Expression of the highly conserved replication-dependent histone gene family increases dramatical... more Expression of the highly conserved replication-dependent histone gene family increases dramatically as a cell enters the S phase of the eukaryotic cell cycle. Requirements for normal histone gene expression in vivo include an element, designated α, located within the protein-encoding sequence of nucleosomal histone genes. Mutation of 5 of 7 nucleotides of the mouse H3.2 α element to yield the sequence found in an H3.3 replication-independent variant abolishes the DNA-protein interaction in vitro and reduces expression fourfold in vivo. A yeast one-hybrid screen of a HeLa cell cDNA library identified the protein responsible for recognition of the histone H3.2 α sequence as the transcription factor Yin Yang 1 (YY1). YY1 is a ubiquitous and highly conserved transcription factor reported to be involved in both activation and repression of gene expression. Here we report that the in vitro histone α DNA-protein interaction depends on YY1 and that mutation of the nucleotides required for t...
The purpose of the present study was to determine the effects of short-term supplementation with ... more The purpose of the present study was to determine the effects of short-term supplementation with the free acid form of β-hydroxy-β-methylbutyrate (HMB-FA) on indices of muscle damage, protein breakdown, recovery and hormone status following a high-volume resistance training session in trained athletes. A total of twenty resistance-trained males were recruited to participate in a high-volume resistance training session centred on full squats, bench presses and dead lifts. Subjects were randomly assigned to receive either 3 g/d of HMB-FA or a placebo. Immediately before the exercise session and 48 h post-exercise, serum creatine kinase (CK), urinary 3-methylhistadine (3-MH), testosterone, cortisol and perceived recovery status (PRS) scale measurements were taken. The results showed that CK increased to a greater extent in the placebo (329 %) than in the HMB-FA group (104 %) (P= 0·004,d= 1·6). There was also a significant change for PRS, which decreased to a greater extent in the place...
Proceedings of the National Academy of Sciences, 1996
The histone gene family in mammals consists of 15-20 genes for each class of nucleosomal histone ... more The histone gene family in mammals consists of 15-20 genes for each class of nucleosomal histone protein. These genes are classified as either replication-dependent or -independent in regard to their expression in the cell cycle. The expression of the replication-dependent histone genes increases dramatically as the cell prepares to enter S phase. Using mouse histone genes, we previously identified a coding region activating sequence (CRAS) involved in the upregulation of at least two (H2a and H3) and possibly all nucleosomal replication-dependent histone genes. Mutation of two seven-nucleotide elements, alpha and omega, within the H3 CRAS causes a decrease in expression in stably transfected Chinese hamster ovary cells comparable with the effect seen upon deletion of the entire CRAS. Further, nuclear proteins interact in a highly specific manner with nucleotides within these sequences. Mutation of these elements abolishes DNA/protein interactions in vitro. Here we report that the i...
Journal of the International Society of Sports Nutrition, 2012
Sikorski et al.: The acute effects of a free acid betahydoxy-beta-methyl butyrate supplement on m... more Sikorski et al.: The acute effects of a free acid betahydoxy-beta-methyl butyrate supplement on muscle damage following resistance training: a randomized, double-blind, placebo-controlled study.
Journal of Strength and Conditioning Research, 2013
Currently no research has investigated the relationship between muscle damage, hormonal status, a... more Currently no research has investigated the relationship between muscle damage, hormonal status, and perceived recovery scale (PRS). Therefore, the purpose of this study was to determine the effects of a high-volume training session on PRS and to determine the relationship between levels of testosterone, cortisol, and creatine kinase (CK) and PRS. Thirty-five trained subjects (21.3 ± 1.9 years) were recruited. All subjects participated in a high-volume resistance training session consisting of 3 sets of full squats, bench press, deadlifts, pullups, dips, bent over rows, shoulder press, and barbell curls and extensions. Pre-PRS and post-PRS measurements (0-10), soreness, CK, cortisol, and testosterone were measured before and 48 hours after training. Perceived recovery scale declined from 8.6 ± 2.3 to 4.2 ± 1.85 (p < 0.05). Leg, chest, and arm soreness increased from pre- to postexercise. Creatine kinase significantly increased from pre- to postworkout (189.4 ± 100.2 to 512 ± 222.7 U/L). Cortisol, testosterone, and free testosterone did not change. There was an inverse relationship between CK and PRS (r = 0.58, p < 0.05). When muscle damage was low before training, cortisol and free and total testosterone were not correlated to PRS. However, when damage peaked at 48 hours postexercise, free, but not total, testosterone showed a low direct relationship with PRS (r = 0.2, p < 0.05). High-volume resistance exercise lowers PRS scores. These changes are partly explained by a rise in serum indices of muscle damage. Moreover, free testosterone seems to have a positive relationship with PRS.
Journal of Strength and Conditioning Research, 2012
Previous research has demonstrated that post-activation potentiation (PAP) increases in an intens... more Previous research has demonstrated that post-activation potentiation (PAP) increases in an intensity-dependent manner. However, these studies did not control for volume loads. The purpose of this study was to investigate the effects of varying intensities and rest period lengths, while controlling for volume load, on vertical jump (VJ) performance. Thirteen men, aged 21 6 3 years with an average relative full squat of 1.7 6 2 times their body weight, were recruited for this study. Participants were assigned to 3 different experimental sessions that required them to perform the back squat at 56% (low intensity), 70% (moderate intensity), and 93% (high intensity) of their 1 repetition maximums. Vertical jump height and power were recorded at 0, 2, 4, 8, and 12 minutes after squat. There was a significant condition by time interaction for VJ height and power, in which both variables did not change in the low-intensity condition, whereas decreasing immediately after squat for both the moderate-and high-intensity conditions. In the moderate-and high-intensity conditions, VJ height and power increased and peaked at minute 4 and returned to baseline by minutes 8 and 12. These results indicate that when controlling for total work, jump performance and power are enhanced similarly by moderate and high squat intensities. However, high-intensity workloads may prolong the duration of PAP. Therefore, athletes may use moderate-and high-intensity loads during warm-ups to improve jump performance and power.
Renal tubular cells elicit adaptive responses following exposure to nephrotoxins, such as cadmium... more Renal tubular cells elicit adaptive responses following exposure to nephrotoxins, such as cadmium. One response is the up-regulation of the 32-kDa redox-sensitive protein, heme oxygenase-1. Exposure of renal proximal tubular epithelial cells to 10 M cadmium demonstrated induction (ϳ20-fold) of heme oxygenase-1 mRNA and protein. Using a 4.5-kb human heme oxygenase-1 promoter construct, the importance of a previously identified cadmium response element (TGCTAGAT) in HeLa cells was verified in renal epithelial cells. Specific protein-DNA interaction with this sequence was demonstrated using nuclear extracts from cadmium-treated cells. Yeast one-hybrid screen of a human kidney cDNA library resulted in the identification of pescadillo, a unique nucleolar, developmental protein, as an interacting protein with the cadmium response element and was confirmed by chromatin immunoprecipitation in vivo and gel shift assays with purified glutathione S-transferase-pescadillo protein in vitro. The specificity of the DNA-protein interaction was verified by the absence of a binding complex when the core sequence of the cadmium response element was mutated or deleted. In addition, B23/nucleophosmin, another nucleolar protein, did not interact with the cadmium response sequence. Overexpression of pescadillo resulted in increased activity of the 4.5-kb human heme oxygenase-1 promoter construct but failed to activate this construct when the cadmium response sequence was mutated. The findings demonstrate the important and previously unrecognized role of pescadillo as a DNA-binding protein interacting specifically with the cadmium response element of the human heme oxygenase-1 gene.
Methods A three-phase double-blind, placebo-and dietcontrolled randomized intervention study was ... more Methods A three-phase double-blind, placebo-and dietcontrolled randomized intervention study was conducted. Phase 1 was an 8-week-periodized resistance-training program; Phase 2 was a 2-week overreaching cycle; and Phase 3 was a 2-week taper. Muscle mass, strength, and power were examined at weeks 0, 4, 8, and 12 to assess the chronic effects of HMB-FA; and assessment of these, as well as cortisol, testosterone, and creatine kinase (cK) was performed at weeks 9 and 10 of the overreaching cycle. Results HMB-FA resulted in increased total strength (bench press, squat, and deadlift combined) over the 12-week training (77.1 ± 18.4 vs. 25.3 ± 22.0 kg, p < 0.001); a greater increase in vertical jump power (991 ± 168 vs. 630 ± 167 W, p < 0.001); and increased lean body mass gain (7.4 ± 4.2 vs. 2.1 ± 6.1 kg, p < 0.001) in HMB-FA-and placebo-supplemented groups,
American Journal of Physiology-Renal Physiology, 2004
Heme oxygenases (HOs) catalyze the rate-limiting step in heme degradation, resulting in the forma... more Heme oxygenases (HOs) catalyze the rate-limiting step in heme degradation, resulting in the formation of iron, carbon monoxide, and biliverdin, the latter of which is subsequently converted to bilirubin by biliverdin reductase. Recent attention has focused on the biological effects of product(s) of this enzymatic reaction, which have important antioxidant, anti-inflammatory, and cytoprotective functions. Two major isoforms of the HO enzyme have been described: an inducible isoform, HO-1, and a constitutively expressed isoform, HO-2. A third isoform, HO-3, closely related to HO-2, has also been described. Several stimuli implicated in the pathogenesis of renal injury, such as heme, nitric oxide, growth factors, angiotensin II, cytokines, and nephrotoxins, induce HO-1. Induction of HO-1 occurs as an adaptive and beneficial response to these stimuli, as demonstrated by studies in renal and non-renal disease states. This review will focus on the molecular regulation of the HO-1 gene in ...
American journal of physiology. Renal physiology, 2003
Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, releasing iron, car... more Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, releasing iron, carbon monoxide, and biliverdin. Induction of HO-1 is an adaptive and beneficial response in renal and nonrenal settings of tissue injury. The purpose of this study was to characterize the regulation of the human HO-1 gene in renal proximal tubule and aortic endothelial cells in response to heme and cadmium. Evaluation of multiple human HO-1 promoter-reporter constructs up to -9.1 kb demonstrated only a partial response to heme and cadmium. In an effort to mimic endogenous stimulus-dependent levels of HO-1 induction, we evaluated the entire 12.5 kb of the human HO-1 gene, including introns and exons, in conjunction with a -4.5-kb human HO-1 promoter and observed significant heme- and cadmium-mediated induction of the reporter gene, suggesting the presence of an internal enhancer. Enhancer function was orientation independent and required a region between -3.5 and -4.5 kb of the human HO-1 p...
Expression of the highly conserved replication-dependent histone gene family increases dramatical... more Expression of the highly conserved replication-dependent histone gene family increases dramatically as a cell enters the S phase of the eukaryotic cell cycle. Requirements for normal histone gene expression in vivo include an element, designated α, located within the protein-encoding sequence of nucleosomal histone genes. Mutation of 5 of 7 nucleotides of the mouse H3.2 α element to yield the sequence found in an H3.3 replication-independent variant abolishes the DNA-protein interaction in vitro and reduces expression fourfold in vivo. A yeast one-hybrid screen of a HeLa cell cDNA library identified the protein responsible for recognition of the histone H3.2 α sequence as the transcription factor Yin Yang 1 (YY1). YY1 is a ubiquitous and highly conserved transcription factor reported to be involved in both activation and repression of gene expression. Here we report that the in vitro histone α DNA-protein interaction depends on YY1 and that mutation of the nucleotides required for t...
The purpose of the present study was to determine the effects of short-term supplementation with ... more The purpose of the present study was to determine the effects of short-term supplementation with the free acid form of β-hydroxy-β-methylbutyrate (HMB-FA) on indices of muscle damage, protein breakdown, recovery and hormone status following a high-volume resistance training session in trained athletes. A total of twenty resistance-trained males were recruited to participate in a high-volume resistance training session centred on full squats, bench presses and dead lifts. Subjects were randomly assigned to receive either 3 g/d of HMB-FA or a placebo. Immediately before the exercise session and 48 h post-exercise, serum creatine kinase (CK), urinary 3-methylhistadine (3-MH), testosterone, cortisol and perceived recovery status (PRS) scale measurements were taken. The results showed that CK increased to a greater extent in the placebo (329 %) than in the HMB-FA group (104 %) (P= 0·004,d= 1·6). There was also a significant change for PRS, which decreased to a greater extent in the place...
Proceedings of the National Academy of Sciences, 1996
The histone gene family in mammals consists of 15-20 genes for each class of nucleosomal histone ... more The histone gene family in mammals consists of 15-20 genes for each class of nucleosomal histone protein. These genes are classified as either replication-dependent or -independent in regard to their expression in the cell cycle. The expression of the replication-dependent histone genes increases dramatically as the cell prepares to enter S phase. Using mouse histone genes, we previously identified a coding region activating sequence (CRAS) involved in the upregulation of at least two (H2a and H3) and possibly all nucleosomal replication-dependent histone genes. Mutation of two seven-nucleotide elements, alpha and omega, within the H3 CRAS causes a decrease in expression in stably transfected Chinese hamster ovary cells comparable with the effect seen upon deletion of the entire CRAS. Further, nuclear proteins interact in a highly specific manner with nucleotides within these sequences. Mutation of these elements abolishes DNA/protein interactions in vitro. Here we report that the i...
Journal of the International Society of Sports Nutrition, 2012
Sikorski et al.: The acute effects of a free acid betahydoxy-beta-methyl butyrate supplement on m... more Sikorski et al.: The acute effects of a free acid betahydoxy-beta-methyl butyrate supplement on muscle damage following resistance training: a randomized, double-blind, placebo-controlled study.
Journal of Strength and Conditioning Research, 2013
Currently no research has investigated the relationship between muscle damage, hormonal status, a... more Currently no research has investigated the relationship between muscle damage, hormonal status, and perceived recovery scale (PRS). Therefore, the purpose of this study was to determine the effects of a high-volume training session on PRS and to determine the relationship between levels of testosterone, cortisol, and creatine kinase (CK) and PRS. Thirty-five trained subjects (21.3 ± 1.9 years) were recruited. All subjects participated in a high-volume resistance training session consisting of 3 sets of full squats, bench press, deadlifts, pullups, dips, bent over rows, shoulder press, and barbell curls and extensions. Pre-PRS and post-PRS measurements (0-10), soreness, CK, cortisol, and testosterone were measured before and 48 hours after training. Perceived recovery scale declined from 8.6 ± 2.3 to 4.2 ± 1.85 (p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). Leg, chest, and arm soreness increased from pre- to postexercise. Creatine kinase significantly increased from pre- to postworkout (189.4 ± 100.2 to 512 ± 222.7 U/L). Cortisol, testosterone, and free testosterone did not change. There was an inverse relationship between CK and PRS (r = 0.58, p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). When muscle damage was low before training, cortisol and free and total testosterone were not correlated to PRS. However, when damage peaked at 48 hours postexercise, free, but not total, testosterone showed a low direct relationship with PRS (r = 0.2, p &amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;amp;lt; 0.05). High-volume resistance exercise lowers PRS scores. These changes are partly explained by a rise in serum indices of muscle damage. Moreover, free testosterone seems to have a positive relationship with PRS.
Journal of Strength and Conditioning Research, 2012
Previous research has demonstrated that post-activation potentiation (PAP) increases in an intens... more Previous research has demonstrated that post-activation potentiation (PAP) increases in an intensity-dependent manner. However, these studies did not control for volume loads. The purpose of this study was to investigate the effects of varying intensities and rest period lengths, while controlling for volume load, on vertical jump (VJ) performance. Thirteen men, aged 21 6 3 years with an average relative full squat of 1.7 6 2 times their body weight, were recruited for this study. Participants were assigned to 3 different experimental sessions that required them to perform the back squat at 56% (low intensity), 70% (moderate intensity), and 93% (high intensity) of their 1 repetition maximums. Vertical jump height and power were recorded at 0, 2, 4, 8, and 12 minutes after squat. There was a significant condition by time interaction for VJ height and power, in which both variables did not change in the low-intensity condition, whereas decreasing immediately after squat for both the moderate-and high-intensity conditions. In the moderate-and high-intensity conditions, VJ height and power increased and peaked at minute 4 and returned to baseline by minutes 8 and 12. These results indicate that when controlling for total work, jump performance and power are enhanced similarly by moderate and high squat intensities. However, high-intensity workloads may prolong the duration of PAP. Therefore, athletes may use moderate-and high-intensity loads during warm-ups to improve jump performance and power.
Renal tubular cells elicit adaptive responses following exposure to nephrotoxins, such as cadmium... more Renal tubular cells elicit adaptive responses following exposure to nephrotoxins, such as cadmium. One response is the up-regulation of the 32-kDa redox-sensitive protein, heme oxygenase-1. Exposure of renal proximal tubular epithelial cells to 10 M cadmium demonstrated induction (ϳ20-fold) of heme oxygenase-1 mRNA and protein. Using a 4.5-kb human heme oxygenase-1 promoter construct, the importance of a previously identified cadmium response element (TGCTAGAT) in HeLa cells was verified in renal epithelial cells. Specific protein-DNA interaction with this sequence was demonstrated using nuclear extracts from cadmium-treated cells. Yeast one-hybrid screen of a human kidney cDNA library resulted in the identification of pescadillo, a unique nucleolar, developmental protein, as an interacting protein with the cadmium response element and was confirmed by chromatin immunoprecipitation in vivo and gel shift assays with purified glutathione S-transferase-pescadillo protein in vitro. The specificity of the DNA-protein interaction was verified by the absence of a binding complex when the core sequence of the cadmium response element was mutated or deleted. In addition, B23/nucleophosmin, another nucleolar protein, did not interact with the cadmium response sequence. Overexpression of pescadillo resulted in increased activity of the 4.5-kb human heme oxygenase-1 promoter construct but failed to activate this construct when the cadmium response sequence was mutated. The findings demonstrate the important and previously unrecognized role of pescadillo as a DNA-binding protein interacting specifically with the cadmium response element of the human heme oxygenase-1 gene.
Methods A three-phase double-blind, placebo-and dietcontrolled randomized intervention study was ... more Methods A three-phase double-blind, placebo-and dietcontrolled randomized intervention study was conducted. Phase 1 was an 8-week-periodized resistance-training program; Phase 2 was a 2-week overreaching cycle; and Phase 3 was a 2-week taper. Muscle mass, strength, and power were examined at weeks 0, 4, 8, and 12 to assess the chronic effects of HMB-FA; and assessment of these, as well as cortisol, testosterone, and creatine kinase (cK) was performed at weeks 9 and 10 of the overreaching cycle. Results HMB-FA resulted in increased total strength (bench press, squat, and deadlift combined) over the 12-week training (77.1 ± 18.4 vs. 25.3 ± 22.0 kg, p < 0.001); a greater increase in vertical jump power (991 ± 168 vs. 630 ± 167 W, p < 0.001); and increased lean body mass gain (7.4 ± 4.2 vs. 2.1 ± 6.1 kg, p < 0.001) in HMB-FA-and placebo-supplemented groups,
American Journal of Physiology-Renal Physiology, 2004
Heme oxygenases (HOs) catalyze the rate-limiting step in heme degradation, resulting in the forma... more Heme oxygenases (HOs) catalyze the rate-limiting step in heme degradation, resulting in the formation of iron, carbon monoxide, and biliverdin, the latter of which is subsequently converted to bilirubin by biliverdin reductase. Recent attention has focused on the biological effects of product(s) of this enzymatic reaction, which have important antioxidant, anti-inflammatory, and cytoprotective functions. Two major isoforms of the HO enzyme have been described: an inducible isoform, HO-1, and a constitutively expressed isoform, HO-2. A third isoform, HO-3, closely related to HO-2, has also been described. Several stimuli implicated in the pathogenesis of renal injury, such as heme, nitric oxide, growth factors, angiotensin II, cytokines, and nephrotoxins, induce HO-1. Induction of HO-1 occurs as an adaptive and beneficial response to these stimuli, as demonstrated by studies in renal and non-renal disease states. This review will focus on the molecular regulation of the HO-1 gene in ...
American journal of physiology. Renal physiology, 2003
Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, releasing iron, car... more Heme oxygenase-1 (HO-1) catalyzes the rate-limiting step in heme degradation, releasing iron, carbon monoxide, and biliverdin. Induction of HO-1 is an adaptive and beneficial response in renal and nonrenal settings of tissue injury. The purpose of this study was to characterize the regulation of the human HO-1 gene in renal proximal tubule and aortic endothelial cells in response to heme and cadmium. Evaluation of multiple human HO-1 promoter-reporter constructs up to -9.1 kb demonstrated only a partial response to heme and cadmium. In an effort to mimic endogenous stimulus-dependent levels of HO-1 induction, we evaluated the entire 12.5 kb of the human HO-1 gene, including introns and exons, in conjunction with a -4.5-kb human HO-1 promoter and observed significant heme- and cadmium-mediated induction of the reporter gene, suggesting the presence of an internal enhancer. Enhancer function was orientation independent and required a region between -3.5 and -4.5 kb of the human HO-1 p...
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Papers by Eric Sikorski