Caffeine Treatments Research Articles

The effects of Red Bull energy drink compared with caffeine on cycling time-trial performance.

This study investigated the ergogenic effects of a commercial energy drink (Red Bull) or an equivalent dose of anhydrous caffeine in comparison with a noncaffeinated control beverage on cycling performance. Eleven trained male cyclists (31.7 ± 5.9 y 82.3 ± 6.1 kg, VO2max = 60.3 ± 7.8 mL · kg-1 · min-1) participated in a double-blind, placebo-controlled, crossover-design study involving 3 experimental conditions. Participants were randomly administered Red Bull (9.4 mL/kg body mass [BM] containing 3 mg/kg BM caffeine), anhydrous caffeine (3 mg/kg BM given in capsule form), or a placebo 90 min before commencing a time trial equivalent to 1 h cycling at 75% peak power output. Carbohydrate and fluid volumes were matched across all trials. Performance improved by 109 ± 153 s (2.8%, P = .039) after Red Bull compared with placebo and by 120 ± 172 s (3.1%, P = .043) after caffeine compared with placebo. No significant difference (P > .05) in performance time was detected between Red Bull and caffeine treatments. There was no significant difference (P > .05) in mean heart rate or rating of perceived exertion among the 3 treatments. This study demonstrated that a moderate dose of caffeine consumed as either Red Bull or in anhydrous form enhanced cycling time-trial performance. The ergogenic benefits of Red Bull energy drink are therefore most likely due to the effects of caffeine, with the other ingredients not likely to offer additional benefit.

Effect of various concentrations of caffeine, pentoxifylline, and kallikrein on hyperactivation of frozen bovine semen.

Caffeine, pentoxifylline, and kallikrein are substances that affect the efficiency of sperms in the fertilization process; however, they have not been adequately studied. The present study aimed to examine the influence of caffeine, kallikrein, and pentoxifylline on sperm motility in bovine as well as investigate their optimum concentrations for increasing the movement of sperms in bovine. Frozen bovine sperms were thawed in universal IVF medium supplemented with 1, 5, and 10 mM caffeine or pentoxifylline or 1, 4, and 8 U/mL kallikrein and were then incubated for 30 min. Treated semen parameters were analyzed using a computer assisted semen analyzer (CASA). Data analysis showed that the mean values concerning progression and motility of sperm increased in caffeine and pentoxifylline treatments when compared with the kallikrein group. The obtained results revealed that kallikrein is not necessary for the improvement of bovine sperm motility. Additionally, our results revealed that 5 mM from caffeine was the best concentration added to the medium, followed by 1 or 5 mM from pentoxifylline. Therefore, it is concluded from the present study that caffeine has hyperactivation efficacy at 5 mM concentration compared to other treatments.

Differential Gene Expression Profiling in Bed Bug (Cimex Lectularius L.) Fed on Ibuprofen and Caffeine in Reconstituted Human Blood

The recent resurgence of the common bed bug (Cimex lectularius L.) infestations worldwide has created a need for renewed research on biology, behavior, population genetics and management practices. Humans serve as exclusive hosts to bed bugs in urban environments. Since a majority of humans consume Ibuprofen (as pain medication) and caffeine (in coffee and other soft drinks) so bug bugs subsequently acquire Ibuprofen and caffeine through blood feeding. However, the effect of these chemicals at genetic level in bed bug is unknown. Therefore, this research was conducted to determine differential gene expression in bed bugs using RNA-Seq analysis at dosages of 200 ppm Ibuprofen and 40 ppm caffeine incorporated into reconstituted human blood and compared against the control. Total RNA was extracted from a single bed bug per replication per treatment and sequenced. Read counts obtained were analyzed using Bioconductor software programs to identify differentially expressed genes, which were then searched against the non-redundant (nr) protein database of National Center for Biotechnology Information (NCBI). Data on comparison of differentially expressed genes between control and Ibuprofen treatments revealed that 659 genes were significantly differentially regulated and 95% of them returned BLAST hits. Heat stress proteins were among the top significantly differentially down regulated genes. Comparison of the control vs caffeine treatments revealed that 2,161 genes were significantly differently regulated (Padj <0.05). Heat shock proteins were among the top ten down regulated genes in both treatments. Finally, using RNAi to identify the exact function of these highly differentially expressed genes and regulating these genes may offer potential for managing bed bug populations.

Caffeine treatment prevented from weight regain after calorie shifting diet induced weight loss.

Low calorie diets are always difficult for obese subjects to follow and lead to metabolic and behavioral adaptation. Therefore, we evaluated the effect of caffeine treatment with calorie shifting diet (CSD) on weight loss. Female subjects (n=60; BMI≥25) completed 4-weeks control diet, 6-weeks CSD (3 repeated phases; each 2-weeks) and 4-weeks follow-up diet, with or without caffeine treatment (5 mg/Kg/day). The first 11 days of each phase included calorie restriction with four meals every day and 4 hours intervals. Significant weight and fat loss were observed after 4-weeks of CSD (5.7 ± 1.24 Kg and 4.84 ± 1.53 Kg) or CSD+Caffeine (7.57 ± 2.33 Kg and 5.24 ± 2.07 Kg) which was consistent for one month of the follow-up (CSD: 5.24 ± 1.83 Kg and 4.3 ± 1.62 Kg, CSD+Caffeine: 12.11 ± 2.31 Kg and 9.85 ± 1.6 Kg, p < 0.05 vs CSD group) and correlated to the restricted energy intake (p < 0.05). During three CSD phases, RMR tended to remain unchanged in both groups.While, CSD or CSD + Caffeine treatments, significantly decreased plasma glucose, total-cholesterol, and triacylglycerol (p < 0.05), even during follow-up period (p < 0.05). HDL-cholesterol was not changed by CSD. Feeling of hunger decreased and subject's satisfaction increased after 4-weeks of CSD (p < 0.05) and remained low to the end of study, while satiety was not affected. Coffeine increased the effect of CSD on feeling of hunger and subject's satisfaction after week 7 (p < 0.05 vs. CSD). These findings indicated that combination of caffeine treatment with CSD could be an effective alternative approach to weight and fat loss with small changes in RMR and improved tolerance of subjects to the new diet.

Caffeine and modafinil promote adult neuronal cell proliferation during 48 h of total sleep deprivation in rat dentate gyrus

It has been established that sleep deprivation (SD) reduces the proliferation of neuronal precursors in the adult hippocampus. It has also been reported that psychostimulant drugs modulate adult neurogenesis. We examined the modulatory role of two psychostimulant drugs modafinil and caffeine on adult neuronal cell proliferation (NCP) during 48h of total SD. A novel automated cage shaking stimulus was used to induce SD based on animal activity. 5-Bromo-2″-deoxyuridine (BrdU; 50mg/kg/day i.p.) was injected at the onset of the light phase for two days. Rats were successfully sleep deprived for 85–94% of total time. Stereological analysis showed that both caffeine and modafinil treatments during SD improved the number of BrdU positive cells as compared to the SD group. Caffeine treatment during SD, significantly increased early proliferative and post-mitotic stages of doublecortin (DCX) positive cells while modafinil treatment during SD, increased intermediate and post-mitotic stages of DCX positive cells compared to SD+Vehicle group. Brain-Derived Neurotrophic Factor (BDNF) expression on BrdU positive cells as well as in the dentate gyrus (DG) region was decreased significantly after sleep deprivation. Both caffeine and modafinil significantly improved BDNF expression in the DG region. Modafinil, but not caffeine, significantly decreased hippocampal adenosine level during SD in comparison to the SD+Vehicle group. It may be concluded that caffeine or modafinil treatment during 48h of SD prevents the SD induced decline in neuronal proliferation and differentiation. Caffeine and modafinil induced alterations of NCP during SD may involve modulation of BDNF and adenosine levels.

QPCA: a scalable assay to measure the perturbation of protein–protein interactions in living cells

One of the most important challenges in systems biology is to understand how cells respond to genetic and environmental perturbations. Here we show that the yeast DHFR-PCA, coupled with high-resolution growth profiling (DHFR-qPCA), is a straightforward assay to study the modulation of protein-protein interactions (PPIs) in vivo as a response to genetic, metabolic and drug perturbations. Using the canonical Protein Kinase A (PKA) pathway as a test system, we show that changes in PKA activity can be measured in living cells as a modulation of the interaction between its regulatory (Bcy1) and catalytic (Tpk1 and Tpk2) subunits in response to changes in carbon metabolism, caffeine and methyl methanesulfonate (MMS) treatments and to modifications in the dosage of its enzymatic regulators, the phosphodiesterases. Our results show that the DHFR-qPCA is easily implementable and amenable to high-throughput. The DHFR-qPCA will pave the way to the study of the effects of drug, genetic and environmental perturbations on in vivo PPI networks, thus allowing the exploration of new spaces of the eukaryotic interactome.

The effects of different doses of caffeine on endurance cycling time trial performance

This study investigated the effects of two different doses of caffeine on endurance cycle time trial performance in male athletes. Using a randomised, placebo-controlled, double-blind crossover study design, sixteen well-trained and familiarised male cyclists (Mean ± s: Age = 32.6 ± 8.3 years; Body mass = 78.5 ± 6.0 kg; Height = 180.9 ± 5.5 cm [Vdot]O2peak = 60.4 ± 4.1 ml · kg−1 · min−1) completed three experimental trials, following training and dietary standardisation. Participants ingested either a placebo, or 3 or 6 mg · kg−1 body mass of caffeine 90 min prior to completing a set amount of work equivalent to 75% of peak sustainable power output for 60 min. Exercise performance was significantly (P < 0.05) improved with both caffeine treatments as compared to placebo (4.2% with 3 mg · kg−1 body mass and 2.9% with 6 mg · kg−1 body mass). The difference between the two caffeine doses was not statistically significant (P = 0.24). Caffeine ingestion at either dose resulted in significantly higher heart rate values than the placebo conditions (P < 0.05), but no statistically significant treatment effects in ratings of perceived exertion (RPE) were observed (P = 0.39). A caffeine dose of 3 mg · kg−1 body mass appears to improve cycling performance in well-trained and familiarised athletes. Doubling the dose to 6 mg · kg−1 body mass does not confer any additional improvements in performance.

The effect of physiological concentrations of caffeine on the power output of maximally and submaximally stimulated mouse EDL (fast) and soleus (slow) muscle

The ergogenic effects of caffeine in human exercise have been shown to improve endurance and anaerobic exercise performance. Previous work has demonstrated that 70 μM caffeine (physiological maximum) can directly increase mouse extensor digitorum longus (EDL) muscle power output (PO) in sprintlike activity by 3%. Our study used the work loop technique on isolated mouse muscles to investigate whether the direct effect of 70 μM caffeine on PO differed between 1) maximally and submaximally activated muscle; 2) relatively fast (EDL) and relatively slow (soleus) muscles; and 3) caffeine concentrations. Caffeine treatment of 70 μM resulted in significant improvements in PO in maximally and submaximally activated EDL and soleus (P < 0.03 in all cases). For EDL, the effects of caffeine were greatest when the lowest, submaximal stimulation frequency was used (P < 0.001). Caffeine treatments of 140, 70, and 50 μM resulted in significant improvements in acute PO for both maximally activated EDL (3%) and soleus (6%) (P < 0.023 in all cases); however, there was no significant difference in effect between these concentrations (P > 0.420 in all cases). Therefore, the ergogenic effects of caffeine on PO were higher in muscles with a slower fiber type (P < 0.001). Treatment with 35 μM caffeine failed to elicit any improvement in PO in either muscle (P > 0.72 in both cases). Caffeine concentrations below the physiological maximum can directly potentiate skeletal muscle PO. This caffeine-induced increase in force could provide similar benefit across a range of exercise intensities, with greater gains likely in activities powered by slower muscle fiber type.

Viability of ICSI oocytes after caffeine treatment and sperm membrane removal with Triton X-100 in pigs

Non-adequate decondensation of injected sperm nucleus is one the main problems of intracytoplasmic sperm injection (ICSI) in porcine. With the aim of improving pronuclear formation, the effects on activation and embryo development rates of 0.1% Triton X-100 (TX) sperm pre-treatment for membrane removal and/or 5 mM Caffeine (CAF) addition in oocyte manipulating and culture medium for 2 h after ICSI or artificial activation were studied. The effects of 4 different Ca2+ concentrations contained in the injection medium on embryo development after sham injection were also analysed. In Experiment 1, no significant effect on cleavage or blastocyst rate was detected independently of Ca2+ concentration contained in the injection medium. In Experiment 2, oocytes injected with TX pre-treated sperm showed a significant higher rate of male pronuclear formation in comparison with oocytes from control group (2PN; 54.1 vs 36.6%). However, no differences on in vitro embryo development, cleavage or blastocyst rates were observed. In Experiment 3, oocytes treated with CAF during and after micromanipulation and injected with sperm pre-treated with TX had a significantly lower oocyte activation rate than any other experimental groups (25.7 vs 56.3–66.3%). No differences were observed in cleavage rates among different experimental groups. However, the CAF group showed a higher blastocyst rate significantly different from TX+CAF group (12.0 vs 1.9%, respectively). In a second approach, the effect of electric field strengths and CAF treatments on oocyte activation was studied. In Experiment 4, oocytes submitted to 0.6 kV/cm showed significant higher activation rates than 1.2 kV/cm ones regardless of the caffeine treatment (83.7 vs 55.9% and 75.7 vs 44.3%; in control and caffeine groups, respectively). No effect of caffeine treatment was observed in any experimental group. In conclusion, TX sperm treatment before ICSI without an additional activation procedure improved male pronuclear formation, but did not improve embryo development until blastocyst stage. No significant effect of caffeine was found when sperm was not treated with TX, although in membrane absence caffeine avoided oocyte activation and embryo development. Finally, caffeine had no effect on female pronuclear formation regardless of electric field strengths applied to the parthenogenetic activation.

Effects of low-dose caffeine supplementation on early morning performance in the standing shot put throw

The purpose of the present investigation was to assess the efficacy of low-dose caffeine use for early morning performance in the shot put event. A double-blind, randomized, crossover design was used to investigate the effects of buccal caffeine supplementation on early morning shot put execution in nine inter-collegiate track and field athletes. In one condition the participants received a piece of caffeinated gum designed to deliver 100 mg of caffeine in a buccal manner, and in a second condition a placebo gum. The gum was chewed for 5 min before being discarded. Participants then completed the first psychomotor vigilance task followed by a series of five warm-up throws, followed by six attempts with a shot put (7.26 kg for males, 4.0 kg for females) measured for distance. The protocol ended with a final psychomotor vigilance task. A repeated-measures analysis of variance (treatment*time) was used to compare performance between the caffeine and placebo treatments over the six measured attempts. A significant difference (treatment×throw) was observed (P=0.030, partial eta-squared = 0.259), indicating that the caffeine treatment produced better performance over the course of the six attempts subsequent to a warm-up. A paired samples t-test (Bonferroni-adjusted for multiple comparisons) revealed that the first attempt in the caffeine treatment (9.62±1.71 m) and in the placebo treatment (9.05±1.69 m) were significantly different (P = 0.050, effect size = 0.996, 95%CI 1.02 to 0.13 m). Repeated-measures analysis of covariance revealed a significant (P=0.016, partial eta-squared = 0.650) interaction effect (treatment×mean reaction time), whereby both at the pre and post time points the mean reaction time on the psychomotor vigilance task was reduced under the caffeine treatment (caffeine: pre 0.306±0.05 s, post 0.316±0.08 s; placebo: pre 0.317±0.06 s, post 0.323±0.06 s). Based on these results, we suggest that caffeine gum can be beneficial for both performance and alertness if used by shot put athletes during early morning sessions.

Focus on therapy of hypnic headache

Hypnic headache (HH) is a primary headache disorder, which occurs exclusively during sleep and usually begins after 50 years of age. There are no controlled trials for the treatment of HH. We reviewed all the available papers, including 119 cases published in literature up to date, reporting the efficacy of the medications used to treat HH. Acute treatment is not recommended, since no drug proved to be clearly effective and also because the intensity and the duration of the attacks do not require the intake of a medication in most cases. As for prevention, a wide variety of medications were reported to be of benefit in HH. The drugs that were found to be effective in at least five cases are: lithium, indomethacin, caffeine and flunarizine. Lithium was the most extensively studied compound and demonstrated to be an efficacious treatment in 32 cases. Unfortunately, despite its efficacy, significant adverse effects and poor tolerability are not rare, mainly in elderly patients. Many patients reported a good response to indomethacin, but some could not tolerate it. Caffeine and melatonin treatments did not yield robust evidence to recommend their use as single preventive agents. Nevertheless, their association with lithium or indomethacin seems to produce an additional therapeutic efficacy. A course of lithium should be tried first, followed 3–4 months later by tapering. If headache recurs during tapering, a longer duration of therapy may be needed. If lithium treatment does not provide a significant response, indomethacin can be commenced as second-line approach. If these treatments prove to be ineffective or poorly tolerated, other agents, such as caffeine and melatonin, can be administered.

Sex-related effects of agmatine on caffeine-induced locomotor activity in Swiss Webster mice

In mammalian brain, agmatine is an endogenous amine that is synthesized through the decarboxylation of l-arginine by arginine decarboxylase. It has been proposed as a new neurotransmitter and/or neuromodulator. It was shown that agmatine had some beneficial effects in animal models of opioid and alcohol addiction. Locomotor stimulant properties of drugs such as ethanol, caffeine, nicotine and amphetamine have been linked to their addictive properties. The present study investigates the effects of agmatine on caffeine-induced locomotor activity both in male and female mice. Adult Swiss Webster mice were used in the study. Locomotor activity was measured for 30 min immediately following caffeine (2.5, 5, 10 and 20 mg/kg, i.p.) or saline treatments. Agmatine (5, 10 and 20 mg/kg, i.p.) were injected 20 min before caffeine (2.5 and 5 mg/kg, i.p.) administration. In both sexes, agmatine (5–20 mg/kg) were also tested for ability to depress or stimulate locomotor activity in the absence of caffeine. Caffeine (5 mg/kg) induced a significant increase in locomotor activity of both male and female mice. There was no significant difference in the locomotor-activating effects of caffeine between male and female mice. Agmatine blocked the caffeine (5 mg/kg)-induced locomotor stimulation dose dependently in male but not female mice. Agmatine had not any effect on the lower dose (2.5 mg/kg) of caffeine in both sexes. These results suggest that agmatine has sex-related inhibitory effects on caffeine-induced locomotor activity in Swiss Webster mice, and male mice are more sensitive than the females to the effect of agmatine.

The temperature and time factors in caffeine potentiation of chemically-induced chromosomal aberrations in root tips of Vicia faba

The temperature and time factors in caffeine potentiation of chemically-induced chromosome damage in bean root tips was studied for 12 chromosome-damaging (inducer) agents. By varying the temperature during the 5-hour post-treatment with caffeine from 10 to 25°C and the duration of the time interval between inducer and caffeine treatments from 0 to 6 hours, the inducer agents could be classified into three groups: (1) potentiation independent of temperature during caffeine exposure and of the time between inducer and caffeine treatments (maleic hydrazide, azaserine, MMS, DMS), (2) potentiation increasing with increasing temperature during caffeine exposure, but independent of the time between inducer and caffeine treatments (EMS, MNU, MNNG, quinacrine mustard), and (3) potentiation dependent on both temperature during caffeine exposure and time between inducer and caffeine treatments (DEB, HN2, mitomycin C, thiotepa). Potentiation appears to be affected by the temperature during the time interval between inducer and caffeine treatments only when it is influenced by the duration of the interval.

Enhancement by methylated oxypurines of the frequency of induced chromosomal aberrations

Two-hour treatments with 5 times 10-3 M caffeine strongly enhanced the frequencies of chromosomal aberrations obtained in root tips of Vicia faba 5 to 6 hours after irradiation with X-rays. The caffeine treatments were effective only when given after X-irradiation and when the time between X-rays and caffeine was less than 60 minutes. In contrast to the frequency of aberrations produced by caffeine alone, which increased when the temperature was lowered, that produced by the combined X-ray-caffeine treatments was higher at 27°C than at 15°C. The aberration frequency produced by X-rays in combination with caffeine was not markedly influenced by sodium azide, an inhibitor of oxidative phosphorylation. A comparison was made between the effects of caffeine, theophylline and 1,3,7,9-tetramethyluric acid alone and in combination with X-rays In unirradiated root-tip cells, 1,3,7,9-tetramethyluric acid produced the highest frequency of aberrations, whereas theophylline was practically inactive. When given as post-treatments to X-rays, caffeine produced the largest and 1,3,7,9-tetramethyluric acid the smallest increase in the frequency of chromosomal aberrations.

Effect of Caffeine on Sport-Specific Endurance Performance: A Systematic Review

Endurance athletes often ingest caffeine because of its reported ergogenic properties. Although there are a vast number of studies quantifying caffeine's effects, many research studies measure endurance performance using a time-to-exhaustion test (subjects exercise at a fixed intensity to volitional exhaustion). Time-to-exhaustion as a performance measure is not ideal because of the high degree of measurement variability between and within subjects. Also, we are unaware of any endurance sports in which individuals win by going a longer distance or for a longer amount of time than their competitors. Measuring performance with a time-trial test (set distance or time with best effort) has high reproducibility and is more applicable to sport. Therefore, the purpose of this review was to critically and objectively evaluate studies that have examined the effect of caffeine on time-trial endurance (>5 minutes) performance. A literature search revealed 21 studies with a total of 33 identifiable caffeine treatments that measured endurance performance with a time-trial component. Each study was objectively analyzed with the Physiotherapy Evidence Database (PEDro) scale. The mean PEDro rating was 9.3 out of 10, indicating a high quality of research in this topic area. The mean improvement in performance with caffeine ingestion was 3.2 +/- 4.3%; however, this improvement was highly variable between studies (-0.3 to 17.3%). The high degree of variability may be dependent on a number of factors including ingestion timing, ingestion mode/vehicle, and subject habituation. Further research should seek to identify individual factors that mediate the large range of improvements observed with caffeine ingestion. In conclusion, caffeine ingestion can be an effective ergogenic aid for endurance athletes when taken before and/or during exercise in moderate quantities (3-6 mg.kg body mass). Abstaining from caffeine at least 7 days before use will give the greatest chance of optimizing the ergogenic effect.

Caffeine Overcomes Genistein-Induced G2/M Cell Cycle Arrest in Breast Cancer Cells

Although inhibition of tumor cell growth by genistein is mediated by different types of cell cycle arrest, its regulation of genes related to the cell cycle is not clear. In this study, genistein caused a concentration-dependent growth inhibition in the hormone-independent cell line MDA-MB-435S. Flow cytometric analysis showed that genistein induced a concentration-dependent accumulation of cells in the G2/M phase of the cell cycle. Caffeine enhanced the inhibition of cell proliferation induced by genistein. Caffeine alone did not have an appreciable effect on the phases of the cell cycle, but caffeine at 3 mM completely eliminated genistein-induced G2/M cell cycle arrest. cDNA microarrays were used to investigate the mechanism of genistein-induced, caffeine-negated G2/M arrest. We identified 12 genes, which had opposite responses to genistein and caffeine treatments. Among these, 5 genes were upregulated by genistein and downregulated by caffeine; 7 genes were downregulated by genistein and upregulated by caffeine. Reversal by caffeine of genistein-induced G2/M phase arrest in breast cancer cell lines could increase their sensitivity to genistein and enhance genistein-induced inhibition of cell growth. Genes that have opposite responses to genistein and caffeine may be involved in regulation of the G2/M phase of the cell cycle.

Effects Of Caffeinated Sports Drinks On Physical/Mental Function During Team Sports Exercise

The ergogenic effects of caffeine ingestion (240–840 mg) on exercise performance is well established, albeit in some cases with negative side effects. However, recent evidence shows that much smaller doses can have beneficial effect on endurance performance as well as on mental function. Less is known about the possible synergistic effects of low dose of caffeine (CAF) and carbohydrate (CHO) feedings, especially as related to team sports performance. PURPOSE: To determine the effects of low dose (100 mg) CAF ingestion along with typically recommended CHO on physical and mental function during intermittent high intensity exercise that mimics the demands of competitive team sports. METHODS: Physically active male (N=6) and female (N=4) low to moderate caffeine users participated in this study. Experimental trials consisted of four ∼22-minute quarters (QTR) of shuttle running. Physical and mental function tests were administered throughout the experimental trial including (20-m sprint, vertical jump, gross motor skill test, force sensation, rating of perceived exertion, profile of mood states and psychomotor vigilance test).On four separate occasions, subjects received 20-oz of flavored water placebo (PLA), CHO, PLA+CAF and CHO+CAF prior to exercise. During exercise, subjects ingest PLA, CHO, PLA and CHO respectively (3.5 ml·kg-1). RESULTS: No difference was found in 20-m sprint time, vertical jump and gross motor skills performance between any treatments. However, both caffeine treatments (PLA+CAF and CHO+CAF) significantly improved psychomotor vigilance and reduced force sensation at HALFTIME (p<0.05). CHO+CAF also improved self-reported perception of fatigue, vigor and overall mood compared to PLA+CAF (p<0.05). Systolic blood pressure was increased with CHO+CAF compare to other three treatments. However, no difference was found in heart rate, core body temperature, plasma volume change, or hydration status between any treatments. CONCLUSION: The data suggest that ingestion of a low dose of caffeine prior to exercise did not add to the expected benefits of CHO feedings alone on physical tasks. However, all of the mental function tasks were significantly enhanced with the combination of CHO plus CAF, with little or no adverse side effects. Funded by Gatorade Sports Science Institute.

Highly efficient induction of triploid Pacific abalone Haliotis discus hannai by caffeine treatment

In order to develop a highly efficient method for mass production of triploid Pacific abalone Haliotis discus hannai, caffeine treatment that is safe and inexpensive was optimized. To suppress the first meiotic division, fertilized eggs were exposed to either a 10- or 15-mM caffeine solution for 24 min beginning at 12 min after fertilization. In most treated batches, the rates of cleaved eggs showed no significant difference from the control batches. However, in most treated batches, the rates of occurrence of normal larvae and the survival rates of the early juveniles were significantly lower than those of the controls. The triploid rates at 6 days to 11 months after settlement in all the treated batches were extremely high (91–100%). There was no significant difference in the mean triploid rates between 10- and 15-mM caffeine treatments. These results suggest that both treatment conditions were conducive to triploid abalone production. One live 2n/3n mosaic specimen was found in the treated batches. However, since the frequency of mosaic was extremely low, the mosaicism would probably not have an adverse effect on the stable production of triploid abalones.

58 EFFECTS OF CAFFEINE AND VANADATE ON NUCLEAR REMODELING AND MICROTUBULE DISTRIBUTION OF PORCINE NUCLEAR TRANSFER EMBRYOS

The present study was conducted to control nuclear remodeling types by treatment with caffeine or vanadate, and to examine the microtubule distribution of nuclear transfer embryos (NTs) after nuclear remodeling control and the mitotic spindle assembly and its morphological changes during the first mitosis of NTs in the pig. Enucleated oocytes were treated with 5 mM caffeine or 0.5 mM sodium orthovanadate (vanadate) for 2.5 or 0.5 h to increase or decrease MPF activity before injection of fetal fibroblast cells. Reconstituted eggs were fused by an electric stimulation (ES, 1.5 kV cm-1), activated by a combination of 2 pulses of ES (1.0 kV cm-1), and cultured for 3 h with 2 mM 6-dimethylaminopurine (6-DMAP) at 1 h after fusion treatment. Some matured oocytes were also treated by the same chemicals before parthenogenetic activation under the same conditions as NTs, and cultured in vitro to evaluate the effects of these chemicals on embryo development. NTs and parthenogenetic embryos were cultured in PZM-3 for 20 h or 6 days at 39�C, 5% CO2 in air, respectively. Nuclear remodeling types of NTs were examined at 1 h after fusion (before activation) by the whole-mount method. At least 3 replicates for each experiment were performed. Microtubules and DNA of NTs that were fixed at 1 h or 20 h after fusion were detected by indirect immunocytochemical technique. Images were captured using laser scanning confocal microscopy. Caffeine and vanadate did not affect the development to the blastocyst stage of porcine parthenogenetic embryos. When a nucleus was exposed to oocyte cytoplasm treated with caffeine, premature chromosome condensation (PCC) occurred at a higher rate (82/98, 83.7%) compared to control (42/73, 57.5%) and vanadate-treated (11/91, 12.1%) groups (P &amp;lt; 0.05). The proportion of embryos that did not undergo nuclear envelope breakdown (NEBD) was higher in the vanadate treatment group (87.9%) compared to the caffeine and control groups (16.3 and 42.5%, respectively; P &amp;lt; 0.05). The frequency of embryos showing a γ-tubulin only and both γ- and β-tubulins were 3.9–9.4% and 21.9–34.6%, respectively, in NTs (total 87 embryos) at 1 h after fusion regardless of caffeine and vanadate treatments. In the majority of NTs (61.5–68.6%), microtubules were not observed. At 20 h after fusion, the frequency of the embryos undergoing normal mitosis was similar in the control (17/45, 37.8%) and caffeine (19/43, 44.2%) groups, but it was significantly lower in the vanadate group (7/37, 18.9%; P &amp;lt; 0.05). The present study demonstrates that the nuclear remodeling type of NTs can be controlled by treatment with MPF regulators, caffeine and vanadate, and such treatment is not related to the microtubule distribution in porcine NTs. The finding, however, that the vanadate can delay the mitotic progression of porcine NTs at the first cell cycle may be due to the lack of NEBD and PCC. This work was supported by the Korea Research Foundation Grant funded by the Korean Government (MOEHRD;KRF-2005-042-F00030).

Atrial natriuretic peptide induces an acrosome reaction in giant panda spermatozoa and enhances their penetration of salt-stored porcine oocytes

Atrial natriuretic peptide (ANP) is a vasodilator peptide primarily produced in the heart. Locally synthesized ANP has been found in reproductive tissues of various mammals and humans, and plays an important role in rat oocyte maturation and human sperm function. The objective of the present study was to determine the effects of ANP on the function (acrosome reaction and zona penetration) of giant panda spermatozoa. In fresh and frozen-thawed spermatozoa that had been preincubated for 2.5 h, treatment with ANP (for 60 min) significantly increased the proportion of acrosome-reacted spermatozoa; maximal response (an acrosome reaction in 18.3 and 21.8% of fresh and frozen-thawed spermatozoa, respectively) was detected at 1 nM ANP. Treatment with C-ANP-(4–23), an analogue of ANP and specific binder to natriuretic peptide receptors-C (NPRC), had no significant effect on the acrosome reaction. However, the cyclic guanosine 5′-monophosphate (cGMP)-dependent protein kinase (PKG) inhibitor KT5823 completely abolished the effect of ANP on acrosome reaction. The effects of ANP, caffeine and heparin on frozen-thawed sperm function were studied by insemination of porcine salt-stored oocytes in a modified Tris-buffered medium (mTBM). The presence of ANP, caffeine or heparin in the insemination medium resulted in a higher proportion ( P < 0.05) of oocytes with spermatozoa in the zona and perivitelline space (PVS), and a higher average number of spermatozoa/oocyte ( P < 0.05) in the zona and PVS. However, in the absence of ANP, caffeine and heparin, there were no oocytes with a spermatozoon in the PVS. There were no differences among ANP, caffeine or heparin treatments for the proportion of oocytes penetrated or average number of spermatozoa/oocyte in the zona and PVS. In conclusion, we inferred that ANP induced the acrosome reaction of preincubated giant panda spermatozoa by a PKG pathway. Furthermore, ANP enhanced the penetrability of porcine salt-stored oocytes by frozen-thawed giant panda spermatozoa.