Caffeine Abstinence in Habituated Users: Cardiovascular Responses to Exercise With Blood Flow Restriction.

CONTEXT: Blood flow restriction resistance exercise studies often require caffeine abstinence to avoid cardiovascular effects that could change the blood flow restriction stimulus. However, effects may be attenuated for habituated users. OBJECTIVE: To compare cardiovascular responses to blood flow restriction resistance exercise when habituated users consume or abstain from caffeine. DESIGN: Thirty participants completed a 3-visit within-subject study beginning with familiarization and caffeine intake questionnaire. METHODS: Visits 2 and 3 consisted of blood flow restriction resistance exercise (3 sets bicep curls to failure, 30% 1-repetition max, 40% arterial occlusion pressure [AOP]), following participants' normal caffeine consumption (CAFF) or abstaining (ABS). AOP, systolic (SBP) and diastolic (DBP) blood pressure, and heart rate were measured preexercise and postexercise. Prevalues and preexercise to postexercise change scores for SBP, DBP, AOP (all millimeters of mercury), heart rate (in beats per minute), and repetitions were compared between conditions. Results are represented as mean (SD). RESULTS: Preexercise AOP was similar for CAFF (137.8 [14.4]) and ABS (137.1 [14.9], BF10 = 0.2), although pre-SBP was higher for CAFF (115.4 [9.8]) than ABS (112.3 [9.4], BF10 = 1.9). Pre-DBP was similar between conditions. The exercise-induced change in AOP was greater for CAFF (18.4 [11.2]) than ABS (13.2 [14.9]), though evidence was anecdotal (BF10 = 0.7). Exercise-induced changes in SBP, DBP, and heart rate were similar between conditions (all BF10 ≤ 0.40). More repetitions were completed for CAFF (63 [26]) than ABS (57 [17], BF10 = 2.1). CONCLUSIONS: The findings of this study suggest that for habituated users, maintaining daily caffeine habits will not have substantial effects on common cardiovascular variables relevant to blood flow restriction.

Affective and Perceptual Responses During an 8-Week Resistance Training to Failure Intervention at Low vs. High Loads in Untrained Women.

ABSTRACT: Anderson, AlOK, Voskuil, CC, Byrd, MT, Garver, MJ, Rickard, AJ, Miller, WM, Bergstrom, HC, and Dinyer McNeely, TK. Affective and perceptual responses during an 8-week resistance training to failure intervention at low vs. high loads in untrained women. J Strength Cond Res 37(3): 546-554, 2023-This study examined the effects of resistance training (RT) to failure on the perceptual and affective responses, intent-to-continue RT to failure in a self-initiated session, and affect-intent relationship. Twenty-three untrained women (mean ± SD : age 21.2 ± 2.2 years; height 167 ± 5.7 cm; body mass, 62.3 ± 16.2 kg) completed an 8-week, full-body RT to failure intervention at a low (30% 1RM; n = 11) or high (80% 1RM; n = 12) load. The Borg's rating of perceived exertion (RPE) scale was used to assess the acute (aRPE) and session (sRPE) RPE immediately after repetition failure and each training session, respectively. Immediately, 15-minute, and 60-minute postsession affective responses were assessed using the feeling scale (FS; -5 to +5), and intent to continue to RT was assessed on a scale of 0-100% intention. During week 4 (W4) and week 8 (W8), aRPE (W4: 18 ± 2, W8: 18 ± 2; p ≤ 0.032) and sRPE (W4: 17 ± 2, W8: 18 ± 1; p ≤ 0.018) were greater than that during week 1 (W1; aRPE: 17 ± 2; sRPE: 16 ± 2). The FS responses increased from immediately to 60-minute postsession during W4 ( p ≤ 0.019) and W8 ( p ≤ 0.049). The correlation between affect and intent-to-continue RT increased from W1 ( r = 0.416) to W8 ( r = 0.777). Regardless of load, untrained women reported similar perceptual, affective, and intention responses. These variables should be considered to improve RT program adoption and adherence in women.

Effects of Maximal vs. Submaximal Isometric Fatiguing Exercise on Subsequent Submaximal Exercise Performance.

Miller, WM, Ye, X, and Jeon, S. Effects of maximal vs. submaximal isometric fatiguing exercise on subsequent submaximal exercise performance. J Strength Cond Res 34(7): 1875-1883, 2020-Exercise-induced muscle fatigue directly influences subsequent exercise performance. Thus, we examined task failure times for submaximal intermittent fatiguing isometric contractions performed after a bout of sustained maximal vs. submaximal isometric fatiguing contractions with the dominant elbow flexors. Twenty physically active individuals 8 men (mean ± SD = 21.4 ± 1.8 years; 80.9 ± 12.5 kg; 180 ± 6.4 cm) and 12 women (mean ± SD = 21.4 ± 2.7; 66.8 ± 15.6 kg; 165.7 ± 7.1 cm) participated in a 3-visit randomized cross-over study. Visit 1 included familiarization, and 2 and 3 were randomized for sustained maximal (until force was below 50% of the maximal force value) or submaximal conditions (50% of maximal force until task failure), followed by submaximal intermittent isometric contractions to task failure. Surface electromyography was recorded through the biceps brachii during all fatiguing contractions. Task failure time was significantly shorter for the maximal compared with submaximal condition, and no significant difference in sex or condition was found for the subsequent fatiguing condition. Electromyography amplitude significantly increased in the submaximal intermittent isometric contractions from prefatigue and first and final postfatigue with no condition or sex differences. Electromyography mean frequency significantly decreased from prefatigue and first and final postfatigue for both sexes, with no condition or sex differences. With both maximal and submaximal exercises inducing the same level of force deficit, our results suggested that both exercises might have imposed a similar burden on the neuromuscular system, thereby not providing differential effects on the subsequent submaximal exercise performance.

The Effects of a Preconditioning Rolling Session on Subsequent Eccentric Exercise-Induced Muscle Damage.

West, JT, Miller, WM, Jeon, S, and Ye, X. The effects of a preconditioning rolling session on subsequent eccentric exercise-induced muscle damage. J Strength Cond Res 34(8): 2112-2119, 2020-The aim of this study was to examine the effects of a preexercise unilateral quadriceps muscle rolling intervention on subsequent ipsilateral (IPSI) or contralateral (CTRA) knee extension eccentric exercise-induced muscle damage. Twenty-seven healthy volunteers (14 men) underwent an eccentric exercise protocol (6 sets of 10 repetitions with 75% of the maximal isometric strength) with a single-leg knee extension machine. Before the eccentric exercise, the subjects were randomly assigned to either (a) IPSI group: rolling the ipsilateral knee extensor muscles, (b) CTRA: rolling the contralateral muscles, or (c) Control: sitting for 6 minutes (same duration as the rolling intervention protocol) relaxed. The muscle soreness, passive knee extension range of motion, and knee extension isometric strength were measured before, immediately, 24 hours, and 48 hours after exercise. The magnitudes of the range of motion decrement were attenuated in both the IPSI (p = 0.031) and CTRA (p = 0.014) groups 24 hours after the eccentric exercise, when compared with the control. Isometric strength (p = 0.783) and muscle soreness (p = 0.586) responses were not significantly different among the 3 groups (time points and sexes merged). Additionally, women displayed an overall faster recovery than men in isometric strength (p = 0.001) and muscle soreness (p = 0.024), evidenced by the measurements at 48 hours after exercise. Our study suggests that unilateral quadriceps rolling intervention before high-intensity muscle-damaging exercise has a beneficial effect on maintaining range of motion in both the ipsilateral and contralateral muscles.

Comparison of Myoelectric Activity During a Suspension-Based and Traditional Split Squat.

Miller, WM, Barnes, JT, Sofo, SS, and Wagganer, JD. Comparison of myoelectric activity during a suspension-based and traditional split squat. J Strength Cond Res 33(12): 3236-3241, 2019-Over several decades, traditional resistance training has incorporated body-mass suspension training (SPT), for example, Total-Body Resistance Exercise (TRX); however, very little research has been completed investigating the effects of lower body SPT. Therefore, the purpose of this study was to assess sex differences in myoelectric activity via electromyography while performing a stable (i.e., bench) traditional split squat (TSS) compared with suspension body-mass split squat (TRXSS). Nineteen recreationally active individuals (mean ± SD = men: n = 9; 21.4 ± 2.1 years; 177.2 ± 9.0 cm; 84.2 ± 8.8 kg; women: n = 10; 21.5 ± 1.6 years; 160.7 ± 8.5 cm; 59.1 ± 7.9 kg) participated in this investigation. Subjects performed 3 training sessions, 1 familiarization and 2 (one for each variation) split squat. The TRXSS was performed by placing the rear foot within the foot cradle of the strap, while the TSS required placement of the rear foot on a stable bench (40.64 cm). The TRXSS resulted in higher peak and average myoelectric activity overall, as well as greater gluteus maximus myoelectric activity. No gender or rectus femoris myoelectric activity differences were found. The increased myoelectric activity of the gluteus maximus during the TRXSS was most likely due to increased medial/lateral stabilization demands on the suspended leg. These findings provide insight into TRX as a useful exercise modality for recreationally active or rehabilitative purposes.

A systematic review of the human health and social well-being outcomes of green infrastructure for stormwater and flood management.

BACKGROUND: The increase in frequency and intensity of urban flooding is a global challenge. Flooding directly impacts residents of industrialized cities with aging combined sewer systems, as well as cities with less centralized infrastructure to manage stormwater, fecal sludge, and wastewater. Green infrastructure is growing in popularity as a sustainable strategy to mimic nature-based flood management. Although its technical performance has been extensively studied, little is known about the effects of green stormwater infrastructure on human health and social well-being. METHODS: We conducted a multidisciplinary systematic review of peer-reviewed and gray literature on the effects of green infrastructure for stormwater and flood management on individuals', households', and communities' a) physical health; b) mental health; c) economic well-being; and d) flood resilience and social acceptance of green infrastructure. We systematically searched databases such as PubMed, Web of Science, and Scopus; the first 300 results in Google Scholar; and websites of key organizations including the United States Environmental Protection Agency. Study quality and strength of evidence was assessed for included studies, and descriptive data were extracted for a narrative summary. RESULTS: Out of 21,213 initial results, only 18 studies reported health or social well-being outcomes. Seven of these studies used primary data, and none allowed for causal inference. No studies connected green infrastructure for stormwater and flood management to mental or physical health outcomes. Thirteen studies were identified on economic outcomes, largely reporting a positive association between green infrastructure and property values. Five studies assessed changes in perceptions about green infrastructure, but with mixed results. Nearly half of all included studies were from Portland, Oregon. CONCLUSIONS: This global systematic review highlights the minimal evidence on human health and social well-being relating to green infrastructure for stormwater and flood management. To enable scale-up of this type of infrastructure to reduce flooding and improve ecological and human well-being, widespread acceptance of green infrastructure will be essential. Policymakers and planners need evidence on the full range of benefits from different contexts to enable financing and implementation of instfrastructure options, especially in highly urbanized, flood-prone settings around the world. Therefore, experts in social science, public health, and program evaluation must be integrated into interdisciplinary green infrastructure research to better relate infrastructure design to tangible human outcomes.

Low-Load vs. High-Load Resistance Training to Failure on One Repetition Maximum Strength and Body Composition in Untrained Women.

Dinyer, TK, Byrd, MT, Garver, MJ, Rickard, AJ, Miller, WM, Burns, S, Clasey, JL, and Bergstrom, HC. Low-load vs. high-load resistance training to failure on one repetition maximum strength and body composition in untrained women. J Strength Cond Res 33(7): 1737-1744, 2019-This study examined the effects of resistance training (RT) to failure at low and high loads on one repetition maximum (1RM) strength and body composition (bone- and fat-free mass [BFFM] and percent body fat [%BF]) in untrained women. Twenty-three untrained women (age: 21.2 ± 2.2 years; height: 167.1 ± 5.7 cm; body mass: 62.3 ± 16.2 kg) completed a 12-week RT to failure intervention at a low (30% 1RM) (n = 11) or high (80% 1RM) (n = 12) load. On weeks 1, 5, and 12, subjects completed 1RM testing for 4 different exercises (leg extension [LE], seated military press [SMP], leg curl [LC], and lat pull down [LPD]) and a dual-energy x-ray absorptiometry scan to assess body composition. During weeks 2-4 and 6-7, the subjects completed 2 sets to failure for each exercise. During weeks 8-11, the subjects completed 3 sets to failure for each exercise. The 1RM strength increased from week 1 to week 5 (LE: 18 ± 16%; SMP: 9 ± 11%; LC: 12 ± 22%; LPD: 13 ± 9%), week 1 to week 12 (LE: 32 ± 24%; SMP: 17 ± 14%; LC: 23 ± 26%; LPD: 25 ± 13%), and week 5 to week 12 (LE: 11 ± 9%; SMP: 7 ± 9%; LC: 10 ± 7%; LPD: 11 ± 11%) in each exercise, with no significant differences between groups. There were no significant changes in BFFM (p = 0.241) or %BF (p = 0.740) for either group. Resistance training to failure at 30% 1RM and 80% 1RM resulted in similar increases in 1RM strength, but no change in BFFM or %BF. Untrained women can increase 1RM strength during RT at low and high loads, if repetitions are taken to failure.

Traditional Versus Suspended Push-up Muscle Activation in Athletes and Sedentary Women.

Syed-Abdul, MM, Soni, DS, Miller, WM, Johnson, RJ, Barnes, JT, Pujol, TJ, and Wagganer, JD. Traditional versus suspended push-up muscle activation in athletes and sedentary women. J Strength Cond Res 32(7): 1816-1820, 2018-Many strength training programs incorporate push-up exercises, which primarily activate upper-body muscles. Past data support the fact that shoulder girdle muscles (i.e., triceps (T) and anterior deltoids [AD]) exhibit greater electromyography (EMG) activity when a push-up is performed on an unstable (i.e., suspended [SP]) vs. stable (i.e., traditional [TD]) surface (). Sixty-nine healthy female volunteers (soccer players [SO], n = 24; gymnasts [GY], n = 21; sedentary [SE], n = 24) performed three TD and three SP push-ups. Muscle activation, expressed as absolute integral (mV), was measured using EMG analysis. Significant increases in muscle activation were exhibited by GY (TD: p < 0.01 and SP: p < 0.001) and SO (TD: p < 0.05 and SP: p < 0.05) compared to SE for the T muscle. Only SO (p < 0.05) exhibited significantly higher muscle activation during the SP versus TD. For the AD, values were significantly higher for SO (TD: p < 0.001 and SP: p < 0.001) and GY (TD: p < 0.01 and SP: p < 0.01) compared to the SE group. In addition, significantly higher values were exhibited by SO compared with GY during TD push-ups (p < 0.01). Both the SO (p < 0.05) and GY (p < 0.05) group exhibited significantly higher values during SP versus TD push-ups. Finally, values were significantly higher for the AD compared to the T muscle only in the SO group during TD (p < 0.01) and SP (p < 0.05) push-ups. Data from this study for trained women (i.e., SO) are consistent with previous studies, whereas for untrained women (i.e., SE) the findings differed during TD and SP push-ups for both muscles. Differences were also observed between female SO and GY are unexplainable and therefore need further investigation.

The Where and How for Reaching Transgender Women and Men Who Have Sex with Men with HIV Prevention Services in Guatemala.

This study aims to describe the transgender women and men who have sex with men (MSM) missed through venue-based sampling and illustrate how data on venues can be used to prioritize service delivery. Respondent-driven sampling (RDS) and time-location sampling (TLS) were used concurrently in 2010 for behavioral surveillance among MSM and transgender women in Guatemala City. RDS recruits who did not frequent venues (n = 106) were compared to TLS recruits (n = 609). TLS participants recruited at different types of venues were compared. RDS recruits who did not frequent venues were less educated, less likely to identify as gay, more likely to have concurrent partners and female sexual partners. Participants recruited at NGOs, saunas, hotels, streets and parks had more partners, were more likely to receive money for sex or have concurrent partners. Prevention programs for MSM and transgender women should characterize social venues and people that frequent them and improve service coverage through venues and social networks.

SIRT1 is a critical regulator of K562 cell growth, survival, and differentiation.

Inhibition of histone deacetylases (HDACi) has emerged as a promising approach in the treatment of many types of cancer, including leukemias. Among the HDACs, Class III HDACs, also known as sirtuins (SIRTs), are unique in that their function is directly related to the cell's metabolic state through their dependency on the co-factor NAD(+). In this study, we examined the relation between SIRTs and the growth, survival, and differentiation of K562 erythroleukemia cells. Using a mass spectrometry approach we previously developed, we show that SIRT expression and deacetylase activity in these cells changes greatly with differentiation state (undifferentiated vs. megakaryocytic differentiation vs. erythroid differentiation). Moreover, SIRT1 is crucially involved in regulating the differentiation state. Overexpression of wildtype (but not deacetylase mutant) SIRT1 resulted in upregulation of glycophorin A, ~2-fold increase in the mRNA levels of α, γ, ε, and ζ-globins, and spontaneous hemoglobinization. Hemin-induced differentiation was also enhanced by (and depended on) higher SIRT1 levels. Since K562 cells are bipotent, we also investigated whether SIRT1 modulation affected their ability to undergo megakaryocytic (MK) differentiation. SIRT1 was required for commitment to the MK lineage and subsequent maturation, but was not directly involved in polyploidization of either K562 cells or an already-MK-committed cell line, CHRF-288-11. The observed blockage in commitment to the MK lineage was associated with a dramatic decrease in the formation of autophagic vacuoles, which was previously shown to be required for K562 cell MK commitment. Autophagy-associated conversion of the protein LC3-I to LC3-II was greatly enhanced by overexpression of wildtype SIRT1, further suggesting a functional connection between SIRT1, autophagy, and MK differentiation. Based on its clear effects on autophagy, we also examined the effect of SIRT1 modulation on stress responses. Consistent with results of prior studies, we found that SIRT1 silencing modestly promoted drug-induced apoptosis, while overexpression was protective. Furthermore, pan-SIRT inhibition mediated by nicotinamide pre-treatment substantially increased imatinib-induced apoptosis. Altogether, our results suggest a complex role for SIRT1 in regulating many aspects of K562 cell state and stress response. These observations warrant further investigation using normal and leukemic primary cell models. We further suggest that, ultimately, a well-defined mapping of HDACs to their substrates and corresponding signaling pathways will be important for optimally designing HDACi-based therapeutic approaches.

Hoxa10 null animals exhibit reduced platelet biogenesis.

The transcription factor HOXA10 is an important regulator of myelopoiesis. Engineered over-expression of Hoxa10 in mice results in a myeloproliferative disorder that progresses to acute myeloid leukaemia (AML) over time, and in humans over-expression is associated with poor outcomes in AML. Here, we report that loss of Hoxa10 expression in mice results in reduced platelet count and platelet production, but does not affect clotting efficiency. About 40% fewer platelets were found in Hoxa10 null animals in comparison to wild type littermates. We found a nearly 50% reduction in the percentage of reticulated platelets in Hoxa10 null mice, suggesting deficient platelet production. Furthermore, Hoxa10 null animals recovered less efficiently from induced thrombocytopenia, supporting our hypothesis of defective platelet production. This also correlated with reduced colony formation potential of stem and progenitor cells seeded in megakaryocyte-enhancing conditions in vitro. Together, our results indicate that HOXA10 is important for megakaryopoiesis and platelet biogenesis.

Megakaryocyte Polyploidization and Proplatelet Formation in Low-Attachment Conditions.

In vitro-derived platelets (PLTs), which could provide an alternative source of PLTs for patient transfusions, are formed from polyploid megakaryocytes (MKs) that extend long cytoplasmic projections, termed proplatelets (proPLTs). In this study, we compared polyploidization and proPLT formation (PPF) of MKs cultured on surfaces that either promote or inhibit protein adsorption and subsequent cell adhesion. A megakaryoblastic cell line exhibited increased polyploidization and arrested PPF on a low-attachment surface. Primary human MKs also showed low levels of PPF on the same surface, but no difference in ploidy. Importantly, both cell types exhibited accelerated PPF after transfer to a surface that supports attachment, suggesting that pre-culture on a non-adhesive surface may facilitate synchronization of PPF and PLT generation in culture.

Separation of in-vitro-derived megakaryocytes and platelets using spinning-membrane filtration.

In-vitro-derived platelets (PLTs) could potentially overcome problems associated with donated PLTs, including contamination and alloimmunization. Although several groups have produced functional PLTs from stem cells in vitro, the challenge of developing this technology to yield transfusable PLT units has yet to be addressed. The asynchronous nature of in vitro PLT generation makes a single harvest point infeasible for collecting PLTs as soon as they are formed. The current standard of performing manual centrifugations to separate PLTs from nucleated cells at multiple points during culture is labor-intensive, imprecise, and difficult to standardize in accordance with current Good Manufacturing Practices (cGMP). In an effort to develop a more effective method, we adapted a commercially-available, spinning-membrane filtration device to separate in-vitro-derived PLTs from nucleated cells and recover immature megakaryocytes (MKs), the precursor cells to PLTs, for continued culture. Processing a mixture of in-vitro-derived MKs and PLTs on the adapted device yielded a pure PLT population and did not induce PLT pre-activation. MKs recovered from the separation process were unaffected with respect to viability and ploidy, and were able to generate PLTs after reseeding in culture. Being able to efficiently harvest in-vitro-derived PLTs brings this technology one step closer to clinical relevance.

Dynamic transcription factor activity profiles reveal key regulatory interactions during megakaryocytic and erythroid differentiation.

The directed differentiation toward erythroid (E) or megakaryocytic (MK) lineages by the MK-E progenitor (MEP) could enhance the ex vivo generation of red blood cells and platelets for therapeutic transfusions. The lineage choice at the MEP bifurcation is controlled in large part by activity within the intracellular signal transduction network, the output of which determines the activity of transcription factors (TFs) and ultimately gene expression. Although many TFs have been implicated, E or MK differentiation is a complex process requiring multiple days, and the dynamics of TF activities during commitment and terminal maturation are relatively unexplored. Herein, we applied a living cell array for the large-scale, dynamic quantification of TF activities during MEP bifurcation. A panel of hematopoietic TFs (GATA-1, GATA-2, SCL/TAL1, FLI-1, NF-E2, PU.1, c-Myb) was characterized during E and MK differentiation of bipotent K562 cells. Dynamic TF activity profiles associated with differentiation towards each lineage were identified, and validated with previous reports. From these activity profiles, we show that GATA-1 is an important hub during early hemin- and PMA-induced differentiation, and reveal several characteristic TF interactions for E and MK differentiation that confirm regulatory mechanisms documented in the literature. Additionally, we highlight several novel TF interactions at various stages of E and MK differentiation. Furthermore, we investigated the mechanism by which nicotinamide (NIC) promoted terminal MK maturation using an MK-committed cell line, CHRF-288-11 (CHRF). Concomitant with its enhancement of ploidy, NIC strongly enhanced the activity of three TFs with known involvement in terminal MK maturation: FLI-1, NF-E2, and p53. Dynamic profiling of TF activity represents a novel tool to complement traditional assays focused on mRNA and protein expression levels to understand progenitor cell differentiation.

Comparing the acute responses between a manual and automated blood flow restriction system.

The purpose of this study was to compare acute responses between manual and automated blood flow restriction (BFR) systems. Methods: A total of 33 individuals completed this study. On visit 1, arterial occlusion pressure (AOP, mm Hg), cardiovascular responses, and discomfort (RPE-D) were measured with each BFR system at rest. On visit 2, unilateral bicep curls were completed [30% one-repetition maximum; 50% AOP] with one system per arm. Muscle thickness (MT, cm) and maximal force (N) were assessed before (pre), immediately (post-0), 5 min (post-5), and 10 min (post-10) post-exercise. Ratings of perceived exertion (RPE-E) and ratings of perceived discomfort (RPE-D) were assessed throughout the exercise. AOP and repetitions were compared with Bayesian paired t-tests. Other outcomes were compared with Bayesian RMANOVAs. BF10 represents the likelihood of the best model vs. the null. The results are presented as mean ± SD. Results: Supine cardiovascular responses and RPE-D were similar for manual and automated (all BF10 ≤ 0.2). Supine AOP for manual (157 ± 20) was higher than that of automated (142 ± 17; BF10 = 44496.0), but similar while standing (manual: 141 ± 17; automated: 141 ± 22; BF10 = 0.2). MT (time, BF10 = 6.047e + 40) increased from Pre (3.9 ± 0.7) to Post-0 (4.4 ± 0.8; BF10 = 2.969e + 28), with Post-0 higher than Post-5 (4.3 ± 0.8) and Post-10 (4.3 ± 0.8; both BF10 ≥ 275.2). Force (time, BF10 = 1.246e + 29) decreased from Pre (234.5 ± 79.2) to Post-0 (149.8 ± 52.3; BF10 = 2.720e + 22) and increased from Post-0 to Post-5 (193.3 ± 72.7; BF10 = 1.744e + 13), with Post-5 to Post-10 (194.0 ± 70.6; BF10 = 0.2) being similar. RPE-E increased over sets. RPE-D was lower for manual than automated. Repetitions per set were higher for manual (Set 1: 37 ± 18; Set 4: 9 ± 5) than automated (Set 1: 30 ± 7; Set 4: 7 ± 3; all BF10 ≥ 9.7). Conclusion: Under the same relative pressure, responses are mostly similar between BFR systems, although a manual system led to lower exercise discomfort and more repetitions.

Post-occlusive reactive hyperemia in habituated caffeine users: Effects of abstaining versus consuming typical doses.

BACKGROUND: Post-occlusive reactive hyperemia (PORH) typically requires caffeine abstinence. For habitual users, it is unknown if abstinence affects PORH. OBJECTIVE: Compare PORH after habitual users consume or abstain from caffeine. METHODS: On separate visits (within-subject), PORH was measured in 30 participants without abstinence from typical caffeine doses (CAFF) and with abstinence (ABS). Measurements included baseline and peak hyperemic velocity, tissue saturation index slopes during ischemia (Slope 1) and following cuff deflation (Slope 2), resting arterial occlusion pressure (AOP), heart rate (HR), systolic (SBP), and diastolic (DBP) blood pressure. All variables were compared using Bayesian paired t-tests. BF10 = likelihood of alternative vs null. Results are mean±SD. RESULTS: Comparing baseline velocity (cm/s) between CAFF (9.3±4.8) and ABS (7.5±4.9) yielded anecdotal evidence (BF10 = 1.0). Peak hyperemic velocity (cm/s) was similar (CAFF = 77.3±16.7; ABS = 77.6±19.0, BF10 = 0.20). For slopes (TSI% /s), CAFF Slope 1 = -0.11±0.04 and Slope 2 = 1.9±0.46 were similar (both BF10≤0.20) to ABS Slope 1 = -0.12±0.03 and Slope 2 = 1.8±0.42. SBP and DBP (mmHg) were both similar (CAFF SBP = 116.0±9.8, DBP = 69.6±5.8; ABS SBP = 115.5±10.7, DBP = 69.5±5.4; both BF10≤0.22). Comparing AOP (mmHg) (CAFF = 146.6±15.0; ABS = 143.0±16.4) yielded anecdotal evidence (BF10 = 0.46). HR (bpm) was similar (CAFF = 66.5±12.3; ABS = 66.9±13.0; BF10 = 0.20). CONCLUSIONS: In habitual users, consuming or abstaining from typical caffeine doses does not appear to affect post-occlusive reactive hyperemia.

Profiling deacetylase activities in cell lysates with peptide arrays and SAMDI mass spectrometry.

The development of arrays that can profile molecular activities in cells is important to understanding signaling pathways in normal and pathological settings. While oligonucleotide arrays are now routinely used to profile global gene expression, there is still a lack of tools for profiling enzyme activities in cell lysates. This paper describes the combination of peptide arrays formed on self-assembled monolayers and mass spectrometry to provide a label-free approach for identifying patterns of enzyme activities in cell lysates. The approach is demonstrated by profiling lysine deacetylase (KDAC) activities in cell lysates of the CHRF megakaryocytic (Mk) cell line. Class-specific deacetylase inhibitors were used to show that terminal Mk differentiation of CHRF cells is marked by a pronounced decrease in sirtuin activity and by little change in activity of KDACs 1-11. This work establishes a platform that can be used to identify changes in global activity profiles of cell lysates for a wide variety of enzymatic activities.

Administration of nicotinamide does not increase platelet levels in mice.

Elucidating ways to enhance megakaryopoiesis in vivo would have therapeutic applications for thrombocytopenia and transfusion medicine. Nicotinamide has been shown to enhance endomitosis in megakaryocytes cultured in vitro, suggesting that it may be beneficial for the production of platelets in culture. We hypothesized that regular injections of nicotinamide in mice would also increase platelets in vivo. However, we found that platelet counts were reduced by about 25% with daily injections of nicotinamide. Altering the schedule, duration, or nicotinamide dose did not improve platelet production. Consistent with lower platelet levels, nicotinamide also tended to decrease megakaryocyte frequency in sternum and spleen sections, as well as colony formation in vitro by bone marrow progenitor cells. However, there was no effect on the fraction or ploidy of CD41(+) cells harvested from bone marrow. Together, our results suggest that, although nicotinamide increases polyploidization of megakaryocytes in culture, it does not have translatable effects in vivo.

Proposed megakaryocytic regulon of p53: the genes engaged to control cell cycle and apoptosis during megakaryocytic differentiation.

During endomitosis, megakaryocytes undergo several rounds of DNA synthesis without division leading to polyploidization. In primary megakaryocytes and in the megakaryocytic cell line CHRF, loss or knock-down of p53 enhances cell cycling and inhibits apoptosis, leading to increased polyploidization. To support the hypothesis that p53 suppresses megakaryocytic polyploidization, we show that stable expression of wild-type p53 in K562 cells (a p53-null cell line) attenuates the cells' ability to undergo polyploidization during megakaryocytic differentiation due to diminished DNA synthesis and greater apoptosis. This suggested that p53's effects during megakaryopoiesis are mediated through cell cycle- and apoptosis-related target genes, possibly by arresting DNA synthesis and promoting apoptosis. To identify candidate genes through which p53 mediates these effects, gene expression was compared between p53 knock-down (p53-KD) and control CHRF cells induced to undergo terminal megakaryocytic differentiation using microarray analysis. Among substantially downregulated p53 targets in p53-KD megakaryocytes were cell cycle regulators CDKN1A (p21) and PLK2, proapoptotic FAS, TNFRSF10B, CASP8, NOTCH1, TP53INP1, TP53I3, DRAM1, ZMAT3 and PHLDA3, DNA-damage-related RRM2B and SESN1, and actin component ACTA2, while antiapoptotic CKS1B, BCL2, GTSE1, and p53 family member TP63 were upregulated in p53-KD cells. Additionally, a number of cell cycle-related, proapoptotic, and cytoskeleton-related genes with known functions in megakaryocytes but not known to carry p53-responsive elements were differentially expressed between p53-KD and control CHRF cells. Our data support a model whereby p53 expression during megakaryopoiesis serves to control polyploidization and the transition from endomitosis to apoptosis by impeding cell cycling and promoting apoptosis. Furthermore, we identify a putative p53 regulon that is proposed to orchestrate these effects.

Effect of repeated eccentric exercise on muscle damage markers and motor unit control strategies in arm and hand muscle.

To examine the contralateral repeated bout effect (CL-RBE) on muscle damage markers and motor unit (MU) control strategies, seventeen healthy adults performed two bouts of 60 eccentric contractions with elbow flexor (EF group; n â€‹= â€‹9) or index finger abductor (IA group; n â€‹= â€‹8) muscles, separated by 1 week. All participants randomly performed eccentric exercise on either the right or left arm or hand muscles, and muscle damage markers and submaximal trapezoid contraction tests were conducted pre, post, 1- and 2-day post eccentric protocol. One week after the first bout, the same exercise protocol and measurements were performed on the contralateral muscles. Surface electromyographic (EMG) signals were collected from biceps brachii (BB) or first dorsal interosseous (FDI) during maximal and submaximal tests. The linear regression analyses were used to examine MU recruitment threshold versus mean firing rate and recruitment threshold versus derecruitment threshold relationships. EMG amplitude from BB (bout 1 vs. bout 2 â€‹= â€‹65.71% â€‹± â€‹22.92% vs. 43.05% â€‹± â€‹18.97%, p â€‹= â€‹0.015, d â€‹= â€‹1.077) and the y-intercept (group merged) from the MU recruitment threshold versus derecruitment threshold relationship (bout 1 vs. bout 2 â€‹= â€‹-7.10 â€‹± â€‹14.20 vs. 0.73 â€‹± â€‹16.24, p â€‹= â€‹0.029, d â€‹= â€‹0.513) at 50% MVIC were significantly different between two bouts. However, other muscle damage markers did not show any CL-RBE in both muscle groups. Therefore, despite changes in muscle excitation and MU firing behavior, our results do not support the existence of CL-RBE on BB and FDI muscles.