Caffeine Response Research Articles

17β-estradiol induces hyperresponsiveness in guinea pig airway smooth muscle by inhibiting the plasma membrane Ca2+-ATPase

High serum estrogen concentrations are associated with asthma development and severity, suggesting a link between estradiol and airway hyperresponsiveness (AHR). 17β-estradiol (E2) has non-genomic effects via Ca2+ regulatory mechanisms; however, its effect on the plasma membrane Ca2+-ATPases (PMCA1 and 4) and sarcoplasmic reticulum Ca2+-ATPase (SERCA) is unknown. Hence, in the present study, we aim to demonstrate if E2 favors AHR by increasing intracellular Ca2+ concentrations in guinea pig airway smooth muscle (ASM) through a mechanism involving Ca2+-ATPases.In guinea pig ASM, Ca2+ microfluorometry, muscle contraction, and Western blot were evaluated. Then, we performed molecular docking analysis between the estrogens and Ca2+ ATPases.In tracheal rings, E2 produced AHR to carbachol. In guinea pig myocytes, acute exposure to physiological levels of E2 modified the transient Ca2+ peak induced by caffeine to a Ca2+ plateau. The incubation with PMCA inhibitors (lanthanum and carboxyeosin, CE) partially reversed the E2-induced sustained plateau in the caffeine response. In contrast, cyclopiazonic acid (SERCA inhibitor), U-0126 (an inhibitor of ERK 1/2), and choline chloride did not modify the Ca2+ plateau produced by E2. The mitochondrial uniporter activity and the capacitative Ca2+ entry were unaffected by E2. In guinea pig ASM, Western blot analysis demonstrated PMCA1 and PMCA4 expression. The results from the docking modeling demonstrate that E2 binds to both plasma membrane ATPases. In guinea pig tracheal smooth muscle, inhibiting the PMCA with CE, induced hyperresponsiveness to carbachol. 17β-estradiol produces hyperresponsiveness by inhibiting the PMCA in the ASM and could be one of the mechanisms responsible for the increase in asthmatic crisis in women.

Characterization of caffeine response regulatory variants in vascular endothelial cells.

Genetic variants in gene regulatory sequences can modify gene expression and mediate the molecular response to environmental stimuli. In addition, genotype-environment interactions (GxE) contribute to complex traits such as cardiovascular disease. Caffeine is the most widely consumed stimulant and is known to produce a vascular response. To investigate GxE for caffeine, we treated vascular endothelial cells with caffeine and used a massively parallel reporter assay to measure allelic effects on gene regulation for over 43,000 genetic variants. We identified 665 variants with allelic effects on gene regulation and 6 variants that regulate the gene expression response to caffeine (GxE, false discovery rate [FDR] < 5%). When overlapping our GxE results with expression quantitative trait loci colocalized with coronary artery disease and hypertension, we dissected their regulatory mechanisms and showed a modulatory role for caffeine. Our results demonstrate that massively parallel reporter assay is a powerful approach to identify and molecularly characterize GxE in the specific context of caffeine consumption.

Effect of Sample Plates and Sample Matrix on the Quantification Capabilities of Surface-Assisted Flowing Atmospheric-Pressure Afterglow Mass Spectrometry (SA-FAPA-MS).

Ambient desorption/ionization mass spectrometry (ADI-MS) has been broadly applied to accomplish direct analysis without sample preparation or separation. However, quantification capabilities and analytical performance are sometimes limited. Here, we report signal enhancement effects and improved quantification capabilities in plasma-based ADI-MS, when a flowing atmospheric-pressure afterglow (FAPA) source is used to probe analytes on tailored thin-layer chromatography (TLC) plates. It was found that quantitative results could be achieved when the TLC plate merely served as a sampling plate without a preceding separation step. Specifically, the dynamic response of caffeine, nicotine, acetaminophen, and progesterone was investigated with FAPA-MS on a variety of different TLC surfaces (normal-phase silica, reversed-phase-modified silica, cyano [CN]-modified silica, and dimethyl [RP2]-modified silica). All analytes were studied as single-analyte standards and in a multianalyte mixture to evaluate the effect of sample plates and sample matrix on analytical performance and competitive ionization processes. Overall, dimethyl (RP2)- and CN-modified silica resulted in superior performance compared to other TLC materials. After careful optimization and without the use of internal standards, linear ranges of five orders of magnitude were accessible for caffeine and nicotine. Limits of detection down to femtomole amounts of analyte were achieved. Quantitation limits using RP2-TLC and FAPA-MS were 0.062, 0.062l, 0.31, and 14 pmol for caffeine, nicotine, progesterone, and acetaminophen, respectively. Interestingly, the presence of nicotine at relatively high amounts reduced the signal of the other analytes, an observation that was found to correlate with the differences in the enthalpy of vaporization (ΔHvap) and proton affinity. To prove the quantitative capabilities, nicotine quantification in a real matrix-heavy e-liquid sample was demonstrated using an isotopically labeled standard. The use of TLC-based surfaces with FAPA-MS can aid in the direct and quantitative mass spectrometric investigation of complex mixtures.

Caffeine activates HOG-signalling and inhibits pseudohyphal growth in Saccharomyces cerevisiae

ObjectiveCaffeine has a wide range of effects in humans and other organisms. Caffeine activates p38 MAPK, the human homolog to the Hog1 protein that orchestrates the high-osmolarity glycerol (HOG) response to osmotic stress in the yeast Saccharomyces cerevisiae. Caffeine has also been used as an inducer of cell-wall stress in yeast via its activation of the Pkc1-mediated cell wall integrity (CWI) pathway. In this study, using immunodetection of phosphorylated Hog1, microscopy to score nuclear localisation of GFP-tagged Hog1 and a pseudohyphal growth assays, the effect of caffeine on the HOG-pathway and filamentous growth in yeast was studied.ResultsIt was found that caffeine causes rapid, strong and transient Hog1 dual phosphorylation with statistically significant increases at 20, 30 and 40 mM caffeine. In response to caffeine treatment Hog1 was also rapidly localized to the nucleus, supporting the caffeine-induced phosphorylation and activation of Hog1. We also found that caffeine inhibited the pseudohyphal/filamentous growth in diploid cells, but had no effect on invasive growth in haploids. Our data thus highlights that the HOG signalling pathway is activated by caffeine, which has implications for interpreting caffeine responses in yeast and fungi.

Probing the Use of Homemade Carbon Fiber Microsensor for Quantifying Caffeine in Soft Beverages.

In the development of electrochemical sensors, carbon micro-structured or micro-materials have been widely used as supports/modifiers to improve the performance of bare electrodes. In the case of carbon fibers (CFs), these carbonaceous materials have received extensive attention and their use has been proposed in a variety of fields. However, to the best of our knowledge, no attempts for electroanalytical determination of caffeine with CF microelectrode (µE) have been reported in the literature. Therefore, a homemade CF-µE was fabricated, characterized, and used to determine caffeine in soft beverage samples. From the electrochemical characterization of the CF-µE in K3Fe(CN)6 10 mmol L-1 plus KCl 100 mmol L-1, a radius of about 6 µm was estimated, registering a sigmoidal voltammetric profile that distinguishes a µE indicating that the mass-transport conditions were improved. Voltammetric analysis of the electrochemical response of caffeine at the CF-µE clearly showed that no effects were attained due to the mass transport in solution. Differential pulse voltammetric analysis using the CF-µE was able to determine the detection sensitivity, concentration range (0.3 to 4.5 µmol L-1), limit of detection (0.13 μmol L-1) and linear relationship (I (µA) = (11.6 ± 0.09) × 10-3 [caffeine, μmol L-1] - (0.37 ± 0.24) × 10-3), aiming at the quantification applicability in concentration quality-control for the beverages industry. When the homemade CF-µE was used to quantify the caffeine concentration in the soft beverage samples, the values obtained were satisfactory in comparison with the concentrations reported in the literature. Additionally, the concentrations were analytically determined by high-performance liquid chromatography (HPLC). These results show that these electrodes may be an alternative to the development of new and portable reliable analytical tools at low cost with high efficiency.

Characterization of caffeine response regulatory variants in vascular endothelial cells

Characterization of caffeine response regulatory variants in vascular endothelial cells

Functional single-nucleotide polymorphism (rs762551) in CYP1A2 gene affects white coffee intake in healthy 20- to 40-year-old adults

Coffee is one of the most consumed beverages in the world, but the extent to which it is consumed is affected by both environmental and genetic factors. Genome-wide association studies and candidate date association studies have identified several gene variants associated with increased consumption of coffee. Functional single-nucleotide polymorphisms in rs762551 (cytochrome P450 1A2 [CYP1A2]) and rs5751876 (adenosine receptor A2A [ADORA2A]) has been linked to individual caffeine response. Coffee intake has been shown to affect lipid metabolism. We thus hypothesize that rs762551 (CYP1A2) A allele carriers consume more coffee than C allele carriers and that rs5751876 (ADORA2A) C allele carriers consume less coffee than T allele carriers. Additionally, we hypothesize that CYP1A2 genotype can modulate serum glucose concentrations and lipid profile. A total of 421 participants aged 20 to 40 years were recruited from 2016 to 2018 in Poznań, Poland. Genotyping of CYP1A2 and ADORA2A was performed using TaqMan probes. Individuals with AA CYP1A2 genotype consumed relatively more coffee with milk (72.81 ± 10.15 mL/1000 kcal vs 43.38 ± 6.42 mL/1000 kcal, P = .008) and with milk or cream than did C allele carriers, whereas the rs5751876 ADORA2A polymorphism was not associated with coffee or tea intake. Additionally, subjects with AA CYP1A2 genotype had 10% higher serum triacylglycerol (TG) concentrations than C allele carriers. This study suggests that CYP1A2 rs762551 polymorphism is associated with coffee intake and serum TG concentrations in healthy 20- to 40-year-old adults.

Caffeine-related genes influence anxiety disorders in children and adults with ADHD

Attention-deficit/hyperactivity disorder (ADHD) and anxiety disorders (AD) frequently co-occur, increasing morbidity and challenging treatment. Caffeine is a central nervous system stimulant and acts in the brain through adenosine receptors, influencing attention, alertness, and anxiety. In the present study, we performed a gene-set analysis to verify if genes related to caffeine response are associated with anxiety disorders in 240 children and 406 adults with ADHD. We demonstrated an association between the gene-set with AD in children (P = 0.0054) and with the number of anxiety disorders in adults (P = 0.0197). In order to test if this effect is a result of anxiety in general or is related to AD comorbid with ADHD, we evaluated the association between caffeine gene-set with AD in an adult control sample. The gene-set was neither associated with the AD presence (P = 0.3008) nor with the number of AD (P = 0.5594) in this control sample. We also test this gene set with ADHD (n = 55,374) and AD (n = 18,186) GWAS summary statistics, and we did not observe significant results with ADHD (P = 0.5587) or AD (P = 0.3930). These findings suggest the caffeine-related genes play a role in the etiology of an anxiety disorder phenotype present in children and adults with ADHD.

Perturbation of Ca2+ stores and store-operated Ca2+ influx by lidocaine in neuronal N2A and NG108-15 cells

In this report we examined the effects of lidocaine on Ca2+ homeostasis of neuronal cells using microfluorimetric measurement of cytosolic Ca2+ with fura 2 as probe. In mouse neuroblastoma N2A cells, 10 mM lidocaine caused Ca2+ release from the cyclopiazonic acid (CPA)-dischargeable pool and abolished ATP-triggered Ca2+ release. Lidocaine-triggered Ca2+ release was not affected by xestospongin C (XeC), an inositol 1,4,5-trisphosphate receptor (IP3R) inhibitor. N2A cells did not have functional ryanodine receptors (RYR) (absence of caffeine response) and we used differentiated NG108-15 cells (presence of caffeine response) for further experiments. Caffeine-triggered Ca2+ release was unaffected by a brief lidocaine exposure, but was eliminated after a prolonged treatment of lidocaine, suggesting lidocaine abolished caffeine action possibly not by interfering caffeine binding but via Ca2+ store depletion. Lidocaine-elicited Ca2+ release was unaffected by XeC or a high concentration of ryanodine, suggesting Ca2+ release was not via IP3R or RYR. Lidocaine did not affect nigericin-dischargeable lysosomal Ca2+ stores. Lastly, we observed that lidocaine suppressed CPA-induced store-operated Ca2+ influx in both N2A cells and differentiated NG108-15 cells. Our results suggest two novel actions of lidocaine in neuronal cells, namely, depletion of Ca2+ store (via an IP3R- and RYR-independent manner) and suppression of store-operated Ca2+ influx.

Stimulant Use as a Fatigue Countermeasure in Aviation

INTRODUCTION: Fatigue is a common problem in aviation. The identification of efficacious fatigue countermeasures is crucial for sustaining flight performance during fatigue-inducing operations. Stimulants are not recommended for consistent use, but are often implemented during flight operations with a high risk of fatigue. As such, it is important to evaluate the efficacy of approved stimulants for sustaining flight performance, alertness, and mood.METHODS: Four electronic databases (PubMed, PsycInfo, SPORTDiscus, Web of Science) were systematically searched to identify research on the effects of caffeine, dextroamphetamine, and modafinil during simulated or in-flight operations.RESULTS: There were 12 studies identified that assessed the effects of at least 1 stimulant. Overall, dextroamphetamine and modafinil were effective for sustaining flight performance and pilot mood during extended wakefulness. Results with caffeine were inconsistent.DISCUSSION: Dextroamphetamine and modafinil appear to sustain flight performance and mood during extended wakefulness. However, most studies have used flight simulators and short operation durations. Additional research is needed in realistic settings and during longer duration operations. Caffeines effects were inconsistent across studies, possibly due to differences in study methodology or individual caffeine responses. Despite fatigue being a common problem in civilian aviation as well, only one study in this review included civil aviators. More research should be conducted on the effects of caffeine during civil operations.CONCLUSION: Dextroamphetamine and modafinil appear to be effective fatigue countermeasures but should be further evaluated in more ecologically valid settings. The effects of caffeine are unclear at this time and should continue to be evaluated.Ehlert AM, Wilson PB. Stimulant use as a fatigue countermeasure in aviation. Aerosp Med Hum Perform. 2021; 92(3):190200.

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.

The effect of caffeine on cognitive performance is influenced by CYP1A2 but not ADORA2A genotype, yet neither genotype affects exercise performance in healthy adults

PurposeTo determine the influence of two commonly occurring genetic polymorphisms on exercise, cognitive performance, and caffeine metabolism, after caffeine ingestion.MethodsEighteen adults received caffeine or placebo (3 mg kg−1) in a randomised crossover study, with measures of endurance exercise (15-min cycling time trial; 70-min post-supplementation) and cognitive performance (psychomotor vigilance test; PVT; pre, 50 and 95-min post-supplementation). Serum caffeine and paraxanthine were measured (pre, 30 and 120-min post-supplementation), and polymorphisms in ADORA2A (rs5751876) and CYP1A2 (rs762551) genes analysed.ResultsCaffeine enhanced exercise performance (P < 0.001), but effects were not different between participants with ADORA2A ‘high’ (n = 11) vs. ‘low’ (n = 7) sensitivity genotype (+ 6.4 ± 5.8 vs. + 8.2 ± 6.8%), or CYP1A2 ‘fast’ (n = 10) vs. ‘slow’ (n = 8) metabolism genotype (+ 7.2 ± 5.9 vs. + 7.0 ± 6.7%, P > 0.05). Caffeine enhanced PVT performance (P < 0.01). The effect of caffeine was greater for CYP1A2 ‘fast’ vs. ‘slow’ metabolisers for reaction time during exercise (− 18 ± 9 vs. − 1.0 ± 11 ms); fastest 10% reaction time at rest (− 18 ± 11 vs. − 3 ± 15 ms) and lapses at rest (− 3.8 ± 2.7 vs. + 0.4 ± 0.9) (P < 0.05). There were no PVT differences between ADORA2A genotypes (P > 0.05). Serum caffeine and paraxanthine responses were not different between genotypes (P > 0.05).ConclusionCaffeine enhanced CYP1A2 ‘fast’ metabolisers’ cognitive performance more than ‘slow’ metabolisers. No other between-genotype differences emerged for the effect of caffeine on exercise or cognitive performance, or metabolism.

Microsaccade rate as a measure of drug response.

In 22 human subjects we measured microsaccade count across 60 brief fixation trials both pre- and post- administration of 300mg of caffeine. There was a statistically significant reduction in average microsaccade count post-caffeine administration, with a moderate effect size (Cohen’s d of 0.42). Microsaccade count was stable within individuals across time points (Pearson’s r of 0.89). Sensitivity analysis suggests that the pre/post caffeine effect size is robust to choice of parameters used to identify microsaccades. Bootstrap resampling suggests that both the pre/post-caffeine difference and the across-time stability within individuals could be reliably assessed with far fewer trials. The results support the use of microsaccade count as both a trait measure of individual differences and a state measure of caffeine response. We discuss the results in the context of the theory that the superior colliculus is central to the generation of microsaccades and hence that microsaccade rate may be a useful assay for at least some drug-induced changes at the level of the colliculus: a potentially useful tool in the development of therapies for disorders that may involve collicular dysfunction such as ADHD.

Evaluating the temperature dependence of heat-transfer based detection: A case study with caffeine and Molecularly Imprinted Polymers as synthetic receptors

Molecularly Imprinted Polymers (MIPs) are synthesized for the selective detection of caffeine. The polymerization process, monomer and crosslinker monomer composition are varied to determine the optimal synthesis procedure via batch rebinding experiments evaluated with optical detection. The selectivity is tested by comparing the response of caffeine to compounds with similar chemical structures (theophylline and theobromine) and dopamine, another neurotransmitter. Subsequently, the MIP polymer particles are integrated into bulk modified MIP screen-printed electrodes (MIP-modified SPEs). The sensors are used to measure caffeine content in various samples employing the Heat-Transfer Method (HTM), a low-cost and simple thermal detection method that is based on differences in thermal resistance at the solid-liquid interface. At first, the noise is minimized by adjusting the settings of temperature feedback loop. Second, the response of the MIP-modified SPE is studied at various temperatures ranging from 37 to 50 and 85 °C. The binding to MIP-modified SPEs has never been studied at elevated temperatures since most biomolecules are not stable at those temperatures. Using caffeine as proof-of-concept, it is demonstrated that at 85 °C the detection limit is significantly enhanced due to higher signal to noise ratios and enhanced diffusion of the biomolecule. Thermal wave transport analysis (TWTA) is also optimized at 85 °C producing a limit of detection of ∼1 nM. Next, MIP-modified SPEs are used to measure the caffeine concentration in complex samples including caffeinated beverages, spiked tap water and waste water samples.The use of MIP-modified SPEs combined with thermal detection provides sensors that can be used for fast and low-cost detection performed on-site, which holds great potential for the determination of contaminants in environmental samples. The platform is generic and by adapting the MIP layer, we can expand to this a range of relevant targets.

An exercise-induced improvement in isolated skeletal muscle contractility does not affect the performance-enhancing benefit of 70 µmol l-1 caffeine treatment.

This study aimed to examine the effects of exercise-induced increases in skeletal muscle contractile performance on isolated skeletal muscle caffeine sensitivity in mice. CD1 mice (n=28; 30 weeks old) either served as controls or underwent 8 weeks of voluntary wheel running. Following the treatment intervention, whole soleus (SOL) or a section of the costal diaphragm (DIA) was isolated from each mouse and tested to determine the effect of 70 µmol l-1 caffeine on work loop power output. Although caffeine elicited a significant increase in power of both the SOL and the DIA relative to levels in a non-caffeine-treated control, the effect was not different between the experimental groups, despite the muscles of the trained group producing significantly greater muscle power. There was no significant relationship between training volume or baseline work loop power and the caffeine response. These results indicate that an exercise-induced increase in muscle performance did not influence the performance-enhancing effects of caffeine.

PO-304 Caffeine Supplementation Altered Metabolic Profiles in High-intensity Interval Training

Objective Caffeine supplementation is a commonly used nutritional practice. Exogenous metabolites from caffeine, such as paraxanthine, theobromine and theophylline, are eventually excreted through urine. Yet, it is less clear whether caffeine would induce endogenous metabolites altered during exercise. Urine metabolomics is non-invasive method, which mainly focus on alterations of endogenous metabolic profiles caused by diseases, drugs, and lifestyle and nutritional interventions as well. Therefore, the purpose of the present study was to examine the effects of supplementation with caffeine in a well-designed high intensity interval training (HITT). We identified significant alterations in urinary metabolite levels and revealed key metabolic pathways involved in caffeine supplementation in HITT.&#x0D; Methods We performed a randomized, double-blind, placebo- controlled crossover study. Twelve women basketball players (age:19.12 ± 2.64 years, mass: 174.73 ± 5.18 cm, height: 62 ± 5.09 kg, with 8.50±2.11 years training period for basketball) were randomized to placebo (PLA) or caffeine (CAF) with dosage of 3mg on the basis of body weight (kg) 45min before a field HITT test. The test was repeated after three days when players were crossed over to the alternate test. The test began with a 30 min warmup, followed by a high intensity intermittent exercise trail with incremental load for about 25min, and a cool-down. Players are familiar with the test program which included 55 sets of dribble shuttle-run, pass, shoot, and rebound with basketball with a distance of 1540m (55 × 28m), the interval between two sets was gradually reduced. Performance (completed time), heart rates immediate (HR0min) and 1 min (HR1min) after test, blood lactate (BLa), proteinuria and ratings of perceived exertion (RPE) were collected during each protocol. Urine samples were obtained before and 1 h after of the test. 1H-NMR spectra (Bruker AVANCE III HD 600MHz) were obtained and then processed by NMR spectra (MestReNova 9.0). The binning values of NMR spectra are imported into MATLAB, and the peaks are aligned with the icoshift algorithm. Then concentrations of the aligned metabolites were calculated by converting the integral area of proton signals with that of the TSP. Pattern recognition was performed to the processed NMR data, including principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA). Characteristic metabolites were identified that contribute most to the metabolic pattern between groups according to the OPLS-DA models. Finally, we analyzed the metabolic pathway by importing characteristic metabolites with concentrations into the Enrichment Analysis (MetaboAnalysis 3.0) to determine the metabolic pathways with the greatest disturbance related to caffeine during exercise. Moreover, the main effects of exercise, caffeine and the interaction between exercise and caffeine were determined by Repeated measure GLM analysis (Spss 22.0).&#x0D; Results (1) Compared with PLA, CAF had no significant difference in the completed time (25.9 min vs. 26.8 min). Repeated measured analysis showed that there was significant overall time effect on the routine training monitoring parameters, while no statistically group differences in HR0min, HR1min, BLa (199.02±21.36 vs.189.00±22.38 bpm; 148.02±12.60 vs.148.02±20.34 bpm, and 8.89±2.23 vs. 9.52±2.91 mmol/L, respectively). For the qualitative indexes, the positive rate of urine ketone bodies was increased, while RPE did not changed. (2) We identified 32 metabolites in urine sample. PCA showed distinct differentiation of metabolic patterns between each two groups in the four groups (PLAbefore, PLApost, CAFbefore, CAFafter). By using OPLS-DA, we found that the urine metabolic profiles were differences in between caffeine supplementation group and placebo group during the test. OPLS-DA revealed the identified metabolites of exercise and caffeine respectively, among them, lactate, butyric acid, isobutyric acid, 3-hydroxybutyric acid and pyruvic acid could be used as metabolic biomarkers in the HITT response. Supplementation of caffeine increased the production of fat metabolites in urine compared to the PLA. Enrichment analysis showed that the disturbed metabolic pathways shared by PLA and CAF were purine metabolism, glycolysis, insulin signal transduction, galactose metabolism, gluconeogenesis, glucose-alanine cycle, sphingolipid metabolism, alanine metabolism and citric acid cycle. Yet, when compared to the PLA, CAF enhanced fat metabolism and increased pyruvate metabolism, cysteine metabolism and mitochondrial electron transport. These results suggest that caffeine could promote fatty acid metabolism and amino acid metabolism to improve aerobic metabolism and to reduce oxidative stress, and thus promote exercise capacity. (3) Covariance analysis showed that there were significant individual-specific effects of caffeine supplementation.&#x0D; Conclusions Caffeine supplementation during HITT promoted the fat metabolism, and upregulated the TCA, pyruvate metabolism and mitochondrial electron transfer. It is suggested that caffeine could, to some extent, promote energy supply shift from anaerobic metabolic to an aerobic manner, and the enhancement of fat oxidation would be beneficial to glycogen storage for intensively long-duration exercise. Moreover, there are obvious individual differences in caffeine response on sports.

Association between adenosine receptor gene polymorphism and response to caffeine citrate treatment in apnea of prematurity; An Egyptian single-center study

BackgroundCaffeine citrate is the methyl-xanthine of choice used in controlling apnea of prematurity (AOP). Caffeine central effect is mediated via non-selective (A1) and selective (A2a) adenosine receptors antagonism. Variability in caffeine response had been frequently noticed in AOP, suggesting underlying genetic predisposition. Aim of the studyWe evaluated the role of adenosine receptor A1 [ADORA1] and adenosine receptor A2a [ADORA2a] gene polymorphisms in the variability of caffeine response among Egyptian preemies with AOP. Patient and methodsIn this case-control study, 43 preterm neonates with AOP were eligible as cases and 43 preterm babies free from apnea were taken as controls. Preterm neonates with AOP were further divided according to response to caffeine treatment into caffeine responder (n = 18) and caffeine non-responders (n = 25). ADORA1 [716 T > G] and ADORA2a [1976C > T] gene polymorphisms were genotyped by mean of PCR-based RFLP-assay. ResultsThere were significant increase in frequency distribution of ADORA2a [1976C > T] CT (62.7% vs 23.3%), TT (14% vs 4.7%) genotypes and T allele (34.3% vs 16.3%) in cases compared to controls with significant increased risk of AOP development with OR (95%CI); P-value of 8.37(3.03–23.1), P = 0.000; 9.3(1.61–53.61), P = 0.005 and 4.27(2.09–8.70), P = 0.000 respectively. Further, caffeine non-responders were associated with significant increase frequency of ADORA2a CT (80% vs 38.9%) and TT (16% vs 11.1%) genotypes and T allele (56% vs 30.6%) with OR (95%CI) and P-value of 21.38 (2.31–197.8), P = 0.001; 18 (1.24–260.9); P = 0.005 and 2.89(1.17–7.13), P = 0.019 respectively, when compared to caffeine responders. Patients with AOP who had ADORA2a CT and TT genotypes were associated with significant increase in duration of hospital stay and poor outcome. Genotype distribution frequency of studied polymorphisms did not deviate from Hardy Weinberg (HW) equilibrium among controls. ConclusionADORA2a [1976C > T] polymorphism has a significant role in AOP development and variation in caffeine response among preterm babies.

Bare carbon electrodes as simple and efficient sensors for the quantification of caffeine in commercial beverages.

Food quality control is a mandatory task in the food industry and relies on the availability of simple, cost-effective and stable sensing platforms. In the present work, the applicability of bare glassy carbon electrodes for routine analysis of food samples was evaluated as a valid alternative to chromatographic techniques, using caffeine as test analyte. A number of experimental parameters were optimized and a differential pulse voltammetry was applied for quantification experiments. The detection limit was found to be 2 × 10−5 M (3σ criterion) and repeatability was evaluated by the relative standard deviation of 4.5%. The influence of sugars, and compounds structurally related to caffeine on the current response of caffeine was evaluated and found to have no significant influence on the electrode performance. The suitability of bare carbon electrodes for routine analysis was successfully demonstrated by quantifying caffeine content in seven commercially available drinks and the results were validated using a standard ultra-high performance liquid chromatography method. This work demonstrates that bare glassy carbon electrodes are a simple, reliable and cost-effective platform for rapid analysis of targets such as caffeine in commercial products and they represent therefore a competitive alternative to the existing analytical methodologies for routine food analysis.

Are the Current Guidelines on Caffeine Use in Sport Optimal for Everyone? Inter-individual Variation in Caffeine Ergogenicity, and a Move Towards Personalised Sports Nutrition

Caffeine use is widespread in sport, with a strong evidence base demonstrating its ergogenic effect. Based on existing research, current guidelines recommend ingestion of 3–9 mg/kg approximately 60 min prior to exercise. However, the magnitude of performance enhancement following caffeine ingestion differs substantially between individuals, with the spectrum of responses ranging between highly ergogenic to ergolytic. These extensive inter-individual response distinctions are mediated by variation in individual genotype, environmental factors, and the legacy of prior experiences partially mediated via epigenetic mechanisms. Here, we briefly review the drivers of this inter-individual variation in caffeine response, focusing on the impact of common polymorphisms within two genes, CYP1A2 and ADORA2A. Contemporary evidence suggests current standardised guidelines are optimal for only a sub-set of the athlete population. Clearer understanding of the factors underpinning inter-individual variation potentially facilitates a more nuanced, and individually and context-specific customisation of caffeine ingestion guidelines, specific to an individual’s biology, history, and competitive situation. Finally, we identify current knowledge deficits in this area, along with future associated research questions.

Novel pathomechanisms of cardiomyocyte dysfunction in a model of heart failure with preserved ejection fraction.

Heart failure with preserved ejection fraction (HFpEF) is increasingly common, but the underlying cellular mechanisms are not well understood. We investigated cardiomyocyte function and the role of SEA0400, an Na(+) /Ca(2+) exchanger (NCX) inhibitor in a rat model of chronic kidney disease (CKD) with HFpEF. Male Wistar rats were subjected to subtotal nephrectomy (NXT) or sham operation (Sham). After 8 and 24 weeks, in vivo (haemodynamics, echocardiography) and in vitro function (LV cardiomyocyte cell shortening (CS), and Ca(2+) transients (CaT)) were determined without and with SEA0400. In a subgroup of rats, SEA0400 or vehicle was given p.o. (1 mg/kg b.w.) between week 8 and 24. NXT resulted in stable compensated CKD and HFpEF [hypertrophied left ventricle, prolonged LV isovolumetric relaxation constant TAU (IVRc TAU), elevated end diastolic pressure (EDP), increased lung weight (pulmonary congestion), and preserved LV systolic function (EF, dP/dt)]. In NXT cardiomyocytes, the amplitude of CS and CaT were unchanged but relaxation and CaT decay were progressively prolonged at 8 and 24 weeks vs. Sham, individually correlating with diastolic dysfunction in vivo. NCX forward mode activity (caffeine response) was progressively reduced, while NCX protein expression was up-regulated, suggesting increased NCX reverse mode activity in NXT. SEA0400 acutely improved relaxation in NXT in vivo and in cardiomyocytes and improved cardiac remodelling and diastolic function when given chronically. This model of renal HFpEF is associated with slowed relaxation of LV cardiomyocytes. Treatment with SEA0400 improved cardiomyocyte function, remodelling, and HFpEF.