Apolipoprotein E deficiency effects on learning in mice are dependent upon the background strain.

Apolipoprotein E (apoE) deficient mice were bred onto the C57BL/6 and FVB/N strain backgrounds. The cognitive behavior of food-restricted apoE-deficient and wildtype male mice from these strains was assessed in an olfactory cued 8-arm radial maze. At 6 weeks of age, all four types of mice improved in maze performance over the course of 5 days. However, at 6 months of age, only the apoE-deficient mice on the C57BL/6 background failed to improve their maze performance over the 5 day course, as gauged by the number of incorrect choices made before retrieving both food rewards. Thus, an age-dependent and strain-specific effect of apoE deficiency on cognitive behavior was observed in these mice. The background strain affected activity levels in the maze, as well as in an open field assay. Plasma corticosterone levels were assessed in control, fasted, and post-restraint stress states. Fasting and restraint stress led to increases in plasma corticosterone levels. Although there were strain specific effects on fasting corticosterone levels, and the effect of apoE deficiency on post-stress corticosterone levels, there was no association between fasted corticosterone levels and impaired cognitive behavior in the 8-arm radial maze assay.

Human apolipoprotein E allele-specific brain expressing transgenic mice.

Transgenic mice were created using human apolipoprotein E2, E3, and E4 gene fragments driven by the human glial acidic fibrillary protein (GFAP) promoter. Founders were obtained and progeny were assayed for transgene expression in the brain by RNase protection, immunohistochemistry, and Western blotting, demonstrating robust apolipoprotein E (apoE) brain expression, with human apoE representing up to approximately 0.2% of total brain protein. Selected lines were bred to apoE-deficient mice yielding mice which expressed the human transgenic apoE isoforms in the absence of endogenous apoE. Immunohistochemistry revealed accumulation of the transgene encoded human apoE throughout the brain. Double immunofluorescence showed co-expression of the apoE transgene with endogenous glial acidic fibrillary protein. Primary astrocyte cultures from the transgenic mice secreted human apoE into the medium. Aged apoE4 transgenic mouse brain failed to demonstrate any evidence of senile plaques. These mice may be useful for elucidation of the mechanism by which apoE4 is associated with Alzheimer's disease.

The atypical codon usage of the plant psbA gene may be the remnant of an ancestral bias.

The psbA gene of the chloroplast genome has a codon usage that is unusual for plant chloroplast genes. In the present study the evolutionary status of this codon usage is tested by reconstructing putative ancestral psbA sequences to determine the pattern of change in codon bias during angiosperm divergence. It is shown that the codon biases of the ancestral genes are much stronger than all extant flowering plant psbA genes. This is related to previous work that demonstrated a significant increase in synonymous substitution in psbA relative to other chloroplast genes. It is suggested, based on the two lines of evidence, that the codon bias of this gene currently is not being maintained by selection. Rather, the atypical codon bias simply may be a remnant of an ancestral codon bias that now is being degraded by the mutation bias of the chloroplast genome, in other words, that the psbA gene is not at equilibrium. A model for the evolution of selective pressure on the codon usage of plant chloroplast genes is discussed.

Dental management of patients with diabetes mellitus.

It is estimated that a total of 16 million Americans are suffering fro m diabetes mellitus (DM). Dental health-care providers are involved in screening for this disease, as well as treating numerous patients with DM. As do all medically complex patients, individuals with DM pose a challenge for dental providers. This article describes the pathogenesis and physiology of DM and recommends guidelines for dental care.

The successful recruitment of elderly black subjects in a clinical trial: the CRISP experience. Cholesterol Reduction in Seniors Program.

This article describes the recruitment of elderly black subjects into the Cholesterol Reduction in Seniors Program (CRISP), a federal, multi-center, randomized, double-masked feasibility study of cholesterol intervention in the elderly. The study tested the feasibility of recruiting significant numbers of hypercholesterolemic black men, black women, and white women over the age of 65, groups previously underrepresented in federal trials. The study involved dietary modification and drug intervention with either 20 mg or 40 mg of lovastatin or placebo. Maximum follow-up was 18 months. Over the 12-month screening and recruitment period, 431 subjects (108% of the recruitment goal) were randomized. A total of 311 (72% of the study cohort) was female; 105 subjects (24% of the total cohort) were minorities. Media sources were most effective in recruiting white subjects. Church screening was an effective strategy in the black community, although such an approach required considerable resources s and time. The CRISP feasibility study demonstrated that a large cohort of elderly black subjects could be recruited in a cholesterol intervention trial, although the use of community-based approaches required substantial resources and staff time.

Laser therapy versus cryotherapy of lentigines: a comparative trial.

BACKGROUND: Lentigines are common sun-induced benign melanocytic proliferations. Many therapies have been advocated, but few have been systemically evaluated. OBJECTIVE: We studied the effectiveness of two laser modalities in comparison with liquid nitrogen cryotherapy for lentigines. METHODS: We conducted a randomized, controlled, prospective trial comparing liquid nitrogen cryotherapy, argon laser light delivered by a Dermascan shuttered delivery system, and low-fluence carbon dioxide laser irradiation in the treatment of solar lentigines at 99 sites in 13 patients. RESULTS: Cryotherapy was more likely to produce substantial lightening than either argon or CO2 laser treatment, which gave similar results (p < 0.05 for both comparisons). The odds of an excellent results were about 50% higher with cryosurgery than with CO2 or argon laser therapy. CONCLUSION: Liquid nitrogen cryotherapy was superior to argon and CO2 laser therapy in the treatment of benign epidermal pigmented lesions. This study demonstrates that comparative rather than uncontrolled studies are needed to judge the relative efficacy of therapies for benign pigmented lesions such as lentigines.

Efficiency of opaque photoprotective agents in the visible light range.

"Opaque" physical sunscreens are important for photoprotection of individuals with visible light and UV-A photosensitivity such as those with porphyria, drug photoallergy, and polymorphous light eruption. Diffuse spectral transmittance of various thicknesses of opaque sunscreen formulations were measured from 350- to 800-nm range using a spectrophotometer equipped with an integrating sphere. Transmission through 20% zinc oxide paste was high and decreased minimally despite large increases in the sunscreen layer thickness. Adding a visible light absorber such as iron oxide to scattering sunscreens, however, substantially lowered transmittance below that predicted by the product of the transmittances for each component alone. Opaque sunscreens protected against hematoporphyrin derivative photosensitization of albino guinea pig skin; these results were quantitatively consistent with the in vitro findings. Poor photoprotection against visible light is obtained with white paste sunscreens, even if thick layers are applied. The addition of pigments to such sunscreens, however, greatly enhances photoprotection and cosmetic acceptability.

In vitro and in vivo neurochemical effects of methylenedioxymethamphetamine on striatal monoaminergic systems in the rat brain.

A single high dose of methylenedioxymethamphetamine, a psychedelic agent, produced a rapid and persistent depletion of striatal indoles similar to that observed following administration of the serotonergic neurotoxin p-chloroamphetamine. The drug had little effect on dopaminergic variables. Like p-chloroamphetamine, methylenedioxymethamphetamine was found to be a relatively selective agent for inducing [3H]serotonin release in vitro. The serotonin uptake inhibitor, citalopram, blocked both [3H]serotonin release in vitro and striatal serotonin depletion in vivo, indicating that both processes were carrier dependent. In vivo comparisons of the stereoisomers of methylenedioxymethamphetamine indicated two phases of serotonin depletion similar to those reported for p-chloroamphetamine. Although both the (+)- and (-)-stereoisomers produced an acute (3 hr) decrease in striatal indoles, the long-term effects of the drug showed stereoselectivity in that the (+)-enantiomer produced the most dramatic serotonin depletion. Comparison of the effects of the stereoisomers of methylenedioxymethamphetamine and its n-desmethyl analog, methylenedioxyamphetamine, on [3H]serotonin and [3H]dopamine release in vitro showed the (+)-enantiomer of both drugs to be the more potent releasing agent. In spite of its reported lack of hallucinogenic activity, (+)methylenedioxyamphetamine was found to be of a potency similar to that of (+)methylenedioxymethamphetamine in inducing [3H]serotonin release in vitro. The results are discussed in terms of the neurochemical similarities between methylenedioxymethamphetamine and p-chloroamphetamine as well as the proposed role of serotonin release in the behavioral effects of methylenedioxymethamphetamine.

Compartmental analysis of the efflux of l-[3H]norepinephrine from isolated perfused rat lungs.

Isolated rat lungs, pretreated with 100 microM pargyline and 100 microM U-0521 (3',4'-dihydroxy-2-methylpropriophenone) to block metabolism of norepinephrine (NE), were perfused with 0.3 microM 3H-labeled l-norepinephrine (1-[3H]-NE) for 30 min. Efflux samples were then collected for 30 min during washout of the tissue with amine-free Krebs solution. Compartmental analysis (nonlinear least-squares regression) of the efflux of tissue l-[3H]NE content vs. time indicates that NE is accumulated in a large slowly equilibrating compartment (t 1/2 = 58.15 +/- 6.84 min) in addition to distribution in the vascular (blue dextran tracer) and extracellular ([3H]sorbitol tracer) fluid compartments of the lung. Pretreatment of the lungs with 100 microM cocaine hydrochloride reduces the total l-[3H]NE space from 7.44 +/- 1.91 to 2.48 +/- 0.23 ml/g (P less than 0.05) by selectively decreasing the size of the slow NE compartment from 6.99 +/- 1.97 to 1.67 +/- 0.14 ml/g (P less than 0.05). The large size, cocaine sensitivity, and long efflux half time of this compartment suggest that neuronal uptake contributes to the pulmonary vascular inactivation of l-[3H]NE.

Succinylcholine-induced increases in plasma catecholamine levels in humans.

Given the hypothesis that interaction of succinylcholine with nicotinic receptors releases endogenous catecholamines, plasma levels of epinephrine and norepinephrine were determined in anesthetized and manually ventilated patients immediately before and 2 min after intravenous administration of succinylcholine. Anesthesia was induced with intravenous thiopental (3-4 mg/kg) followed by the administration of nitrous oxide and oxygen (1:1) and 0.5-1.0% halothane. Stimulation of the patients was avoided. Succinylcholine (1 mg/kg) or metocurine (0.3 mg/kg) was injected intravenously and ventilation was controlled without intubation. Plasma norepinephrine levels increased from 301 pg/ml to 491 pg/ml (SEM = +/- 19 pg/ml, P less than 0.01, N = 5) 2 min after the injection of succinylcholine; the increase in plasma epinephrine was not statistically significant. The time course of catecholamine elevation was studied in three additional patients. The increase of norepinephrine occurred immediately after the injection of succinylcholine, peaked (647 +/- 67 pg/ml) around the third minute, and disappeared by the 10th min. The increase in epinephrine was less marked. Plasma levels of catecholamines did not change after the injection of metocurine (N = 2). The possibility that succinylcholine stimulates nicotinic receptors on the postganglionic sympathetic terminals is discussed. We propose that the elevation of plasma norepinephrine might contribute to the development of early adverse cardiovascular reactions to succinylcholine.

Two mechanisms of 3H-catecholamine accumulation in rabbit aorta media differentiated by sensitivity to temperature and corticosterone.

To characterize the extraneuronal accumulation of catecholamines (CA) in the media of rabbit aorta pieces of isolated media were incubated with 3H-CA (norepinephrine, isoproterenol or normetanephrine) plus 14C-sorbitol (to estimate the extracellular space) for varying periods at 37 or 0 degree C. The 3H-CA accumulation was 1.3 ml/g at 0 degree C and 2.2 ml/g at 37 degrees C. The sorbitol space was 0.6 ml/g at both temperatures. CA accumulation at 0 and 37 degrees C is significantly different from each other and from sorbitol accumulation. Corticosterone and phenoxybenzamine inhibit the temperature-sensitive component of CA accumulation. Accumulation of norepinephrine at 0 degree C is unaffected by corticosterone, phenoxybenzamine or oxytetracycline. The initial rate of NE accumulation at 37 degrees C, from steady state accumulation at 0 degree C, and the initial rate of accumulation at 0 degree C were linear functions of NE concentration between 10(-7) and 10(-2) M. Based on differences in sensitivity to corticosterone and temperature, we conclude that CA accumulation at 0 degree C is different from the accumulation at 37 degrees C.

Effect of inhibitors of neuronal and extraneuronal uptake on the accumulation and metabolism of 3H-l-norepinephrine in rabbit aorta.

In the isolated adventitia of rabbit aorta, blockade of neuronal uptake decreased the accumulation and deamination of 3H-l-norepinephrine (3H-l-NE) whereas blockade of extraneuronal uptake decreased O-methylation. Thus most of the O-methylation occurs in the extraneuronal elements. In the isolated media, blockade of extraneuronal uptake decreased metabolism but not accumulation of 3H-l-NE. If NE metabolism was prevented by pretreatment with pargyline and U-0521, 3H-l-NE accumulation in the isolated media increased markedly and now inhibitors of extraneuronal uptake could decrease this accumulation of 3H-l-NE by the isolated media. Thus when MAO and COMT are intact, NE does not accumulate in the extraneuronal compartment containing these enzymes, probably because it is metabolized as fast as it enters the compartment. Since the level of 3H-l-NE in the isolated media exceeded the sorbitol space when MAO and COMT are intact, some 3H-l-NE must have accumulated in a compartment which does not contain these enzymes.

The effects of 6-hydroxydopamine on the disposition of norepinephrine in the rabbit aorta.

Pretreatment of rabbits with 6-hydroxydopamine (6-OHDA; 25 mg/kg i.v. on day 0 plus 50 mg/kg i.v. on day 1, sacrifice on day 4) depleted 95% of the endogenous catecholamines from the rabbit thoracic aorta and heart. A partial recovery of endogenous catecholamines occurred over the next 2 weeks. 6-OHDA pretreatment decreased the cocaine-sensitive accumulation and the binding of l-[3H]norepinephrine (l-[3H]NE) by 85% in the rabbit thoracic aorta. Thus, this pretreatment with 6-OHDA produced an adequate chemical sympathectomy of the rabbit thoracic aorta. The effects of 6-OHDA pretreatment on the accumulation ane metabolism of l-[3H]NE were examined in the isolated adventitia and isolated media of the rabbit aorta to determine the origin of the O-methylated metabolites formed by the isolated adventitia. In the isolated adventitia, 6-OHDA pretreatment decreased the accumulation of l-[3H]NE and the formation of deaminated metabolites by 80%, but the formation of [3H]normetanephrine was increased more than 2-fold. 6-OHDA produced only a slight decrease in the formation of one of the deaminated metabolites of l-[3H]NE by the isolated media. These results are interpreted to mean that in the isolated adventitia most of the accumulation and deamination of l-[3H]NE occurs in the adrenergic nerve terminals whereas most of the O-methylation occurs in the extraneuronal elements of the isolated adventitia.

The effect of monoamine oxidase and catechol O-methyltransferase inhibitors on the accumulation and metabolism of [l-3H] norepinephrine by the adventitia and media of rabbit aorta.

The effects of catechol O-methyltransferase (COMT) and monoamine oxidase (MAO) inhibitors on the accumulation and metabolism of [l-3H] norepinephrine ([l-3H]NE) were studied in the isolated adventitia and media of rabbit aorta. The COMT inhibitors produced a greater inhibition of O-methylation in the isolated media than in the isolated adventitia. In addition, exposure to these drugs was associated with an increased level of [l-3H]NE and increased formation of the catechol deaminated metabolites by the isolated media but not by the isolated adventitia. The effects of these drugs on the balance of the oxidized vs. reduced forms of the deaminated metabolites was interpreted to mean that 3-methoxy-4-hydroxyphenylethylglycol is formed from normetanephrine rather than from 3,4-dihydroxyphenylethylglycol whereas 3-methoxy-4-hydroxymandelic acid is formed from 3,4-dihydroxymandelic acid. 3,4-Dihydroxyphenylethylglycol does not seem to serve as a substrate for 3-methoxy-4-hydroxyphenylethylglycol becuase it rapidly passes out of the tissue after it is formed. Parglyline and iproniazid markedly decreased deamination of [l-3H]NE and increased the tissue level of [l-3H]NE. However, these drugs produced little if any compensatory increase in O-methylation of [l-3H]NE. The effects of these drugs are compatible with the hypothesis that adrenergic nerves contain MAO and vascular smooth muscle contains MAO and COMT. Harmaline seemed to block both MAO and COMT. Based on the effects of clorgyline and deprenyl and the other MAO inhibitors tested, the isolated adventitia seems to contain primarily type A MAO and the isolated media seems to contain primarily type B MAO.