Gene expression profile of neurodegeneration induced by alpha1B-adrenergic receptor overactivity: NMDA/GABAA dysregulation and apoptosis.

The alpha1-adrenergic receptors (alpha1ARs) play an important role in mediating sympathetic neurotransmission in peripheral organ systems; however, central alpha1ARs are not well characterized. Additionally, due to the lack of sufficiently subtype-selective drugs or high avidity antibodies, the contribution of each alpha1AR subtype to various central functions is currently unclear. Transcription regulation through alpha1AR subtypes in the CNS is also unknown. Of interest, transgenic mice that systemically overexpress the alpha1BAR show central symptoms that include age-progressive impaired mobility, neurodegeneration and susceptibility to epileptic seizure. To investigate the molecular basis of this phenotype, oligonucleotide microarray studies of whole brains of various ages were carried out to compare gene expression profiles between transgenic and normal brains. The results indicated changes in expression of apoptotic, calcium regulatory, neurodegenerative and genes involved in neurotransmission. Defects in regulation of intracellular calcium are known to play a role in cell death; thus, these genes may provide clues as to the molecular basis of alpha1BAR-induced neurodegeneration. Epilepsy is a disorder that can be caused by an imbalance between excitatory (e.g. glutamate) and inhibitory (e.g. GABA) signals. Microarray analysis of transgenic brains showed increased N-methyl-d-aspartate (NMDA) receptors and decreased GABAA, which were confirmed with immunohistochemistry, western blot and radioligand binding studies. The alpha1BAR also co-localized with the glutamatergic distribution, suggesting a glutamate imbalance as a molecular rationale for the epileptic seizures.

Regulation of the cellular localization and signaling properties of the alpha(1B)- and alpha(1D)-adrenoceptors by agonists and inverse agonists.

The regulation of the cellular distribution and intracellular signaling properties of the alpha(1B)- and alpha(1D)- adrenoceptor (alpha(1)-AR) subtypes was examined in stably transfected Rat 1 fibroblasts. In unstimulated cells, alpha(1B)-AR expression was noted primarily on the cell surface. Treatment with phenylephrine induced internalization of the alpha(1B)-AR and promoted association with arrestin 2. The internalized alpha(1B)-AR colocalized with the transferrin receptor, an endosomal marker. In unstimulated fibroblasts, the alpha(1D)-AR was detected in a perinuclear orientation and was colocalized with arrestin 2 in a compartment also containing the transferrin receptor. After treatment with prazosin, which exhibits inverse agonist properties, the alpha(1D)-AR was redistributed from intracellular sites to the cellular periphery and was no longer associated with the transferrin receptor or arrestin 2. alpha(1D)-AR-expressing cells exhibited a high degree of basal activity for both inositol phosphate formation and extracellular signal regulated kinase (ERK), which was reduced by treatment with prazosin. In these cells, phenylephrine induced a dose-dependent increase in inositol phosphate formation but had no effect on ERK activity. In alpha(1B) -AR-expressing cells, phenylephrine stimulated both inositol phosphate formation and ERK activity. These data show that: 1) there are differences in the cellular localization of the alpha(1)-AR subtypes; 2) the alpha(1B)-AR exhibits expected G protein-coupled receptor activity regarding cellular localization, agonist-mediated internalization, and coupling to second messengers; and 3) the alpha(1D)-AR is constitutively active and, as a result, is localized to intracellular compartments involved in receptor recycling.

Expression of multiple alpha1-adrenoceptors on vascular smooth muscle: correlation with the regulation of contraction.

Previous work has shown that the genes encoding each alpha1-adrenoceptor subtype are coexpressed throughout the peripheral vascular system. We have evaluated subtype-selective antibodies as tools to determine the extent of protein expression in arteries. The alpha1A-, alpha1B-, and alpha1D-adrenoceptors were detected in the medial layer of the aorta, caudal, femoral, iliac, renal, superior mesenteric, and mesenteric resistance arteries. In Rat1 fibroblasts expressing each subtype, immunoreactivity was noted both on the cell surface and in a perinuclear orientation. Intense alpha1B-adrenoceptor immunostaining was similarly localized in cultured femoral and renal vascular smooth muscle cells. Although the cellular localization appeared to be the same, immunoreactivity obtained with alpha1A- and alpha1D-adrenoceptors was much less intense than that with the alpha1B-adrenoceptor. The alpha1A-adrenoceptor selective agonist A-61603 was 22-fold more potent in activating renal artery contraction when compared with the femoral artery. The expression of each alpha1-adrenoceptor was significantly decreased by in vivo application of antisense oligonucleotides targeted against each subtype. Inhibition of the expression of only one, the alpha1A in renal and the alpha1D in femoral arteries, reduced the contractile response to naphazoline. The results show: 1) subtype-selective antibodies can be used in tissues and cell culture to localize the alpha1-adrenoceptor subtypes, 2) in addition to expression on the cell surface, the alpha1-adrenoceptors are expressed intracellularly, and 3) despite expression of all adrenoceptors, a single subtype mediates the contractile response in the femoral and renal arteries.

Be a leader of C-H-A-N-G-E.

Changes in policy, technology, and procedures have made healthcare what it is today. Change is also necessary. And, change will occur with or without you. This article discusses the six actions a practice should follow when taking on the challenge of CHANGE.

Precipitation and the transfer of water, nutrients and pollutants in tree canopies.

The functioning of a forest community depends upon the routes, rates and transformations of material flowing through it. The canopy's interaction with precipitation can result in the interception of rainfall and the scavenging of water from clouds and fog. But this movement of water through the canopy also mediates the transfer of mineral nutrients and the deposition of some atmospheric pollutants. The past decade has seen the development of increasingly complex models of transfer processes in the canopy and new methodologies for studying them. One impulse for these advances has been the need to understand not only the movement of water, nutrients and pollutants but also their reciprocal effects. Another has been the need to evaluate the nature of pollutant-induced effects with respect to the costs and effectiveness of possible remedies.

Response of photosynthesis and cellular antioxidants to ozone in populus leaves.

Atmospheric ozone causes formation of various highly reactive intermediates (e.g. peroxyl and superoxide radicals, H(2)O(2), etc.) in plant tissues. A plant's productivity in environments with ozone may be related to its ability to scavenge the free radicals formed. The effects of ozone on photosynthesis and some free radical scavengers were measured in the fifth emergent leaf of poplars. Clonal poplars (Populus deltoides x Populus cv caudina) were fumigated with 180 parts per billion ozone for 3 hours. Photosynthesis was measured before, during, and after fumigation. During the first 90 minutes of ozone exposure, photosynthetic rates were unaffected but glutathione levels and superoxide dismutase activity increased. After 90 minutes of ozone exposure, photosynthetic rates began to decline while glutathione and superoxide dismutase continued to increase. Total glutathione (reduced plus oxidized) increased in fumigated leaves throughout the exposure period. The ratio of GSH/GSSG also decreased from 12.8 to 1.2 in ozone exposed trees. Superoxide dismutase levels increased twofold in fumigated plants. After 4 hours of ozone exposure, the photosynthetic rate was approximately half that of controls while glutathione levels and superoxide dismutase activity remained above that of the controls. The elevated antioxidant levels were maintained 21 hours after ozone exposure while photosynthetic rates recovered to about 75% of that of controls. Electron transport and NADPH levels remained unaffected by the treatment. Hence, elevated antioxidant metabolism may protect the photosynthetic apparatus during exposure to ozone.

Effects of airborne saline particles on vegetation in relation to variables of exposure and other factors.

Saline particles are a heterogeneous group of chloride(Cl)-containing airborne materials of natural as well as anthropogenic origins. They are usually a local problem of air pollution in terms of source and dispersion, but within these areas their effects on agricultural, ornamental or natural species of plants can be of substantial practical concern. These effects include the accumulation of Cl, the production of foliar lesions, and changes in the plant's levels of mineral nutrients and metabolites, physiological processes, and growth and reproduction. Some quantitative exposure-effect relationships have been formulated for foliar Cl, foliar lesions, and changes in growth and yield. These relationships are sensitive to various factors, such as flux, duration and frequency of exposure, species and stage of development of the plant, size and chemical composition of the particle, and light, temperature, relative humidity and precipitation during or after exposure. The interactions of these factors affect the response of the plant to saline particles by determining three major sets of processes: collection and retention of particles by the foliage; penetration of material from superficial deposits into the foliar tissue; translocation of absorbed Cl (or other ionic components) and susceptibility of tissue to it within the leaf.

Responses of plants to simulated saline drift as affected by species and conditions of exposure.

In exposures to simulated saline drift generated from a 0.6% (w/w) chloride (Cl)-solution, under controlled environmental conditions, the median effective doses for the occurrence of any salt-induced foliar injury (expressed as microgCl cm(-2) deposited in six hours) were: 2.9 for Canada hemlock (Tsuga canadensis [L.] Carr.); 10.3 for white flowering dogwood (Cornus florida L.); 43.5 for potato (Solanum tubersum L. cv Superior); 44.2 for northern red oak (Quercus rubra L.); 65.1 for sweet corn (Zea mays L. cv Golden Cross Bantam); and, 123 for bush bean (Phaseolus vulgaris L. cv Pinto). Response of bean was a function of total deposition and independent of its rate with multiple (one per day) 6-h exposures but not with exposures less than 6 h where toxicity (per mass of Cl) increased with an increase in the rate of deposition. Toxicity of particles increased with an increase in the concentration of Cl in the solution (1.6 or 5.0% w/w) from which they were generated with bush bean but not with hemlock. Post-exposure periods that cycled between 50 and 855% relative humidity (RH) produced a greater incidence of salt-induced foliar injury than did a regime of constant 85% RH.

Reliability of a modification of the health-related physical fitness test for use with physical education majors.

A modified form of the AAHPERD Health-Related Physical Fitness Test was administered to a sample of 200 college physical education majors. The skinfold measures were changed from the original test, and the flexed-arm hang was added to the test battery. The validity and reliability of the test battery has been established for middle school boys and girls. This study determined the multivariate reliability of the modified test battery using a canonical correlation model. The univariate intraclass reliabilities of the test items ranged from .91 to .99. The total redundancy for the modified test battery was .87.

Acid-soluble nucleotides of pinto bean leaves at different stages of development.

Acid-soluble nucleotides of unifoliate leaves of Pinto bean plants (Phaseolus vulgaris L.) were determined at young, mature, and senescent stages of development. At least 25 components could be distinguished on the basis of inorganic phosphorus determinations and 37 or more fractions on the basis of (32)P labeling, with adenosine di- and triphosphates accounting for 60% of the total moles of nucleotide. The total nucleotide P and inorganic P, on a fresh weight basis, decreased about 44% between each stage of leaf development, but decrements in the levels of individual nucleotides varied from this over-all pattern.Minor changes in the relative abundance of the individual nucleotides accompanied aging although the percentage of purine-containing nucleotides decreased with age. Total (32)P activity per leaf in the nucleotide pool increased about 3-fold between the young and mature leaves and decreased slightly as leaves became senescent. In general, the specific activities of the nucleotides increased with increased age and adenosine-, guanosine-, uridine-, and cytidine triphosphates and adenosine diphosphate accounted for approximately 90% of the total activity. The changes in the relative sizes and energy status of the nucleotide pools were not so obvious as the changes in other metabolites that have been reported to accompany aging in leaf tissue.