Biomedical subjects

J M Hopkins

Publications and source records attributed to J M Hopkins.

12 recordsLinked to original sources

PubMed1992

Synaptic contacts of a two-cone flat bipolar cell in a primate retina.

The Golgi-stained dendrites of a two-cone bipolar cell of a vervet monkey were serially sectioned for electron microscopy and shown to make basal synapses with two neighboring cones. The synapses were all on the cone pedicle membrane adjacent to the bipolar processes invaginating to form the triads. This is the characteristic position for flat midget bipolar cells. The numbers of triads in nine adjacent cone pedicles were not significantly different from those in the cones contacted by the bipolar. Based on this measure the two-cone bipolar does not contact a special population of cones. The advantages and disadvantages of using vertical or horizontal sections for the determination by electron microscopy of the connectivity of the dendrites of Golgi-stained flat bipolar cells are discussed.

Animals↗

PubMed1991

Regeneration of axons from adult rat retinal ganglion cells on cultured Schwann cells is not dependent on basal lamina.

The ability of sciatic nerve grafts to support in vivo regeneration of retinal ganglion cell axons in the adult rat raises the question of which peripheral nerve constituents may be required to promote this unexpected central regenerative response. Prime candidates for this role include the surface of the Schwann cell and components of extracellular matrix present in peripheral nerve trunks. To determine the relative importance of Schwann cells and their basal lamina in promoting retinal ganglion cell axon regeneration in the mammalian visual system, we have used an in vitro model. This approach allowed analysis of the abilities of defined peripheral nerve constituents to promote in vitro outgrowth of neurites from explants of adult rat retina harvested 7 to 10 days after in vivo optic nerve crush. Neurite outgrowth was assessed by neurofilament immunofluorescence after 3 to 20 days in vitro. Culture substrata, consisting of isolated Schwann cells (SC), Schwann cells with their assembled extracellular matrix (SC + ECM), or isolated extracellular matrix from which the Schwann cells had been removed (ECM), were prepared by first co-culturing rat Schwann cells with embryonic dorsal root ganglion neurites on a layer of type I collagen, and then manipulating the cultures to produce the desired substrata. Type I collagen alone did not support neurite growth from adult rat retina. SC and SC + ECM supported regeneration of axons from retinal explants at average growth rates of 18 and 30 microns/h, respectively. Isolated ECM was a poor substrate for retinal neurite growth; the few neurites that gained access to this material grew at rates averaging less than 3 microns/h. These observations suggest that regeneration of adult mammalian retinal ganglion cell axons through peripheral nerve grafts (in vivo) is primarily dependent on neurite-promoting factors present on the surface of Schwann cells and does not require organized extracellular matrix.

Animals↗

PubMed1991

In vitro myelination of regenerating adult rat retinal ganglion cell axons by Schwann cells.

Schwann cell cultures provide a highly favorable substrate for retinal ganglion cell (RGC) survival and axon growth in vitro (Bähr and Bunge, Exp Neurol 106:27, 1989; Hopkins and Bunge, Glia 4:46, 1991). In this report we have extended former studies to obtain axon regeneration, long-term survival, and myelination of adult rat RGC axons in co-cultures of retinal explants with purified Schwann cells. By using modified co-culture conditions, we observed myelination of regenerating adult RGC axons by Schwann cells after 3-4 weeks in vitro. Myelination was associated with a one-to-one Schwann cell-axon relationship, characteristic of the formation of peripheral myelin. Under culture conditions that supported myelination, long-term survival (more than 12 weeks) of a small population of RGCs was observed. These findings highlight the remarkable ability of Schwann cells to support long-term survival of adult rat RGCs in the absence of either central nervous system (CNS) target tissue or other peripheral nervous system (PNS) components. This tissue culture system may serve as a model for the systematic study of the molecular mechanisms which are involved in axon regeneration and myelination of adult CNS neurons.

Animals↗

PubMed1991

Regeneration of axons from adult human retina in vitro.

In an effort to establish an in vitro model of regenerating adult human central nervous system (CNS) neurons, we have investigated the potential for neurite growth from explants prepared from normal adult human retina. Eyes (donated for corneal transplantation) were removed within 2.0 h postmortem and stored on ice for 1.5 to 7.0 days. Retinal explants (1 mm2) were prepared and cultured at 37 degrees C on cellular or acellular substrata in an oxygen-rich, humidified atmosphere. Neurite outgrowth, visualized by neurofilament immunofluorescence, was observed only in the presence of Schwann cells, after a quiescent period of approximately 6 days in vitro. Of 50 explants cultured for 7 days or more on substantia containing Schwann cells, 43 showed evidence of viability in vitro and 28 extended neurites onto Schwann cell surfaces. Estimated rates of neurite growth on Schwann cell substrata reached a maximum of 0.22 mm/day. Neurites did not grow beyond the explant border onto culture substrata composed of either polylysine, laminin, type-I collagen, or monolayers of adult human retinal glia. These results demonstrate that under selected conditions, explants prepared from adult human retina harbor viable neurons and that Schwann cells promote and support regeneration of neurites from these neurons in vitro, allowing systematic analysis of conditions favorable to axonal regeneration from adult human CNS neurons.

Axons↗

PubMed1990

Enhanced resistance to cefotaxime and imipenem associated with outer membrane protein alterations in Enterobacter aerogenes.

Mutants exhibiting enhanced resistance to cefotaxime and imipenem were selected by plating a strain of Enterobacter aerogenes, which already produced chromosomal beta-lactamase constitutively, on to varying concentrations of different beta-lactam antibiotics. Frequencies of mutation varied from 10(-5) to 10(-8), depending upon the particular antibiotic and concentration used for selection. Only minor variations in beta-lactamase specific activities were observed and these could not be directly correlated with changes in resistance when compared with the original strain. In the majority of mutants, the selection of an enhanced level of resistance to cefotaxime was associated with a significant increase in resistance to imipenem, but no increase in resistance to the non-beta-lactam antibiotics tested was observed. Examination of outer membrane protein profiles revealed a number of complex changes in the mutants when directly compared to the original strain. In one mutant imipenem/cefotaxime resistance was directly associated with almost total loss of a 42K protein which was non-covalently associated with peptidoglycan and therefore possibly a porin protein.

Cefotaxime↗

PubMed1988

Long-term analysis of organelle translocation in isolated axoplasm of Myxicola infundibulum.

Moving intra-axonal organelles demonstrate frequent variations in speed when viewed over several seconds. To evaluate these and other motion variations, a long-term analysis of organelle motion in isolated axoplasm of Myxicola infundibulum was carried out using differential interference contrast optics and analog and digital image enhancement techniques. Motion characteristics of individual organelles were analyzed for periods of up to 58 minutes. Three principle observations on organelle motion were made: 1) Classes of organelles of the same size demonstrated a 5- to 25-fold variation of speed, with the slowest speeds occurring most frequently; 2) organelle speeds over individual translocations (motion without stopping) are inversely proportional to their size, but the speeds calculated for the long-term analysis of organelle motion (total distance travelled/total observation time, including pauses) did not reflect this observation; and 3) organelles displayed variable trip lengths, durations, mean speeds, and pause durations, and the relationships between these variations showed no repetitive patterns. In contrast to reported observations of uniform velocities of organelles moving on isolated microtubule preparations, these observations suggest that a variety of factors must play a role in organelle translocation in Myxicola axoplasm.

Animals↗

PubMed1987

Cone connections of the horizontal cells of the rhesus monkey's retina.

The presence in the rhesus monkey's retina of a second morphological type of horizontal cell (H2), described by Kolb et al. (1980), is confirmed. Both types of cell are here further described. Their cone connections are quantified and compared with those of mammals and other vertebrates. The dendrites and axons of the H2 type of cell contact only cones as do the dendrites of the H1 cell (originally described by Polyak (1941)) which has an axon contacting only rods. The dendrites of foveal H2 cells contact between 11 and 14 cones; those of H1 contact 7. The number of cones that each type of cell contacts increases with increasing distance from the fovea, so that, by 5-6 mm eccentricity, H2-type cells synapse with between 20 and 30 cones, and the H1 cells with 12-15. The qualitatively estimated coverage factors of each are 3 or 4; every cone synapses with more than one of both types. Neither type of horizontal cell makes chromatically specific connections that are anatomically recognizable, unlike the situation in some teleostean and turtle retinae. Individual horizontal cells, particularly those connected to foveal cones, may have different ratios of chromatic input. At equivalent eccentricities, up to about 6 mm from the fovea, the dendritic fields of H2 horizontal cells are about twice the size of H1 cells and contact about twice the number of cones. These relative differences are closely similar to those of the cat's horizontal cells and it is suggested that they are a basic feature of most placental mammals. The organization of foveal cone fibres within Henle's layer is described. The distribution of primate cone telodendria, gap junctions and synapses in the outer plexiform layer are briefly reviewed and compared with those of other vertebrate retinae.

Animals↗

PubMed1979

Subcellular distribution of radio-labelled iron during intestinal absorption in guinea-pig enterocytes with special reference to the mitochondrial localization of the iron.

1. 59Fe-labelled ferric chloride was introduced into tied loops of guinea-pig jejunum. 2. After 5--30 min the loop was removed, the enterocytes were isolated, homogenized and subjected to analytical subcellular fractionation. 3. Uptake of 59Fe was extremely rapid and after 5 min 45% of the radioactivity sedimented in the mitochondrial fraction. 4. Density gradient centrifugation indicated that approximately 80% of this radioactivity was associated with the mitochondria themselves; the remainder was in brush-border fragments. 5. Selective disruption of the mitochondria demonstrates that the iron is localized to the matrix and the inner membrane, indicating transport of the absorbed iron into the organelle. 6. It is suggested that mitochondria are actively implicated at an early stage in the intestinal transport of iron.

Animals↗

PubMed1977

New culture medium for the presumptive identificaion of Candida albicans and Cryptococcus neoformans.

A new medium composed of Tween 80, oxgall, caffeic acid, and Davis agar (TOC) that provides for the rapid presumptive identification of Candida albicans and Cryptococcus neoformans is described herein. C. albicans is differentiated from other yeasts by the sequential production of germ tubes and chlamydospores. In a comparison with cormeal agar control plates, there was an increase of chlamydospore-forming strains of C. albicans (97.1% versus 87.2%) and a decrease in the time required for chlamydospore formation (24 h versus 48 h). C. neoformans produced a brown pigment of TOC, which is specific for its identification, thus differentiating it from the other yeasts. A comparison of 24-h pigment production by C. neoformans on TOC with that of birdseed agar showed a dark, coffee brown color in the former cultures and a light brown color in the latter. The change in pigmentation of C. neoformans, as well as morphological changes in C. albicans, can be induced within 3 to 12 h and in not more than 24 h on the TOC medium.

Agar↗

PubMed1977

Rapid method for determining nitrate utilization by yeasts.

A test for the nitrate-reductase activity in yeasts has been developed, in which the reaction may be read after only 10 min of incubation. The rapidity of the test is due to the optimization of pH, substrate concentration, and temperature for the reaction.

Culture Media↗

PubMed1975

Improved auxanographic method for yeast assimilations: a comparison with other approaches.

An improved pour-plate auxanographic method has been developed for determining the assimilation of 14 different carbohydrates by medically important yeasts. Reduction of a dye incorporated into the agar has been correlated with the growth and carbohydrate assimilation of the yeasts, allowing the speciation of many yeasts within 24 to 48 h. This technique has been found to compare more than favorably with existing yeast assimilation techniques, in terms of rapid identification, total cost, and technician time in preparing and inoculating the plates. The dye pour-plate auxanographic technique provides an easier-to-interpret, rapid, and reproducible method of mycological identification for small clinical laboratories.

Agar↗

PubMedDate not supplied

Cone bipolar cells and cone synapses in the primate retina.

Primate retinal bipolar cells synapsing with two adjacent cones (2C bipolars) are further described. Their synaptic contacts are either as the central (invaginating) component of the cone triads or as basal (flat) contacts on the membrane of the cone pedicle base. Correspondingly, their axons end either in the b (inner half or in the a (outer) half of the inner plexiform layer. The shape and size of the axon terminals of 2C bipolars are indistinguishable from those of adjacent midget bipolars. Therefore 2C bipolars, like midget bipolars, probably synapse with midget ganglion cells. Two C bipolars have not been identified as connected to foveal cones. But they are not restricted to the retinal periphery, as has previously been supposed, since they occur, mixed with midget (single cone) bipolars, throughout all parts of the retina from about 2.5 mm to at least 10.0 mm from the fovea. It is likely that 2C bipolars are a variant of the midget bipolars; and that they contact some members of the same population of cones, instead of the midgets. This paper briefly reviews, and raises some new, problems concerning our current understanding of the synaptic connectivity patterns of the midget, 2C, and diffuse cone bipolar cells.

Animals↗