Low Doses Of Caffeine Research Articles

Effect of different maintenance doses of caffeine citrate on ventilator weaning in very preterm infants with respiratory distress syndrome: a prospective randomized controlled trial.

To study the effect of different maintenance doses of caffeine citrate on the success rate of ventilator weaning in very preterm infants (gestational age of ≤32 weeks) with respiratory distress syndrome (RDS). A total of 162 preterm infants with RDS who were admitted to the hospital from January 2016 to December 2018 were enrolled in this prospective trial. These infants had a gestational age of ≤32 weeks and required invasive mechanical ventilation. They were randomly divided into a high-dose caffeine group and a low-dose caffeine group, with 81 infants in each group. Within 6 hours after birth, both groups were given caffeine at a dose of 20 mg/kg. After 24 hours, the high- and low-dose caffeine groups were given caffeine at a maintenance dose of 10 mg/kg and 5 mg/kg, respectively. The two groups were compared in terms of re-intubation rate within 48 hours after ventilator weaning, durations of ventilation and oxygen therapy, enteral feeding, weight gain, and the incidence rates of complications and adverse reactions during hospitalization. The high-dose caffeine group had a significantly lower re-intubation rate within 48 hours after ventilator weaning than the low-dose caffeine group (P<0.05), with frequent apnea as the main reason for failed ventilator weaning in both groups. The high-dose caffeine group had significantly shorter durations of mechanical ventilation and oxygen therapy than the low-dose caffeine group (P<0.05). There were no significant differences between the two groups in the time to total enteral feeding, average daily weight gain, body weight at discharge, and the incidence rates of complications (bronchopulmonary dysplasia, retinopathy of prematurity, necrotizing enterocolitis, and intracranial hemorrhage) and adverse reactions (tachycardia, hypertension, and feeding intolerance) (P>0.05). A high maintenance dose of caffeine can safely and effectively reduce the incidence rate of apnea after ventilator weaning and the failure rate of ventilator weaning in RDS preterm infants with a gestational age of ≤32 weeks, and therefore, it holds promise for clinical application.

Caffeine‐induced neurotoxicity mediated by Nrf2 pathway in PC12 cells and zebrafish larvae

Caffeine is one of the most widely used psychostimulants in the world and possesses central excitative, anti-depressive, and neuroprotective properties. However, excessive ingestion or abuse of caffeine can lead to intoxication. Many toxic effects are attributed to oxidative damage, and nuclear factor erythroid 2-related factor 2 (Nrf2) is a critical intracellular regulator of the oxidative stress response. Here, we investigated the neurotoxicity of caffeine in rat pheochromocytoma PC12 cells and zebrafish larvae. It was found that caffeine inhibited the viability of PC12 cells in a dose- and time-dependent manner. Furthermore, it induced PC12 cell apoptosis and elevated reactive oxygen species (ROS) production. Quantitative polymerase chain reaction (qPCR) and western blotting revealed that caffeine also inhibited the expression levels of Nrf2 mRNA and protein and its target genes (e.g., NADPH quinone oxidoreductase 1 [NQO1]). Furthermore, Nrf2 silencing attenuated the toxic effects of caffeine. In addition, zebrafish larvae were treated with different doses of caffeine. Behavioral experiments showed that a low dose of caffeine (0.05 to 0.3 mM) increased the average distance of movement and promoted excitation. Survivorship curves showed that caffeine (0.2 to 1.5 mM) caused lethality. Finally, qPCR revealed that a higher dose of caffeine inhibited mRNA levels in the Nrf2 pathway. Based on these results, this study identified for the first time that overuse of caffeine can induce neurotoxicity by inhibiting the Nrf2 pathway. These results will provide a new perspective for studies on caffeine toxicity.

Acute cognitive performance and mood effects of coffee berry and apple extracts: A randomised, double blind, placebo controlled crossover study in healthy humans

ABSTRACT Background: Polyphenols from coffee berry (chlorogenic acid) and apple (flavanol) have been shown to improve mood and increase cerebral blood flow in healthy humans. These effects may underpin the cognitive effects of polyphenols seen previously. Objective: The aim of the present paper was to extend previous research by investigating the effects of coffee berry at high and low doses when combined with apple extract on cognitive performance and mood. Design: This randomised, double-blind, placebo controlled, crossover trial included 46 healthy males and females,18–49 years of age (mean age 23 years),consuming: 1100 mg coffee berry extract, 1100 mg coffee berry extract plus 275 mg apple extract, 100 mg coffee berry extract plus 275 mg apple extract or placebo on 4 separate occasions, completing cognitive and mood assessments pre-dose and then again at 1-, 3- and 6 hrs post-dose. Results: Analysis revealed a consistent pattern of alerting effects following 1100 mg coffee berry extract. Limited effects on cognitive function were observed. Specifically, faster peg and ball performance (executive function) was observed following 1100 mg coffee berry plus apple extract and accuracy on the Rapid Visual Information Processing (RVIP) task increased on the third of four repetitions following 1100 mg coffee berry alone. Interestingly, more false alarms on RVIP were observed following the same intervention. Conclusions: In line with previous findings, 1100 mg coffee berry engendered increased arousal. The absence of effects on mood when an apple extract was added, and the potential for the low dose of caffeine within the coffee berry to act synergistically with polyphenols, raise interesting future avenues of research. Abbreviations: Cognitive demand battery (CDB), Profile Of Mood States (POMS), Visual Analogue Scale (VAS), Rapid Visual Information Processing (RVIP)

Local recovery of cardiac calcium-induced calcium release interrogated by ultra-effective, two-photon uncaging of calcium.

Key points In cardiac myocytes, subcellular local calcium release signals, calcium sparks, are recruited to form each cellular calcium transient and activate the contractile machinery.Abnormal timing of recovery of sparks after their termination may contribute to arrhythmias.We developed a method to interrogate recovery of calcium spark trigger probabilities and their amplitude over time using two‐photon photolysis of a new ultra‐effective caged calcium compound.The findings confirm the utility of the technique to define an elevated sensitivity of the calcium release mechanism in situ and to follow hastened recovery of spark trigger probabilities in a mouse model of an inherited cardiac arrhythmia, which was used for validation.Analogous methods are likely to be applicable to investigate other microscopic subcellular signalling systems in a variety of cell types. In cardiac myocytes Ca2+‐induced Ca2+ release (CICR) from the sarcoplasmic reticulum (SR) through ryanodine receptors (RyRs) governs activation of contraction. Ca2+ release occurs via subcellular Ca2+ signalling events, Ca2+ sparks. Local recovery of Ca2+ release depends on both SR refilling and restoration of Ca2+ sensitivity of the RyRs. We used two‐photon (2P) photolysis of the ultra‐effective caged Ca2+ compound BIST‐2EGTA and laser‐scanning confocal Ca2+ imaging to probe refractoriness of local Ca2+ release in control conditions and in the presence of cAMP or low‐dose caffeine (to stimulate CICR) or cyclopiazonic acid (CPA; to slow SR refilling). Permeabilized cardiomyocytes were loaded with BIST‐2EGTA and rhod‐2. Pairs of short 2P photolytic pulses (1 ms, 810 nm) were applied with different intervals to test Ca2+ release amplitude recovery and trigger probability for the second spark in a pair. Photolytic and biological events were distinguished by classification with a self‐learning support vector machine (SVM) algorithm. In permeabilized myocytes data recorded in the presence of CPA showed a lower probability of triggering a second spark compared to control or cAMP conditions. Cardiomyocytes from a mouse model harbouring the arrhythmogenic RyRR420Q mutation were used for further validation and revealed a higher Ca2+ sensitivity of CICR. This new 2P approach provides composite information of Ca2+ release amplitude and trigger probability recovery reflecting both SR refilling and restoration of CICR and RyR Ca2+ sensitivity. It can be used to measure the kinetics of local CICR recovery, alterations of which may be related to premature heart beats and arrhythmias.

Low but not moderate amounts of caffeine increase co-consumption of ethanol in C57BL/6J mice

The increasingly popular combination of “energy drinks” containing high amounts of caffeine and alcohol has been shown to induce a stimulated, rather than sedated, state which may result in increased binge drinking and increased risk for alcohol-attributable accidents. We sought to examine consumption patterns of and withdrawal from alcohol and caffeine using a voluntary co-consumption animal model. Male and female adult C57BL/6J mice were given access to increasing doses of caffeine (0.01–0.05%) and/or alcohol (3–20%) in a two-bottle choice, intermittent access voluntary paradigm with fluid consumption recorded daily. Anxiety-like behavior during withdrawal was assessed via elevated plus maze or open field test in experiment 2. Increasing both alcohol and caffeine simultaneously in Experiment 1 resulted in no significant changes in co-consumption compared to mice given access to only alcohol or caffeine. Experiment 2 held caffeine concentration steady while slowly increasing alcohol content and resulted in mice consuming more alcohol when it was consumed in tandem with low dose caffeine. Both male and female mice consumed more caffeine when it was paired with alcohol; however, no significant differences were observed during withdrawal behavior. These results suggest that caffeine may dose-dependently positively influence alcohol consumption in mice and echo clinical literature suggesting that caffeine and alcohol together may result in a heightened state of stimulation and lead to further binge drinking. The intermittent access paradigm affords increased translational validity regarding investigations of alcohol and caffeine co-consumption and may be useful in identifying the neurobiological mechanisms concerning co-consumption of such substances.

Low, but not moderate, caffeine increases alcohol intake in male and female C57 mice

Caffeine and alcohol are two of the most popular and legal drugs around the world. In America, both drugs are easily accessible, and this has led to the two drugs being used in frequent combination with one another. Previous studies have revealed much about the mechanisms of each drug, but the effects of co-consumption are not yet fully understood. The two drugs appear to have opposing effects, as caffeine is a stimulant and alcohol is a depressant, but more research is needed on their effects when combined. One main concern is that caffeine could reduce how intoxicated a person feels, leading to increased alcohol consumption or other risky behaviors. Mice have previously been used to study alcohol, caffeine, and their combination. This previous research is not conclusive, but does suggest the potential for dose-dependent effects of caffeine when combined with alcohol. This experiment intended to compare the consumption rates of caffeine and alcohol and to observe the withdrawal effects of these two drugs. Specifically, varying levels of caffeine and alcohol were examined to see if the effects of the combination is dependent upon dose. C57BL/6J mice (n=48) were provided with caffeine (0.015% or 0.03%) and/or alcohol (3-20%) in a two-bottle intermittent access voluntary paradigm. Fluid consumption was recorded daily for five weeks and mice were then individually tested in an elevated plus maze (EPM) to assess anxiety-like behaviors during withdrawal. Although small amounts of caffeine alone were consumed throughout the study, the amount of alcohol consumed steadily increased until the highest concentration (20%) was presented. Both sexes consumed the most alcohol when it was paired with the lower dose of caffeine (0.015%) compared to either alcohol alone or alcohol in combination with the higher dose of caffeine (0.03%) and consumed more caffeine at each concentration when it was paired with alcohol. Overall, less consistent consumption patterns were observed in female mice. The results from the EPM revealed no significant difference between experimental groups and no differences between male and female mice. These results suggest that a specific amount of caffeine may be necessary to impact alcohol consumption in mice. More research is necessary to determine the psychological effects of withdrawal from these drugs.

Retinoic Acid Signaling Plays a Crucial Role in Excessive Caffeine Intake-Disturbed Apoptosis and Differentiation of Myogenic Progenitors.

Whether or not the process of somitogenesis and myogenesis is affected by excessive caffeine intake still remains ambiguous. In this study, we first showed that caffeine treatment results in chest wall deformities and simultaneously reduced mRNA expressions of genes involved in myogenesis in the developing chicken embryos. We then used embryo cultures to assess in further detail how caffeine exposure affects the earliest steps of myogenesis, and we demonstrated that the caffeine treatment suppressed somitogenesis of chicken embryos by interfering with the expressions of crucial genes modulating apoptosis, proliferation, and differentiation of myogenic progenitors in differentiating somites. These phenotypes were abrogated by a retinoic acid (RA) antagonist in embryo cultures, even at low caffeine doses in C2C12 cells, implying that excess RA levels are responsible for these phenotypes in cells and possibly in vivo. These findings highlight that excessive caffeine exposure is negatively involved in regulating the development of myogenic progenitors through interfering with RA signaling. The RA somitogenesis/myogenesis pathway might be directly impacted by caffeine signaling rather than reflecting an indirect effect of the toxicity of excess caffeine dosage.

Effects of sub-chronic caffeine ingestion on memory and the hippocampal Akt, GSK-3β and ERK signaling in mice

Caffeine, one of the most widely consumed psychoactive substance in the world, has been shown to affect mood, memory, alertness, and cognitive performance. This study aimed to assess the effect of sub-chronic oral gavage of caffeine on memory and the phosphorylation levels of hippocampal Akt (protein kinase B), GSK-3β (Glycogen Synthase Kinase-3beta) and ERK (extracellular signal-regulated kinase) in mice. Adult male NMRI mice were administered with caffeine at the doses of 0.25, 0.5, 0.75 and 1.5 mg/kg/oral gavage for 10 days before behavioral assessments. Upon completion of the behavioral tasks, the hippocampi were isolated for western blot analysis to detect the phosphorylated and total levels of Akt, GSK-3β and ERK proteins. The results showed that sub-chronic caffeine ingestion at the dose of 0.5 mg/kg improves memory in mice both in passive avoidance and novel object recognition tasks. Furthermore, this memory enhancing dose of caffeine elevated the ratios of phosphorylated to total contents of hippocampal Akt, GSK-3β and ERK. This study suggests that sub-chronic low dose of caffeine improves memory and increases the phosphorylation of hippocampal Akt, GSK-3β and ERK proteins.

Stimulatory, but not anxiogenic, doses of caffeine act centrally to activate interscapular brown adipose tissue thermogenesis in anesthetized male rats

The role of central orexin in the sympathetic control of interscapular brown adipose tissue (iBAT) thermogenesis has been established in rodents. Stimulatory doses of caffeine activate orexin positive neurons in the lateral hypothalamus, a region of the brain implicated in stimulating BAT thermogenesis. This study tests the hypothesis that central administration of caffeine is sufficient to activate BAT. Low doses of caffeine administered either systemically (intravenous [IV]; 10 mg/kg) and centrally (intracerebroventricular [ICV]; 5–10 μg) increases BAT thermogenesis, in anaesthetised (1.5 g/kg urethane, IV) free breathing male rats. Cardiovascular function was monitored via an indwelling intra-arterial cannula and exhibited no response to the caffeine. Core temperature did not significantly differ after administration of caffeine via either route of administration. Caffeine administered both IV and ICV increased neuronal activity, as measured by c-Fos-immunoreactivity within subregions of the hypothalamic area, previously implicated in regulating BAT thermogenesis. Significantly, there appears to be no neural anxiety response to the low dose of caffeine as indicated by no change in activity in the basolateral amygdala. Having measured the physiological correlate of thermogenesis (heat production) we have not measured indirect molecular correlates of BAT activation. Nevertheless, our results demonstrate that caffeine, at stimulatory doses, acting via the central nervous system can increase thermogenesis, without adverse cardio-dynamic impact.

An Apple Extract Beverage Combined with Caffeine Can Improve Alertness, Mental Fatigue, and Information Processing Speed

The psychological effects of low-dose caffeine combined with polyphenols from apples have rarely been explored scientifically yet synergistic effects are plausible. A randomized, double-blind, placebo-controlled cross-over experiment was used to test the psychological effects of apple extract beverages combined with 10, 20, 37.5, and 75 mg caffeine. Comparisons were made to both a placebo drink that was artificially sweetened and colored to mimic the test beverages and a positive control drink with 75 mg caffeine but without apple extract. Compared to placebo, it was hypothesized that dose-dependent improvements in cognitive performance, mood, and motivation would be realized after consuming the beverage with apple extract containing added caffeine. Outcomes were assessed before, 60 to 110, and 125 to 175 min post-beverage. The positive control beverage resulted in more serial seven subtractions, greater motivation to perform cognitive tasks, and reduced feelings of fatigue (all p < .005). The study found that psychological effects (i) were not observed for beverages containing apple extract and 10 or 20 mg caffeine, (ii) of the apple extract beverage containing 75 mg caffeine generally mimicked the effects of the positive control drink and significantly increased serial seven processing speed, and (iii) of the apple extract beverage containing 37.5 mg improved feelings of alertness and mental fatigue. In sum, effects of apple extract combined with caffeine were not dose-dependent; the apple extract beverage containing 75 mg caffeine improved information processing speed and the apple extract beverage with 37.5 mg caffeine improved feelings of alertness and mental fatigue.

Effects of Repeated Low-dose Caffeine Ingestion During a Night of Total Sleep Deprivation on Physical Performance and Psychological State in Young Recreational Runners

Background: Caffeine is frequently consumed by athletes to maintain alertness in conditions of sleep loss. However, the effect of caffeine ingestion during total sleep deprivation during aerobic exercise remains questionable. Objectives: The present study aimed to assess the effects of repeated intake of low-dose caffeine during a night of total sleep deprivation on endurance performance and psychological states. Methods: Twelve recreational runners performed four test sessions in a double-blind randomized order after a placebo or 6 mg/kg of caffeine ingestion after a baseline night (BN) or three doses of 2 mg/kg of caffeine during a night of total sleep deprivation (TSD). At each session, they completed an exhaustive run around a 400 m athletics track and performed the Feeling Scale test. ANOVA with repeated measures followed by paired t-tests was used to analyze the data. Results: In comparison with BN, the TSD condition significantly impaired running performance (P &lt; 0.001). On the contrary, caffeine administration improved endurance running performance (P &lt; 0.001) and increased feeling of well-being (P &lt; 0.05) after TSD night as compared to placebo. Conclusions: Repeated ingestion of low-dose caffeine is an effective way to mitigate the negative effects of total sleep deprivation on endurance performance and mood states.

Perceived Consumption of a High-Dose Caffeine Drink Delays Neuromuscular Fatigue.

Elhaj, HM, Imam, O, Page, BW, Vitale, JM, and Malek, MH. Perceived consumption of a high-dose caffeine drink delays neuromuscular fatigue. J Strength Cond Res 36(5): 1185-1190, 2022-The placebo effect is a concept in which a desired outcome arises, mainly from the belief that the treatment (i.e., supplement or drug) was beneficial although no active ingredient was given. The results of studies related to the placebo effect primarily examine functional performance. What remains unanswered, however, is whether these changes in performance are associated with neuromuscular alterations in the exercised muscles. The purpose of the study, therefore, was to determine the influence of the placebo effect on the physical working capacity fatigue threshold (PWCFT) for a continuous exercise paradigm. To achieve this aim, subjects were told that they were participating in a study to determine the dosage response (low or high) of caffeine on neuromuscular fatigue when in fact no caffeine was given during the experiment. We hypothesized that the perceived consumption of the high-dose caffeine drink would result in a higher PWCFT than the perceived consumption of the low-dose caffeine drink and placebo. Secondarily, we hypothesized that the perceived consumption of the high-dose caffeine drink would result in a higher power output than the perceived consumption of the placebo. Nine healthy college-aged men (mean ± SEM: age, 25.7 ± 1.3 years; body mass, 84.4 ± 3.1 kg; and height: 1.82 ± 0.02 m) volunteered to be in the study. For each of the visits, subjects were given an 8 oz. bottle of water with dissolved crystal light. After the drink was consumed, subjects rested in the laboratory for 1 hour before performing the incremental single-leg knee-extensor ergometry. Immediately after the termination of the incremental single-leg knee-extensor ergometry, the subject was asked which caffeine dose (placebo, low, or high) they believed they consumed for that visit. There were no significant mean differences for maximal power output for the 3 perceived conditions (placebo: 62 ± 3, low-dose caffeine: 62 ± 4, and high-dose caffeine: 65 ± 3 W). When the subjects perceived consuming the high-dose caffeine drink, there were significant mean differences (all p-values < 0.01), for PWCFT, between the other conditions (mean ± SEM: placebo: 23 ± 3 W, low-dose caffeine: 26 ± 2 W, and high-dose caffeine: 42 ± 3 W). This corresponded to a significant mean difference (all p-values < 0.01) when the PWCFT was presented as a percentage of the maximal power output (mean ± SEM: placebo: 37 ± 5%, low-dose caffeine: 42 ± 3%, and high-dose caffeine: 64 ± 3%). The application of our results may indicate that the subject's expectancy, to caffeine consumption, plays a critical role in delaying the onset of neuromuscular fatigue despite not receiving any caffeine in their drinks.

Effects of sub‐chronic oral caffeine consumption on memory and the hippocampal Akt, GSK‐3β and ERK in mice

AbstractBackgroundCaffeine, one of the most widely consumed psychoactive substance in the world, has been shown to affect mood, memory, alertness, and cognitive performance. However, the literature about the effects of caffeine intake on memory is equivocal. This study aimed to assess the effect of sub‐chronic oral gavage of caffeine on memory and the phosphorylation levels of hippocampal Akt (protein kinase B), GSK‐3β (Glycogen Synthase Kinase‐3beta) and ERK (extracellular signal‐regulated kinase) in mice.MethodAdult male NMRI mice were administered with sub‐chronic caffeine at the doses of 0.25, 0.5, 0.75 and 1.5 mg/kg/oral gavage for 10 days before training which continued until the retention day of passive avoidance task. Upon completion of the behavioral assessments, the hippocampi were isolated for western blot analysis to detect the phosphorylated and total levels of Akt, GSK‐3β and ERK proteins.ResultThe results showed that sub‐chronic caffeine ingestion at the dose of 0.5 mg/kg improves memory in mice. Furthermore, the memory enhancing dose of caffeine (0.5 mg/kg) elevated the phosphorylated to total ratios of hippocampal Akt, GSK‐3β and ERK.ConclusionThis study suggests that sub‐chronic low dose of caffeine improves memory and increases the phosphorylation of hippocampal Akt, GSK‐3β and ERK proteins.

Caffeine-related effects on cognitive performance: Roles of apoptosis in rat hippocampus following sleep deprivation

Caffeine is a common stimulant widely existed in food and has stimulatory effects on the central nervous system, shift-work individuals often rely on caffeine to maintain attention and keep awake. Although sleep deprivation (SD) is widely considered as an independent risk factor for cognition retardations, however, little is well understood about the synergistic role of caffeine dosage and SD for cognitive performance. This research intended to investigate the underlying molecular mechanism of varying caffeine doses on cognitive function after sleep deprivation. The results revealed that SD attenuated the cognitive dysfunction, associated with ultrastructure damage and pyramidal neuron loss in the hippocampus, decreased in the level of VIP and AVP. SD also significantly accelerated the neuropeptide-associated apoptosis in the hippocampus, which may modulate via the cAMP-PKA-CREB signal path axis and activation of the downstream apoptosis genes. Additionally, the data indicated that low-dose caffeine (LC) contributed to cognitive enhancement, and high-dose caffeine (HC) aggravated cognitive impairment by modulating hippocampal neuronal apoptosis. Our studies suggest that caffeine, particularly in high dosage, may be a potential factor to influence the neurocognitive outcome caused by sleep loss, and the appropriate amount of caffeine ingested after sleep deprivation deserves serious consideration.

Effects of a Low Dose of Caffeine Alone or as Part of a Green Coffee Extract, in a Rat Dietary Model of Lean Non-Alcoholic Fatty Liver Disease without Inflammation.

Non-alcoholic fatty liver disease is a highly prevalent condition without specific pharmacological treatment, characterized in the initial stages by hepatic steatosis. It was suggested that lipid infiltration in the liver might be reduced by caffeine through anti-inflammatory, antioxidative, and fatty acid metabolism-related mechanisms. We investigated the effects of caffeine (CAF) and green coffee extract (GCE) on hepatic lipids in lean female rats with steatosis. For three months, female Sprague-Dawley rats were fed a standard diet or a cocoa butter-based high-fat diet plus 10% liquid fructose. In the last month, the high-fat diet was supplemented or not with CAF or a GCE, providing 5 mg/kg of CAF. Plasma lipid levels and the hepatic expression of molecules involved in lipid metabolism were determined. Lipidomic analysis was performed in liver samples. The diet caused hepatic steatosis without obesity, inflammation, endoplasmic reticulum stress, or hepatic insulin resistance. Neither CAF nor GCE alleviated hepatic steatosis, but GCE-treated rats showed lower hepatic triglyceride levels compared to the CAF group. The GCE effects could be related to reductions of hepatic (i) mTOR phosphorylation, leading to higher nuclear lipin-1 levels and limiting lipogenic gene expression; (ii) diacylglycerol levels; (iii) hexosylceramide/ceramide ratios; and (iv) very-low-density lipoprotein receptor expression. In conclusion, a low dose of CAF did not reduce hepatic steatosis in lean female rats, but the same dose provided as a green coffee extract led to lower liver triglyceride levels.

Detection of caffeine intake by means of EC-SERS spectroscopy of human saliva.

This work presents the application of EC-SERS spectroscopy for the detection of caffeine consumption from human saliva. Caffeine and paraxanthine as the major metabolite of caffeine were tested. Model samples of saliva spiked with caffeine were investigated, and detection of caffeine in real-life saliva samples was tested in order to ensure the viability of the method for clinical applications. Two doses of caffeine (2 mg/kg and 3.5 mg/kg) were ingested by volunteers, and their saliva samples were taken at different time periods ranging from 1 h to 10 h after the consumption. Density functional theory calculations of caffeine and paraxanthine adsorbed on the silver surface were performed in order to better understand the adsorption of the investigated molecules and to make a correct assignment of the experimental spectral bands of the EC-SERS spectra. It was determined that a low dose caffeine consumption can be detected by the appearance of the SERS spectral marker band of caffeine and paraxanthine at 692 cm-1. The intensity of this band is mostly reasoned by the paraxanthine concentration since the intensity changes of the band over time correlates to the concentration changes of paraxanthine determined by the pharmacokinetic studies of paraxanthine and caffeine in the human saliva. It was found that the limit of detection paraxanthine in saliva by means of EC-SERS is as low as 15 μM and can be further improved.

Caffeine Compromises Proliferation of Human Hippocampal Progenitor Cells.

The age-associated reduction in the proliferation of neural stem cells (NSCs) has been associated with cognitive decline. Numerous factors have been shown to modulate this process, including dietary components. Frequent consumption of caffeine has been correlated with an increased risk of cognitive decline, but further evidence of a negative effect on hippocampal progenitor proliferation is limited to animal models. Here, we used a human hippocampal progenitor cell line to investigate the effects of caffeine on hippocampal progenitor integrity and proliferation specifically. The effects of five caffeine concentrations (0 mM = control, 0.1 mM ∼ 150 mg, 0.25 mM ∼ 400 mg, 0.5 mM ∼ 750 mg, and 1.0 mM ∼ 1500 mg) were measured following acute (1 day) and repeated (3 days) exposure. Immunocytochemistry was used to quantify hippocampal progenitor integrity (i.e., SOX2- and Nestin-positive cells), proliferation (i.e., Ki67-positive cells), cell count (i.e., DAPI-positive cells), and apoptosis (i.e., CC3-positive cells). We found that progenitor integrity was significantly reduced in supraphysiological caffeine conditions (i.e., 1.0 mM ∼ 1500 mg), but relative to the lowest caffeine condition (i.e., 0.1 mM ∼ 150 mg) only. Moreover, repeated exposure to supraphysiological caffeine concentrations (i.e., 1.0 mM ∼ 1500 mg) was found to affect proliferation, significantly reducing % Ki67-positive cells relative to control and lower caffeine dose conditions (i.e., 0.1 mM ∼ 150 mg and 0.25 mM ∼ 400 mg). Caffeine treatment did not influence apoptosis and there were no significant differences in any measure between lower doses of caffeine (i.e., 0.1 mM, 0.25 mM, 0.5 mM) – representative of daily human caffeine intake – and control conditions. Our study demonstrates that dietary components such as caffeine can influence NSC integrity and proliferation and may be indicative of a mechanism by which diet affects cognitive outcomes.

Effects of Various Doses of Caffeine Ingestion on Intermittent Exercise Performance and Cognition.

To date, no study has examined the effects of caffeine on prolonged intermittent exercise performance that imitates certain team-sports, and the suitable concentration of caffeine for improved intermittent exercise performance remains elusive. The purpose of the present cross-over, double-blind preliminary study was to investigate effects of low, moderate, and high doses of caffeine ingestion on intermittent exercise performance and cognition. Ten males performed a familiarization session and four experimental trials. Participants ingested capsules of placebo or caffeine (3, 6, or 9 mg/kg) at 1 h before exercise, rested quietly, and then performed cycling for 2 × 30 min. The cycling protocol consisted of maximal power pedaling for 5 s (mass × 0.075 kp) every minute, separated by unloaded pedaling for 25 s and rest for 30 s. At pre-ingestion of capsules, 1 h post-ingestion, and post-exercise, participants completed the Stroop task. The mean power-output (MPO), peak power-output (PPO), and response time (RT) in the Stroop task were measured. Only 3 mg/kg of caffeine had positive effects on the mean PPO and MPO; 3 mg/kg caffeine decreased RTs significantly in the incongruent and congruent conditions. These results indicate that the ingestion of low-dose caffeine had greater positive effects on the participants’ physical strength during prolonged intermittent exercise and cognition than moderate- or high-dose caffeine.

Low caffeine dose improves intermittent sprint performance in hot and humid environments.

While the effects of caffeine have been evaluated in relation to endurance exercise, few studies have assessed the ergogenic effects of low caffeine doses on intermittent exercise performance in hot and humid environments. Thus, we aimed to determine the effects of low-dose caffeine supplementation on intermittent exercise performance under these conditions. Eight male soccer players (age, 19.9±0.3 years; height, 173.7±6.3cm; body mass, 65.1±5.5kg; V˙O2max, 50.0±3.1mL ⋅ kg-1⋅ min-1) participated in this double-blind, randomized, cross-over study. Caffeine was orally administered at 60min before exercise (dosage, 3mg ⋅kg-1). The participants completed a 90-min intermittent sprint cycling protocol under two conditions (after receiving caffeine and placebo) at 32°C and at 70% relative humidity. A significant improvement in the total amount of work was observed in the caffeine condition compared to the placebo condition (155.0±15.8 vs 150.8±14.5kJ, respectively; p<0.05, d=0.28). In contrast, the rectal temperature measured at the end of exercise showed no significant difference between the conditions (38.9±0.4°C and 38.7±0.5°C in the caffeine and placebo conditions, respectively; p>0.05, d=0.57). Other thermal responses, such as the mean skin temperature, heart rate, or sweat volume, were not significantly different between these conditions. These results suggested that a low caffeine dose improved the intermittent sprint performance and the reasons could be explained by the fact that a low caffeine dose ingestion did not affect the thermoregulatory responses compared to the placebo condition and, thus, did not attenuate its ergogenic effect on exercise in hot and humid environments.

Cognition and Brain Activation in Response to Various Doses of Caffeine: A Near-Infrared Spectroscopy Study.

Caffeine, which is widely used for enhancing athletic performance, has been suggested to have a positive impact on cognition via stimulating the brain. However, no study published to date has explored the effects of different doses of caffeine ingestion on brain activation via cortical hemodynamics. The purpose of the present crossover, double-blind study was to investigate the effects of low, moderate, and high doses of caffeine ingestion on cognitive performance and brain activation. Ten healthy male subjects ingested placebo or caffeine (3, 6, or 9 mg/kg body mass). The effects of each treatment condition were evaluated by Stroop tasks before and 60 min after the ingestion of caffeine. Reaction time (RT) and accuracy of responses to congruent and incongruent stimuli were assessed. As an index of brain activation with cognition, levels of oxygenated hemoglobin (HbO) were measured via near-infrared spectroscopy. A 4 × 2 mixed ANOVA revealed that there were significant interaction effects for RT in both incongruent and congruent conditions (P < 0.01, Pη2 = 0.384; P < 0.05, Pη2 = 0.259; and P < 0.05, Pη2 = 0.309). Both 3 and 6 mg/kg of caffeine ingestion significantly decreased RT to incongruent stimuli. The only dose of caffeine to decrease RT in response to congruent stimuli was 3 mg/kg. None of the doses of caffeine administered affected accuracy of responses to incongruent or congruent stimuli. Under the congruent stimulus condition, ingestion of 3 mg/kg of caffeine significantly increased mean HbO in the dorsolateral prefrontal cortex, frontal pole area, ventrolateral prefrontal cortex (P < 0.01, Pη2 = 0.319; P < 0.05, Pη2 = 0.263; and P < 0.05, Pη2 = 0.259, respectively). None of the doses of caffeine investigated affected HbO under the incongruent stimulus condition. Ingestion of low-dose caffeine has greater effects on cognition and brain activation than moderate and high doses of caffeine, suggesting that low-dose caffeine may be a selective supplement in enhancing executive function and prefrontal activities.