Histone H3K18 & H3K23 acetylation directs establishment of MLL-mediated H3K4 methylation.

In an unmodified state, positively charged histone N-terminal tails engage nucleosomal DNA in a manner which restricts access to not only the underlying DNA, but also key tail residues subject to binding and/or modification. Charge-neutralizing modifications, such as histone acetylation, serve to disrupt this DNA-tail interaction, facilitating access to such residues. We previously showed that a polyacetylation-mediated chromatin "switch" governs the read-write capability of H3K4me3 by the MLL1 methyltransferase complex. Here, we discern the relative contributions of site-specific acetylation states along the H3 tail and extend our interrogation to other chromatin modifiers. We show that the contributions of H3 tail acetylation to H3K4 methylation by MLL1 are highly variable, with H3K18 and H3K23 acetylation exhibiting robust stimulatory effects, and that this extends to the related H3K4 methyltransferase complex, MLL4. We show that H3K4me1 and H3K4me3 are found preferentially co-enriched with H3 N-terminal tail proteoforms bearing dual H3K18 and H3K23 acetylation (H3{K18acK23ac}). We further show that this effect is specific to H3K4 methylation, while methyltransferases targeting other H3 tail residues (H3K9, H3K27, & H3K36), a methyltransferase targeting the nucleosome core (H3K79), and a kinase targeting a residue directly adjacent to H3K4 (H3T3) are insensitive to tail acetylation. Together, these findings indicate a unique and robust stimulation of H3K4 methylation by H3K18 and H3K23 acetylation and provide key insight into why H3K4 methylation is often associated with histone acetylation in the context of active gene expression.

Cyclic AMP signaling in Dictyostelium promotes the translocation of the copine family of calcium-binding proteins to the plasma membrane.

BACKGROUND: Copines are calcium-dependent phospholipid-binding proteins found in many eukaryotic organisms and are thought to be involved in signaling pathways that regulate a wide variety of cellular processes. Copines are characterized by having two C2 domains at the N-terminus accompanied by an A domain at the C-terminus. Six copine genes have been identified in the Dictyostelium genome, cpnA - cpnF. RESULTS: Independent cell lines expressing CpnA, CpnB, CpnC, CpnE, or CpnF tagged with green fluorescent protein (GFP) were created as tools to study copine protein membrane-binding and localization. In general, the GFP-tagged copine proteins appeared to localize to the cytoplasm in live cells. GFP-tagged CpnB, CpnC, and CpnF were also found in the nucleus. When cells were fixed or when live cells were treated with calcium ionophore, the GFP-tagged copine proteins were found associated with the plasma membrane and vesicular organelles. When starved Dictyostelium cells were stimulated with cAMP, which causes a transitory increase in calcium concentration, all of the copines translocated to the plasma membrane, but with varying magnitudes and on and off times, suggesting each of the copines has distinct calcium-sensitivities and/or membrane-binding properties. In vitro membrane binding assays showed that all of the GFP-tagged copines pelleted with cellular membranes in the presence of calcium; yet, each copine displayed distinct calcium-independent membrane-binding in the absence of calcium. A lipid overlay assay with purified GFP-tagged copine proteins was used to screen for specific phospholipid-binding targets. Similar to other proteins that contain C2 domains, GFP-tagged copines bound to a variety of acidic phospholipids. CpnA, CpnB, and CpnE bound strongly to PS, PI(4)P, and PI(4,5)P2, while CpnC and CpnF bound strongly to PI(4)P. CONCLUSIONS: Our studies show that the Dictyostelium copines are soluble cytoplasmic and nuclear proteins that have the ability to bind intracellular membranes. Moreover, copines display different membrane-binding properties suggesting they play distinct roles in the cell. The transient translocation of copines to the plasma membrane in response to cAMP suggests copines may play a specific role in chemotaxis signaling.

Risk factors for closed-head injuries during military airborne operations.

INTRODUCTION: One of the more serious adverse outcomes of military airborne operations is closed-head injuries. This investigation examined risk factors for closed-head injuries in an Army airborne infantry unit. METHODS: Closed-head injuries were defined as energy exchanges in which the head contacted the ground or an object and the soldier sought medical care for external trauma, headache, loss of consciousness, or altered mental state. Injury data were obtained by investigators in the drop zone and diagnoses were confirmed by a physician. Operational data on potential injury risk factors were obtained from routine reports published by the infantry unit. Weather data were obtained using a pocket weather tracker. RESULTS: There were 96,132 jumps resulting in 310 closed-head injuries for a crude incidence of 3.22/1000 jumps. In 98% of known cases the injury was associated with parachute landings. Multivariate logistic regression revealed that independent risk factors for closed-head injuries included night jumps, combat loads, higher wind speeds, higher temperatures, enlisted rank, and entanglements. CONCLUSIONS: This study identifies the current incidence and extrinsic risk factors for closed-head injuries during military airborne operations. Although it is necessary to conduct airborne training under realistic conditions, trainers and commanders should be aware of these risks and plan airborne operations accordingly.

H3K18 & H3K23 acetylation directs establishment of MLL-mediated H3K4 methylation.

In an unmodified state, positively charged histone N-terminal tails engage nucleosomal DNA in a manner which restricts access to not only the underlying DNA, but also key tail residues subject to binding and/or modification. Charge-neutralizing modifications, such as histone acetylation, serve to disrupt this DNA-tail interaction, facilitating access to such residues. We previously showed that a polyacetylation-mediated chromatin "switch" governs the read-write capability of H3K4me3 by the MLL1 methyltransferase complex. Here, we discern the relative contributions of site-specific acetylation states along the H3 tail and extend our interrogation to other chromatin modifiers. We show that the contributions of H3 tail acetylation to H3K4 methylation by MLL1 are highly variable, with H3K18 and H3K23 acetylation exhibiting robust stimulatory effects, and that this extends to the related H3K4 methyltransferase complex, MLL4. We show that H3K4me1 and H3K4me3 are found preferentially co-enriched with H3 N-terminal tail proteoforms bearing dual H3K18 and H3K23 acetylation (H3{K18acK23ac}). We further show that this effect is specific to H3K4 methylation, while methyltransferases targeting other H3 tail residues (H3K9, H3K27, & H3K36), a methyltransferase targeting the nucleosome core (H3K79), and a kinase targeting a residue directly adjacent to H3K4 (H3T3) are insensitive to tail acetylation. Together, these findings indicate a unique and robust stimulation of H3K4 methylation by H3K18 and H3K23 acetylation and provide key insight into why H3K4 methylation is often associated with histone acetylation in the context of active gene expression.

A prospective investigation of injury incidence and injury risk factors among Army recruits in military police training.

BACKGROUND: United States Army military police (MP) training is a 19-week course designed to introduce new recruits to basic soldiering skills, Army values and lifestyle, and law enforcement skills and knowledge. The present investigation examined injury rates and injury risk factors in MP training. METHODS: At the start of training, 1,838 male and 553 female MP recruits were administered a questionnaire containing items on date of birth, height, weight, tobacco use, prior physical activity, injury history, and menstrual history. Injuries during training were obtained from electronic medical records and the training units provided data on student graduation and attrition. RESULTS: Successfully graduating from the course were 94.3% of the men and 83.7% of the women. Experiencing at least one injury during training were 34.2% of the men and 66.7% of the women (risk ratio (women/men) = 1.95, 95% confidence interval = 1.79-2.13). Recruits were at higher injury risk if they reported that they were older, had smoked in the past, or had performed less frequent exercise or sports prior to MP training. Men were at higher injury risk if they reported a prior injury and women were at higher risk if they reported missing at least six menstrual cycles in the last year or had previously been pregnant. CONCLUSION: The present investigation was the first to identify injury rates and identify specific factors increasing injury risk during MP training.

Single-molecule analysis of transcription activation: dynamics of SAGA co-activator recruitment.

Transcription activators are said to stimulate gene expression by "recruiting" coactivators to promoters, yet this term fits several different kinetic models. To directly analyze dynamics of activator-coactivator interactions, single-molecule microscopy was used to image promoter DNA, a transcription activator, and the Spt-Ada-Gcn5 Acetyltransferase (SAGA) complex within nuclear extract. SAGA readily, but transiently, binds nucleosome-free DNA without activator, while chromatin template association occurs nearly exclusively when activator is present. On both templates, activator increases SAGA association rates by up to an order of magnitude, and dramatically extends its dwell times. These effects reflect direct interactions with the transactivation domain, as VP16 or Rap1 activation domains produce different SAGA dynamics. Despite multiple bromodomains, acetyl-CoA or histone H3/H4 tail acetylation only modestly improves SAGA binding. Unexpectedly, histone acetylation more strongly affects activator residence. Our studies thus reveal two modes of SAGA interaction with the genome: a short-lived activator-independent interaction with nucleosome-free DNA, and a state tethered to promoter-bound transcription activators that can last up to several minutes.

A systematic review of the effects of physical training on load carriage performance.

Soldiers are often required to carry heavy loads during military operations. This article reports on a systematic literature review examining the influence of physical training on load carriage performance. Several literature databases, reference lists, and other sources were explored to find studies that quantitatively examined the effects of physical training on the time taken for individuals to complete a set distance carrying an external load, with the majority of the load contained in a backpack. Effect sizes (Cohen's d statistic) were used in meta-analyses to examine the changes in load carriage performance after various modes of physical training. Effect sizes quantified training-related changes in terms of SD units. Ten original research studies met the review criteria. Meta-analysis indicated that large training effects (≥0.8SD units) were apparent when progressive resistance training was combined with aerobic training and when that training was conducted at least 3 times per week, over at least 4 weeks. When progressive load-carriage exercise was part of the training program, much larger training effects were evident (summary effect size [SES] = 1.7SD units). Field-based training that combined a wide variety of training modes and included progressive load-carriage exercise was also very effective in improving load carriage performance (SES = 1.1SD units). Aerobic training alone or resistance training alone had smaller and more variable effects, depending on the study. This review indicates that combinations of specific modes of physical training can substantially improve load carriage performance.

Injury rates and injury risk factors among Federal Bureau of Investigation new agent trainees.

BACKGROUND: A one-year prospective examination of injury rates and injury risk factors was conducted in Federal Bureau of Investigation (FBI) new agent training. METHODS: Injury incidents were obtained from medical records and injury compensation forms. Potential injury risk factors were acquired from a lifestyle questionnaire and existing data at the FBI Academy. RESULTS: A total of 426 men and 105 women participated in the project. Thirty-five percent of men and 42% of women experienced one or more injuries during training. The injury incidence rate was 2.5 and 3.2 injuries/1,000 person-days for men and women, respectively (risk ratio (women/men) = 1.3, 95% confidence interval = 0.9-1.7). The activities most commonly associated with injuries (% of total) were defensive tactics training (58%), physical fitness training (20%), physical fitness testing (5%), and firearms training (3%). Among the men, higher injury risk was associated with older age, slower 300-meter sprint time, slower 1.5-mile run time, lower total points on the physical fitness test (PFT), lower self-rated physical activity, lower frequency of aerobic exercise, a prior upper or lower limb injury, and prior foot or knee pain that limited activity. Among the women higher injury risk was associated with slower 300-meter sprint time, slower 1.5-mile run time, lower total points on the PFT, and prior back pain that limited activity. CONCLUSION: The results of this investigation supported those of a previous retrospective investigation emphasizing that lower fitness and self-reported pain limiting activity were associated with higher injury risk among FBI new agents.

The Advanced Tactical Parachute System (T-11): injuries during basic military parachute training.

BACKGROUND: Since the 1950s, the standard U.S. military troop parachute system has been the T-10. TheT-10 is currently being replaced by the newer T-11 system. PURPOSE: This investigation compared injury incidence between the T-10 and T-11 military parachute systems. METHODS: Participants were students in basic parachute training at the U.S. Army Airborne School (USAAS). Students performed their first parachute jumps with the T-11 and subsequent jumps with the T-10. Injury data were collected from routine reports produced by the USAAS. Combat loaded jumps and night jumps were excluded from the analysis since these were only conducted with the T-10. RESULTS: There were a total of 76 injuries in 30,755 jumps for an overall cumulative injury incidence of 2.5/1000 jumps. With the T-10 parachute, there were 61 injuries in 21,404 jumps for a cumulative injury incidence of 2.9/1000 jumps; with the T-11 parachute there were 15 injuries in 9351 jumps for a cumulative injury incidence of 1.6/1000 jumps [risk ratio (T10/T11) = 1.78, 95% confidence interval = 1.01-3.12, P = 0.04]. DISCUSSION: Limitations to this analysis included the fact that the T-11 was only used on the first jumps among students who had likely never previously performed a parachute jump and that aircraft exit procedures differed very slightly for the two parachutes. Nonetheless, the data suggest that injury incidence is lower with the T-11 parachute than with the T-10 parachute when airborne training operations are conducted during the day without combat loads.

Military parachuting injuries, associated events, and injury risk factors.

INTRODUCTION: The purpose of this investigation was to examine injury incidence, events associated with injury, and injury risk factors during parachuting in an Army airborne infantry unit. METHODS: Injury data were obtained by the investigators on the drop zone and confirmed by a physician. Operational data (potential injury risk factors) were obtained from routine reports published by the infantry unit. Weather data were obtained using a Kestrel Model 4500 pocket weather tracker. RESULTS: There were a total of 23,031 jumps resulting in 242 injured soldiers for a crude injury incidence of 10.5 per 1000 jumps. Parachute entanglement incidence was 0.5 per 1000 jumps. Where an event associated with the injury could be determined (67% of cases), these included ground impact (75%), static line problems (11%), tree landings (4%), entanglements (4%), and aircraft exits (3%). Univariate analysis showed that higher injury risk was associated with night jumps (versus day jumps), combat loads (versus unloaded jumps), higher wind speeds, higher dry bulb temperatures, higher humidity, C17 Globemaster or C130 Hercules aircrafts (compared to the other aircraft), exits through aircraft side doors (versus tailgates), and entanglements. Multivariate analysis indicated that independent risk factors for injuries included night jumps, combat loads, higher wind speeds, higher dry bulb temperatures, and entanglements. DISCUSSION: This investigation provided injury incidence, events associated with injury, and quantitative assessments of injury risk factors and their interactions during military parachuting. An appreciation of these subjects can assist medical and operational planners in further reducing the incidence of injury during airborne operations.

A prospective investigation of injury incidence and risk factors among army recruits in combat engineer training.

BACKGROUND: United States Army combat engineer (ENG) training is an intense 14-week course designed to introduce new recruits to basic soldiering activities, Army values and lifestyle, and engineering skills and knowledge. The present investigation examined injury rates and injury risk factors in ENG training. METHODS: At the start of their training, 1,633 male ENG recruits were administered a questionnaire containing items on date of birth, height, weight, tobacco use, prior physical activity, and injury history. Injuries during training were obtained from electronic medical records and the training units provided data on student graduation and attrition. Risk factors were identified using Cox regression. RESULTS: Ninety-two percent of the recruits successfully graduated from the course and 47% of the recruits experienced one or more injuries during training. Univariate Cox regression demonstrated that recruits were at higher injury risk if they reported that they were older, had a higher or lower body mass index, had smoked in the past, had performed less exercise (aerobic or muscle strength) or sports prior to ENG training, had experienced a previous time-loss lower limb injury (especially if they had not totally recovered from that injury), or had a lower educational level. CONCLUSIONS: The present investigation was the first to identify injury rates and identify specific factors increasing injury risk during ENG training. The identified risk factors provide a basis for recommending future prevention strategies.

Activities associated with injuries in initial entry training.

Previous studies have not reported activities associated with injuries in initial entry training (IET) because these data were seldom available in medical records and not contained in electronic databases. This investigation obtained activities associated with outpatient encounters in IET recorded by primary medical care providers at Fort Leonard Wood, Missouri. Data were entered into a standard database that included fields for diagnosis and activity associated with the injury. Fifty percent of the new injury encounters (i.e., exclusive of follow-ups) were not associated with a specific event but were reported as having a gradual onset. Other activities included physical training (16%), road marching (15%), confidence/obstacle courses (5%), and barracks activities (3%). Risks per unit of training time were estimated at 13, 62, and 97 injuries per hour for physical training, road marching, and the confidence/obstacle courses, respectively. The most frequently recorded diagnoses were joint pain (27%), strains (15%), blisters (14%), sprains (13%), and tendonitis (12%). The types of injuries and their anatomical locations were similar to those reported in other IET investigations, although blister-related encounters were higher. This investigation identifies activities with the highest risk of injury in IET and those that should be targeted for injury prevention efforts.