East Indian sandalwood (Santalum album L.) oil confers neuroprotection and geroprotection in Caenorhabditis elegans via activating SKN-1/Nrf2 signaling pathway.

East Indian Sandalwood Oil (EISO) has diverse beneficial effects and has been used for thousands of years in traditional folk-medicine for treatment of different human ailments. However, there has been no in-depth scientific investigation to decipher the neuroprotective and geroprotective mechanism of EISO and its principle components, α- and ß-santalol. Hence the current study was undertaken to assess the protective effects of EISO, and α- and ß-santalol against neurotoxic (6-OHDA/6-hydroxydopamine) and proteotoxic (α-synuclein) stresses in a Caenorhabditis elegans model. Initially, we found that EISO and its principle components exerted an excellent antioxidant and antiapoptotic activity as it was able to extend the lifespan, and inhibit the ROS generation, and germline cell apoptosis in 6-OHDA-intoxicated C. elegans. Further, we showed that supplementation of EISO, and α- and ß-santalol reduced the 6-OHDA and α-synuclein-induced Parkinson's disease associated pathologies and improved the physiological functions. The genetic and reporter gene expression analysis revealed that an EISO, or α- and ß-santalol-mediated protective effect does not appear to rely on DAF-2/DAF-16, but selectively regulates SKN-1 and its downstream targets involved in antioxidant defense and geroprotective processes. Together, our findings indicated that EISO and its principle components are worth exploring further as a candidate redox-based neuroprotectant for the prevention and management of age-related neurological disorders.

Suppression of lipopolysaccharide-stimulated cytokine/chemokine production in skin cells by sandalwood oils and purified α-santalol and ß-santalol.

Medicinally, sandalwood oil (SO) has been attributed with antiinflammatory properties; however, mechanism(s) for this activity have not been elucidated. To examine how SOs affect inflammation, cytokine antibody arrays and enzyme-linked immunosorbent assays were used to assess changes in production of cytokines and chemokines by co-cultured human dermal fibroblasts and neo-epidermal keratinocytes exposed to lipopolysaccharides and SOs from Western Australian and East Indian sandalwood trees or to the primary SO components, α-santalol and ß-santalol. Lipopolysaccharides stimulated the release of 26 cytokines and chemokines, 20 of which were substantially suppressed by simultaneous exposure to either of the two sandalwood essential oils and to ibuprofen. The increased activity of East Indian SO correlated with increased santalol concentrations. Purified α-santalol and ß-santalol equivalently suppressed production of five indicator cytokines/chemokines at concentrations proportional to the santalol concentrations of the oils. Purified α-santalol and ß-santalol also suppressed lipopolysaccharide-induced production of the arachidonic acid metabolites, prostaglandin E2, and thromboxane B2, by the skin cell co-cultures. The ability of SOs to mimic ibuprofen non-steroidal antiinflammatory drugs that act by inhibiting cyclooxygenases suggests a possible mechanism for the observed antiinflammatory properties of topically applied SOs and provides a rationale for use in products requiring antiinflammatory effects.

Neuropeptide Y modulates excitatory synaptic transmission in the olfactory bulb.

Although the olfactory bulb contains one of the highest concentrations of neuropeptide Y in the CNS, its function in the bulb remains unclear. In this study, we used whole-cell electrophysiological, molecular, and primary culture techniques to investigate neuropeptide Y gene expression and neuromodulatory actions of neuropeptide Y on rat olfactory bulb neurons. Northern analysis showed that neuropeptide Y mRNA increases with animal age or time in culture, in a parallel manner. In electrophysiology experiments, agonists that activate neuropeptide Y receptors (whole neuropeptide Y) and the Y2 receptor subtype (neuropeptide Y 13-36) reduced spontaneous excitatory activity in bulb interneurons. In investigating potential presynaptic effects, both agonists reduced the amplitude of calcium channel currents in the presynaptic (mitral/tufted) cell. Also consistent with a presynaptic effect, both agonists reduced the frequency but not the amplitude of miniature excitatory postsynaptic currents (or "minis") in interneurons. In examining potential postsynaptic effects, both agonists slightly increased membrane resistance but had no effect on currents evoked by glutamate. Together, these data suggest that neuropeptide Y inhibits excitatory neurotransmission between olfactory bulb neurons via a presynaptic effect on transmitter (glutamate) release.

Tumor suppressor protein p53 mRNA and subcellular localization are altered by changes in cellular copper in human Hep G2 cells.

Copper toxicity causes hepatic damage that can lead to the development of hepatocarcinoma. Similarly, copper deficiency has been reported to increase hepatocyte tumorigenesis. Thus, the objective of this work was to explore the role of copper toxicity and deficiency in the regulation of the tumor suppressor protein p53. Using Northern analysis, Western analysis, immunocytochemistry and the human hepatoma cell line Hep G2, this work showed that elevations in hepatocyte copper consistent with Wilson's disease (5.7-fold increase) induced p53 mRNA and confirmed that copper toxicity is correlated with apoptotic cell death. However, Western analysis and immunocytochemistry showed that post-transcriptional mechanisms are a significant part of the process, with p53 translocation from the cytosol into the nucleus of copper-treated cells. Treatment of Hep G2 cells with increasing concentrations of the copper chelator tetraethylenepentamine (TEPA, 0-50 micromol/L, 48 h) reduced cellular copper and increased mean p53 mRNA abundance by over fourfold with nuclear translocation of the wild-type protein. However, TEPA treatment did not result in a loss of cell viability or appear to induce apoptosis.

An evaluation of I Can Cope. Insights into program design and implementation.

The I Can Cope program is an educational program developed to provide information on how to promote physical and emotional well-being to individuals with cancer, their families, and friends. Through focus group and individual interviews and a mailed survey, the effectiveness of the selection, marketing, implementation, and support processes associated with implementing the program in the central Arizona and greater Las Vegas, Nevada, regions was evaluated. Findings, recommendations, lessons learned, and use of findings are discussed.

Abg1: a novel gene up-regulated by abscisic acid in guard cells of Vicia faba L.

A novel gene (abg1) was isolated by differential display RT-PCR from guard cells of Vicia faba L. Abg1 transcript accumulated in guard cells that were incubated with 5 microM S(+)-ABA for 1 h. The full-length abg1 cDNA was 753 bp, which included a 513 bp coding region. The deduced 17.8 kDa protein shared sequence similarity with several desiccation-related proteins reported in plants.

Licania michauxii Prance root extract induces hsp 70 mRNA and necrotic cell death in cultured human hepatoma and colon carcinoma cell lines.

An extract of Licania michauxii Prance root was found to be cytotoxic to cultured human hepatoma (HepG2) and colon carcinoma (Caco-2) cells. Morphological and nuclear characteristics of treated cells were consistent with necrotic death. Increases in the chaperone protein hsp 70 and hsp 70 mRNA were dose dependent reaching peak mRNA levels (40-fold above control) at 6 h. Increases in nuclear localization of hsp 70 was also observed with treatment. Heat treatment of cells for 45 min to induce hsp 70 prior to treatment with the extract provided transient protection from the necrotic response.

Effect of altered thyroid hormone status on rat brain ferritin H and ferritin L mRNA during postnatal development.

The iron binding protein ferritin is a heterogeneous mix of 24 heavy (H) and light (L) subunits. The H subunit is associated with iron utilization, while the L subunit is responsible for iron storage. Examination of the developmental pattern of mRNA abundance in rat brain revealed that ferritin L mRNA is highest at birth and declines during the first postnatal week. A similar decline was seen in ferritin H mRNA, but was followed by an increase in ferritin H mRNA in the second postnatal week which continued through postnatal day 21. The pattern of H mRNA regulation is similar to that in previous reports of total ferritin protein in the developing rat brain and is consistent with the fact that brain ferritin is predominately ferritin H. The effect of thyroid hormone on the developmental regulation of ferritin mRNAs was examined by the subcutaneous injection of a single dose of exogenous thyroxine (T(4); 2 microg/g) on postnatal day 1. Hypothyroidism was induced in pregnant dams with propylthiouracil (PTU; 0.05% in drinking water) from gestational day 7. Northern analysis from postnatal days 2-21 showed that T(4) increased ferritin H mRNA throughout development, while ferritin L mRNA was decreased compared to age-matched controls. PTU treatment decreased ferritin H and increased L mRNA in the later stages (days 14-21) of development. Given the distinct functions of ferritin H and L this suggests a role for thyroid hormone in the ability of the brain to regulate stored vs. utilizable iron during critical periods of development.

Self-selection of copper-containing diets by copper-deficient and overloaded rats.

To determine the effect of copper status on the preference for copper-containing food, male Sprague-Dawley rats were weaned to a copper-deficient, copper-adequate or high-copper diet. Four weeks later, alterations in copper status were confirmed by measurement of liver copper concentrations. Rats (n=10) were then given the choice between a copper-adequate or a copper-restricted diet of similar composition. Preference scores indicated that control rats preferred copper-deficient food. Preferences of rats on the high-copper diet were not different from control rats, suggesting that copper overload did not alter the pattern of selection. However, 7 of the 10 copper-deficient rats ate 80% or more of their intake as copper-adequate food resulting in preference scores that were significantly different from controls (p<0.03). This alteration in preference was corrected within 1 day of access to copper-adequate food.

Dopamine transporters participate in the physiological regulation of prolactin.

Three populations of hypothalamic neuroendocrine dopaminergic (NEDA) neurons, arising from the arcuate and periventricular nuclei of the hypothalamus release dopamine (DA) that acts at the pituitary gland to regulate the secretion of PRL. It is generally accepted that NEDA neurons lack functional DA transporters (DATs), which are responsible for uptake of DA from the synaptic cleft into the presynaptic axon terminal. This study localized DATs to the hypothalamo-pituitary axis and evaluated the effect of DAT blockade on the hypothalamo-pituitary regulation of PRL. After 7 days of treatment with cocaine (a nonspecific amine transporter blocker) or mazindol (a specific DAT blocker), the relative abundance of PRL messenger RNA (mRNA) in the anterior lobe (AL) of OVX rats was significantly decreased, whereas the relative abundance of tyrosine hydroxylase mRNA in the hypothalamus was significantly increased. The effect of cocaine or mazindol administration on DA turnover and serum PRL concentration was examined in estradiol (E2)-treated OVX rats. E2 administration (i.v.) resulted in a significant increase in serum PRL within 4 h; however, cocaine or mazindol administration abolished the E2-induced increase of PRL. Cocaine or mazindol significantly increased the concentration of DA at the site of the axon terminals within the median eminence (ME), intermediate lobe (IL) and neural lobe (NL), indicating blockade of uptake. Because formation of DOPAC requires uptake of DA, concentrations of DOPAC in the ME, IL and NL decreased following treatment with either cocaine or mazindol. These data, together with the presence of immunopositive DAT in the ME, pituitary stalk, IL, and NL, suggest that a functional DAT system is present within all three populations of NEDA neurons. Moreover, similarity between the effects of cocaine and mazindol treatment indicate that blockade of the DAT, but not other amine transporters, is responsible for suppression of PRL gene expression and secretion. Blockade of DATs prevent uptake of DA into NEDA neurons and consequently increases the amount of DA that diffuses into the portal vasculature and reaches the AL. These data provide evidence that DATs play a physiological role in the regulation of DA release from and TH expression in NEDA neurons and consequently PRL secretion and PRL gene expression and further support our previous observation that the regulation of PRL secretion involves all three populations of NEDA neurons.

Regulation of mitochondrial cytochrome b mRNA by copper in cultured human hepatoma cells and rat liver.

Copper overload and deficiency are known to cause morphological and functional mitochondrial abnormalities. The reverse transcriptase-polymerase chain reaction (RT-PCR)-based method of differential display of mRNA was used to identify genes with altered expression in cultured human hepatoma cells (Hep G2) exposed to increasing concentrations of copper (0-100 microM, 24 h). Copper regulation of a cloned PCR product, identified as the gene for the mitochondrially encoded cytochrome b, was confirmed by Northern analysis and in situ hybridization. Copper toxicity increased cytochrome b mRNA abundance up to 3.6-fold, and copper chelation reduced it by 50%. Hepatic cytochrome b mRNA was also increased in rats fed a high-copper diet. Thapsigargin treatment resulted in a significant increase in cytochrome b mRNA, suggesting that an increase in intracellular calcium may be involved in the mechanism of copper action. Furthermore, although cyclohexamide (CHX) alone did not increase cytochrome b mRNA, the addition of CHX and copper resulted in a sixfold increase. These data suggest a role for cytochrome b in the response to increases or decreases in hepatic copper.

Regulation of metallothionein-3 mRNA by thyroid hormone in developing rat brain and primary cultures of rat astrocytes and neurons.

Metallothionein-3 (MT-3) is a brain specific member of the MT family. Unlike other members of this family, MT-3 has been shown to act as a neuronal growth inhibitory factor. MT-3 mRNA abundance increases throughout the developmental period, reaching adult levels by postnatal day 21. The role of thyroid hormone in the developmental regulation of MT-3 mRNA was tested because thyroid hormone is known to regulate brain gene expression. Furthermore, gestational hypothyroidism results in developmental brain abnormalities. Hypothyroidism was induced in pregnant dams by the administration of PTU from gestational day 7, resulting in a 4- to 6-fold increase in pup MT-3 mRNA abundance on the day of birth (day 0) and on postnatal day 3. Normal pups did not reach this level of brain MT-3 mRNA until postnatal day 21. Administration of thyroxine (T(4), 2 microg/g) to pups on postnatal day 1 or day 20 resulted in a decrease in MT-3 mRNA abundance on postnatal day 21, regardless of when the injection was given. Furthermore, addition of T(4) to primary cultures of brain (olfactory bulb) astrocytes and neurons from 4-day-old rats resulted in a significant decrease in MT-3 mRNA in 24 h. Given the neuronal growth inhibitory function of MT-3, these data suggest that MT-3 may play a role in the CNS-related consequences of hypo- and hyperthyroidism during development.

Developmental regulation of hepatic ceruloplasmin mRNA and serum activity by exogenous thyroxine and dexamethasone.

Ceruloplasmin (Cp) is a copper-dependent oxidase with roles that include the regulation of iron metabolism, participation in the acute-phase response to inflammation, and antioxidant systems. Although developmental increases in hepatic Cp gene expression and serum activity have been described, the molecular mechanisms that are responsible for this regulation are not fully understood. The studies described here explored the possible role of glucocorticoids and thyroxine (T4) in the early neonatal development of Cp by the administration of these hormones on postnatal Day 1 (24 hr after birth), and the measurement of both hepatic Cp mRNA and serum activity through postnatal Day 10. Administration of the synthetic glucocorticoid hormone, dexamethasone (2 micrograms/g body wt), resulted in an increase in Cp mRNA on Days 3-7 that was accompanied by an increase in serum Cp activity that reached statistical significance at Day 10. Exogenous T4 (2 micrograms/g body wt) significantly increased Cp mRNA 24 hr after administration. Serum Cp activity was also significantly elevated by the early neonatal administration of T4. Furthermore, gestational hypothyroidism resulted in a significant decrease in Cp activity after postnatal Day 3. These data suggest a role for thyroid hormone and possibly glucocorticoids in the normal developmental regulation of Cp.

Exercise--it's never too late: the strong-for-life program.

OBJECTIVES: This investigation determined whether an in-home resistance training program achieved health benefits in older adults with disabilities. METHODS: A randomized controlled trial compared the effects of assigning 215 older persons to either a home-based resistance exercise training group or a waiting list control group. Assessments were conducted at baseline and at 3 and 6 months following randomization. The program consisted of videotaped exercise routines performed with elastic bands of varying thickness. RESULTS: High rates of exercise adherence were achieved, with 89% of the recommended exercise sessions performed over 6 months. Relative to controls, subjects who participated in the program achieved statistically significant lower extremity strength improvements of 6% to 12%, a 20% improvement in tandem gait, and a 15% to 18% reduction in physical and overall disability at the 6-month follow-up. No adverse health effects were encountered. CONCLUSIONS: These findings provide important evidence that home-based resistance exercise programs designed for older persons with disabilities hold promise as an effective public health strategy.

Regulation of neuropeptide Y mRNA and peptide concentrations by copper in rat olfactory bulb.

Neuropeptide Y is highly abundant in both the peripheral and central nervous systems and is known to have diverse functions including regulation of feeding behavior, blood pressure, circadian rhythms, reproductive behavior and the response to stress. Northern analysis showed that copper deficiency increased brain NPY mRNA abundance particularly in the olfactory bulb (OB). These increases were not accompanied by alterations in food intake or blood pressure. After 4 weeks of a copper-restricted diet, OB copper concentrations decreased to 44% of control and NPY mRNA increased 1.5-fold. Addition of a copper chelator to the restricted diet, resulted in a two-fold increase in OB NPY mRNA over copper adequate controls. These results were confirmed in primary cultures of OB neurons suggesting that the regulation of NPY mRNA is at the level of the bulb rather than by a hormonal or other copper-regulated factor external to the OB. Immunoreactive NPY (IR-NPY) levels were not, however, increased following the 4 weeks of copper deficiency. Addition of the chelator resulted in a 1.4-fold increase in IR-NPY that, while statistically significant, was not proportional to the two-fold increase in NPY mRNA in the same study. This may suggest that copper deficiency inhibits the translational mechanisms responsible for the synthesis of NPY or that NPY is exported from the bulb in copper deficiency.

Free zinc increases at the site of injury after cortical stab wounds in mature but not immature rat brain.

The accumulation of free zinc (Zn2+) appears to play a role in the neuronal degeneration that occurs after brain injury. Given that neonates respond to brain injury with increased plasticity compared to adults, this study compared the effect of age on free Zn2+ and the Zn2+-binding protein metallothionein-3 (MT-3) after injury. Unilateral cortical stab wounds were produced in 3-day-old and adult rats. Four weeks later, brains were removed for in situ visualization of free Zn2+ and measurement of MT-3 mRNA. Free Zn2+ and MT-3 mRNA accumulated after 4 weeks at the site of injury site when injury occurred in adults. However, 4 weeks after neonatal injury there was no increase in free Zn2+ or MT-3 mRNA in or around the site of injury.

Home-based resistance training: predictors of participation and adherence.

This study identified factors associated with exercise participation and adherence in a sample of 102 sedentary, functionally limited, community-dwelling adults aged 60 to 94 years who participated in a home-based resistance training program. Stepwise regression analyses revealed that baseline physical factors (i.e., higher levels of mobility, weaker muscle strength, and fewer numbers of new medical conditions) were associated with higher rates of participation in the home program. Positive attitudes and a sense of control toward exercise, lower levels of confusion and depressive moods, and the development of fewer new medical problems during the program were related to higher levels of adherence to the program. Findings revealed that although physical health variables were the primary indicators of an older person's overall participation in the program, it was the psychological factors that were most important to adherence to this home-based program.

Mechanisms of copper conservation in organs.

Organ copper is conserved in response to dietary copper restriction. In organs such as brain and heart, conservation is highly efficient, resulting in the loss of little organ copper. In contrast, conservation of copper in liver is induced only after a significant amount of organ copper is lost. Thus, the conservation of copper during dietary restriction is highly organ specific. Although the long-term pattern of organ copper conservation in rats has now been described through use of the continuous feeding of a single stable isotope, the mechanisms responsible for this conservation have not been identified or studied. These mechanisms may include copper-regulated changes in gene expression as well as other mechanisms. We now have the molecular tools that will permit the isolation of copper-regulated genes that may play a role in the conservation of organ copper. Identification of these mechanisms will allow the exploration of the effects of mild short-term and long-term copper deficiency and the role of copper in other physiologic and biochemical systems.

Response of rat adrenal neuropeptide Y and tyrosine hydroxylase mRNA to acute stress is enhanced by long-term voluntary exercise.

Neuropeptide Y (NPY) and catecholamines are synthesized in response to stress. Adrenal NPY mRNA and tyrosine hydroxylase (TH) mRNA were measured by Northern analysis 2 h after a single 20 min bout of shaker stress in exercised and sedentary male Sprague-Dawley rats. Long-term exercise (18 weeks of voluntary wheel running) alone did not significantly alter adrenal NPY mRNA or TH mRNA levels. However, increases in stress-induced NPY and TH mRNA abundances were significantly enhanced by long-term exercise (P < 0.01). These results suggest that long-term physical activity may enhance the ability to synthesize NPY and catecholamines under conditions of stress.

Regulation of ceruloplasmin by retinoic acid in the developing rat.

Ceruloplasmin (Cp), the major copper-binding protein in the plasma, is an acute phase protein with ferrioxidase activity. Both its oxidase activity and hepatic mRNA abundance increase during the developmental period. To test the possible role of retinoic acid, a derivative of vitamin A known to participate in cellular differentiation and development, on the developmental regulation of Cp, neonatal rat pups were injected with 2 micrograms 13-cis-retinoic acid (RA)/g body weight on postnatal day 1. Serum Cp activity and hepatic Cp mRNA were measured over the next 3 weeks in RA-treated and vehicle-treated controls. Serum Cp activity increased 2.5-fold 24 h after RA administration. However, hepatic Cp mRNA abundance was not elevated during this time period, suggesting that the action of RA on Cp activity was the result of post-transcriptional changes.