CiteULike: Tag caffeine

Relationship between Caffeine-Induced Changes in Resting Cerebral Perfusion and Blood Oxygenation Level-Dependent Signal

Paul Laurienti — 2006-10-02T14:59:24-00:00

AJNR Am J Neuroradiol, Vol. 24, No. 8. (1 September 2003), pp. 1607-1611.

BACKGROUND AND PURPOSE: Recent interest has emerged in the use of pharmacologic methods to maximize blood oxygenation level-dependent (BOLD) signal intensity changes in functional MR imaging (fMRI). Adenosine antagonists, such as caffeine and theophylline, have been identified as potential agents for this purpose. The present study was designed to determine whether caffeine-induced decreases in cerebral perfusion result in enhanced BOLD responses to visual and auditory stimuli. METHODS: MR imaging was used to measure resting cerebral perfusion and stimulus-induced BOLD signal intensity changes in 19 patients. We evaluated the relationship between resting cerebral perfusion and the magnitude of BOLD signal intensity induced by visual and auditory stimulation under caffeine and placebo conditions. RESULTS: The data showed that changes in resting cerebral perfusion produced by caffeine are not a consistent predictor of BOLD signal intensity magnitude. Although all cerebral perfusion was reduced in all study participants in response to caffeine, only 47% of the participants experienced BOLD signal intensity increase. This finding was independent of the participants' usual caffeine consumption. CONCLUSION: The data presented herein show that the relationship between resting cerebral perfusion and the magnitude of BOLD signal intensity is complex. It is not possible to consistently enhance BOLD signal intensity magnitude by decreasing resting perfusion with caffeine. Future studies aimed at evaluating the relationship between perfusion and BOLD signal intensity changes should seek a means to selectively modulate known components of the neural and vascular responses independently.

Dopaminergic effects of caffeine in the human striatum and thalamus.

V Kaasinen — 2007-09-21T20:00:50-00:00

Neuroreport, Vol. 15, No. 2. (9 February 2004), pp. 281-285.

Epidemiological studies have provided evidence that caffeine, an adenosine receptor antagonist, reduces the risk for Parkinson's disease. There are indications of specific interactions between striatal adenosine A(2A) and dopamine D(2) receptors, but the in vivo effects of caffeine on human dopamine system have not been investigated. In the present study, the dopaminergic effects of caffeine were examined with [(11)C]raclopride positron emission tomography (PET) in eight healthy habitual coffee drinkers after 24 h caffeine abstinence. Compared to oral placebo, 200 mg oral caffeine induced a 12% decrease in midline thalamic binding potential (p < 0.001). A trend-level increase in ventral striatal [(11)C]raclopride binding potential was seen with a correlation between caffeine-related arousal and putaminal dopamine D(2) receptor binding (r = -0.81, p = 0.03). The findings indicate that caffeine has effects on dopaminergic neurotransmission in the human brain, which may be differential in the striatum and the thalamus.

Dopamine: the salient issue.

MA Ungless — 2005-03-14T18:35:22-00:00

Trends Neurosci, Vol. 27, No. 12. (December 2004), pp. 702-706.

There is general agreement that midbrain dopamine neurons play key roles in reward processing. What is more controversial is the role they play in processing salient stimuli that are not rewarding. This controversy has arisen for three main reasons. First, salient sensory stimuli such as tones and lights, which are assumed not to be rewarding, increase dopamine neuron activity. Second, aversive stimuli increase firing in a minority of putative dopamine neurons. Third, dopamine release is increased following aversive stimuli. Consequently, it has been suggested that these midbrain dopamine neurons are activated by all salient stimuli, rather than specifically by rewards. However, reconsideration of these issues, in light of new findings, suggests this controversy can be resolved in favour of reward theories.

Distinguishing Whether Dopamine Regulates Liking, Wanting, and/or Learning About Rewards.

S Robinson — 2005-03-14T18:23:04-00:00

Behav Neurosci, Vol. 119, No. 1. (February 2005), pp. 5-15.

To determine whether dopamine regulates liking, wanting, and/or learning about rewards during goal-directed behavior, the authors tested genetically engineered dopamine-deficient (DD) mice for acquisition of an appetitive T-maze task with and without endogenous dopamine signaling. Experiment 1 established that DD mice treated with L-dihydroxyphenylalanine (L-dopa [LD]) perform similarly to controls on a T-maze task designed to measure liking, wanting, and learning about rewards. Experiment 2, which tested saline-, caffeine-, and LD-treated DD mice on the T maze, separated performance factors from cognitive processes and revealed that dopamine is not necessary for mice to like or learn about rewards but is necessary for mice to seek (want) rewards during goal-directed behavior. ((c) 2005 APA, all rights reserved).

Espresso reward learning, hold the dopamine: theoretical comment on robinson et Al. (2005).

KC Berridge — 2005-03-14T18:22:15-00:00

Behav Neurosci, Vol. 119, No. 1. (February 2005), pp. 336-341.

Question: Is dopamine needed for reward learning? Answer: No--at least, not in the brain of a caffeinated dopamine-deficient (DD) mutant mouse. That is the conclusion of an important paper in this issue by S. Robinson, S. M. Sandstrom, V. H. Denenberg, and R. D. Palmiter (see record 2005-01705-001). Those authors demonstrate that reward learning can proceed normally in the brains of DD mice, even though they contain no dopamine at the time of learning, if the mice are given caffeine just before learning. Caffeine activates the DD mice by a nondopaminergic mechanism, allowing them to learn where to obtain food reward in a T-maze runway. Their reward-learning-without-dopamine is revealed on a subsequent test day, when dopamine function is restored by L-dopa administration. Robinson et al. conclude that dopamine is not needed for normal learning about rewards, or for hedonic "liking" of rewards during learning, but rather specifically for a motivational "wanting" component of reward, such as incentive salience. ((c) 2005 APA, all rights reserved).

The appetite-suppressant effect of nicotine is enhanced by caffeine

A Jessen — 2005-06-14T18:59:21-00:00

Diabetes, Obesity and Metabolism, Vol. 7, No. 4. (July 2005), pp. 327-333.

A2A adenosine receptor protects tumors from antitumor T cells.

A Ohta — 2006-09-22T05:44:58-00:00

Proc Natl Acad Sci U S A, Vol. 103, No. 35. (29 August 2006), pp. 13132-13137.

The A2A adenosine receptor (A2AR) has been shown to be a critical and nonredundant negative regulator of immune cells in protecting normal tissues from inflammatory damage. We hypothesized that A2AR also protects cancerous tissues by inhibiting incoming antitumor T lymphocytes. Here we confirm this hypothesis by showing that genetic deletion of A2AR in the host resulted in rejection of established immunogenic tumors in approximately 60% of A2AR-deficient mice with no rejection observed in control WT mice. The use of antagonists, including caffeine, or targeting the A2 receptors by siRNA pretreatment of T cells improved the inhibition of tumor growth, destruction of metastases, and prevention of neovascularization by antitumor T cells. The data suggest that effects of A2AR are T cell autonomous. The inhibition of antitumor T cells via their A2AR in the adenosine-rich tumor microenvironment may explain the paradoxical coexistence of tumors and antitumor immune cells in some cancer patients (the "Hellstrom paradox"). We propose to target the hypoxia-->adenosine-->A2AR pathway as a cancer immunotherapy strategy to prevent the inhibition of antitumor T cells in the tumor microenvironment. The same strategy may prevent the premature termination of immune response and improve the vaccine-induced development of antitumor and antiviral T cells. The observations of autoimmunity during melanoma rejection in A2AR-deficient mice suggest that A2AR in T cells is also important in preventing autoimmunity. Thus, although using the hypoxia-->adenosine-->A2AR pathway inhibitors may improve antitumor immunity, the recruitment of this pathway by selective drugs is expected to attenuate the autoimmune tissue damage.

Stimulus functions of caffeine in humans: relation to dependence potential

SJ Heishman — 2005-10-24T18:31:44-00:00

Neuroscience And Biobehavioral Reviews, Vol. 16, No. 3. ( 1992), pp. 273-287.

The interoceptive stimulus functions common to drugs of dependence include positive subjective effects, discriminative functions, and reinforcing functions. Data from studies measuring these stimulus functions constitute the objective assessment of a drug's dependence potential. This paper reviews the subjective effects, discriminative stimulus, and reinforcing stimulus functions of caffeine in humans to assess the dependence potential of caffeine. The stimulus effects of caffeine are compared with those of d-amphetamine, a prototypic CNS stimulant that has been studied under similar conditions, to evaluate the relative dependence potential of caffeine. Finally, caffeine's effects are evaluated in terms of generally accepted criteria for defining drug dependence. It is concluded that caffeine partially meets the primary criteria of drug dependence: 1) the majority of caffeine use is highly controlled, but not compulsive; 2) caffeine is psychoactive; and 3) caffeine functions as a reinforcer under certain conditions in humans, but not in animals. Caffeine thus has limited dependence potential. Additionally, although caffeine shares stimulus functions with d-amphetamine, it does so under limited conditions and should be considered to have a relatively lower dependence potential. [Journal Article, Review; 181 Refs; In English; United States]

Regulation of G proteins and adenylyl cyclase in brain regions of caffeine-tolerant and -dependent mice

KA Leite-Morris — 2005-10-24T18:16:35-00:00

Brain Research, Vol. 804, No. 1. (31 August 1998), pp. 52-62.

Regulation of post-receptor signaling provides a mechanism of adaptation to chronic psychotropic drug treatment. In this study, the regulation of guanine nucleotide binding proteins (G proteins) and G protein-stimulated adenylyl cyclase activity was examined in brain regions of caffeine-tolerant and -dependent mice. Chronic caffeine doses were administered via mini-osmotic pumps over 7 days at 0, 42, 85 and 125 mg kg-1 day-1. These chronic caffeine doses were linearly correlated with plasma caffeine concentrations. In behavioral studies, the stimulant effects of acute caffeine on motor activity were significantly diminished in a dose-dependent manner after chronic caffeine, suggesting the development of tolerance. Abrupt discontinuation of chronic caffeine treatment (at 85 and 125 mg kg-1 day-1) produced a dose-dependent and reversible reduction in motor activity 24 h later, suggestive of a caffeine withdrawal syndrome. Utilizing quantitative immunoblotting methods, we found that hippocampal Gi1,2 and Gi3 subunits were significantly reduced by 20.2% and 11.1%, respectively, in caffeine tolerant/dependent mice (caffeine 125 mg kg-1 day-1 vs. vehicle controls). Decreases in inhibitory G protein subunit concentrations in hippocampus were accompanied by a significant increase (by 21%) in hippocampal G protein function, as measured by guanine nucleotide-stimulated adenylyl cyclase activity, in caffeine-treated mice. This same caffeine treatment also produced significant decreases in cortical Gs subunits of 14.0%. Since short-term caffeine treatment has been shown to reduce adenylyl cyclase activity, chronic caffeine treatment could produce adaptive increases in G protein-stimulated adenylyl cyclase to oppose this effect via G protein regulation.

Role of adenosine receptors in caffeine tolerance

SG Holtzman — 2005-10-24T18:37:24-00:00

The Journal Of Pharmacology And Experimental Therapeutics, Vol. 256, No. 1. (January 1991), pp. 62-68.

Caffeine is a competitive antagonist at adenosine receptors. Receptor up-regulation during chronic drug treatment has been proposed to be the mechanism of tolerance to the behavioral stimulant effects of caffeine. This study reassessed the role of adenosine receptors in caffeine tolerance. Separate groups of rats were given scheduled access to drinking bottles containing plain tap water or a 0.1% solution of caffeine. Daily drug intake averaged 60-75 mg/kg and resulted in complete tolerance to caffeine-induced stimulation of locomotor activity, which could not be surmounted by increasing the dose of caffeine. 5'-N-ethylcarboxamidoadenosine (0.001-1.0 mg/kg) dose dependently decreased the locomotor activity of caffeine-tolerant rats and their water-treated controls but was 8-fold more potent in the latter group. Caffeine (1.0-10 mg/kg) injected concurrently with 5-N-ethylcarboxamidoadenosine antagonized the decreases in locomotor activity comparably in both groups. Apparent pA2 values for tolerant and control rats also were comparable: 5.05 and 5.11. Thus, the adenosine-antagonist activity of caffeine was undiminished in tolerant rats. The effects of chronic caffeine administration on parameters of adenosine receptor binding and function were measured in cerebral cortex. There were no differences between brain tissue from control and caffeine-treated rats in number and affinity of adenosine binding sites or in receptor-mediated increases (A2 adenosine receptor) and decreases (A1 adenosine receptor) in cAMP accumulation. These results are consistent with theoretical arguments that changes in receptor density should not affect the potency of a competitive antagonist. Experimental evidence and theoretical considerations indicate that up-regulation of adenosine receptors is not the mechanism of tolerance to caffeine-induced stimulation of locomotor activity. [Journal Article; In English; United States]

Caffeine therapy for apnea of prematurity.

B Schmidt — 2007-09-15T17:03:07-00:00

N Engl J Med, Vol. 354, No. 20. (18 May 2006), pp. 2112-2121.

BACKGROUND: Methylxanthines reduce the frequency of apnea of prematurity and the need for mechanical ventilation during the first seven days of therapy. It is uncertain whether methylxanthines have other short- and long-term benefits or risks in infants with very low birth weight. METHODS: We randomly assigned 2006 infants with birth weights of 500 to 1250 g during the first 10 days of life to receive either caffeine or placebo, until drug therapy for apnea of prematurity was no longer needed. We evaluated the short-term outcomes before the first discharge home. RESULTS: Of 963 infants who were assigned to caffeine and who remained alive at a postmenstrual age of 36 weeks, 350 (36 percent) received supplemental oxygen, as did 447 of the 954 infants (47 percent) assigned to placebo (adjusted odds ratio, 0.63; 95 percent confidence interval, 0.52 to 0.76; P<0.001). Positive airway pressure was discontinued one week earlier in the infants assigned to caffeine (median postmenstrual age, 31.0 weeks; interquartile range, 29.4 to 33.0) than in the infants in the placebo group (median postmenstrual age, 32.0 weeks; interquartile range, 30.3 to 34.0; P<0.001). Caffeine reduced weight gain temporarily. The mean difference in weight gain between the group receiving caffeine and the group receiving placebo was greatest after two weeks (mean difference, -23 g; 95 percent confidence interval, -32 to -13; P<0.001). The rates of death, ultrasonographic signs of brain injury, and necrotizing enterocolitis did not differ significantly between the two groups. CONCLUSIONS: Caffeine therapy for apnea of prematurity reduces the rate of bronchopulmonary dysplasia in infants with very low birth weight. (ClinicalTrials.gov number, NCT00182312.).

Long-Term Effects of Caffeine Therapy for Apnea of Prematurity

Barbara Schmidt — 2008-04-10T20:13:34-00:00

N Engl J Med, Vol. 357, No. 19. (8 November 2007), pp. 1893-1902.

Background Methylxanthine therapy is commonly used for apnea of prematurity but in the absence of adequate data on its efficacy and safety. It is uncertain whether methylxanthines have long-term effects on neurodevelopment and growth. Methods We randomly assigned 2006 infants with birth weights of 500 to 1250 g to receive either caffeine or placebo until therapy for apnea of prematurity was no longer needed. The primary outcome was a composite of death, cerebral palsy, cognitive delay (defined as a Mental Development Index score of <85 on the Bayley Scales of Infant Development), deafness, or blindness at a corrected age of 18 to 21 months. Results Of the 937 infants assigned to caffeine for whom adequate data on the primary outcome were available, 377 (40.2%) died or survived with a neurodevelopmental disability, as compared with 431 of the 932 infants (46.2%) assigned to placebo for whom adequate data on the primary outcome were available (odds ratio adjusted for center, 0.77; 95% confidence interval [CI], 0.64 to 0.93; P=0.008). Treatment with caffeine as compared with placebo reduced the incidence of cerebral palsy (4.4% vs. 7.3%; adjusted odds ratio, 0.58; 95% CI, 0.39 to 0.87; P=0.009) and of cognitive delay (33.8% vs. 38.3%; adjusted odds ratio, 0.81; 95% CI, 0.66 to 0.99; P=0.04). The rates of death, deafness, and blindness and the mean percentiles for height, weight, and head circumference at follow-up did not differ significantly between the two groups. Conclusions Caffeine therapy for apnea of prematurity improves the rate of survival without neurodevelopmental disability at 18 to 21 months in infants with very low birth weight. (ClinicalTrials.gov number, NCT00182312 .) 10.1056/NEJMoa073679

Coffee, tea, colas, and risk of epithelial ovarian cancer.

YJ Song — 2008-04-10T20:12:35-00:00

Cancer epidemiology, biomarkers & prevention : a publication of the American Association for Cancer Research, cosponsored by the American Society of Preventive Oncology, Vol. 17, No. 3. (March 2008), pp. 712-716.

Associations of coffee, tea, and other caffeinated beverages with ovarian cancer risk remain uncertain. In a population-based study in Washington State, 781 women with epithelial ovarian cancer diagnosed in 2002 to 2005 and 1,263 controls completed self-administered questionnaires detailing consumption of caffeinated and noncaffeinated coffee, teas, and colas and in-person interviews regarding reproductive and hormonal exposures. We assessed risk associated with coffee, tea, and cola drinking and with total caffeine consumption using logistic regression to calculate odds ratios and 95% confidence intervals. Neither caffeinated nor decaffeinated coffees were associated with ovarian cancer risk; also, we observed no association of total caffeine with risk using a combined index that summed intake from coffee, tea, and carbonated soft drinks. Among teas, neither herbal/decaffeinated nor black teas were associated with risk; however, women who reported drinking >/=1 cup/d of green tea had a 54% reduction in risk (P(trend) = 0.01). Associations of green tea with risk were similar when invasive and borderline cases were considered separately and when Asian women were excluded from analysis. Green tea, which is commonly consumed in countries with low ovarian cancer incidence, should be further investigated for its cancer prevention properties. (Cancer Epidemiol Biomarkers Prev 2008;17(3):712-6).

Effect of green tea and (-)-epigallocatechin gallate on ethanol-induced toxicity in HepG2 cells.

Sang Il Lee — 2008-04-10T20:12:05-00:00

Phytotherapy research : PTR (18 March 2008)

Despite the continuing reports supporting the hepatoprotective effects of green tea against ethanol intoxication, there remain controversies regarding the active compound(s) and molecular mechanism. These issues were addressed in the present study using cultured HepG2 cells exposed to a lethal dose of ethanol. Gamma-glutamyl transferase (GGT) was chosen as a marker of ethanol toxicity because it is widely used in clinics. When the cells were treated with ethanol at various concentrations, there was a dose-dependent increase of GGT activity in the culture media and loss of cell viability. Pretreatment of the cells with green tea extract attenuated the changes significantly. Among the green tea constituents, (-)-epigallocatechin gallate (EGCG) attenuated the ethanol cytotoxicity effectively, whereas l-theanine and caffeine had no effects. The ethanol cytotoxicity was also attenuated by alcohol dehydrogenase inhibitor 4-methyl pyrazol and GGT inhibitor acivicin as well as by thiol modulators such as S-adenosyl-l-methionine, N-acetyl-l-cysteine and glutathione. EGCG failed to prevent the intracellular glutathione loss caused by ethanol, but it appeared to be a strong GGT inhibitor. Therefore the cytoprotective effects of green tea could be attributed to the inhibition of GGT activity by EGCG. This study suggests that GGT inhibitors including EGCG may provide a novel strategy for attenuating ethanol-induced liver damage. Copyright (c) 2008 John Wiley & Sons, Ltd.

Caffeine blocks disruption of blood brain barrier in a rabbit model of Alzheimer's disease

Xuesong Chen — 2008-04-03T15:41:42-00:00

Journal of Neuroinflammation, Vol. 5, No. 1. (2008)

High levels of serum cholesterol and disruptions of the blood brain barrier (BBB) have all been implicated as underlying mechanisms in the pathogenesis of Alzheimer's disease. Results from studies conducted in animals and humans suggest that caffeine might be protective against Alzheimer's disease but by poorly understood mechanisms. Using rabbits fed a cholesterol-enriched diet, we tested our hypothesis that chronic ingestion of caffeine protects against high cholesterol diet-induced disruptions of the BBB. New Zealand rabbits were fed a 2 cholesterol-enriched diet, and 3 mg caffeine was administered daily in drinking water for 12 weeks. Total cholesterol and caffeine concentrations from blood were measured. Olfactory bulbs (and for some studies hippocampus and cerebral cortex as well) were evaluated for BBB leakage, BBB tight junction protein expression levels, activation of astrocytes, and microglia density using histological, immunostaining and immunoblotting techniques. We found that caffeine blocked high cholesterol diet-induced increases in extravasation of IgG and fibrinogen, increases in leakage of Evan's blue dye, decreases in levels of the tight junction proteins occludin and ZO-1, increases in astrocytes activation and microglia density where IgG extravasation was present. Chronic ingestion of caffeine protects against high cholesterol diet-induced increases in disruptions of the BBB, and caffeine and drugs similar to caffeine might be useful in the treatment of Alzheimer's disease.

Effects of green tea fractions on oxygen-induced retinal neovascularization in the neonatal rat.

Y Saito — 2008-04-10T20:10:26-00:00

Journal of clinical biochemistry and nutrition, Vol. 41, No. 1. (July 2007), pp. 43-49.

This study aimed to investigate the preventive effects of green tea fractions (GTFs) on rat model of oxygen-induced retinopathy (OIR). Neonatal Sprague-Dawley rats were exposed to daily cycles of 80% oxygen (20.5 h), ambient air (0.5 h), and progressive return to 80% oxygen (3 h) until postnatal day 12 (P12), then the rats were placed in ambient air until P18. The green tea was fractionated by DM-A50, DM-W, M-B, and M-W. The rats were treated once daily from P6 to P17 by gastric gavage of GTFs (0.05 or 0.01 g/ml) or distilled water (DW) at 50 microl/10 g body weight. On P18, the rats were sacrificed and the retinal samples were collected. The retinal neovascularization (NV) was scored and avascular areas (AVAs) were measured as a % of total retinal area (%AVAs) in ADPase stained retinas. The NV scores in 0.01 g/ml M-W were significantly lower than those in DW. The %AVAs in 0.05 g/ml DM-A50 and in 0.05 g/ml and 0.01 g/ml M-W were significantly lower than those in DW. There were less catechins, and less caffeine in M-W fraction compared with other GTFs, suggesting components of green tea except for catechins and caffeine might suppress the neovascularization in rat model of OIR.

Caffeine prevents age-associated recognition memory decline and changes brain-derived neurotrophic factor and tirosine kinase receptor (TrkB) content in mice.

M S Costa — 2008-04-30T21:15:57-00:00

Neuroscience (22 March 2008)

The beneficial effects of caffeine on cognition are controversial in humans, whereas its benefit in rodents had been well characterized. However, most studies were performed with acute administration of caffeine and the tasks used to evaluate cognition had aversive components. Here, we evaluated adulthood administration of caffeine up to old age on recognition memory in mice using the object recognition task (ORT) and on brain-derived neurotrophic factor (BNDF) and tirosine kinase receptor (TrkB) immunocontent in the hippocampus. Adult mice (6 months old) received either drinking water or caffeine (1 mg/mL) during 12 months. At 18 months of age both groups were tested for ORT. Our results showed that aged mice exhibited lower performance in the recognition memory compared with adults (6 months old). Furthermore, caffeine-treated mice showed similar performance to adult mice in the ORT and an improvement compared with their age-matched control mice. Caffeine also counteracted the age-related increase in BDNF and TrkB immunocontent. Our results corroborate with other studies and reinforce that caffeine consumed in adulthood may prevent recognition memory decline with aging. This preventive effect may involve a decrease in the hippocampal BDNF and TrkB immunocontent.

Caffeine Ingestion Is Associated With Reductions in Glucose Uptake Independent of Obesity and Type 2 Diabetes Before and After Exercise Training

Sojung Lee — 2008-07-11T16:28:32-00:00

Diabetes Care, Vol. 28, No. 3. (1 March 2005), pp. 566-572.

OBJECTIVE--We investigated the effect of caffeine ingestion on insulin sensitivity in sedentary lean men (n = 8) and obese men with (n = 7) and without (n = 8) type 2 diabetes. We also examined whether chronic exercise influences the relationship between caffeine and insulin sensitivity in these individuals. RESEARCH DESIGN AND METHODS--Subjects underwent two hyperinsulinemic-euglycemic clamp procedures, caffeine (5 mg/kg body wt) and placebo, in a double-blind, randomized manner before and after a 3-month aerobic exercise program. Body composition was measured by magnetic resonance imaging. RESULTS--At baseline, caffeine ingestion was associated with a significant reduction (P < 0.05) in insulin sensitivity by a similar magnitude in the lean (33%), obese (33%), and type 2 diabetic (37%) groups in comparison with placebo. After exercise training, caffeine ingestion was still associated with a reduction (P < 0.05) in insulin sensitivity by a similar magnitude in the lean (23%), obese (26%), and type 2 diabetic (36%) groups in comparison with placebo. Exercise was not associated with a significant increase in insulin sensitivity in either the caffeine or placebo trials, independent of group (P > 0.10). CONCLUSIONS--Caffeine consumption is associated with a substantial reduction in insulin-mediated glucose uptake independent of obesity, type 2 diabetes, and chronic exercise. 10.2337/diacare.28.3.566

Caffeine Impairs Glucose Metabolism in Type 2 Diabetes

James Lane — 2008-07-11T16:29:54-00:00

Diabetes Care, Vol. 27, No. 8. (1 August 2004), pp. 2047-2048.

10.2337/diacare.27.8.2047

Relation of caffeine intake and blood caffeine concentrations during pregnancy to fetal growth: prospective population based study.

DG Cook — 2007-03-23T16:19:45-00:00

BMJ, Vol. 313, No. 7069. (30 November 1996), pp. 1358-1362.

OBJECTIVES: To examine the association of plasma caffeine concentrations during pregnancy with fetal growth and to compare this with relations with reported caffeine intake. DESIGN: Prospective population based study. SETTING: District general hospital, inner London. SUBJECTS: Women booking for delivery between 1982 and 1984. Stored plasma was available for 1,500 women who had provided a blood sample on at least one occasion and for 640 women who had provided a sample on all three occasions (at booking, 28 weeks, and 36 weeks). MAIN OUTCOME MEASURE: Birth weight adjusted for gestational age, maternal height, parity, and sex of infant. The exposures of interest were reported caffeine consumption and blood caffeine concentration. Cigarette smoking was assessed by blood cotinine concentration. RESULTS: Caffeine intake showed no changes during pregnancy, but blood caffeine concentrations rose by 75%. Although caffeine intake increased steadily with increasing cotinine concentration above 15 ng/ml, blood caffeine concentrations fell. Caffeine consumption was inversely related to adjusted birth weight, the estimated effect being a 1.3% fall in birth weight for a 1,000 mg per week increase in intake (95% confidence interval 0.5% to 2.1%). The apparent caffeine effect was confined to cigarette smokers, among whom the estimated effect was-1.6%/1000 mg a week (-2.9% to -0.2%) after adjustment for cotinine and -1.3% (-2.7% to 0.1%) after further adjustment for social class and alcohol intake. Adjusted birth weight was unrelated to blood caffeine concentrations overall (P = 0.09, but a positive coefficient), after adjustment for cotinine (P = 0.73), or among current smokers (P = 0.45). CONCLUSIONS: Smokers consume more caffeine than non-smokers. Blood caffeine concentrations during pregnancy are not related to fetal growth, but caffeine intake is negatively associated with birth weight, with this effect being apparent only in smokers. The effect remains of borderline significance after adjustment for other factors. Prudent advice for pregnant women would be to reduce caffeine intake in conjunction with stopping smoking.

Metabolic and hormonal effects of caffeine: randomized, double-blind, placebo-controlled crossover trial.

T Mackenzie — 2007-12-02T23:16:04-00:00

Metabolism, Vol. 56, No. 12. (December 2007), pp. 1694-1698.

In short-term studies, caffeine has been shown to increase insulin levels, reduce insulin sensitivity, and increase cortisol levels. However, epidemiological studies have indicated that long-term consumption of beverages containing caffeine such as coffee and green tea is associated with a reduced risk of type 2 diabetes mellitus. There is a paucity of randomized studies addressing the metabolic and hormonal effects of consuming caffeine over periods of more than 1 day. We evaluated the effect of oral intake of 200 mg of caffeine taken twice a day for 7 days on glucose metabolism, as well as on serum cortisol, dehydroepiandrosterone (DHEA), and androstenedione, and on nighttime salivary melatonin. A double-blind, randomized, placebo-controlled crossover study with periods of 7 days and washouts of 5 days comparing caffeine with placebo capsules was conducted. Participants were 16 healthy adults aged 18 to 22 years with a history of caffeine consumption. Blood samples from each subject were assayed for glucose, insulin, serum cortisol, DHEA, and androstenedione on the eighth day of each period after an overnight fast. Nighttime salivary melatonin was also measured. Insulin levels were significantly higher (by 1.80 muU/mL; 95% confidence interval, 0.33-3.28) after caffeine intake than after placebo. The homeostasis model assessment index of insulin sensitivity was reduced by 35% (95% confidence interval, 7%-62%) by caffeine. There were no differences in glucose, DHEA, androstenedione, and melatonin between treatment periods. This study provides evidence that daily caffeine intake reduces insulin sensitivity; the effect persists for at least a week and is evident up to 12 hours after administration.

Influence of Caffeine and other Methylxanthines on Mechanical Properties of Isolated Mammalian Heart Muscle: EVIDENCE FOR A DUAL MECHANISM OF ACTION

John Blinks — 2007-03-31T08:52:41-00:00

Circ Res, Vol. 30, No. 4. (1 April 1972), pp. 367-392.

Caffeine, theophylline, and theobromine have similar effects on the contractions of kitten atrial and papillary muscle preparations in vitro. In concentrations between 2 and 20 mM they both intensify and prolong the active state, as indicated by isometric and delayed-release isotonic contractions; contracture is not normally produced. Instantaneous force-velocity curves are shifted approximately symmetrically by caffeine; force-velocity curves derived from simple afterloaded contractions are misleading because of the great prolongation of activity. After the addition of caffeine the onset of the increased degree of activation is more rapid than that of the prolongation of activity; procaine antagonizes the prolongation of activity but not the intensification. In the presence of the methylxanthines, the duration of contraction is no longer abbreviated by isoproterenol, though it is still readily influenced by changes in frequency. The prolongation of activity by Sr2^plus; differs in significant respects from that induced by methylxanthines. The results suggest that the methylxanthines exert two effects on excitation-contraction coupling. One of these is presumed to be the inhibition of calcium sequestration by the sarcoplasmic reticulum; the other may be an effect on the cell membrane that leads to increased calcium entry. Most of the features of the altered mechanical response can be explained on this basis if it is assumed that intracellular calcium stores available for release are depleted as a result of the process that impairs calcium sequestration.

Ryanodine and caffeine prevent ventricular arrhythmias during acute myocardial ischemia and reperfusion in rat heart.

FT Thandroyen — 2007-03-31T08:31:48-00:00

Circ Res, Vol. 62, No. 2. (February 1988), pp. 306-314.

This study investigates the possible role of oscillatory release of calcium from sarcoplasmic reticulum in the genesis of ventricular arrhythmias during acute myocardial ischemia and reperfusion in isolated rat hearts. We used ryanodine and caffeine, which are known to modulate the oscillatory release of calcium from sarcoplasmic reticulum. During 30 minutes of left main coronary artery ligation, all 13 control hearts developed ventricular premature beats (number of beats, 225 +/- 51) and ventricular tachycardia (duration, 123 +/- 21 seconds); five hearts developed ventricular fibrillation. In a separate series of experiments, reperfusion after 15 minutes of coronary artery ligation caused ventricular fibrillation to occur within 15 seconds in all 12 hearts. Ryanodine (10(-9) to 10(-7) M) abolished ventricular arrhythmias during coronary artery ligation and prevented reperfusion ventricular fibrillation. Ryanodine (10(-9), 10(-8), and 10(-7) M) caused 15%, 23%, and 74% decreases in the maximal rate of rise of left ventricular pressure development and 20%, 32%, and 85% decreases in the maximal rate of fall of left ventricular pressure development, respectively, prior to coronary artery ligation. During acute myocardial ischemia, ryanodine 10(-9) M maintained and 10(-8) M impaired left ventricular function; 10(-7) M caused left ventricular failure. Coronary perfusion rate did not increase during ischemia. Antiarrhythmic activity occurred independent of preservation of high energy phosphates, reduction in tissue lactate, or tissue cyclic adenosine monophosphate in the ischemic myocardium. Caffeine 10(2) M decreased the incidence of ventricular arrhythmias during ischemia and upon reperfusion; protection occurred coincident with development of diastolic contracture. Caffeine increased ischemic tissue cyclic adenosine monophosphate content and worsened tissue energy status.(ABSTRACT TRUNCATED AT 250 WORDS)

The relationship between caffeine contracture of intact muscle and the effect of caffeine on reticulum.

A Weber — 2007-03-31T08:31:03-00:00

J Gen Physiol, Vol. 52, No. 5. (November 1968), pp. 750-759.

Fast Release of Calcium from Sarcoplasmic Reticulum Vesicles Monitored by Chlortetracycline Fluorescence

Kazuo Nagasaki — 2007-03-31T08:10:22-00:00

J Biochem (Tokyo), Vol. 94, No. 4. (1 October 1983), pp. 1101-1109.

Rapid Ca2+ release rate from sarcoplasmic reticulum vesicles was determined by the stopped flow method in terms of chlortetracycline fluorescence. Intensity of chlortetracycline fluorescence was proportional to the intravesicular free. Ca2+ concentration. Ca2+ efflux was activated by extravesicular Ca2+ with an apparent dissociation constant of 25 microM and was inhibited with an inhibition constant of 120 microM in the absence of Mg2+. Caffeine enhanced the Ca2+ release rate by increasing only the affinity of Ca2+ for the activation site. Mg2+ reduced the Ca2+ release rate by competitive binding to the activation site. ATP increased the Ca2+ release rate very much without changing the affinities of Ca2+ for the activation and inhibition sites, i.e., ATP seems to increase the pore radius or number of the Ca2+ channels without affecting the gating mechanism of the channel. These results are consistent with those reported in skinned muscle sarcoplasmic reticulum. The maximum rate of Ca2+ release in the presence of ATP reached 80 s-1. This value is considered to be sufficient to cause muscular contraction.

Activation of the S phase DNA damage checkpoint by mitomycin C.

E Mladenov — 2007-08-25T17:45:30-00:00

J Cell Physiol, Vol. 211, No. 2. (May 2007), pp. 468-476.

We have studied the rate of DNA synthesis, cell cycle distribution, formation of gamma-H2AX, and Rad51 nuclear foci and association of Rad51 with the nuclear matrix after treatment of HeLa cells with the interstrand crosslinking agent mitomycin C (MMC) in the presence of the kinase inhibitors caffeine and wortmannin. The results showed that MMC treatment arrested the cells in S-phase and induced the appearance of gamma-H2AX and Rad51 nuclear foci, accompanied with a sequestering of Rad51 to the nuclear matrix. These effects were abrogated by caffeine, which inhibits the Ataxia-telangiectasia mutated (ATM) and ATM- and Rad3-related (ATR) kinases. However, wortmannin at a concentration that inhibits ATM, but not ATR did not affect cell cycle progression, damage-induced phosphorylation of H2AX and Rad51 foci formation, and association with the nuclear matrix, suggesting that the S-phase arrest induced by MMC is ATR-dependent. These findings were confirmed by experiments with ATR-deficient and AT cells. They indicate that the DNA damage ATR-dependent S-phase checkpoint pathway may regulate the spatiotemporal organization of the process of repair of interstrand crosslinks.

Caffeine Reduces the Activation Extent and Contrast-to-Noise Ratio of the Functional Cerebral Blood Flow Response but not the BOLD Response

Joy Liau — 2008-05-26T16:40:11-00:00

NeuroImage, Vol. In Press, Accepted Manuscript

Effects of Coffee Bean Aroma on the Rat Brain Stressed by Sleep Deprivation: A Selected Transcript- and 2D Gel-Based Proteome Analysis

Han-Seok Seo — 2008-06-13T13:58:01-00:00

J. Agric. Food Chem. (3 June 2008)

Abstract: The aim of this study was 2-fold: (i) to demonstrate influences of roasted coffee bean aroma on rat brain functions by using the transcriptomics and proteomics approaches and (ii) to evaluate the impact of roasted coffee bean aroma on stress induced by sleep deprivation. The aroma of the roasted coffee beans was administered to four groups of adult male Wistar rats: 1, control group; 2, 24 h sleep deprivation-induced stress group (the stress group); 3, coffee aroma-exposed group without stress (the coffee group); and 4, the stress with coffee aroma group (the stress with coffee group). Reverse transcriptase-polymerase chain reaction (RT-PCR) analysis of some known genes responsive to aroma or stress was performed using total RNA from these four groups. A total of 17 selected genes of the coffee were differently expressed over the control. Additionally, the expression levels of 13 genes were different between the stress group and the stress with coffee group: Up-regulation was found for 11 genes, and down-regulation was seen for two genes in the stress with coffee group. We also looked to changes in protein profiles in these four samples using two-dimensional (2D) gel electrophoresis; 25 differently expressed gel spots were detected on 2D gels stained by silver nitrate. Out of these, a total of nine proteins were identified by mass spectrometry. Identified proteins belonged to five functional categories: antioxidant; protein fate; cell rescue, defense, and virulence; cellular communication/signal transduction mechanism; and energy metabolism. Among the differentially expressed genes and proteins between the stress and the stress with coffee group, NGFR, trkC, GIR, thiol-specific antioxidant protein, and heat shock 70 kDa protein 5 are known to have antioxidant or antistress functions. In conclusion, the roasted coffee bean aroma changes the mRNA and protein expression levels of the rat brain, providing for the first time clues to the potential antioxidant or stress relaxation activities of the coffee bean aroma. Keywords: Brain; coffee aroma; proteomic analysis; sleep deprivation; transcriptomics analysis.

Inhibition of ATM and ATR kinase activities by the radiosensitizing agent, caffeine.

JN Sarkaria — 2006-08-09T21:48:09-00:00

Cancer Res, Vol. 59, No. 17. (1 September 1999), pp. 4375-4382.

Caffeine exposure sensitizes tumor cells to ionizing radiation and other genotoxic agents. The radiosensitizing effects of caffeine are associated with the disruption of multiple DNA damage-responsive cell cycle checkpoints. The similarity of these checkpoint defects to those seen in ataxia-telangiectasia (A-T) suggested that caffeine might inhibit one or more components in an A-T mutated (ATM)-dependent checkpoint pathway in DNA-damaged cells. We now show that caffeine inhibits the catalytic activity of both ATM and the related kinase, ATM and Rad3-related (ATR), at drug concentrations similar to those that induce radiosensitization. Moreover, like ATM-deficient cells, caffeine-treated A549 lung carcinoma cells irradiated in G2 fail to arrest progression into mitosis, and S-phase-irradiated cells exhibit radioresistant DNA synthesis. Similar concentrations of caffeine also inhibit gamma- and UV radiation-induced phosphorylation of p53 on Ser15, a modification that may be directly mediated by the ATM and ATR kinases. DNA-dependent protein kinase, another ATM-related protein involved in DNA damage repair, was resistant to the inhibitory effects of caffeine. Likewise, the catalytic activity of the G2 checkpoint kinase, hChk1, was only marginally suppressed by caffeine but was inhibited potently by the structurally distinct radiosensitizer, UCN-01. These data suggest that the radiosensitizing effects of caffeine are related to inhibition of the protein kinase activities of ATM and ATR and that both proteins are relevant targets for the development of novel anticancer agents.

Metabolic and physiological effects of ingesting extracts of bitter orange, green tea and guarana at rest and during treadmill walking in overweight males

C Sale — 2006-01-18T16:32:12-00:00

International Journal of Obesity, Vol. aop, No. current.

The effects of caffeine and expectancy on attention and memory.

A Oei — 2006-02-04T16:12:16-00:00

Hum Psychopharmacol, Vol. 20, No. 3. (April 2005), pp. 193-202.

The present study contrasted caffeine's effects on individuals who expect caffeine to stimulate them and those who do not. Secondly, whether a message that caffeine rather than placebo was administered would also affect these two groups of subjects differently was investigated. The study was conducted single-blind in a 2x2x2 mixed design. The between subjects factor was whether they expected caffeine to stimulate them (E+) or not (E-) according to their self reports obtained before the experiment began. The within subjects factors were message (told caffeine vs told placebo) and beverage type (given caffeine vs placebo). Sixteen subjects in each group (n=32) performed on signal detection, memory scanning and delayed free recall tasks following ingestion of either caffeinated or decaffeinated coffee on two sessions each, a total of four experimental sessions. On each session, subjects were given a message regarding their drink (told caffeine vs told placebo). However, on two sessions there was a mismatch between the message and drink given. For signal detection, performance under caffeine was better than placebo in the E+ but not the E- group. However, subjects in the E+ group did not benefit more than the E- group in either message condition. On memory scanning, detections and false alarms did not differ for either beverage, nor was there a differential finding in the E+ and E- groups. However, reaction time under caffeine condition was shorter. No effects of message were found. Caffeine and message also did not have any effect on performance on the delayed free recall task. The hypothesis that caffeine and message would affect E+ and E- subjects differentially was partly supported.

Regulation of calcium release by calcium inside the sarcoplasmic reticulum in ventricular myocytes

Valeriy Lukyanenko — 2008-07-01T01:55:01-00:00

Pflügers Archiv European Journal of Physiology, Vol. 432, No. 6. (1996), pp. 1047-1054.

Abstract To study the effects of changes in sarcoplasmic reticulum (SR) intraluminal Ca2+ on the Ca2+ release mechanism, we correlated the activity of single cardiac ryanodine receptor (RyR) channels, monitored in planar bilayers, with the properties of spontaneous elementary Ca2+ release events (sparks) in intact ventricular myocytes, monitored by scanning confocal microfluorimetry. Under both normal conditions and Ca2+ overload, induced by elevation of extracellular [Ca2+], Ca2+ sparks represented single populations of events. During Ca2+ overload, the frequency of sparks increased from 0.8 to 3.1 events per second per 100 μm line scanned, and their amplitude increased from 100 nM to 400 nM. The duration of the Ca2+ sparks, however, was not altered. Changes in the properties of Ca2+ sparks were accompanied by only an ≈ 30% increase in the SR Ca2+ content, as determined by emptying the intracellular Ca2+ stores using caffeine. When single Ca2+ release channels were incorporated into lipid bilayers and activated by cytoplasmic Ca2+ (≈ 100 nM) and ATP (3 mM), elevation of Ca2+ on the luminal side from 20 μM to 0.2–20 mM resulted in a 1.2-fold to 7-fold increase, respectively, in open probability (P o). This potentiation of P o was due to an increase in mean open time and frequency of events. The relative effect of luminal Ca2+ was greater at low levels of cytoplasmic [Ca2+] than at high levels of cytoplasmic [Ca2+], and no effect of luminal Ca2+ was observed to occur in channels activated by 0.5–50 μM cytoplasmic Ca2+ in the absence of ATP. Our results suggest that SR Ca2+ release channels are modulated by SR intraluminal Ca2+. These alterations in properties of release channels may account for, or contribute to, the mechanism of spontaneous Ca2+ release in cardiac myocytes

Calcium handling by sarcoplasmic reticulum of neonatal swine cardiac myocytes

CM Hohl — 2008-06-11T23:47:31-00:00

Am J Physiol Heart Circ Physiol, Vol. 273, No. 1. (1 July 1997), pp. H192-199.

Dynamic regulation of sarcoplasmic reticulum Ca(2+) content and release by luminal Ca(2+)-sensitive leak in rat ventricular myocytes.

V Lukyanenko — 2007-05-08T00:42:27-00:00

Biophys J, Vol. 81, No. 2. (August 2001), pp. 785-798.

In cardiac muscle, excitation-contraction (E-C) coupling is determined by the ability of the sarcoplasmic reticulum (SR) to store and release Ca(2+). It has been hypothesized that the Ca(2+) sequestration and release mechanisms might be functionally linked to optimize the E-C coupling process. To explore the relationships between the loading status of the SR and functional state of the Ca(2+) release mechanism, we examined the effects of changes in SR Ca(2+) content on spontaneous Ca(2+) sparks in saponin-permeabilized and patch-clamped rat ventricular myocytes. SR Ca(2+) content was manipulated by pharmacologically altering the capacities of either Ca(2+) uptake or leak. Ca(2+) sparks were recorded using a confocal microscope and Fluo-3 and were quantified considering missed events. SR Ca(2+) content was assessed by application of caffeine. Exposure of permeabilized cells to anti-phospholamban antibodies elevated the SR Ca(2+) content and increased the frequency of sparks. Suppression of the SR Ca(2+) pump by thapsigargin lowered [Ca(2+)](SR) and reduced the frequency of sparks. The ryanodine receptor (RyR) blockers tetracaine and Mg(2+) transiently suppressed the frequency of sparks. Upon washout of the drugs, sparking activity transiently overshot control levels. Low doses of caffeine transiently potentiated sparking activity upon application and transiently depressed the sparks upon removal. In patch-clamped cardiac myocytes, exposure to caffeine produced only a transient increase in the probability of sparks induced by depolarization. We interpret these results in terms of a novel dynamic control scheme for SR Ca(2+) cycling. A central element of this scheme is a luminal Ca(2+) sensor that links the functional activity of RyRs to the loading state of the SR, allowing cells to auto-regulate the size and functional state of their SR Ca(2+) pool. These results are important for understanding the regulation of intracellular Ca(2+) release and contractility in cardiac muscle.

The effects of thapsigargin on [Ca2+]i in isolated rat mesenteric artery vascular smooth muscle cells

I Baró — 2008-05-29T22:11:35-00:00

Pflügers Archiv European Journal of Physiology, Vol. 420, No. 1. (1 January 1992), pp. 115-117.

The effects of thapsigargin were studied on single cells isolated from side branches of the rat mesenteric artery. Thapsigargin (150 nM) produced a transient increase of [Ca2+]i. This transient rise of [Ca2+]i was unaffected by removing external Ca2+ ions. This suggests that thapsigargin is releasing Ca2+ ions from an intracellular store. In the absence of thapsigargin both noradrenaline and caffeine also produced a transient increase of [Ca2+]i. These increases were abolished by prior exposure to thapsigargin. Correspondingly, the effects of thapsigargin were abolished by prior exposure to caffeine. These results show that thapsigargin releases Ca2+ from the noradrenaline and caffeine-sensitive stores.

Modulation of cytosolic and intra-sarcoplasmic reticulum calcium waves by calsequestrin in rat cardiac myocytes

Zuzana Kubalova — 2008-06-13T03:30:23-00:00

J Physiol, Vol. 561, No. 2. (1 December 2004), pp. 515-524.

Waves of Ca2+-induced Ca2+ release occur in various cell types and are involved in the pathology of certain forms of cardiac arrhythmia. These arrhythmias include catecholaminergic polymorphic ventricular tachycardia (CPVT), certain cases of which are associated with mutations in the cardiac calsequestrin gene (CASQ2). To explore the mechanisms of Ca2+ wave generation and unravel the underlying causes of CPVT, we investigated the effects of adenoviral-mediated changes in CASQ2 protein levels on the properties of cytosolic and sarcoplasmic reticulum (SR) Ca2+ waves in permeabilized rat ventricular myocytes. The free [Ca2+] inside the sarcoplasmic reticulum ([Ca2+]SR) was monitored by fluo-5N entrapped into the SR, and cytosolic Ca2+ was imaged using fluo-3. Overexpression of CASQ2 resulted in significant increases in the amplitude of Ca2+ waves and interwave intervals, whereas reduced CASQ2 levels caused drastic reductions in the amplitude and period of Ca2+ waves. CASQ2 abundance had no impact on resting diastolic [Ca2+]SR or on the amplitude of the [Ca2+]SR depletion signal during the Ca2+ wave. However, the recovery dynamics of [Ca2+]SR following Ca2+ release were dramatically altered as the rate of [Ca2+]SR recovery increased [~]3-fold in CASQ2-overexpressing myocytes and decreased to 30% of control in CASQ2-underexpressing myocytes. There was a direct linear relationship between Ca2+ wave period and the half-time of basal [Ca2+]SR recovery following Ca2+ release. Loading the SR with the low affinity exogenous Ca2+ buffer citrate exerted effects quantitatively similar to those observed on overexpressing CASQ2. We conclude that free intra-SR [Ca2+] is a critical determinant of cardiac Ca2+ wave generation. Our data indicate that reduced intra-SR Ca2+ binding activity promotes the generation of Ca2+ waves by accelerating the dynamics of attaining a threshold free [Ca2+]SR required for Ca2+ wave initiation, potentially accounting for arrythmogenesis in CPVT linked to mutations in CASQ2. 10.1113/jphysiol.2004.073940

Modulation of focal and global Ca2+ release in calsequestrin-overexpressing mouse cardiomyocytes

Wei Wang — 2008-07-15T03:42:53-00:00

J Physiol, Vol. 524, No. 2. (15 April 2000), pp. 399-414.

Potentiation of fractional sarcoplasmic reticulum calcium release by total and free intra-sarcoplasmic reticulum calcium concentration.

TR Shannon — 2007-04-29T05:12:55-00:00

Biophys J, Vol. 78, No. 1. (January 2000), pp. 334-343.

Our aim was to measure the influence of sarcoplasmic reticulum (SR) calcium content ([Ca](SRT)) and free SR [Ca] ([Ca](SR)) on the fraction of SR calcium released during voltage clamp steps in isolated rabbit ventricular myocytes. [Ca](SRT), as measured by caffeine application, was progressively increased by conditioning pulses. Sodium was absent in both the intracellular and in the extracellular solutions to block sodium/calcium exchange. Total cytosolic calcium flux during the transient was inferred from I(Ca), [Ca](SRT), [Ca](i), and cellular buffering characteristics. Fluxes via the calcium current (I(Ca)), the SR calcium pump, and passive leak from the SR were evaluated to determine SR calcium release flux (J(rel)). Excitation-contraction (EC) coupling was characterized with respect to both gain (integral J(rel)/integral I(Ca)) and fractional SR calcium release. Both parameters were virtually zero for a small, but measurable [Ca](SRT). Gain and fractional SR calcium release increased steeply and nonlinearly with both [Ca](SRT) and [Ca](SR). We conclude that potentiation of EC coupling can be correlated with both [Ca](SRT) and [Ca](SR). While fractional SR calcium release was not linearly dependent upon [Ca](SR), intra-SR calcium may play a crucial role in regulating the SR calcium release process.

Changes of intracellular [Ca2+] during refilling of sarcoplasmic reticulum in rat ventricular and vascular smooth muscle.

I Baro — 2008-06-09T03:03:24-00:00

J Physiol, Vol. 465, No. 1. (1 June 1993), pp. 21-41.

1. Intracellular calcium concentration ([Ca2+]i) was measured in single myocytes isolated from either the cardiac ventricle or the mesenteric artery of the rat. 2. In both cardiac and smooth muscle, the application of caffeine produced an increase of [Ca2+]i which spontaneously decayed back to resting levels. In vascular smooth muscle cells, removal of caffeine produced a transient fall of [Ca2+]i to below the resting level. [Ca2+]i then returned to control levels. A transient undershoot of [Ca2+]i on removal of caffeine was also sometimes seen in cardiac cells. When the undershoot was absent in cardiac cells it could be induced by elevating [Ca2+]o. 3. In vascular smooth muscle cells noradrenaline increased [Ca2+]i and an undershoot of [Ca2+]i could be produced by its removal. In cardiac cells a small undershoot could sometimes be seen following the systolic Ca2+ transient produced by electrical stimulation. 4. In both cardiac and vascular cells the time constant of decay of the caffeine response (tau caff) was less than that of the recovery from the undershoot (tau us). On average the ratio tau us:tau caff was about 5 in smooth muscle. In cardiac cells the recovery of the undershoot was also considerably slower than that of the caffeine response. 5. If caffeine was removed before the rise of [Ca2+]i had fully decayed spontaneously then the magnitude of the undershoot was reduced. 6. It is suggested that the undershoot of [Ca2+]i on removal of caffeine results from refilling of the SR decreasing [Ca2+]i. The data from vascular cells can be fitted by this model if the dissociation constant, Kd, of the surface membrane Ca2+ pump for [Ca2+]i is about 1 microM. 7. Using the model, it is concluded from the ratio of the time constants shown above that the caffeine releasable content of the sarcoplasmic reticulum constitutes about 80% of total cellular calcium in both cardiac and smooth muscle.

Ca2+ Scraps: Local Depletions of Free [Ca2+] in Cardiac Sarcoplasmic Reticulum During Contractions Leave Substantial Ca2+ Reserve

Thomas Shannon — 2008-07-06T20:44:16-00:00

Circ Res, Vol. 93, No. 1. (11 July 2003), pp. 40-45.

Free [Ca2+] inside the sarcoplasmic reticulum ([Ca2+]SR) is difficult to measure yet critically important in controlling many cellular systems. In cardiac myocytes, [Ca2+]SR regulates cardiac contractility. We directly measure [Ca2+]SR in intact cardiac myocytes dynamically and quantitatively during beats, with high spatial resolution. Diastolic [Ca2+]SR (1 to 1.5 mmol/L) is only partially depleted (24% to 63%) during contraction. There is little temporal delay in the decline in [Ca2+]SR at release junctions and between junctions, indicating rapid internal diffusion. The incomplete local Ca2+ release shows that the inherently positive feedback of Ca2+-induced Ca2+ release terminates, despite a large residual driving force. These findings place stringent novel constraints on how excitation-contraction coupling works in heart and also reveal a Ca2+ store reserve that could in principle be a therapeutic target to enhance cardiac function in heart failure. 10.1161/01.RES.0000079967.11815.19

Single cardiac sarcoplasmic reticulum Ca2+-release channel: activation by caffeine

E Rousseau — 2008-07-23T03:59:56-00:00

Am J Physiol Heart Circ Physiol, Vol. 256, No. 2. (1 February 1989), pp. H328-333.

An estimate of the calcium content of the sarcoplasmic reticulum in rat ventricular myocytes

A Varro — 2008-06-11T23:27:28-00:00

Pflügers Archiv European Journal of Physiology, Vol. 423, No. 1. (1 April 1993), pp. 158-160.

The aim of this paper was to estimate the Ca content of the sarcoplasmic reticulum (s.r.) and to compare this with the amount of Ca which enters the cell via the calcium current in systole. The s.r. Ca content was measured electrophysiologically in voltage-clamped rat ventricular myocytes. Rapid application of caffeine produced a transient increase of [Ca2+]i which was accompanied by a transient inward Na-Ca exchange current. The integral of this current gives a measure of the Ca2+ pumped out of the cell by Na-Ca exchange. Ni2+ (5 mM) inhibited the current and decreased the rate of fall of [Ca2+]i to 32% of the control suggesting that Na-Ca exchange is responsible for 68% of Ca removal from the cytoplasm following the addition of caffeine. Correcting for the Na-Ca independent Ca removal suggests that the s.r. Ca content is equivalent to about 120 µmol per litre cell. Furthermore we estimate that, during systole, Ca entry into the cell via the sarcolemmal calcium current is equal to about 6 % of the Ca content of the s.r.

Modulation of CICR has no maintained effect on systolic Ca2+: simultaneous measurements of sarcoplasmic reticulum and sarcolemmal Ca2+ fluxes in rat ventricular myocytes

AW Trafford — 2008-06-09T00:55:27-00:00

J Physiol, Vol. 522, No. 2. (15 January 2000), pp. 259-270.

Luminal Ca2+ Controls Termination and Refractory Behavior of Ca2+-Induced Ca2+ Release in Cardiac Myocytes

Dmitry Terentyev — 2008-07-11T03:26:37-00:00

Circ Res, Vol. 91, No. 5. (6 September 2002), pp. 414-420.

Despite extensive research, the mechanisms responsible for the graded nature and early termination of Ca2+-induced Ca2+ release (CICR) from the sarcoplasmic reticulum (SR) in cardiac muscle remain poorly understood. Suggested mechanisms include cytosolic Ca2+-dependent inactivation/adaptation and luminal Ca2+-dependent deactivtion of the SR Ca2+ release channels/ryanodine receptors (RyRs). To explore the importance of cytosolic versus luminal Ca2+ regulatory mechanisms in controlling CICR, we assessed the impact of intra-SR Ca2+ buffering on global and local Ca2+ release properties of patch-clamped or permeabilized rat ventricular myocytes. Exogenous, low-affinity Ca2+ buffers (5 to 20 mmol/L ADA, citrate or maleate) were introduced into the SR by exposing the cells to "internal" solutions containing the buffers. Enhanced Ca2+ buffering in the SR was confirmed by an increase in the total SR Ca2+ content, as revealed by application of caffeine. At the whole-cell level, intra-SR [Ca2+] buffering dramatically increased the magnitude of Ca2+ transients induced by ICa and deranged the smoothly graded ICa-SR Ca2+ release relationship. The amplitude and time-to-peak of local Ca2+ release events, Ca2+ sparks, as well as the duration of local Ca2+ release fluxes underlying sparks were increased up to 2- to 3-fold. The exogenous Ca2+ buffers in the SR also reduced the frequency of repetitive activity observed at individual release sites in the presence of the RyR activator Imperatoxin A. We conclude that regulation of RyR openings by local intra-SR [Ca2+] is responsible for termination of CICR and for the subsequent restitution behavior of Ca2+ release sites in cardiac muscle. 10.1161/01.RES.0000032490.04207.BD

Cardiac Neuronal Nitric Oxide Synthase Isoform Regulates Myocardial Contraction and Calcium Handling

Claire Sears — 2008-06-11T23:19:44-00:00

Circ Res, Vol. 92, No. 5. (21 March 2003), pp. e52-59.

A neuronal isoform of nitric oxide synthase (nNOS) has recently been located to the cardiac sarcoplasmic reticulum (SR). Subcellular localization of a constitutive NOS in the proximity of an activating source of Ca2+ suggests that cardiac nNOS-derived NO may regulate contraction by exerting a highly specific and localized action on ion channels/transporters involved in Ca2+ cycling. To test this hypothesis, we have investigated myocardial Ca2+ handling and contractility in nNOS knockout mice (nNOS-/-) and in control mice (C) after acute nNOS inhibition with 100 micromol/L L-VNIO. nNOS gene disruption or L-VNIO increased basal contraction both in left ventricular (LV) myocytes (steady-state cell shortening 10.3+/-0.6% in nNOS-/- versus 8.1+/-0.5% in C; P<0.05) and in vivo (LV ejection fraction 53.5+/-2.7 in nNOS-/- versus 44.9+/-1.5% in C; P<0.05). nNOS disruption increased ICa density (in pA/pF, at 0 mV, -11.4+/-0.5 in nNOS-/- versus -9.1+/-0.5 in C; P<0.05) and prolonged the slow time constant of inactivation of ICa by 38% (P<0.05), leading to an increased Ca2+ influx and a greater SR load in nNOS-/- myocytes (in pC/pF, 0.78+/-0.04 in nNOS-/- versus 0.64+/-0.03 in C; P<0.05). Consistent with these data, [Ca2+]i transient (indo-1) peak amplitude was greater in nNOS-/- myocytes (410/495 ratio 0.34+/-0.01 in nNOS-/- versus 0.31+/-0.01 in C; P<0.05). These findings have uncovered a novel mechanism by which intracellular Ca2+ is regulated in LV myocytes and indicate that nNOS is an important determinant of basal contractility in the mammalian myocardium. The full text of this article is available at http://www.circresaha.org. 10.1161/01.RES.0000064585.95749.6D

Increasing Ryanodine Receptor Open Probability Alone Does Not Produce Arrhythmogenic Calcium Waves: Threshold Sarcoplasmic Reticulum Calcium Content Is Required

Luigi Venetucci — 2008-06-09T00:45:36-00:00

Circ Res, Vol. 100, No. 1. (5 January 2007), pp. 105-111.

Diastolic waves of Ca2+ release have been shown to activate delayed afterdepolarizations as well as some cardiac arrhythmias. The aim of this study was to investigate whether increasing ryanodine receptor open probability alone or in the presence of beta-adrenergic stimulation produces diastolic Ca release from the sarcoplasmic reticulum (SR). When voltage-clamped rat ventricular myocytes were exposed to caffeine (0.5 to 1.0 mmol), diastolic Ca2+ release was seen to accompany the first few stimuli but was never observed in the steady state. We attribute the initial phase of diastolic Ca2+ release to a decrease in the threshold SR Ca2+ content required to activate Ca2+ waves and its subsequent disappearance to a decrease of SR content below this threshold. Application of isoproterenol (1 micromol/L) increased the amplitude of the systolic Ca2+ transient and also the SR Ca2+ content but did not usually produce diastolic Ca2+ release. Subsequent addition of caffeine, however, resulted in diastolic Ca2+ release. We estimated the time course of recovery of SR Ca2+ content following recovery from emptying with a high (10 mmol/L) concentration of caffeine. Diastolic Ca2+ release recommenced only when SR content had increased back to its final level. We conclude that increasing ryanodine receptor open probability alone does not produce arrhythmogenic diastolic Ca2+ release because of the accompanying decrease of SR Ca2+ content. beta-Adrenergic stimulation increases SR content and thereby allows the increased ryanodine receptor open probability to produce diastolic Ca2+ release. The implications of these results for arrhythmias associated with abnormal ryanodine receptors are discussed. 10.1161/01.RES.0000252828.17939.00