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Biomedical subjects

E W Johnson

Publications and source records attributed to E W Johnson.

At least 19 recordsLinked to original sources
PubMed1992

Calbindin-like immunoreactivity in two peripheral chemosensory tissues of the rat: taste buds and the vomeronasal organ.

In the rat, calbindin-like immunoreactivity was observed at both the light and electron microscopic levels within the chemoreceptor neurons of the vomeronasal organ (VNO) and both intragemmal cells and associated nerve fibers of the circumvallate taste buds. All VNO neurons were immunoreactive. Only a subset of intragemmal taste cells was immunoreactive; associated immunoreactive nerve fibers were apposed to both labeled and unlabeled cells but no synaptic contacts were observed.

Animals↗
PubMed1992

Expression and function of insulin-like growth factor receptors on anti-CD3-activated human T lymphocytes.

The pattern of expression of receptors for insulin-like growth factors (IGF-I and IGF-II) and insulin was studied on monocyte-depleted human peripheral blood T cells activated via anti-CD3. Binding assays demonstrated the sequential appearance of receptors for IGF-I, IGF-II, and insulin on activated T cells. IGF-IR appeared early, their expression reaching maximum levels at or before the peak of cellular proliferation. IGF-IIR expression generally followed that of the IGF-IR and was more transient, with increases and decreases in expression paralleling the rise and decline of cellular proliferation. Insulin receptor expression remained low throughout the activation time course. The identity of the IGFR on anti-CD3-activated T cells was confirmed in affinity cross-linking experiments. These data demonstrated a 135,000 Mr peptide that specifically binds radiolabeled IGF-I and corresponds to the alpha subunit of the type I IGF-IR, and a 260,000 Mr peptide that specifically binds radiolabeled IGF-II and corresponds to the type II IGFR. We have additionally found that IGF-I and IGF-II (in nanomolar concentrations) produce as much as a threefold enhancement of T cell proliferation early in the activation process, correlating with the early appearance of IGF-IR. The effect of both IGF appeared to be mediated through the type I receptor, since an antibody (alpha IR3), which blocks binding to the alpha subunit of this receptor, inhibited enhancement by up to 83%. Furthermore, we have found expression of IGF-IR on T cells after activation to be associated with both CD4+ and CD8+ T cell subpopulations. These observations provide a foundation for investigating the contribution of IGF in regulating T cell proliferation, differentiation, and effector function.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1992

SPECT imaging of the benzodiazepine receptor: autoradiographic comparison of receptor density and radioligand distribution.

SPECT imaging of benzodiazepine (BZ) receptor using the antagonist [123I]iomazenil in nonhuman primates was correlated with ex vivo autoradiography and in vitro receptor autoradiography of postmortem tissue sections taken at the time of maximal brain uptake of the radiotracer. Cryostat sections prepared from brain tissue blocks at the orientation of the SPECT imaging plane showed high activity in gray matter regions, with gray to white matter ratios of greater than 30:1. After at least one week decay of 123I, these same tissue sections were used to localize and quantify the distribution of BZ receptors with the standard technique of in vitro receptor autoradiography using [125I]iomazenil. The ex vivo autoradiographic distribution of activity was highly correlated (r = 0.89) with the distribution of BZ receptors, although a few brain regions showed reproducible discrepancies between ex vivo and in vitro results. The ex vivo autoradiograms provided quantitative data from a realistic "biological phantom," which may be used to assess the accuracy of image reconstruction and to investigate differences between the distribution of an intravenously (i.v.) administered tracer and that of its target in brain tissue.

Animals↗
PubMed1992

Serial cultivation of normal human keratinocytes: a defined system for studying the regulation of growth and differentiation.

We have developed a defined method for human epidermal keratinocyte culture. The minimally supplemented basal medium supported establishment of primary cultures from neonatal foreskin in a defined environment. It also supported serial cultivation and rapid expansion of cell number. Casein replaced serum for defined cryopreservation. Cells were serially cultivated in medium containing 0.08 mM calcium. The rate of cell division however remained high after addition of 1.8 mM calcium. The particulate transglutaminase activity of the cultures was low at confluence, even in the presence of 1.88 mM calcium, indicating an enrichment of the basal cell population. Culture with small amounts (0.3%) of chelated serum increased particulate transglutaminase activity approximately 2.2-fold in low calcium cultures and approximately 3.5-fold in high calcium cultures. A gradual reduction in growth rate of serum-treated cultures upon serial cultivation also indicated a depletion of cells with basal cell character. Bovine hypothalamic extract and cholera toxin were able to avert, in part, the differentiation-promoting effects of serum. Keratinocytes serially cultivated in the defined medium maintained the ability to develop normally into a morphologically differentiated epidermis.

Blood Proteins↗
PubMed1992

"Central" and "peripheral" benzodiazepine receptors: opposite changes in human epileptogenic tissue.

We measured the density of two benzodiazepine (BZ) receptor subtypes in neurosurgically obtained hippocampal tissue from the seizure focus of patients with temporal lobe epilepsy (TLE) showing mesial temporal sclerosis, the most common pathologic finding in TLE. We performed quantitative in vitro receptor autoradiography with [125I]Ro 16-0154, a probe for the central-type BZ receptor and with [3H]PK 11195, a probe for the peripheral-type BZ receptor. In comparison with autopsy and neurosurgical control groups, patients with mesial temporal sclerosis had regionally selective decreased central-type and increased peripheral-type BZ receptors. These changes paralleled regional losses of neurons and proliferation of glia. Decreases of the inhibitory central-type BZ receptor may be a component of the enhanced excitability of the seizure focus and also may allow localization of the focus by in vivo neuroreceptor imaging. Single photon emission computed tomography (SPECT) imaging of two TLE patients with [123I]Ro 16-0154 suggests that this technique may provide a more sensitive means of localizing the seizure focus than current imaging methods relying on changes in blood flow or glucose metabolism.

Adult↗
PubMed1991

Epidermis generated in vitro: practical considerations and applications.

The technology for culture of epidermis is one of the most advanced to date for generation of a tissue in vitro. Cultured epidermis is already used for a number of applications ranging from use as a permanent skin replacement to use as an organotypic culture model for toxicity testing and basic research. While simple epidermal sheets have been grafted successfully, more advanced models for skin replacement consisting of both dermal and epidermal components are in development and being tested in a number of laboratories. One of the most advanced in vitro models is the living skin equivalent, an organotypic model consisting of a collagen lattice contracted and nourished by dermal fibroblasts overlaid with a fully formed epidermis.

Culture Techniques↗
PubMed1991

Calibration of [123I]iodine-labeled tissue standards for autoradiographic studies.

Autoradiographic images can be analyzed with computer-assisted microdensitometry relative to radioactive reference standards to provide quantitative measurements of regional radioactivity concentrations. [123I]Iodine containing sections of brain paste have been calibrated relative to plastic-embedded tritium(3H) and 125Iodine standards. For exposure times of 8, 12 and 24 h, plastic-embedded standards covered a range concentrations of [123I]iodine from 1000 to 100,000 dpm/mg wet weight. Iodine-123 radioactivity was linear with section thickness from 5 to 30 microns. These studies confirmed the feasibility of using commercially available longer-lived reference standards to provide [123I]iodine equivalent values in absolute units of dpm (or microCi) per mg tissue.

Animals↗
PubMed1991

SPECT imaging of the benzodiazepine receptor: feasibility of in vivo potency measurements from stepwise displacement curves.

Iodine-123-labeled Ro 16-0154 is a high affinity, reversibly binding radiotracer for the benzodiazepine (BZ) receptor. Brain uptake of this radioligand was relatively stable and showed high ratios of specific-to-nonspecific uptake, with greater than 90% displaced by intravenous administration of BZ receptor agents. Repeated injections of increasing doses of each of five BZ drugs (Ro 16-0154, Ro 15-1788, clonazepam, alprazolam, and diazepam) yielded stepwise displacement curves, which were analyzed to measure the in vivo potencies of these agents. The relatively long half-life of 123I and the stable biologic uptake of the radiotracer allowed such potency estimations in just one experiment following a single injection of radioligand. The in vivo potencies of these five agents were highly correlated with their affinities for the BZ receptor determined with in vitro homogenate binding. A single crystal probe provided potency measurements virtually identical to simultaneously performed SPECT imaging studies. In conclusion, stepwise displacement by agents administered following the injection of the radioligand 123I-Ro 16-0154 provided a reliable means of measuring the in vivo potencies of BZ receptor agents. This in vivo determination may predict the clinical potency of BZ drugs than in vitro homogenate estimations, because the in vivo measure provides the summed effects of receptor affinity, plasma protein binding, penetration of the blood-brain barrier, and metabolism of the displacing agent.

Alprazolam↗
PubMed1991

Asthmatic amyotrophy. Three cases.

Asthmatic amyotrophy is becoming a well established clinical phenomenon. A polio-like syndrome, it is characterized by an asymmetrical lower motor neuron paralysis after an acute asthmatic episode. We are presenting three additional cases of this syndrome to add to the literature describing this phenomenon. Our cases are similar to the 18 previously reported cases showing an asymmetrical flaccid paralysis accompanied by normal sensation. The prognosis for full recovery of function is poor. Anterior horn cell damage is postulated because no sensory loss has been found in any patients.

Asthma↗
PubMed1990

Receptor binding characterization of the benzodiazepine radioligand 125I-Ro16-0154: potential probe for SPECT brain imaging.

The binding of an iodinated benzodiazepine (BZ) radioligand has been characterized, particularly in regard to its potential use as a neuroreceptor brain imaging agent with SPECT (Single Photon Emission Computed Tomography). Ro16-0154 is an iodine-containing BZ antagonist and a close analog of Ro15-1788. In tissue homogenates prepared from human and monkey brain, the binding of 125I-labeled Ro16-0154 was saturable, of high affinity (Kd = 0.5 nM at 37 degrees C), and had high ratios of specific to non-specific binding (approximately 40:1). Physiological concentrations of NaCl (150 mM) enhanced specific binding approximately 15% compared to buffer without this salt. Kinetic studies of association and dissociation demonstrated a temperature dependent decrease in affinity with increasing temperature. Drug displacement studies confirmed that 125I-Ro16-0154 binds to the "central" type BZ receptor: binding is virtually identical to that of 3H-Ro15-1788 except that 125I-Ro16-0154 shows an almost 10 fold higher affinity at 37 degrees C. These in vitro results suggest that 123I-labeled Ro16-0154 shows promise as a selective, high affinity SPECT probe of the brain's BZ receptor.

Animals↗