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1.
Annu Int Conf IEEE Eng Med Biol Soc ; 2022: 629-632, 2022 07.
Article in English | MEDLINE | ID: mdl-36086013

ABSTRACT

Various studies have looked at the efficiency of artificial vessel and tissue networks in the study of photoplethysmography (PPG) in an effort to better understand the origin of various morphological features present in the signal. Whilst there are all reasonable attempts made to replicate geometrical features such as vessel depth, vessel wall thickness and diameter etc., not many studies have attempted to replicate the mechanical properties such as vessel elasticity and tissue compressibility. This study reports two methods for tissue mechanical testing for the analysis of vessel elasticity and tissue compressibility. A two-part polydimethylsiloxane (PDMS) was used as a base material for both tissue and vessel construction, and the properties altered by changing the curing component ratio. Tissue compression properties were investigated using an industrially calibrated materials testing device using the protocol from the ASTM 0575-91 testing method. Vessel elasticity was investigated using a custom method and apparatus to report vessel diameter and length change simultaneously. Tissue compressive properties proved reasonably easy to replicate through catalyst alteration, however the vessel elasticity properties were found to be higher than expected at all reasonable catalyst ratios. The property of hyper-elasticity was observed in the artificial vessels though, leading to the conclusion that alternative material recipes or construction methods may be needed to correctly replicate the expected mechanical characteristics. Clinical Relevance- The latest generation of health monitoring devices, especially those that are wearable and used widely by individuals wishing to monitor their health daily are becoming smarter and more sophisticated in their functionality. The majority of such devices use photoplethysmography (PPG) as their primary monitoring technique. Being able to replicate the PPG in a phantom allows the continued study and development of devices, and to improve their functionality without the continued need for extensive user-testing.


Subject(s)
Mechanical Tests , Photoplethysmography , Elasticity , Humans , Phantoms, Imaging , Photoplethysmography/methods
2.
Neurobiol Dis ; 158: 105486, 2021 10.
Article in English | MEDLINE | ID: mdl-34450329

ABSTRACT

Non-convulsive epileptiform activity is a common and under-studied comorbidity of Alzheimer's disease that may significantly contribute to onset of clinical symptoms independently of other neuropathological features such as ß-amyloid deposition. We used repeated treatment with low dose kainic acid (KA) to trigger sub-threshold epileptiform activity in young (less than 6 months) wild-type (WT) and APP/PSEN1 mice to test the role of disruption to the glutamatergic system in epileptiform activity changes and the development of memory deficits. Short-term repeated low-dose KA (five daily treatments with 5 mg/kg, IP) impaired long-term potentiation in hippocampus of APP/PSEN1 but not WT mice. Long-term repeated low-dose KA (fourteen weeks of bi-weekly treatment with 7.5-10 mg/kg) led to high mortality in APP/PSEN1 mice. KA treatment also impaired memory retention in the APP/PSEN1 mice in a Morris water maze task under cognitively challenging reversal learning conditions where the platform was moved to a new location. Four weeks of bi-weekly treatment with 5 mg/kg KA also increased abnormal spike activity in APP/PSEN1 and not WT mice but did not impact sleep/wake behavioral states. These findings suggest that hyperexcitability in Alzheimer's disease may indeed be an early contributor to cognitive decline that is independent of heavy ß-amyloid-plaque load, which is absent in APP/PSEN1 mice under 6 months of age.


Subject(s)
Amyloid beta-Protein Precursor/genetics , Cognitive Dysfunction/genetics , Cognitive Dysfunction/metabolism , Glutamic Acid/metabolism , Homeostasis/physiology , Presenilin-1/genetics , Animals , Electroencephalography , Epilepsy/chemically induced , Epilepsy/genetics , Female , Hippocampus/drug effects , Hippocampus/metabolism , Kainic Acid , Long-Term Potentiation , Maze Learning/drug effects , Memory Disorders/chemically induced , Memory Disorders/psychology , Mice , Mice, Inbred C57BL , Plaque, Amyloid/pathology
3.
Physiol Meas ; 35(10): 1961-73, 2014 Oct.
Article in English | MEDLINE | ID: mdl-25229803

ABSTRACT

Photoplethysmography (PPG) signals have been investigated at a new anatomical site, the anterior fontanelle (ANTF), on the hypothesis that blood supply at this location is preferentially preserved during cases of poor peripheral circulation which might cause the commercial pulse oximeters to fail to estimate accurately arterial blood oxygen saturation (SpO2). Two custom built reflectance PPG sensors have been developed, one for placement on the fontanelle and one on the periphery (foot). A PPG processing system and software were also developed to process the raw PPG signals and to estimate SpO2. A pilot study on sixteen babies, (9 male, 7 female) with a median age of 15.5 d (interquartile range = 46.8 d) and a median weight of 3.15 kg (SD = 0.93 kg), on a neonatal intensive care unit (NICU) has been carried out. PPG signals from the ANTF were of good quality and high signal-to-noise ratio. The amplitudes of the ANTF PPGs were found to be sensitive to changes in amplitude when amplitudes were observed at the reference PPG site. Bland-Altman analysis of the gold standard blood gas analysis reveals that all three sensors are inaccurate at SaO2 < 85-90 %, but the ANTF sensor shows better mean difference than the commercial device.


Subject(s)
Cranial Fontanelles/blood supply , Photoplethysmography/methods , Female , Humans , Infant, Newborn , Male , Oxygen/blood , Photoplethysmography/instrumentation , Pilot Projects , Signal Processing, Computer-Assisted
4.
Article in English | MEDLINE | ID: mdl-25570393

ABSTRACT

Lithium medication is the gold standard of treatment in Bipolar Disorder patients, preventing and reducing mood swings and suicidality. However, despite its effectiveness, it is a potentially hazardous drug requiring regular monitoring of blood levels to ensure toxic levels are not reached. This paper describes the first steps towards developing a new portable device that can be used by Bipolar Disorder patients to facilitate the analysis of lithium blood levels at home. Solutions of lithium carbonate have been optically fingerprinted using a high-end spectrophotometer. Preliminary measurements indicate that while the visible to near infrared region of the absorption spectra fall heavily within the water band, measurements in the Ultraviolet region show a strong distinction between different lithium concentrations. The optical spectra of Lithium in the 220 nm to 230 nm region demonstrated the ability to differentiate between concentrations representing those found in patients.


Subject(s)
Bipolar Disorder/blood , Lithium Carbonate/analysis , Lithium/blood , Optics and Photonics/methods , Optics and Photonics/statistics & numerical data , Absorption, Physicochemical , Calibration , Humans , Lithium Carbonate/chemistry , Water/chemistry
5.
Article in English | MEDLINE | ID: mdl-24110041

ABSTRACT

Pulse oximetry is being used in everyday clinical practice in anesthesia utilizing peripheral saturation sensors. However, it may be unreliable in certain clinical situations such as peripheral hypoperfusion. Similar situations occur in burns patients and more importantly burns to extremities which limit the sites available for measurement of peripheral oxygen saturation (SpO2). To overcome these limitations, the esophagus has been investigated as an alternative measurement site, as perfusion may be preferentially preserved centrally. A miniaturized reflectance esophageal saturation (SpO2 probe has been constructed utilizing infrared and red photodiodes and a photodetector. Our case study was aimed at evaluating the reliability of esophageal pulse oximetry in a major burns infant. Measurable photoplethysmographic (PPG) traces and SpO2 values were obtained in the neonatal esophagus. It was found that the esophageal pulse oximeter results were in good agreement with oxygen saturation measurements obtained by a commercial ear lobe pulse oximeter. This study suggests that the esophagus can be used as an alternative site for monitoring arterial blood oxygen saturation by pulse oximetry in burned infants.


Subject(s)
Burns/diagnosis , Burns/blood , Esophagus/physiology , Humans , Infant , Oximetry/methods , Oxygen/blood , Reproducibility of Results
6.
Article in English | MEDLINE | ID: mdl-24110273

ABSTRACT

There is a need for more reliable, non-invasive and alternative measurement sites for the monitoring of arterial blood oxygen saturation in critically ill newborns at times of peripheral compromise. A pilot investigation on 14 Intensive Care Unit (ICU) newborns was conducted utilizing custom-made reflectance photoplethysmographic (PPG) sensors placed at the fontanelle and foot. The results suggest that the fontanelle is sensitive to changes in saturation, where saturation values obtained from the custom sensor were compared against commercial pulse oximeter values and results from a blood gas analyzer, however careful placement of the sensor at the fontanelle is an issue that needs further investigation.


Subject(s)
Arteries/physiology , Cranial Fontanelles/physiology , Oxygen/blood , Photoplethysmography/methods , Signal Processing, Computer-Assisted , Female , Humans , Infant , Infant, Newborn , Infrared Rays , Male , Partial Pressure , Pilot Projects
7.
Article in English | MEDLINE | ID: mdl-23366216

ABSTRACT

There is a need for a more reliable, non-invasive and alternative measurement site for the monitoring of arterial blood oxygen saturation in critically ill neonates when peripheral perfusion is poor. The anterior fontanelle, a unique anatomical feature of the neonate, has been presented as an alternative site for the estimation of arterial blood oxygen saturation (SpO(2)). A new fontanelle photoplethysmographic sensor and processing system has been developed to investigate fontanelle photoplethysmographic (PPG) signals and estimate SpO(2) values at this anatomical location. Preliminary clinical trials have shown that good quality PPG signals with large amplitudes and high signal to noise ratio can be obtained from the neonatal fontanelle. The estimation of SpO(2) values from the fontanelle were in broad agreement with a commercial foot pulse oximeter.


Subject(s)
Cranial Fontanelles/metabolism , Oxygen/blood , Photoplethysmography/methods , Biosensing Techniques , Blood Gas Analysis , Foot , Humans , Infant, Newborn
8.
Article in English | MEDLINE | ID: mdl-22254240

ABSTRACT

There is a need for more reliable, non-invasive and alternative measurement sites for the monitoring of arterial blood oxygen saturation in critically ill newborns at times of peripheral compromise. The anterior fontanelle, a unique anatomical feature of the newborn, has been presented as an alternative site for the estimation of oxygen saturation. A multi-wavelength non-invasive optoelectronic sensor has been designed and developed for the investigation of photoplethysmographic (PPG) signals and blood oxygen saturation values from the fontanelle. In vivo thermal tests of the optical sensor show that under normal operating conditions the heating at the skin surface was negligible (<0.1 °C). Good quality PPGs with large amplitudes and high signal to noise ratio were recorded at all three (red, infrared and green) wavelengths prior to clinical measurements.


Subject(s)
Blood Volume Determination/instrumentation , Cranial Fontanelles/physiology , Optical Devices , Oximetry/instrumentation , Photoplethysmography/instrumentation , Transducers , Cranial Fontanelles/blood supply , Equipment Design , Equipment Failure Analysis , Humans , Infant, Newborn , Reproducibility of Results , Sensitivity and Specificity
9.
Free Radic Biol Med ; 49(5): 821-9, 2010 Sep 01.
Article in English | MEDLINE | ID: mdl-20541602

ABSTRACT

The sodium-dependent vitamin C transporter (SVCT2) is responsible for the transport of vitamin C into cells in multiple organs, from either the blood or the cerebrospinal fluid. Mice null for SVCT2 (SVCT2(-/-)) do not survive past birth but the cause of death has not yet been ascertained. After mating of SVCT2(+/-) males and SVCT2(+/-) females, fewer SVCT2(-/-) and SVCT2(+/-) progeny were observed than would be expected according to Mendelian ratios. Vitamin C levels in SVCT2(-/-), SVCT2(+/-), and SVCT2(+/+) were genotype-dependent. SVCT2(-/-) fetuses had significantly lower vitamin C levels than littermates in placenta, cortex, and lung, but not in liver (the site of vitamin C synthesis). Low vitamin C levels in placenta and cortex were associated with elevations in several markers of oxidative stress: malondialdehyde, isoketals, F(2)-isoprostanes, and F(4)-neuroprostanes. Oxidative stress was not elevated in fetal SVCT2(-/-) lung tissue despite low vitamin C levels. In addition to the expected severe hemorrhage in cortex, we also found hemorrhage in the brain stem, which was accompanied by cell loss. We found evidence of increased apoptosis in SVCT2(-/-) mice and disruption of the basement membrane in fetal brain. Together these data show that SVCT2 is critical for maintaining vitamin C levels in fetal and placental tissues and that the lack of SVCT2, and the resulting low vitamin C levels, results in fetal death and, in SVCT2(-/-) mice that survive the gestation period, in oxidative stress and cell death.


Subject(s)
Ascorbic Acid/blood , Organic Anion Transporters, Sodium-Dependent/genetics , Oxidative Stress/genetics , Symporters/genetics , Animals , Brain/embryology , Brain/metabolism , Brain/pathology , Cell Death/genetics , Down-Regulation/genetics , Female , Genotype , Male , Malondialdehyde/metabolism , Mice , Mice, Inbred C57BL , Mice, Transgenic , Organic Anion Transporters, Sodium-Dependent/metabolism , Organic Anion Transporters, Sodium-Dependent/physiology , Placenta/metabolism , Placenta/pathology , Pregnancy , Sodium-Coupled Vitamin C Transporters , Symporters/metabolism , Symporters/physiology , Tissue Distribution , Up-Regulation/genetics
10.
Pharmacol Biochem Behav ; 94(4): 543-52, 2010 Feb.
Article in English | MEDLINE | ID: mdl-19941887

ABSTRACT

Vitamin C is a powerful antioxidant and its levels are decreased in Alzheimer's patients. Even sub-clinical vitamin C deficiency could impact disease development. To investigate this principle we crossed APP/PSEN1 transgenic mice with Gulo knockout mice unable to synthesize their own vitamin C. Experimental mice were maintained from 6 weeks of age on standard (0.33 g/L) or reduced (0.099 g/L) levels of vitamin C and then assessed for changes in behavior and neuropathology. APP/PSEN1 mice showed impaired spatial learning in the Barnes maze and water maze that was not further impacted by vitamin C level. However, long-term decreased vitamin C levels led to hyperactivity in transgenic mice, with altered locomotor habituation and increased omission errors in the Barnes maze. Decreased vitamin C also led to increased oxidative stress. Transgenic mice were more susceptible to the activity-enhancing effects of scopolamine and low vitamin C attenuated these effects in both genotypes. These data indicate an interaction between the cholinergic system and vitamin C that could be important given the cholinergic degeneration associated with Alzheimer's disease.


Subject(s)
Amyloid beta-Protein Precursor/genetics , Ascorbic Acid Deficiency/genetics , Ascorbic Acid Deficiency/psychology , Presenilin-1/genetics , Amyloid/metabolism , Animals , Ascorbic Acid/metabolism , Ascorbic Acid Deficiency/mortality , Brain/metabolism , Brain/pathology , Exploratory Behavior/drug effects , Exploratory Behavior/physiology , Female , L-Gulonolactone Oxidase/genetics , Lipid Peroxidation , Male , Maze Learning/physiology , Mice , Mice, Transgenic , Oxidative Stress/physiology , Survival Rate
11.
Acta Anaesthesiol Scand ; 54(3): 385-8, 2010 Mar.
Article in English | MEDLINE | ID: mdl-19878099

ABSTRACT

A 72 year-old woman with cholangiocarcinoma presented for endoscopic retrograde cholangio pancreatography (ERCP) for diagnostic intraductal endoscopy under GETA. During the technically difficult procedure the patient became suddenly hypoxic, hypotensive, bradycardic, and progressed to PEA code (ETCO2 5 mmHg). ACLS was initiated. Transesophageal echo demonstrated massive right heart air accumulation; abdominal X-Ray showed air filled bile ducts. Central access was obtained, a pulmonary artery catheter floated, and 30 ml of air aspirated from the RV. Within 5 minutes pulses returned; the patient was transferred to the ICU. MRI revealed two watershed infarcts in the right frontal lobe. The patient fully recovered and returned a month later for an uneventful ERCP.


Subject(s)
Cholangiopancreatography, Endoscopic Retrograde/adverse effects , Embolism, Air/complications , Embolism, Air/etiology , Intraoperative Complications/etiology , Shock/etiology , Aged , Anesthesia, General , Carbon Dioxide/blood , Cardiopulmonary Resuscitation , Echocardiography, Transesophageal , Female , Heart Arrest/etiology , Humans , Hypotension/etiology , Hypoxia/etiology , Respiration, Artificial
12.
Nutr Neurosci ; 12(5): 203-18, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19761651

ABSTRACT

The present study investigated the relationships among oxidative stress, beta-amyloid and cognitive abilities in the APP/PSEN1 double-transgenic mouse model of Alzheimer's disease. In two experiments, long-term dietary supplements were given to aged APP/PSEN1 mice containing vitamin C alone (1 g/kg diet; Experiment 1) or in combination with a high (750 IU/kg diet, Experiments 1 and 2) or lower (400 IU/kg diet, Experiment 2) dose of vitamin E. Oxidative stress, measured by F(4)-neuroprostanes or malondialdehyde, was elevated in cortex of control-fed APP/PSEN1 mice and reduced to wild-type levels by vitamin supplementation. High-dose vitamin E with C was less effective at reducing oxidative stress than vitamin C alone or the low vitamin E+C diet combination. The high-dose combination also impaired water maze performance in mice of both genotypes. In Experiment 2, the lower vitamin E+C treatment attenuated spatial memory deficits in APP/PSEN1 mice and improved performance in wild-type mice in the water maze. Amyloid deposition was not reduced by antioxidant supplementation in either experiment.


Subject(s)
Alzheimer Disease/metabolism , Antioxidants/administration & dosage , Memory , Oxidative Stress , Aging , Amyloid beta-Protein Precursor/deficiency , Amyloid beta-Protein Precursor/genetics , Analysis of Variance , Animals , Antioxidants/adverse effects , Antioxidants/analysis , Ascorbic Acid/administration & dosage , Ascorbic Acid/adverse effects , Ascorbic Acid/analysis , Behavior, Animal/physiology , Brain Chemistry , Cognition , Diet , Disease Models, Animal , Female , Liver/chemistry , Male , Mice , Mice, Transgenic , Plaque, Amyloid/metabolism , Presenilin-1/deficiency , Presenilin-1/genetics , Presenilin-1/physiology , Vitamin E/administration & dosage , Vitamin E/adverse effects , Vitamin E/analysis
13.
Behav Brain Res ; 205(2): 550-8, 2009 Dec 28.
Article in English | MEDLINE | ID: mdl-19703495

ABSTRACT

Vitamin C (ascorbate) has important antioxidant functions that can help protect against oxidative stress in the brain and damage associated with neurodegenerative disorders such as Alzheimer's disease. When administered parenterally ascorbate can bypass saturable uptake mechanisms in the gut and thus higher tissue concentrations can be achieved than by oral administration. In the present study we show that ascorbate (125 mg/kg) administered intraperitoneally (i.p.) 1-h before testing, partially attenuated scopolamine-induced (1 mg/kg i.p.) cognitive deficits in Morris water maze performance in young mice. Cumulative search error, but not escape latency nor path length, was significantly improved during acquisition in ascorbate plus scopolamine-treated mice although performance did not equal that of control mice. During the probe trial, scopolamine led to increased search error and chance level of time spent in the platform quadrant, whereas mice pre-treated with ascorbate prior to scopolamine did not differ from control mice on these measures. Ascorbate had no effect on unimpaired, control mice and neither did it reduce the peripheral, activity-increasing effects of scopolamine. Ascorbate alone increased acetylcholinesterase activity in the medial forebrain area but had no effect in cortex or striatum. This change, and its action against the amnestic effects of the muscarinic antagonist scopolamine, suggest that ascorbate may be acting in part via altered cholinergic signaling. However, further investigation is necessary to isolate the cognition-enhancing effects of ascorbate.


Subject(s)
Ascorbic Acid/pharmacology , Learning Disabilities/drug therapy , Maze Learning/drug effects , Nootropic Agents/pharmacology , Space Perception/drug effects , Acetylcholinesterase/metabolism , Animals , Ascorbic Acid/blood , Ascorbic Acid/metabolism , Cerebral Cortex/drug effects , Cerebral Cortex/enzymology , Cerebral Cortex/metabolism , Corpus Striatum/drug effects , Corpus Striatum/enzymology , Corpus Striatum/metabolism , Escape Reaction/drug effects , Female , Learning Disabilities/chemically induced , Liver/drug effects , Liver/metabolism , Locomotion/drug effects , Male , Mice , Mice, Inbred Strains , Prosencephalon/drug effects , Prosencephalon/enzymology , Prosencephalon/metabolism , Scopolamine , Time Factors
14.
Pharmacol Biochem Behav ; 93(4): 443-50, 2009 Oct.
Article in English | MEDLINE | ID: mdl-19539642

ABSTRACT

Alzheimer's disease is a progressive and fatal neurodegenerative disease characterized by a build up of amyloid beta (Abeta) deposits, elevated oxidative stress, and deterioration of the cholinergic system. The present study investigated short-term cognitive-enhancing effects of acute intraperitoneal (i.p.) Vitamin C (ascorbate) treatment in APP/PSEN1 mice, a mouse model of Alzheimer's disease. Middle-aged (12 months) and very old (24 months) APP/PSEN1 bigenic and wild-type mice were treated with ascorbate (125 mg/kg i.p.) or the vehicle 1 h before testing on Y-maze spontaneous alternation and Morris water maze tasks. Very old mice performed more poorly on cognitive tasks than middle-aged mice. Ascorbate treatment improved Y-maze alternation rates and swim accuracy in the water maze in both wild-type and APP/PSEN1 mice. Abeta deposits and oxidative stress both increased with age, and acetylcholinesterase (AChE) activity was significantly reduced in APP/PSEN1 compared to wild-type mice. However, the short course of acute ascorbate treatment did not alter Alzheimer-like neuropathological features of plaque deposition, oxidative stress, or AChE activity. These data suggest that ascorbate may have noötropic functions when administered parenterally in high doses and that the mode of action is via an acute, pharmacological-like mechanism that likely modulates neurotransmitter function.


Subject(s)
Aging/psychology , Amyloid beta-Peptides/genetics , Antioxidants/pharmacology , Ascorbic Acid/pharmacology , Maze Learning/drug effects , Presenilin-1/genetics , Acetylcholinesterase/metabolism , Amyloid beta-Peptides/physiology , Animals , Brain Chemistry/drug effects , Malondialdehyde/metabolism , Maze Learning/physiology , Mice , Mice, Transgenic , Oxidative Stress/drug effects , Oxidative Stress/physiology , Presenilin-1/physiology
15.
FEBS Lett ; 508(3): 489-92, 2001 Nov 23.
Article in English | MEDLINE | ID: mdl-11728478

ABSTRACT

The selenoenzyme thioredoxin reductase (TR) can recycle ascorbic acid, which in turn can recycle alpha-tocopherol. Therefore, we evaluated the role of selenium in ascorbic acid recycling and in protection against oxidant-induced loss of alpha-tocopherol in cultured liver cells. Treatment of HepG2 or H4IIE cultured liver cells for 48 h with sodium selenite (0-116 nmol/l) tripled the activity of the selenoenzyme TR, measured as aurothioglucose-sensitive dehydroascorbic acid (DHA) reduction. However, selenium did not increase the ability of H4IIE cells to take up and reduce 2 mM DHA, despite a 25% increase in ascorbate-dependent ferricyanide reduction (which reflects cellular ascorbate recycling). Nonetheless, selenium supplements both spared ascorbate in overnight cultures of H4IIE cells, and prevented loss of cellular alpha-tocopherol in response to an oxidant stress induced by either ferricyanide or diazobenzene sulfonate. Whereas TR contributes little to ascorbate recycling in H4IIE cells, selenium spares ascorbate in culture and alpha-tocopherol in response to an oxidant stress.


Subject(s)
Ascorbic Acid/metabolism , Hepatocytes/metabolism , Oxidative Stress , Sodium Selenite/pharmacology , Thioredoxin-Disulfide Reductase/metabolism , alpha-Tocopherol/metabolism , Animals , Culture Media , Dehydroascorbic Acid/metabolism , Diazonium Compounds/pharmacology , Ferricyanides/metabolism , Ferricyanides/pharmacology , Hepatocytes/drug effects , Humans , Oxidation-Reduction , Oxidoreductases/metabolism , Rats , Selenocysteine/pharmacology , Sulfanilic Acids/pharmacology , Thioredoxins/pharmacology , Tumor Cells, Cultured
16.
Biochim Biophys Acta ; 1528(2-3): 159-66, 2001 Oct 03.
Article in English | MEDLINE | ID: mdl-11687303

ABSTRACT

Vitamin C, or ascorbic acid, is efficiently recycled from its oxidized forms by human erythrocytes. In this work the dependence of this recycling on reduced glutathione (GSH) was evaluated with regard to activation of the pentose cycle and to changes in pyridine nucleotide concentrations. The two-electron-oxidized form of ascorbic acid, dehydroascorbic acid (DHA) was rapidly taken up by erythrocytes and reduced to ascorbate, which reached intracellular concentrations as high as 2 mM. In the absence of D-glucose, DHA caused dose-dependent decreases in erythrocyte GSH, NADPH, and NADH concentrations. In the presence of 5 mM D-glucose, GSH and NADH concentrations were maintained, but those of NADPH decreased. Reduction of extracellular ferricyanide by erythrocytes, which reflects intracellular ascorbate recycling, was also enhanced by D-glucose, and ferricyanide activated the pentose cycle. Diethylmaleate at concentrations up to 1 mM was found to specifically deplete erythrocyte GSH by 75-90% without causing oxidant stress in the cells. Such GSH-depleted erythrocytes showed parallel decreases in their ability to take up and reduce DHA to ascorbate, and to reduce extracellular ferricyanide. These results show that DHA reduction involves GSH-dependent activation of D-glucose metabolism in the pentose cycle, but that in the absence of D-glucose DHA reduction can also utilize NADH.


Subject(s)
Ascorbic Acid/metabolism , Erythrocytes/metabolism , Dehydroascorbic Acid/pharmacology , Glucose/pharmacology , Glutathione , Humans , Maleates/pharmacology , NAD/metabolism , NADP/metabolism , Oxidation-Reduction , Pentose Phosphate Pathway , Substrate Cycling
17.
Biochem Pharmacol ; 62(7): 873-81, 2001 Oct 01.
Article in English | MEDLINE | ID: mdl-11543722

ABSTRACT

Ascorbic acid may be involved in the defense against oxidant stress in endothelial cells. Such a role requires that the cells effectively recycle the vitamin from its oxidized forms. In this work, we studied the ability of cultured bovine aortic endothelial cells (BAECs) to take up and reduce dehydroascorbic acid (DHA) to ascorbate, as well as the dependence of ascorbate recycling on intracellular GSH. BAECs took up and reduced DHA to ascorbate much more readily than they took up ascorbate. Although BAECs in culture did not contain ascorbate, ascorbate accumulated to concentrations of 2-3 mM in BAECs following incubation with 400 microM DHA. Extracellular ferricyanide oxidized intracellular ascorbate, which was recycled by the cells. Reduction of DHA, either when added to the cells or when generated in response to ferricyanide, caused significant decreases in intracellular GSH concentrations. Depletion of intracellular GSH with 1-chloro-2,4-dinitrobenzene, diethylmaleate, and diamide almost abolished the ability of the cells to reduce DHA to ascorbate. DHA reduction by thioredoxin reductase was evident in dialyzed cell extracts, but occurred at rates far lower than direct GSH reduction of DHA. These results suggest that maximal rates of DHA reduction, and thus recycling of ascorbate from DHA, are dependent upon GSH in these cells.


Subject(s)
Ascorbic Acid/metabolism , Endothelium, Vascular/metabolism , Glutathione/metabolism , Animals , Biological Transport , Cattle , Dehydroascorbic Acid/metabolism , Humans
18.
Am J Physiol Endocrinol Metab ; 281(1): E72-80, 2001 Jul.
Article in English | MEDLINE | ID: mdl-11404224

ABSTRACT

To use primary cultures of human skeletal muscle cells to establish defects in glucose metabolism that underlie clinical insulin resistance, it is necessary to define the rate-determining steps in glucose metabolism and to improve the insulin response attained in previous studies. We modified experimental conditions to achieve an insulin effect on 3-O-methylglucose transport that was more than twofold over basal. Glucose phosphorylation by hexokinase limits glucose metabolism in these cells, because the apparent Michaelis-Menten constant of coupled glucose transport and phosphorylation is intermediate between that of transport and that of the hexokinase and because rates of 2-deoxyglucose uptake and phosphorylation are less than those of glucose. The latter reflects a preference of hexokinase for glucose over 2-deoxyglucose. Cellular NAD(P)H autofluorescence, measured using two-photon excitation microscopy, is both sensitive to insulin and indicative of additional distal control steps in glucose metabolism. Whereas the predominant effect of insulin in human skeletal muscle cells is to enhance glucose transport, phosphorylation, and steps beyond, it also determines the overall rate of glucose metabolism.


Subject(s)
Glucose/metabolism , Muscle, Skeletal/metabolism , 3-O-Methylglucose/metabolism , Adult , Aged , Biological Transport, Active , Cell Differentiation , Cells, Cultured , Female , Hexokinase/metabolism , Humans , Hypoglycemic Agents/pharmacology , Insulin/pharmacology , Kinetics , Male , Microscopy, Fluorescence , Middle Aged , Muscle, Skeletal/cytology , NADP/metabolism , Phosphorylation , Pyruvic Acid/pharmacology , Stimulation, Chemical
19.
Free Radic Biol Med ; 31(1): 117-24, 2001 Jul 01.
Article in English | MEDLINE | ID: mdl-11425497

ABSTRACT

Reduction of the ascorbate free radical (AFR) at the plasma membrane provides an efficient mechanism to preserve the vitamin in a location where it can recycle alpha-tocopherol and thus prevent lipid peroxidation. Erythrocyte ghost membranes have been shown to oxidize NADH in the presence of the AFR. We report that this activity derives from an AFR reductase because it spares ascorbate from oxidation by ascorbate oxidase, and because ghost membranes decrease steady-state concentrations of the AFR in a protein- and NADH-dependent manner. The AFR reductase has a high apparent affinity for both NADH and the AFR (< 2 microM). When measured in open ghosts, the reductase is comprised of an inner membrane activity (both substrate sites on the cytosolic membrane face) and a trans-membrane activity that mediates extracellular AFR reduction using intracellular NADH. However, the trans-membrane activity constitutes only about 12% of the total measured in ghosts. Ghost AFR reductase activity can also be differentiated from NADH-dependent ferricyanide reductase(s) by its sensitivity to the detergent Triton X-100 and insensitivity to enzymatic digestion with cathepsin D. This NADH-dependent AFR reductase could serve to recycle ascorbic acid at a crucial site on the inner face of the plasma membrane.


Subject(s)
Ascorbic Acid/metabolism , Erythrocyte Membrane/metabolism , Electron Spin Resonance Spectroscopy , Ferricyanides , Free Radicals , Humans , In Vitro Techniques , NAD/metabolism , NADH, NADPH Oxidoreductases/metabolism , Oxidation-Reduction
20.
J Nutr ; 131(6): 1798-802, 2001 Jun.
Article in English | MEDLINE | ID: mdl-11385070

ABSTRACT

Selenium and vitamin E deficiencies were studied as part of an evaluation of oxidant defenses in guinea pigs. Male guinea pigs (100-120 g) were fed a control diet (C) or the diet without selenium (0 Se), without vitamin E (0 E), or without either selenium or vitamin E (0 Se-0 E). Between d 30 and 35, 7 of 13 guinea pigs fed the 0 Se-0 E diet were euthanized because of severe weakness of their extremities. No guinea pigs in the other diet groups developed weakness. Guinea pigs from each group were killed on d 37. Selenium deficiency and vitamin E deficiency were verified by measurement of glutathione peroxidase and alpha-tocopherol. Creatine phophokinase (CPK) activity was greater than controls in both groups fed vitamin E-deficient diets, but the increase was greater in the 0 Se-0 E group than in the 0 E group. Muscle F(2)-isoprostanes were greater than controls in both groups fed vitamin E-deficient diets with the level in the 0 Se-0 E group greater than that in the 0 E group. Histologic muscle necrosis was severe in the 0 Se-0 E group, minimal in the 0 E group and absent from other groups. The diets used in this study induced selenium and vitamin E deficiencies in guinea pigs. The study demonstrates that combined selenium and vitamin E deficiency results in a fatal myopathy in guinea pigs that is associated with lipid peroxidation in the affected muscle. This nutritional myopathy is much more severe than the myopathy that occurs with vitamin E deficiency alone.


Subject(s)
Muscle, Skeletal/physiopathology , Muscular Diseases/etiology , Selenium/deficiency , Vitamin E Deficiency/metabolism , Animals , Body Weight , Creatine Kinase/analysis , Creatine Kinase/blood , Diet , Dinoprost/analogs & derivatives , Dinoprost/analysis , F2-Isoprostanes , Guinea Pigs , Liver/metabolism , Male , Muscular Diseases/blood , Muscular Diseases/physiopathology , Necrosis , Survival Analysis , Vitamin E/analysis , Vitamin E/blood , Vitamin E Deficiency/blood
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