonstrating rhabdomyoblasts in epoxy-embedded tumors.

To facilitate the survey of actomyosin-containing cells in soft tissue sarcomas with skeletal muscle differentiation, the tannic acid-phosphomolybdic acid-Levanol fast cyanine 5RN procedure was applied to plastic-free (deresined) thick sections (2-3 microns) obtained from epoxy blocks conventially prepared for transmission electron microscopy. By light microscopy, rhabdomyoblasts of varying configuration revealed intense dark blue staining reaction. The stained sections were used for orientation and direct interpretation of the ultrastructural material.

Azo Compounds↗
PubMed2004

Prediction of scratch resistance of cobalt chromium alloy bearing surface, articulating against ultra-high molecular weight polyethylene, due to third-body wear particles.

The entrapment of abrasive particles within the articulation between a cobalt chromium alloy (CoCrMo) femoral component and an ultra-high molecular weight polyethylene (UHMWPE) cup of artificial hip joints or tibial inserts of artificial knee joints usually scratches the metallic bearing surface and consequently increases the surface roughness. This has been recognized as one of the main causes of excessive polyethylene wear, leading to osteolysis and loosening of the prosthetic components. The purpose of this study was to use the finite element method to investigate the resistance of the cobalt chromium alloy bearing surface to plastic deformation, as a first approximation to causing scratches, due to various entrapped debris such as bone, CoCrMo and ZrO2 (contained in radiopaque polymethyl methacrylate cement). A simple axisymmetric micro contact mechanics model was developed, where a spherical third-body wear particle was indented between the two bearing surfaces, modelled as two solid cylinders of a given diameter, under the contact pressure determined from macro-models representing either hip or knee implants. The deformation of both the wear particle and the bearing surfaces was modelled and was treated as elastic-plastic. The indented peak-to-valley height on the CoCrMo bearing surface from the finite element model was found to be in good agreement with that reported in a previous study when the third-body wear particle was assumed to be rigid. Under the physiological contact pressure experienced in both hip and knee implants, ZrO2 wear particles were found to be fully embedded within the UHMWPE bearing surface, and the maximum von Mises stresses within the CoCrMo bearing surface reached the corresponding yield strength. Consequently, the CoCrMo bearing surface was deformed plastically and the corresponding peak-to-valley height (surface roughness) was found to increase with both the hardness and the size of the wear particle. Even in the case of CoCrMo wear particles, with similar mechanical properties to those of the CoCrMo bearing surface, a significant plastic deformation of the bearing surface was also noted; this highlighted the importance of considering the deformation of the wear particles. These findings support the hypotheses made by clinical studies on the contribution of entrapped debris to increased surface roughness of CoCrMo femoral bearing surfaces.

Compressive Strength↗
PubMed2005

A microfabricated capillary electrophoresis chip with multiple buried optical fibers and microfocusing lens for multiwavelength detection.

We present a new microfluidic device utilizing multiwavelength detection for high-throughput capillary electrophoresis (CE). In general, different fluorescent dyes are only excited by light sources with appropriate wavelengths. When excited by an appropriate light source, a fluorescent dye emits specific fluorescence signals of a longer wavelength. This study designs and fabricates plastic micro-CE chips capable of performing multiple-wavelength fluorescence detection by means of multimode optic fiber pairs embedded downstream of the separation channel. For detection purposes, the fluorescence signals are enhanced by positioning microfocusing lens structures at the outlets of the excitation fibers and the inlets of the detection fibers, respectively. The proposed device is capable of detecting multiple samples labeled with different kinds of fluorescent dyes in the same channel in a single run. The experimental results demonstrate that various proteins, including bovine serum albumin and beta-casein, can be successfully injected and detected by coupling two light sources of different wavelengths to the two excitation optic fibers. Furthermore, the proposed device also provides the ability to measure the speed of the samples traveling in the microchannel. The developed multiwavelength micro-CE chip could have significant potential for the analysis of DNA and protein samples.

Electrophoresis, Capillary↗
PubMed1998

Solid-phase extraction-high-performance liquid chromatography determination of verapamil and norverapamil enantiomers in urine.

A simple, rapid, sensitive and selective method has been developed for the stereospecific determination of verapamil and its metabolite, norverapamil in urine. For sample preparation we utilized a membrane-based solid-phase extraction (SPE) disk consisting of a thin, particle-loaded membrane inserted in a plastic syringe-like barrel. The particles, which may be C8 or C18 bonded phase (C8 in this work), are embedded within a matrix of PTFE (Teflon) fibrils. Overall analyte recoveries were above 85%, even at low concentration of 3.0 ng/ml with reproducibilities (C.V. values) below 13.1%. This method of extraction has the advantage of speed and considerable reduction in solvent volumes compared to conventional SPE and solvent extraction. The separation of all the enantiomers was achieved using a single chiral stationary phase column, the cellulose-based reversed-phase, Chiralcel OD-R. Analyte concentrations of less than 3.0 ng/ml could be quantitated with C.V. values below 14%. Calibration curves were linear in the range 2.5-300 ng/ml. Intra-day and inter-day reproducibilities were 10.5-14.2% at 3 ng/ml, 4.8-9.3% at 138.5 ng/ml and 7.8-10.1% at 280 ng/ml level, respectively, for all the enantiomers.

Adult↗
PubMedDate not supplied

Ultrastructure of normal epithelial cells in Papanicolaou-stained cervical smears. An application of a modified open-face embedding technique for transmission electron microscopy.

The results of an open-face embedding technique applied to Papanicolaou-stained uterine cervical smears are presented. By this technique, cells are smeared on a plastic sheet, fixed in 1.5% buffered glutaraldehyde and stained by the Papanicolaou method; areas of special interest are then selected by light microscopy for electron microscopic observations. Thus, a comparison is possible between the light microscopic characteristics of cells in smear preparations and their ultrastructural counterparts. The good ultrastructural preservation allowed detailed study of nuclei and cytoplasm. The endocervical cells had intact nuclei with nuclear envelope, Golgi apparatus, well-preserved mitochondria and two types of secretory material. The superficial and intermediate squamous cells showed loss of desmosomes; dense, fine cytoplasmic fibrils; small, fatty droplets; and degenerated nuclei with loss of the nuclear envelope. Compared with ultrastructural studies on cervical tissues, the intermediate cells in cervical smears were more degenerated, presumably because predominantly degenerated intermediate cells exfoliate. This study indicates that the light microscopic impression of the nuclear appearance is misleading: in the routine smear the nuclei of superficial and intermediate cells may look "intact" and "well preserved" whereas ultrastructurally the nuclei are degenerated.

Cytological Techniques↗
PubMed1979

Structure of Retzius lines in partially demineralized human enamel.

Human third molars were partially demineralized in an acid-alcohol solution and embedded in Epon 812. Six-micron sections were cut from regions of the cervical enamel exhibiting prominent Retzius lines. The plastic was removed from the specimens by microincineration and were examined with the scanning electron microscope. The most prominent structural feature seen along the Retzius lines was the cervical translocation of some of the prisms. The scanning electron microscopic images also suggested that prisms were translocating in the transverse plane of the tooth. A series of pores, which appeared to be empty, were observed in association with the translocations occurring along the Retzius lines.

Acids↗
PubMed2002

Air-coupled ultrasound inspection of various materials.

Conventional ultrasound inspection, a standard non-destructive testing method, uses a coupling medium (e.g. water) because of impedance mismatch. This liquid contact is a drawback because it prevents inspection of many materials. There is a need, then, for air-coupled ultrasound testing, which is now feasible because of low impedance focused narrow band transducers and sensitive electronics, both of which improve the signal-to-noise ratio. We present results obtained on fibre-reinforced plastics, water sensitive materials (e.g. reinforced ceramics), and "shape adaptive" structures to reveal delaminations, impacts, and growth of internal defects. Actuators embedded in "adaptive" structures are used as transmitters while the receiver records the signals. Thus it is possible to image defect areas and non-linear behaviour of potential defects.

Journal Article↗
PubMed1998

Electron microscopy: use of transmission and scanning electron microscopy to study cells in culture.

Standard techniques for electron microscopy were developed for tissues dissected from animals. Optimal methods for electron microscopy of cells in culture are different. This chapter describes methods for processing cells grown on plastic or in suspension. Both transmission and scanning electron microscopy are discussed. The focus is on the procedures for fixation, dehydration, embedding, and staining, which will help the reader to obtain superior electron micrographs of cultured cells.

Animals↗