Biomedical subjects

J A Joseph

Publications and source records attributed to J A Joseph.

At least 37 records · Page 2Linked to original sources

PubMed2001

Antioxidant and pro-oxidant capacity of catecholamines and related compounds. Effects of hydrogen peroxide on glutathione and sphingomyelinase activity in pheochromocytoma PC12 cells: potential relevance to age-related diseases.

The antioxidant and pro-oxidant capacity of catecholamines (CA) and related compounds were analyzed using the oxygen radical absorbance capacity (ORAC) assay. In the assay 2,2'-azobis (2-amidino-propane) dihydrochloride (AAPH), a peroxyl radical generator, ROO*; H2O2-Cu2+, mainly a hydroxyl radical generator, *OH; and Cu2+ a transition metal were used. The antioxidant effect of CA and its related compounds were in the order: neurotransmitters: dopamine (DA), norepinephrine (NE) > metabolites > amino acid precursors as measured by using AAPH. The antioxidant effect of CA and related compounds as measured by using AAPH were linearly correlated with concentration, while the antioxidant effect of CA in scavenging *OH produced by H2O2-Cu2+ increased proportionally to concentration at low concentration, but after reaching a maximum declined with increasing concentration. In the presence of Cu2+, CA acted as pro-oxidant. Glutathione (GSH) acted as a pro-oxidant when H2O2-Cu2+ or when Cu2+ alone was used as an oxidant and showed much higher pro-oxidant effect than DA, which could have relevance in the vulnerability of dopaminergic neurons to oxidative stress in the aging and aging related diseases. The antioxidant capacity of CA and many related compounds seems to be correlated with the numbers of hydroxyl groups and their position on the benzoic ring. The O-methylation and sulfate conjugation of the hydroxyl substitution inactivates both the antioxidant and pro-oxidant activities of CA. Our results show that oxidative stress induced by low (5 microM) or high (300 microM) doses H2O2 in pheochromocytoma PC12 cells significantly up-regulate the activity of Mg-dependent neutral sphingomyelinase (Sase), and significantly decreased GSH.

Animals↗

PubMed2001

Automated measurement of age-related changes in the locomotor response to environmental novelty and home-cage activity.

The likelihood to explore in an open-field environment decreases with age. Older animals tend to be less active and explore less both in novel and home-cage environments. The locomotor performance (fine movements, ambulatory movements, and rearing) of male Fischer 344 (F344) rats that were 6 (n=6) or 22 (n=6) months of age was evaluated by continuous automated counting of photobeam interruptions, every 30 min, during 60 consecutive hours, in standard polycarbonate cages. Novel environment performance was determined by photobeam interruption counting during the first hour in the new cage. The remaining 59 h were evaluated as home-cage activity. A significant age-related decrease in ambulatory and fine motor activity was seen during the first hour of testing (novel environment). In addition, aged rats showed a decreased number of ambulatory and fine movements in home-cage activity, predominantly during the dark portion of the light cycle and during or around both light-switch periods (05:00 and 17:00). No differences were seen in rearing behavior. These findings provide a more detailed analysis and additional evidence of the activity decreases and rhythmic changes seen in aged F344 rats under uninterrupted testing conditions.

Aging↗

PubMed2001

Effect of age on object exploration, habituation, and response to spatial and nonspatial change.

To measure their ability to detect novel arrangements in a given environment, young (6 months old) and senescent (22-24 months old) male F344 rats were repeatedly exposed to a given spatial configuration of objects contained in an open field. After the rats were habituated to the novel environment (1 trial with no objects, followed by 3 trials with 5 salient objects), the spatial arrangement of the objects was modified (2 trials), and object novelty was tested (2 trials) by substituting a familiar object with a new one at the same location (nonspatial change). The results indicated that the senescent rats explored old objects less than young rats, particularly on Trial 2. On the 1st trial with displaced objects (Trial 5), the senescent rats explored the displaced objects less than the young rats. However, when a new object was placed in the field (Trials 7-8), there were no age differences in new object exploration. These results suggest that senescent rats have decrements in the ability to build spatial representations of the environment and to use this information to detect such changes, even though object recognition is not impaired with age.

Aging↗

PubMed2001

Effects of exposure to 56Fe particles on the acquisition of a conditioned place preference in rats.

Exposure to low doses of 56Fe particles produces changes in neural function and behavior. The present experiments were designed to examine the effects of irradiation on the acquisition of a dopamine-mediated conditioned place preference (CPP). In the CPP procedure, rats are given an injection of the dopamine agonist amphetamine in one distinctive compartment and a saline injection in a different compartment of a three-compartment apparatus. Control rats develop a preference for the amphetamine-paired compartment. In contrast, rats exposed to 1 Gy of 56Fe particles fail to develop a similar preference. The results of the experiment indicate that exposure to low doses of heavy particles can disrupt the neural mechanisms that mediate the reinforcement of behavior.

Amphetamine↗

PubMed2000

Polyphenolics enhance red blood cell resistance to oxidative stress: in vitro and in vivo.

In this study we investigated the potential antioxidant properties of blueberry polyphenolics in vitro and vivo, using red blood cell (RBC) resistance to reactive oxygen species (ROS) as the model. In vitro incubation with anthocyanins or hydroxycinnamic acids (HCA) (0.5 and 0.05 mg/ml) was found to enhance significantly RBC resistance to H2O2 (100 microM) induced ROS production. This protection was also observed in vivo following oral supplementation to rats at 100 mg/ml. However, only anthocyanins were found to afford protection at a significant level, this at 6 and 24 h post supplementation. This protection was not consistent with the measured plasma levels of anthocyanins. Indeed, plasma polyphenolic concentrations were highest after 1 h, declining considerably after 6 h and not detected after 24 h. The difference in absorption between anthocyanins and HCA is likely to have contributed to the observed difference in their abilities to afford protection to RBC. This protection represents a positive role following dietary consumption of polyphenolics from blueberries, against ROS formation within RBC in vivo.

Animals↗

PubMed2000

Oxidative stress protection and vulnerability in aging: putative nutritional implications for intervention.

Research indicates that vulnerability to oxidative stress (OSV) may increase in aging, suggesting that age-related neurodegenerative diseases such as Alzheimer's disease (AD) or vascular dementia (VAD) may be superimposed upon a vulnerable neuronal environment. Determinations in cell models have suggested that the enhanced OSV may be the result of, (a) increases in membrane lipids, especially sphingomyelin and the sphingomyelin metabolite, sphingosine-1-phosphate, (b) decreases in glutathione, and (c) CNS distribution of OS-sensitive neuronal muscarinic receptor subtypes (e.g. M1, M2 and M4). These changes appear to enhance, (a) decrements in cellular calcium buffering following KCl-induced depolarization, and (b) cell death under OS conditions. Among the most effective agents that antagonized cellular OSV were the combination of polyphenolics found in fruits (e.g. blueberry extract) with high antioxidant activity. Subsequent experiments using dietary supplementation with fruit (strawberry) or vegetable (spinach) extracts have shown that such extracts are also effective in forestalling and reversing the deleterious effects of behavioral aging in F344 rats. Thus, it appears that the beneficial effects of the polyphenolics found in fruits and vegetables in neuronal aging and behavior may be similar to those seen with respect to carcinogenesis and cardiovascular disease.

Aging↗

PubMed2000

Incorporation of the elderberry anthocyanins by endothelial cells increases protection against oxidative stress.

The objective of this study was to investigate the ability of endothelial cells (EC) to incorporate anthocyanins and to examine their potential benefits against various oxidative stressors. Endothelial dysfunction has been proposed to play an important role in the initiation and development of vascular disease, with studies having shown that administration of antioxidants improves endothelial function. Elderberry extract contains 4 anthocyanins, which where incorporated into the plasma membrane and cytosol of EC following 4 h incubation at 1 mg.ml(-1). However, incorporation within the cytosol was considerably less than that in the membrane. Uptake within both regions appeared to be structure dependent, with monoglycoside concentrations higher than that of the diglucosides in both compartments. The enrichment of EC with elderberry anthocyanins conferred significant protective effects in EC against the following oxidative stressors: hydrogen peroxide (H(2)O(2)); 2, 2'-azobis(2-amidinopropane) dihydrochloride (AAPH); and FeSO(4)/ascorbic acid (AA). These results show for the first time that vascular EC can incorporate anthocyanins into the membrane and cytosol, conferring significant protective effects against oxidative insult. These findings may have important implications on preserving EC function and preventing the initiation of EC changes associated with vascular diseases.

Amidines↗

PubMed2000

Mechanisms of hydrogen peroxide-induced calcium dysregulation in PC12 cells.

The mechanisms of H(2)O(2)-induced elevated calcium baselines in PC12 cells were investigated in the present study by using fura-2-fluorescent image analysis. The results showed that the calcium comes from both intracellular and extracellular sources. Although the major intracellular source was mitochondria, only the extracellular calcium influx was responsible for the sustained post-H(2)O(2)-exposure increases. This calcium influx was partially blocked by calcium channel antagonists [verapamil (L-type) or mibefradil (nonselective)] and was more effectively blocked by the sodium channel antagonist, tetrodotoxin (TTX). Membrane depolarization following H(2)O(2) exposure contributed to the opening of the ion channels. The H(2)O(2)-induced calcium influx was blocked by TTX even in a sodium-free buffer, indicating that calcium directly fluxed through sodium channels. Sodium-calcium exchangers (NCX) on the plasma membrane did not play a role, because use of a specific reverse mode NCX inhibitor, No. 7943, was ineffective in blocking the influx. The H(2)O(2)-induced calcium influx was mimicked by using a thiol-selective oxidizing reagent, 2', 2'-dithiodipyridine, and in both situations, the calcium levels were completely reversed by a thiol-selective reducing reagent, dithiothreitol. Our results indicated that mechanisms of oxidant-induced elevated calcium baselines in PC12 cells involved calcium influx through sodium and calcium channels that may be directly or indirectly attributed to thiol oxidation.

2,2'-Dipyridyl↗

PubMed2000

CNS-induced deficits of heavy particle irradiation in space: the aging connection.

Our research over the last several years has suggested that young (3 mo) rats exposed to whole-body 56Fe irradiation show neuronal signal transduction alterations and accompanying motor behavioral changes that are similar to those seen in aged (22-24 mo) rats. Since it has been postulated that 1-2% of the composition of cosmic rays contain 56Fe particles of heavy particle irradiation, there may be significant CNS effects on astronauts on long-term space flights which could produce behavioral changes that could be expressed during the mission or at some time after the return. These, when combined with other effects such as weightlessness and exposure to proton irradiations may even supercede mutagenic effects. It is suggested that by determining mechanistic relationships that might exist between aging and irradiation it may be possible to determine the common factor(s) involved in both perturbations and develop procedures to offset their deleterious effects. For example, one method that has been effective is nutritional modification.

Aging↗

PubMed2000

Effects of exposure to heavy particles on a behavior mediated by the dopaminergic system.

The effects of exposure to heavy particles on behaviors mediated by the central nervous system (CNS) are qualitatively different than the effects produced by exposure to other types of radiation. One behavior mediated by the CNS is the amphetamine-induced taste aversion, which is produced by pairing a novel tasting solution with injection of amphetamine. When the conditioning day is three days following irradiation, exposing rats to low doses of 56Fe particles (600 MeV/n or 1 GeV/n) eliminates the taste aversion produced by injection of amphetamine, which is dependent upon the integrity of the central dopaminergic system, but has no effect on the aversion produced by injection of lithium chloride which is mediated by the gastrointestinal system. In contrast to the effects obtained using heavy particles, exposing rats to 60CO gamma rays or to fission spectrum neutrons has no selective effect upon the acquisition of either amphetamine- or lithium chloride-induced taste aversions. When the conditioning day occurs four months following exposure to 1 GeV/n 56Fe particles, there is an enhancement of the amphetamine-induced taste aversion. The implications of these findings for approaches to risk assessment are considered.

Amphetamine↗

PubMed2000

Effect of fruits, vegetables, or vitamin E--rich diet on vitamins E and C distribution in peripheral and brain tissues: implications for brain function.

Age-related neurodegenerative conditions are the principal cause of declining cognitive and motor function during aging. Evidence support that fruits and vegetables containing generous amounts of antioxidant nutrients are important for neurological function. We investigated the effect of diets enriched with fruits or vegetables but low in vitamin E and a diet high in vitamin E on the distribution of vitamins C and E in the brain and dopamine release of Fischer 344 rat model, over an 8-month period. The low-vitamin E diet resulted in lowered alpha-tocopherol levels in brain and peripheral tissues, whereas the animals that received a diet enriched in vitamin E showed a significant increase, between 500-900%. Vitamin C concentration in plasma, heart, and liver was reduced in the vitamin E-supplemented group. It is concluded that supplementation or depletion of alpha-tocopherol for 8 months results in marked changes in vitamin E levels in brain tissue and peripheral tissues, and varied distribution of alpha-tocopherol throughout the different brain regions examined. In addition, compared to control group, rats supplemented with strawberry, spinach, or vitamin E showed a significant enhancement in striatal dopamine release. These findings suggest that other nutrients present in fruits and vegetables, in addition to the well-known antioxidants, may be important for brain function.

Aging↗

PubMed2000

Arson. Tracking the culprit in Alzheimer's disease.

By focusing on the lesions in Alzheimer's disease, and regarding them as either critical or irrelevant, researchers may have missed much regarding the origin and pathogenesis of this disease. In this article we consider that the lesions are so obvious not only because they are pathognomonic for Alzheimer's disease, but also because they represent a major departure from normal physiology. We suggest that these myriad pathological changes are homeostatic compensatory mechanisms to aging.

Aging↗

PubMed2000

Spatial learning and memory deficits induced by exposure to iron-56-particle radiation.

It has previously been shown that exposing rats to particles of high energy and charge (HZE) disrupts the functioning of the dopaminergic system and behaviors mediated by this system, such as motor performance and an amphetamine-induced conditioned taste aversion; these adverse behavioral and neuronal effects are similar to those seen in aged animals. Because cognition declines with age, spatial learning and memory were assessed in the Morris water maze 1 month after whole-body irradiation with 1.5 Gy of 1 GeV/nucleon high-energy (56)Fe particles, to test the cognitive behavioral consequences of radiation exposure. Irradiated rats demonstrated cognitive impairment compared to the control group as seen in their increased latencies to find the hidden platform, particularly on the reversal day when the platform was moved to the opposite quadrant. Also, the irradiated group used nonspatial strategies during the probe trials (swim with no platform), i.e. less time spent in the platform quadrant, fewer crossings of and less time spent in the previous platform location, and longer latencies to the previous platform location. These findings are similar to those seen in aged rats, suggesting that an increased release of reactive oxygen species may be responsible for the induction of radiation- and age-related cognitive deficits. If these decrements in behavior also occur in humans, they may impair the ability of astronauts to perform critical tasks during long-term space travel beyond the magnetosphere.

Analysis of Variance↗

PubMed1999

Effect of vitamin E intake on levels of vitamins E and C in the central nervous system and peripheral tissues: implications for health recommendations.

Vitamin E (alpha-gamma-tocopherol) is an important component in biological membranes. A decrease in its concentration imposes structural and functional damage to the cells. The object of this study was to assess the effect of a graded dietary vitamin E (E) intake on E concentration in specific regions of the brain, and its influence on vitamin C levels and neurological function. Following a 2-month period, rats supplemented with 5, 30, 60, 250 or 500 mg all-rac-alpha-tocopherol-acetate/kg diet (mg E/kg diet) exhibited a significant increase of E concentration in brain and peripheral tissues. However, while blood and liver showed a dose response increase in E concentration which correlated well with the different levels of E in the diet, the central nervous system (CNS) followed the same pattern of increase of vitamin E in brain tissue only when the diet was supplemented with 5, 30, or 60 mg E/kg diet. No further increase in E concentration was observed when the diet was supplemented with 250 or 500 mg E/kg diet. Similarly, the heart tissue showed a significant increase in its E concentration when the was enriched with 5, 30, or 60 mg E/kg diet, with no further increases at 250 or 500 mg. Vitamin C concentration in brain cortex and cerebellum, plasma, liver, and heart was reduced in the groups receiving 250 or 500 mg E/kg diet. Compared to the low E group, rats supplemented with the 60, 250 or 500 mg E/kg diet showed a significant enhancement in striatal dopamine (DA) release, but no differences were observed among the latter three groups.

Animals↗

PubMed1999

Reversals of age-related declines in neuronal signal transduction, cognitive, and motor behavioral deficits with blueberry, spinach, or strawberry dietary supplementation.

Ample research indicates that age-related neuronal-behavioral decrements are the result of oxidative stress that may be ameliorated by antioxidants. Our previous study had shown that rats given dietary supplements of fruit and vegetable extracts with high antioxidant activity for 8 months beginning at 6 months of age retarded age-related declines in neuronal and cognitive function. The present study showed that such supplements (strawberry, spinach, or blueberry at 14.8, 9.1, or 18.6 gm of dried aqueous extract per kilogram of diet, respectively) fed for 8 weeks to 19-month-old Fischer 344 rats were also effective in reversing age-related deficits in several neuronal and behavioral parameters including: oxotremorine enhancement of K(+)-evoked release of dopamine from striatal slices, carbachol-stimulated GTPase activity, striatal Ca(45) buffering in striatal synaptosomes, motor behavioral performance on the rod walking and accelerod tasks, and Morris water maze performance. These findings suggest that, in addition to their known beneficial effects on cancer and heart disease, phytochemicals present in antioxidant-rich foods may be beneficial in reversing the course of neuronal and behavioral aging.

Aging↗

PubMed1999

The effects of dietary antioxidants on psychomotor performance in aged mice.

Male C57BL/6NIA mice were provided one of six different antioxidant diets: vitamin E, glutathione, vitamin E plus glutathione, melatonin, strawberry extract, or control, beginning at 18 months of age. A battery of motor tests--rod walk, wire hang, plank walk, and inclined screen-was administered either: 1) before dietary treatment and then 6 months later at 24 months of age: or 2) only after 6 months of dietary treatment at age 24 months. An untreated group of 4-month-old mice served as young controls. Psychomotor performance was lower in 18-month-old mice compared with 4-month-old mice in the rod walk, wire hang, and inclined screen tests; however, no further decline was seen from 18 to 24 months on any measure. Chronic dietary antioxidant treatments were not effective in reversing age-related deficits in psychomotor behavior, except for the glutathione diet on inclined screen performance. It seems that motor performance deteriorates profoundly with age, because deficits at 18 months of age were as severe as they were at 24 months, and these age-associated motor deficits may be difficult to reverse, even with antioxidant treatment.

Aging↗

PubMed1999

Quantifying cellular oxidative stress by dichlorofluorescein assay using microplate reader.

Oxidative stress (OS) has been implicated in various degenerative diseases in aging. In an attempt to quantify OS in a cell model, we examined OS induced by incubating for 30 min with various free radical generators in PC12 cells by using the dichlorofluorescein (DCF) assay, modified for use by a fluorescent microplate reader. The nonfluorescent fluorescin derivatives (dichlorofluorescin, DCFH), after being oxidized by various oxidants, will become DCF and emit fluorescence. By quantifying the fluorescence, we were able to quantify the OS. Our results indicated that the fluorescence varied linearly with increasing concentrations (between 0.1 and 1 mM) of H2O2 and 2,2'-azobios(2-amidinopropane) dihydrochloride (AAPH; a peroxyl radical generator). By contrast, the fluorescence varied as a nonlinear response to increasing concentrations of 3-morpholinosydnonimine hydrochloride (SIN-1; a peroxynitrite generator), sodium nitroprusside (SNP; a nitric oxide generator), and dopamine. Dopamine had a biphasic effect; it decreased the DCF fluorescence, thus acting as an antioxidant, at concentrations <500 microM in cells, but acted as a pro-oxidant by increasing the fluorescence at 1 mM. While SNP was not a strong pro-oxidant, SIN-1 was the most potent pro-oxidant among those tested, inducing a 70 times increase of fluorescence at a concentration of 100 microM compared with control. Collectively, due to its indiscriminate nature to various free radicals, DCF can be very useful in quantifying overall OS in cells, especially when used in conjunction with a fluorescent microplate reader. This method is reliable and efficient for evaluating the potency of pro-oxidants and can be used to evaluate the efficacy of antioxidants against OS in cells.

Amidines↗

PubMed1999

Structure-activity relationships of quercetin in antagonizing hydrogen peroxide-induced calcium dysregulation in PC12 cells.

Oxidative stress can induce neurotoxic insults by increasing intracellular calcium (Ca2+), which has been implicated in various neurodegenerative diseases in aging. Previously, we showed that hydrogen peroxide induced calcium dysregulation in PC12 cells, as evidenced by (i) an increase in calcium baselines, (ii) a decrease in depolarization-induced calcium influx, and (iii) a failure to recover the Ca2+ levels. In the present experiments, we investigated whether a dietary flavonoid, quercetin, can antagonize the effects of hydrogen peroxide in the same cell model. We also investigated the possible structure-activity relationships of quercetin by comparing the results with four other flavonoids, each having a slightly different structure from quercetin. Our results indicated that two structural components, including (i) 3', 4'-hydroxyl (OH) groups in the B ring and (ii) a 2,3-double bond in conjugation with a 4-oxo group in the C ring, along with the polyphenolic structures were crucial for the protection. These structural components are found in quercetin, and this compound was also the most efficacious in reducing both the H2O2-induced Ca2+ dysregulation in cells and oxidative stress assessed via the dichlorofluorescein assay. Collectively, these data indicated that the particular polyphenolic structural components of quercetin provided its strong antioxidant property of protecting cells against H2O2-induced oxidative stress and calcium dysregulation.

Animals↗