Search PubMed⌕ Search

Biomedical subjects

J M Ruijter

Publications and source records attributed to J M Ruijter.

At least 37 records · Page 2Linked to original sources

Densitometric measurements of the mandible: accuracy and validity of intraoral versus extraoral radiographical techniques in an in vitro study.

In this study a comparison is made between intraoral and extraoral radiographical techniques for quantitative measurements of bone density in the edentulous mandible. A phantom model, composed of a dry mandible surrounded by a soft tissue substitute was used. Radiographs were made of aluminium implants of known volumes. Intra- and extraorally placed reference wedges of aluminium or a combination of aluminium and plexiglas were used for the measurement of the aluminium volume of the above-mentioned implants and compared with the real volume. The IBAS image analysis system was used for image processing and measurements. A systematic deficit in the measured volume of 15 to 29% was found. This error was irrespective of the radiographical technique and indicates a constant underestimation of the aluminium volumes. The differential influence of the secondary radiation on the images of the jaw and the wedge is proposed to be a possible cause of this constant underestimation. The intraoral and extraoral techniques do not display significant statistical differences in the levels of validity or accuracy. Obtaining serial intraoral radiographs of adequate quality in the atrophic edentulous mandible can be problematic. It is concluded from this in vitro study that in these cases Oblique Lateral Cephalometric Radiographs may provide a valuable alternative for quantitative image analysis.

Absorptiometry, Photon↗

Clearance of neurotransmitter from the cone synaptic cleft in goldfish retina.

Dendrites of bipolar and horizontal cells protrude deeply into the synaptic terminals of cones in goldfish retina. This arrangement gives the impression that the cone synaptic terminal surrounds a morphologically shielded compartment, the cone synaptic cleft, from which clearance of neurotransmitter by diffusion is limited. In this study the time constant of this clearance has been approached in two ways: (1) the morphological parameters determining the clearance (extracellular synaptic volume and leak area), were estimated using morphometric methods. These data were introduced into a diffusion model of the cone pedicle, yielding a time constant for the clearance of < 1 msec; (2) the time constant of the light onset response when the glutamate transporter in the cone was blocked with DL-threo-beta-hydroxyaspartate or dihydrokainate, was interpreted as the time constant of the clearance, yielding values of almost 650 msec compared to around 90 msec in control conditions. The decay time-constant of the Ca-dependent tail-currents in cones was used, as an approximation of the dynamics of the intracellular Ca-concentration and thus of the glutamate release by the cones. The decay time constant was about 800 msec. This suggests that the intracellular Ca-concentration in the synaptic terminal and hence the glutamate release by the cones drops with a similar large time constant. These results indicate that the cone pedicle in goldfish does not limit the clearance of neurotransmitters from the synaptic cleft and that the fast light onset response of horizontal cells under control conditions is due to activation of glutamate transporters by hyperpolarization of the cone membrane potential while the glutamate release drops slowly. The slow horizontal cell light onset response in beta-hydroxyaspartate or dihydrokainate may be due to a slow reduction of the glutamate release by the cones at light onset.

Animals↗

Synergistic action of heparin and serum on basic fibroblast growth factor-modulated DNA synthesis and mitochondrial activity of cultured bovine corneal endothelial cells.

Basic fibroblast growth factor (bFGF) is a major mitogen and chemoattractant for many cell types. The synergistic role of fetal bovine serum (FBS) and heparin on the modulation of tissue-cultured bovine corneal endothelial cells by bFGF was studied. Cell modulation was assessed by DNA synthesis ([3H]thymidine incorporation), mitochondrial activity (MTT assay), mitochondrial volume (transmission electron microscope study), and cell shape in mitotically competent cells and cells mitotically inhibited by 5-fluorouracil, mitomycin C, and irradiation. The dose-dependent stimulation of bFGF was found to be different for DNA synthesis and mitochondrial activity, the maximal effective dose being 1 and 10 ng/ml, respectively. Supplementation of bFGF with FBS enhanced both DNA synthesis and mitochondrial activity and caused a shift in the dose response to lower bFGF concentrations for the DAN synthesis, but not for the mitochondrial activity. Supplementation of bFGF with heparin resulted in an additional response for the mitochondrial activity, but not for the DNA synthesis. In addition, bFGF increased mitochondrial volume and induced elongation of the cells. These processes were further enhanced by the addition of heparin to bFGF. These results suggest that serum and heparin have a differential effect on mitogenic and non-mitogenic cells processes modulated by bFGF.

Animals↗

Outflow characteristics of isolated anterior segments of human eyes.

PURPOSE: To evaluate the relationship between intraocular pressure (IOP) and flow through the trabecular meshwork in isolated anterior segments of human eyes. Outflow facility (C) is thought to decrease proportionally with increasing IOP in human eyes in vivo and in vitro. METHODS: Twenty-nine eviscerated human anterior segments were perfused with ascending and descending pressure sequences in a stepwise fashion (range, 4 to 40 mm Hg); 11 of the 29 eyes were treated similarly after 20 hours of perfusion at 12 mm Hg. Pressure-flow sequences of individual eyes were evaluated with a linear (constant C) and a nonlinear regression method (C decreasing with increasing IOP). In addition, in eight intact postmortem eyes, pressure-flow characteristics were determined, followed by perfusion of their isolated anterior segments. RESULTS: Pressure-flow sequences as determined by linear regression had an average correlation coefficient of 0.99. Average C (slope of the plot) was 0.26 +/- 0.03 microliter minute-1 mm Hg-1. There was no influence of direction of pressure sequence or time on C. To test for linearity, the hypothetical outflow obstruction coefficient (Q) was determined for each plot. Median Q of 29 eyes was 0.003 mm Hg-1, and in 45% (13 eyes) Q was negative, suggesting facilitation instead of obstruction. This indicates that the outflow obstruction coefficient is not a physiological parameter in isolated anterior segments. CONCLUSIONS: The relationship between IOP and flow through perfused isolated anterior segments of human eyes is linear between 4 and 40 mm Hg, indicating that within this range outflow facility is constant and does not decrease with increasing IOP.

Adult↗

Nitric oxide induces vascular permeability changes in the guinea pig conjunctiva.

The role of nitric oxide (NO) as an inflammatory mediator in the mechanism of increased microvascular permeability was examined in a guinea pig model of allergic conjunctivitis. Topical challenge with antigen, compound 48/80, histamine or platelet activating factor (PAF) resulted in a marked increase of the conjunctival vascular permeability. Vascular permeability was determined by measuring the albumin content in the lavage fluid of the challenged eyes after 30 min. Pretreatment with NG-nitro-L-arginine methyl ester (L-NAME) eyedrops caused a significant inhibition of the clinical score and the vascular permeability after challenge with either antigen, histamine or PAF. Aminoguanidine prophylaxis also resulted in a significant inhibition of both the clinical score and vascular permeability in response to all the used provocative agents except PAF. Our observations indicate that NO is an important factor in the induction of the vascular permeability provoked by histamine but seems to play no role in the mechanism by which PAF exerts increased vascular permeability in the conjunctiva.

Albumins↗

Repeated administration of a selective dopamine D2 receptor agonist to 6-OHDA-lesioned rats does not affect the survival and outgrowth of intrastriatal fetal mesencephalic grafts.

The objective of the present study was to investigate the effects of chronic activation of dopamine D2 receptors on the development of grafted fetal rat mesencephalic dopaminergic neurons. Therefore, unilaterally 6-hydroxydopamine - lesioned rats received intrastriatal mesencephalic cell suspension grafts and were subsequently chronically treated with the selective dopamine D2 receptor agonist LY 171555 (Quinpirole). After treatment for 6 consecutive weeks, the rats were processed for tyrosine-hydroxylase immunocytochemistry to assess the survival and outgrowth from grafted dopaminergic neurons. morphological analysis revealed that, like the volume and morphology of the graft, neither the number nor the cell area of grafted dopaminergic neurons was significantly different between vehicle- and LY 171555-treated animals. To obtain a quantitative estimate of the graft-derived dopaminergic reinnervation, a computerized image analysis system was used. Using this procedure, which was based on the densitometric measurement of tyrosine hydroxylase immunoreactivity in the area adjacent to the grafted tissue, it was found that the extent of graft-derived outgrowth also appeared to be unaffected upon chronic treatment with LY 171555. It is concluded that long-term concurrent administration of a dopamine D2 receptor agonist for 6 consecutive weeks does not impair the survival and outgrowth of grafted rat fetal mesencephalic dopaminergic neurons.

Animals↗

Effects of spontaneous bioelectric activity and gangliosides on cell survival in vitro.

Chronic suppression of spontaneous bioelectric activity in spinal cord explants in the presence of tetrodotoxin (TTX) during network formation caused a large reduction in cell number (lowered DNA levels). The addition of gangliosides failed to protect against this cell loss. Conversely, the omission of galactose from the growth medium had no effect on DNA levels. It was concluded that the presence or absence of afferent selectivity is unlikely to require the survival of a regionally specific subpopulation of preferred dorsal root ganglion target cells. Neocortical explants also showed a large reduction in DNA levels following chronic TTX treatment, and morphometric analysis confirmed that neuronal survival was affected to the same degree. Chronic ganglioside supplementation failed to influence DNA and cell counts in either control or TTX-treated explants, but one of the added gangliosides (GD1a) stimulated extensive neuritic outgrowth in electrically silenced cultures. Particular ganglioside species, therefore, may exert a growth stimulating influence that can partially compensate for the absence of bioelectric self-stimulation during early development.

Animals↗

Reduction in bioelectric GABA and NMDA responses in organotypic neocortical explants by chronic elevation of potassium.

Long-term cultures of organotypic neonatal rat neocortex slices were maintained in a serum-free medium supplemented with 25 mM potassium. Such cultures continue to be bioelectrically silent upon return to a 5 mM potassium medium. The absence of responses to pressure ejected gamma-aminobutyric acid (GABA) and N-methyl-D-aspartate (NMDA) from neurons in treated explants suggests that one consequence of chronic depolarization is a reduction in the density of postsynaptic transmitter receptors. [3H]Muscimol binding to neocortical membrane preparations shows a large reduction in the binding of this agonist to GABAA receptors. These data show that the quantitative expression of at least one neurotransmitter receptor, the GABAA receptor, relies on voltage-dependent activity in developing neocortical neurons in vitro.

Action Potentials↗

Effect of chronic exposure to high magnesium on neuron survival in long-term neocortical explants of neonatal rats in vitro.

In order to assess the effect of elevated magnesium, neuronal morphology and physiology was studied in chronically cultured organotypic neonatal rat occipital neocortex. Explants grown in 10 mM magnesium were found to experience an approximate 30% cell loss (as shown by cell count and DNA-protein analysis), while 12.5 and 15 mM magnesium showed ca. 47 and 60% cell losses, respectively. Intracellular recording from 10 mM magnesium explants revealed that measurable postsynaptic potentials and action potentials could occur, apparently depending on the type of cell examined. All post-synaptic activities ceased in 12.5 mM magnesium cultures, though action potentials could be elicited by current stimulation. The effects of known depolarizing agents, viz. potassium and N-methyl-D-aspartate, on 12.5 mM magnesium-grown explants were also examined. Explants grown in the presence of 12.5 mM magnesium plus 10 mM potassium showed a dramatic increase in the loss of neurons. The simultaneous addition of 6,7-dinitro-quinoxaline-2,3-dione showed this to be due to an increase in non-N-methyl-D-aspartate mediated cell death in response to glutamate release brought about by the depolarizing effects of the potassium. The addition of 10 microM N-methyl-D-aspartate to 12.5 mM magnesium-grown cultures, on the other hand, improved cell survival to control levels. The mechanism of this reciprocal neuroprotective effect of N-methyl-D-aspartate against magnesium has yet to be elucidated. We conclude that these findings are consistent with regard to the opposing actions of N-methyl-D-aspartate and magnesium on calcium influx and various metabolic processes within the explants.

Animals↗

Chronic blockade of bioelectric activity in neonatal rat neocortex in vitro: physiological effects.

We have examined what effect the loss of spontaneous bioelectric activity has on neural network formation in organotypic rat neocortical explants grown under serum-free culture conditions. Explants were taken from dorsal midline (presumptive visual) and lateral (presumptive auditory) occipital cortex and chronically exposed to tetrodotoxin which blocked all measurable bioelectric activity between change of medium. Extracellular recordings revealed complex, rhythmic spontaneous and evoked multiunit discharges in all explants examined (following tetrodotoxin washout in the experimental group). Control auditory explants had significantly more sites from which electric activity could be recorded compared with control visual explants. Auditory cultures showed no effect of the tetrodotoxin treatment, whereas visual explants showed significant increases over control values, equalling the auditory values. This increased level of spontaneous bioelectric activity was maintained for at least 10 days following transfer of the cultures to control growth medium. There was no significant difference between control visual and auditory explants regarding the number of sites from which evoked activity was seen. Nor did either cortex group show an effect of tetrodotoxin on the number of sites from which evoked activity was seen. The frequency with which spontaneous bioelectric discharges occurred per site increased with age in auditory vs visual cortex. These differences, however, were abolished in the tetrodotoxin-treated groups. It was concluded that neocortical explants which have experienced chronic suppression of spontaneous electric activity did not suffer deficits in neural network formation, though there is an effect on the incidence and frequency with which such activity is given.

Action Potentials↗

Chronic blockade of bioelectric activity in neonatal rat cortex grown in vitro: morphological effects.

Culture thickness, numerical density of neurons and neuronal survival were studied in timed series of control and tetrodotoxin-silenced neocortical cultures to provide information on the role of bioelectric activity on neuronal development. In control cultures, culture thickness and number of surviving neurons decrease during the first weeks in vitro, but remain constant between 2 and 3 weeks indicating that the cultures are essentially mature. In the 4th week in vitro a further decrease in surviving neurons was observed. In tetrodotoxin-treated cultures the number of surviving neurons decreased significantly between 1 and 2 weeks in vitro, to remain constant thereafter. However, culture thickness significantly increased at 3 and 4 weeks in vitro after an initial drop between 1 and 2 weeks. Compared to age-matched controls at 2 and 3 weeks in vitro, only ca 50% of the neurons survived the loss of bioelectric activity. Similar differences were present between 1 and 2 weeks. Thus, the loss of all measurable bioelectric activity induces neuronal death in neocortical explants, but promotes neuropil formation by the surviving cells.

Animals↗

Elevated potassium prevents neuronal death but inhibits network formation in neocortical cultures.

Chronic depolarization is inimical to neuronal growth and synaptogenesis so that spontaneous action potential generation appears to be required for the normal cytomorphological maturation of neocortical networks. The efficacy of 25 mM K in suppressing spontaneous bioelectric activity was monitored by extra- and intracellular recording from the explants. Intracellular recording from individual neurons showed that membrane potentials were reduced to ca -30 mV in potassium cultures but rapidly repolarized to ca -50 mV when returned to normal growth medium. Though action potentials could be readily evoked from these explants, spontaneous discharges and postsynaptic potentials were absent from potassium-treated cultures. Both spontaneous bioelectric activity and postsynaptic potentials returned to the cultures by 5 days after returning the explants to normal growth medium. Extracellular recordings also showed that the explants were bioelectrically silent in the presence of 25 mM K or 25 mM K plus tetrodotoxin. In contrast to tetrodotoxin alone, bioelectric activity was absent when the cultures (with or without tetrodotoxin) were returned to normal growth medium. The explants gradually began to evince spontaneous bioelectric activity between 3 and 5 days after being returned to normal growth medium. Massive cell death induced by chronic exposure to tetrodotoxin was totally prevented by concomitant addition of 25 mM potassium, though these explants were significantly thinner than controls due to a large decrease in neuropil. We conclude that chronic depolarization of neonatal cortical explants by potassium results in a delayed return of spontaneous bioelectric discharges. Chronic depolarization results in a retardation of network formation in these explants apparently due to a lack of neurite and/or synapse formation.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗

The effects of potassium-induced depolarization, glutamate receptor antagonists and N-methyl-D-aspartate on neuronal survival in cultured neocortex explants.

The effects of elevating the potassium concentration of the growth medium of neocortical explants was studied. Under control conditions, 10 mM potassium resulted in ca 20% decrease in the number of surviving neurons. The same potassium concentration, however, was clearly neurotrophic in tetrodotoxin-grown cultures: tetrodotoxin-induced neuronal death was significantly reduced. Both effects could be mimicked by the addition of 10 microM N-methyl-D-aspartate (NMDA); lower concentrations were without effect; higher concentrations were neurotoxic under both control and tetrodotoxin conditions. The neurotoxic, as well as the neurotrophic effects of 10 mM potassium appear to be mediated through depolarization-induced glutamate release since they could be influenced by the application of glutamate receptor antagonists. The addition of the NMDA receptor antagonist D-2-amino-7-phosphonoheptanoate (APH) blocked the trophic effect of 10 mM potassium in tetrodotoxin-grown cultures, resulting in low survival. On the other hand, the addition of the non-NMDA antagonist 6,7-dinitroquinoxaline-2,3-dione (DNQX) resulted in neuronal survival similar to control cultures, indicating that it blocked the toxic effects of glutamate, leaving the trophic effects on the NMDA receptor untouched. Under control (non-TTX) conditions, neither DNQX nor APH showed significant effects on 10 mM potassium-induced cell death, indicating that stimulation of the non-NMDA, as well as the NMDA receptors is neurotoxic. This differential effect of NMDA receptor stimulation on neuronal survival is discussed with respect to the maturational and/or functional state of the neurons in the culture.

2-Amino-5-phosphonovalerate↗

Immunocytochemical indications for neuronal co-localization of GABA and aspartate in cultured neocortex explants.

The application of postembedding immunocytochemistry on serial semithin plastic sections, revealed the presence of gamma-aminobutyric acid (GABA)-positive and aspartate-positive neurons in cultured neocortex explants. GABA-positive neurons were found in all layers of the cultured cortex, whereas aspartate-positive neurons were mainly restricted to layer 6. Although exact quantification was not carried out because of the variability in aspartate immunoreactivity, colocalization of GABA and aspartate immunoreactivity could unambiguously be observed in a few neurons.

Animals↗

Electrophysiological properties of neurons in neonatal rat occipital cortex slices grown in a serum-free medium.

The electrophysiological properties of individual neurons within organotypic explants of neonatal rat cortex were examined via intracellular recordings. The explants were grown for two weeks in a serum-free medium. The electrophysiological properties of the neurons within these explants were similar to those reported for both adult cortex in vivo and short-term in vitro slice preparations. The results of the present study show that cortical explants grown under serum-free conditions can serve as a useful model for long-term developmental studies associated with the physiological basis of neural network formation.

Action Potentials↗

Calculation of survival curves and statistical comparison of two censored populations: a computer program in BASIC.

In experimental aging research the handling of censored data imposes a serious problem in the calculation of survival curves of partly censored populations and the statistical comparison of such populations. The presented computer program combines a method to calculate a corrected survival curve and a test to compare two censored populations. In both methods, the proportion of the animals that survive or die during an interval and the number at risk at the start of that interval is used in the calculation of relative survival or death during that interval. The program is written in an elementary BASIC and will run on practically every computer programmable in BASIC. The presented method can be used for testing the difference between survival characteristics of two animal populations but might also be employed in evaluating the effect of a medical treatment.

Aging↗