Cimetidine has been shown to protect against acetaminophen-mediated hepatotoxicity in both rats and mice. In contrast to cimetidine, ranitidine recently has been determined to potentiate the hepatotoxic action of acetaminophen in Fischer 344 rats. The present studies were designed to characterize this ranitidine-acetaminophen interaction. Acetaminophen administration (750 mg per kg, p.o.) to F344 rats produced maximal hepatic necrosis, 24 hr after treatment, as assessed by SGPT activity and histopathology. Ranitidine pretreatment 30 min prior to acetaminophen treatment increased the toxicity but did not alter its course. Ranitidine administration (50 mg per kg) enhanced acetaminophen hepatotoxicity throughout the toxic dose range of acetaminophen (600 to 1,000 mg per kg) and potentiation of acetaminophen hepatotoxicity by ranitidine was dose-dependent. Maximal increases were observed at 50 mg per kg ranitidine whereas, doses of ranitidine greater than 100 mg per kg inhibited acetaminophen toxicity. SGPT data were corroborated by histopathologic evaluation. Ranitidine was not hepatotoxic when administered alone (500 mg per kg), or following glutathione depletion, or after induction of hepatic mixed-function oxidase activity. The results obtained in these studies support the suggestion that, at high doses (greater than 100 mg per kg), ranitidine reduces acetaminophen hepatotoxicity by reducing metabolic activation, while at lower doses ranitidine potentiates acetaminophen hepatotoxicity. Inhibition by ranitidine of acetaminophen conjugation is proposed as a possible mechanism of this potentiation.

Acetaminophen↗

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Image analysis of helical objects: the Brandeis Helical Package.

Although the fundamental steps in Fourier-based image analysis of electron micrographs of helical structures have not changed significantly since the techniques were developed 30 years ago, there have been developments which aid both the analysis itself and the interpretation of results. Increases in computational resources have allowed the automation of many of the repetitive steps in image processing. We describe here a set of computer programs which have been developed at Brandeis University over the past 10 or more years. These programs, referred to as the Brandeis Helical Package, are designed to operate independently of each other with a simple and more uniform protocol for the flow of information between them. The programs are now easier to understand and to use and therefore represent a good tool for the analysis of helical structures.

Actins↗

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Divergent changes in D-1 and D-2 dopamine binding sites in human brain during aging.

The density of D-1 and D-2 dopamine receptors in human caudate nucleus and putamen, obtained postmortem, were studied throughout the adult lifespan using [3H]fluphenazine as the dopamine receptor ligand. The D-1 subtype increased progressively with age in both regions, while the D-2 subtype declined in caudate nucleus. The ratio of D-1/D-2 Bmax in both regions increased from approximately 1 at age 20 to 2 by age 75. The dopamine content in putamen declined with age and was inversely correlated with D-1 receptor density. We suggest that D-1 receptor density is up-regulated by loss of dopamine during aging. The D-2 receptor density in caudate nucleus was positively correlated with choline acetyltransferase activity, suggesting that loss of intrastriatal neurons with age may contribute to the decrease in D-2 sites. These divergent changes in dopamine receptor subtypes with age result in an altered complement of dopamine receptors in older humans and may provide a basis for selective pharmacotherapy in disorders of the basal ganglia.

Adolescent↗

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Considerations in the treatment of neurological disorders with trophic factors.

The rationale for the therapeutic administration of NGF in Alzheimer's disease cannot be based purely on the premise of replacement therapy, and must instead consider the general anabolic properties of this trophic substance. In addition, there are potentially deleterious side effects of such therapy which should be evaluated in clinical trials. The selectivity of NGF for cholinergic neurons (in CNS) compared to the more generalized degeneration found in Alzheimer's disease raises the possibility of using trophic factors supporting a broader variety of neuron classes (FGF, ILGF).

Alzheimer Disease↗

PubMedDate not supplied

Age-related changes in glial fibrillary acidic protein mRNA in the mouse brain.

Several RNA sequences were tested for age-related changes in prevalence levels in the mouse cerebral cortex, hippocampus, and cerebellum. In all three regions, there were increased levels of RNA for glial fibrillary acidic protein, an astrocyte-specific protein, by RNA gel-blot analysis and by a solution hybridization assay. There was no change in glutamine synthetase mRNA level, another glial protein. The only other mRNA sequence which changed was Thy-1 antigen, a neuronal protein, which decreased slightly in the hippocampus. We conclude that with age there is an age-related increase in glial fibrillary acidic protein RNA prevalence potentially reflecting an increase in the size, number, and/or fibrous character of astrocytes.

Aging↗

PubMedDate not supplied

Patch averaging of electron images of GP3*I crystals with variable thickness.

The combination of Fourier and correlation averaging techniques with multivariate statistical analysis and classification, a method known as patch averaging, is used to analyze untilted and tilted images of negatively stained GP32*I crystals, which exhibit variable thicknesses in a single crystal. Within a single image, coherent areas of the same apparent thickness can be distinguished from areas of differing thicknesses. Analysis using the phase relationships among symmetry-related reflections from reconstituted images obtained from untilted micrographs confirms the ability of the method to classify these variable thicknesses properly. Furthermore, the phases from some of the reconstituted images obtained from both untilted and tilted micrographs were found to match well with the phases in a previously determined three-dimensional data set of this crystal with pg symmetry along the crystallographic b axis. These results indicate the utility of the patch averaging procedures in the structural determination of protein crystals with different thicknesses.

Crystallization↗

PubMedDate not supplied

New approaches to the study of central nervous system function. Immune-nervous system interactions and cell culture.

The paper by Lal and Forster is discussed with reference to future experiments which might provide insight into mechanisms regarding their exciting data that circulating brain reactive antibodies may cause learning deficits. The paper by Azmitia et al. on cell culture techniques is discussed with respect to the types of studies in which culture systems have proven most valuable in the past, and should continue to do so in the future.

Aging↗

PubMedDate not supplied

Combined in situ hybridization and immunocytochemistry in the assay of pharmacological effects on tyrosine hydroxylase mRNA concentration.

An assay for tyrosine hydroxylase (TH) mRNA by in situ hybridization in combination with immunocytochemistry (ICC) for TH on the same section is described. The in situ hybridization protocol was optimized for [35S]cRNA (complementary RNA, i.e. anti-sense strand) probe concentration and time of hybridization. The specificity of hybridization was measured by several critera. The advantage of measuring grain density versus grains per cell is discussed for quantitation of in situ autoradiography. Finally, the reserpine-induced increase in adrenal TH mRNA was used to validate quantitative aspects of the in situ hybridization technique by comparison with blot hybridization. In contrast to the adrenal, reserpine did not increase TH mRNA in substantia nigra (s. nigra) neurons as measured by either technique.

Animals↗

PubMedDate not supplied

Development and psychometric evaluation of the Environment-Behavior Interaction Code (EBIC).

BACKGROUND: Although the behavioral changes with progressive dementia are seen to increasingly depend on the environmental context until late stage disease, measurement has not reflected this interaction in real time to allow examination of antecedents to disruptive behavior. OBJECTIVES: To develop and evaluate the psychometric properties of the Environment-Behavior Interaction Code (EBIC) for use in dementia care research with either sequential or nonsequential analyses of behavioral data. METHOD: Development of the computer-based (sequential event format) EBIC provided an observational coding system to classify all behavior and environmental context in real time, so that the probability of social environmental antecedents to resident disruptive behavior could be estimated. A checklist (interval format) EBIC, based on the same behavioral taxonomy, was developed for clinical outcome research. A total of 158 elderly residents of dementia care units were purposively selected from three large long-term care facilities for the psychometric study components. RESULTS: Psychometric results indicated significant (p < 0.01) known-groups validity for the disruptive behavior construct, which was defined as a composite of aversive, harmful, and high intensity neutral behavior. Interrater agreement for the event format of the EBIC was estimated by average kappa (0.65) and percentage agreement (78%). For the interval format, the mean interrater kappa was 0.80 with 96% agreement. Stability of the event format using a 2-week retest interval ranged from r= .50 (positive behavior) to r = 0.73 (negative behavior, defined as aversive + harmful). On replication with a new sample, stability was higher for positive (r = 0.92) and negative (r = 0.95) components, and for composite scores of nondisruptive (positive + low intensity neutral; r = 0.65) and disruptive (r= 0.85) behavior. CONCLUSION: This research provided support for the reliability and validity of both event and interval EBIC formats. Measurement using the EBIC taxonomy has applicability to dementia care research questions that call for either sequential analysis of social interactions or nonsequential analysis of behavioral outcomes in intervention studies.

Aged↗

PubMedDate not supplied

The application of toxicokinetic data to dosage selection in toxicology studies.

Appropriate dosage selection is a key element in the design of toxicology studies and, hence, is the first step in the process of evaluating the safety of a new chemical or pharmaceutical agent. This demands careful consideration of exposure to the drug or chemical under investigation in relation to the pharmacological or toxicological effects it evokes in an experimental animal. Toxicokinetic data provide this perspective, but they should not be considered exclusively of other data which reflect the specific activity, potency, or metabolism of the drug or chemical in each individual test species. It is equally inappropriate to base dosage selection in toxicology studies exclusively on functional or morphological endpoints that cause effects outside the range which can be accommodated by homeostatic mechanisms and repair processes. Finally, extrapolation of toxicokinetic data across species lines can lead to serious miscalculations with respect to both dosage selection and the process of risk assessment. In each case, decisions should be based on the integration of toxicokinetic data with other measures and endpoints of biological and toxicological effect.

Animals↗

PubMedDate not supplied

Mice transgenic for a human amyloid precursor protein promoter-lacZ reporter construct.

Transgenic mouse lines were generated that expressed a 2-kb amyloid precursor protein (APP) promoter/beta-galactosidase reporter gene construction. In brain, hippocampal pyramidal neurons, neurons in the deeper layers of cerebral cortex, and neurons in several thalamic nuclei were heavily labeled by beta-galactosidase histochemistry. In general, molecular layers and white matter regions did not express the reporter gene. When compared with in situ hybridization for endogenous murine APP RNA, the striatum and outer layers of cerebral cortex had little reporter expression. Thus, the match between reporter expression and endogenous APP expression in brain was not perfect. A similar mismatch between the relative expression of the reporter gene and endogenous APP RNA distribution was found in homogenates from several organs. Although prior work in transgenic mice found similar mismatches in reporter gene distribution, none had tested the APP promoter construct in response to neuronal injury. Kainic acid injections successfully increased murine APP expression in the transgenic mice, but had no effect on the reporter gene expression. Based on these data and those collected by others, we conclude that the 2-kb region upstream of the APP transcription initiation site contains some elements responsible for the tissue-specific expression of this gene, but does not contain all the cis-acting elements sufficient for either the differential tissue distribution of this gene or the regulation of this gene subsequent to neural damage.

Amyloid beta-Protein Precursor↗