Rachel K. Evans

Integrins in immunity

A successful immune response depends on the capacity of immune cells to travel from one location in the body to another–these cells are rapid migrators, travelling at speeds of μm/minute. Their ability to penetrate into tissues and to make contacts with other cells depends chiefly on the β2 integrin known as LFA-1. For this reason, we describe the control of its activity in some detail. For the non-immunologist, the fine details of an immune response often seem difficult to fathom. However, the behaviour of immune cells, known as leukocytes (Box 1), is subject to the same biological rules as many other cell types, and this holds true particularly for the functioning of the integrins on these cells. In this Commentary, we highlight, from a cell-biology point of view, the integrin-mediated immune-cell migration and cell-cell interactions that occur during the course of an immune response.

Influence of exercise mode and osteogenic index on bone biomarker responses during short-term physical training

Prescribing exercise based on intensity, frequency, and duration of loading may maximize osteogenic responses in bone, but a model of the osteogenic potential of exercise has not been established in humans. In rodents, an osteogenic index (OI) has been used to predict the osteogenic potential of exercise. The current study sought to determine whether aerobic, resistance, or combined aerobic and resistance exercise programs conducted over eight weeks and compared to a control group could produce changes in biochemical markers of bone turnover indicative of bone formation. We further sought to determine whether an OI could be calculated for each of these programs that would reflect observed biochemical changes. We collected serum biomarkers [bone-specific alkaline phosphatase (BAP), osteocalcin, tartrate-resistant acid phosphatase (TRAP), C-terminal telopeptide fragment of type I collagen (CTx), deoxypyridinoline (DPD), 25-hydroxy vitamin D (25(OH)D), and parathyroid hormone (PTH)] in 56 women (20.3+/-1.8 years) before, during and after eight weeks of training. We also measured bone mineral density (BMD) at regional areas of interest using DXA and pQCT. Biomarkers of bone formation (BAP and osteocalcin) increased in the Resistance and Combined groups (p<0.05), while biomarkers of bone resorption (TRAP and DPD) decreased and increased, respectively, after training (p<0.05) in all groups. Small changes in volumetric and areal BMD (p<0.05) were observed in the distal tibia in the Aerobic and Combined groups, respectively. Mean weekly OIs were 16.0+/-1.9, 20.6+/-2.2, and 36.9+/-5.2 for the Resistance, Aerobic, and Combined groups, respectively. The calculated osteogenic potential of our programs did not correlate with the observed changes in biomarkers of bone turnover. The results of the present study demonstrate that participation in an eight week physical training program that incorporates a resistance component by previously inactive young women results in alterations in biomarkers of bone remodeling indicative of increased formation without substantial alterations in markers of resorption.

Chronic lymphocytic leukemia cells induce defective LFA-1-directed T cell motility by altering Rho GTPase signaling that is reversible with lenalidomide

T lymphocytes have an essential role in adaptive immunity and rely on the activation of integrin lymphocyte function-associated antigen-1 (LFA-1) to mediate cell arrest and migration. In cancer, malignant cells modify the immune microenvironment to block effective host antitumor responses. We show for the first time that CD4 and CD8 T cells from patients with chronic lymphocytic leukemia (CLL) exhibit globally impaired LFA-1-mediated migration and that this defect is mediated by direct tumor cell contact. We show that following the coculture of previously healthy T cells with CLL cells, subsequent LFA-1 engagement leads to altered Rho GTPase activation signaling by downregulating RhoA and Rac1, while upregulating Cdc42. Of clinical relevance, repair of this T-cell defect was demonstrated using the immunomodulatory drug lenalidomide, which completely rescued adhesion and motility function by restoring normal Rho GTPase activation signaling. Our report identifies a novel cancer immune evasion mechanism whereby tumor cells induce Rho GTPase signaling defects in T cells that prevent appropriate LFA-1 activation and motility. We believe these findings identify important biomarkers and highlight the clinical utility of immunotherapy to rescue normal T-cell function in CLLs that are likely to have relevance in other cancers.

Effects of a 4-Month Recruit Training Program on Markers of Bone Metabolism

UNLABELLED Stress fracture susceptibility results from accelerated bone remodeling after onset of novel exercise and may be reflected in bone turnover changes. It is unknown if the bone turnover response to exercise is different between sexes. PURPOSE To assess disparity between sexes in bone metabolism markers during military recruit training and to evaluate relationships between bone turnover markers and factors that may affect bone metabolism. METHODS Volunteers were age-matched men (n = 58) and women (n = 199), 19 yr old, entering gender-integrated combat training. Blood was collected at 0, 2, and 4 months and anthropometric and fitness measures at 0 and 4 months. Serum was analyzed for biomarkers reflecting bone formation (bone alkaline phosphatase and procollagen I N-terminal peptide), bone resorption (C-telopeptide cross-links of type I collagen and tartrate-resistant acid phosphatase), endocrine regulation (parathyroid hormone, calcium, and 25(OH)D), and inflammation (interleukin 1B, interleukin 6, and tumor necrosis factor alpha). Data were analyzed using ANOVA, correlation, and regression analyses. RESULTS Bone turnover markers were higher in men (P < 0.01) and increased similarly for both sexes from 0 to 2 months (P < 0.01). Independent of gender, VO2max (R = 0.477) and serum calcium (R = 0.252) predicted bone formation activity (bone alkaline phosphatase) at baseline (P < 0.01). Serum calcium and parathyroid hormone decreased (2.0 and 6.4%, respectively) from 0 to 2 months (P < 0.001), returning to baseline at 4 months for both sexes. Men exhibited a decrease in 25(OH)D from 0 to 4 months (P = 0.007). Changes in endocrine regulators were significantly correlated with changes in bone turnover markers. Inflammatory markers did not differ between sexes and did not increase. CONCLUSION Military training increased bone formation and resorption markers in 2 months, suggesting rapid onset of strenuous exercise accelerates bone turnover similarly in men and women. Although bone turnover markers were higher in men than women, bone formation status may be related to aerobic fitness and serum calcium independent of gender and may be affected by small changes in endocrine regulators related to nutrition.

Imaging of Lower Extremity Stress Fracture Injuries

Stress reactions and stress fractures in the lower extremities occur frequently in military and athletic populations. As the clinical symptoms of stress fracture may mimic other less severe musculoskeletal injuries, the diagnosis of stress fracture can often be delayed. The following article reviews the characteristics, advantages and disadvantages of the various imaging tools available to detect stress fracture of the lower limbs in order to clarify their utility when diagnosing this condition. Plain radiography, the primary imaging tool for diagnosing suspected stress injuries, may not detect stress fracture injury until fracture healing is well underway. In some cases of suspected stress fracture, this delay in diagnosis can lead to catastrophic fracture and surgical intervention. Bone scintigraphy has long been recommended for the diagnosis of stress fracture, claiming that skeletal scintigraphy is 100% sensitive for the detection of stress fracture. However, there is a potential for a false negative examination and findings might be nonspecific as tumours or infections may mimic stress injury. In addition, bone scintigraphy involves ionizing radiation and it should not be used whenever there is an alternative. Computed tomography (CT) provides exquisitely fine osseous detail, but should be reserved only for specific indications because it also involves ionizing radiation. Magnetic resonance (MR) imaging, which is noninvasive, has no ionizing radiation, is more rapidly performed than bone scintigraphy, and should be the method of choice for stress fracture diagnosis whenever it is available. However, using MR imaging demands an experienced diagnostician in order to decrease reported false-positive injuries. The ultrasonography technique, which is being used increasingly in the evaluation of the musculoskeletal system has recently been shown to have some potential in the diagnosis of stress fracture; however, currently the imaging modalities are insufficient. The peripheral quantitative CT (pQCT) device, which has been developed to specifically assess skeletal status of the extremities, provides data on bone geometry, strength and density. However, the pQCT needs further evaluation prior to being considered for use in diagnosis stress changes in bone. This article reviews the utility of each of the imaging modalities currently available to detect stress fracture injuries of the lower extremities, as well as other utilization factors, which include exposure to ionizing radiation, the ability to detect early- and late-stage reactions in the bone and surrounding soft tissues, and the ability to differentiate between different types of bone lesions.

Effects of warm-up before eccentric exercise on indirect markers of muscle damage.

PURPOSE To test whether active and passive warm-up conducted before eccentric exercise attenuates clinical markers of muscle damage. METHODS Untrained subjects were exposed to one of five conditions: low-heat passive warm-up (N = 10), high-heat passive warm-up (N = 4), or active warm-up (N = 9), preceding eccentric exercise; eccentric exercise without warm-up (N = 10); or high-heat passive warm-up without eccentric exercise (N = 10). Passive warm-up of the elbow flexors was achieved using pulsed short-wave diathermy, and active warm-up was achieved by concentric contraction. Creatine kinase (CK) activity, strength, range of motion, swelling, and muscle soreness were observed before treatment (baseline) and 24, 48, 72, and 168 h after treatment. RESULTS High-heat passive warm-up without eccentric exercise did not affect any marker of muscle damage and was used as our control group. Markers of muscle damage were not different between groups that did or did not conduct warm-up before eccentric exercise. The active warm-up and eccentric groups exhibited a greater circumferential increase than controls (P < 0.0002), however, that was not observed after passive warm-up. Additionally, the active warm-up group exhibited a greater CK response than controls at 72 h (P < 0.05). The high-heat passive warm-up before eccentric exercise group exhibited significant change from controls at the least number of time points, but due to a small sample size (N = 4), these data should be viewed as preliminary. CONCLUSION Our observations suggest that passive warm-up performed before eccentric exercise may be more beneficial than active warm-up or no warm-up in attenuating swelling but does not prevent, attenuate, or resolve more quickly the other clinical symptoms of eccentric muscle damage as produced in this study.

Biological constraints that limit compensation of a common skeletal trait variant lead to inequivalence of tibial function among healthy young adults.

Having a better understanding of how complex systems like bone compensate for the natural variation in bone width to establish mechanical function will benefit efforts to identify traits contributing to fracture risk. Using a collection of pQCT images of the tibial diaphysis from 696 young adult women and men, we tested the hypothesis that bone cells cannot surmount the nonlinear relationship between bone width and whole bone stiffness to establish functional equivalence across a healthy population. Intrinsic cellular constraints limited the degree of compensation, leading to functional inequivalence relative to robustness, with slender tibias being as much as two to three times less stiff relative to body size compared with robust tibias. Using Path Analysis, we identified a network of compensatory trait interactions that explained 79% of the variation in whole‐bone bending stiffness. Although slender tibias had significantly less cortical area relative to body size compared with robust tibias, it was the limited range in tissue modulus that was largely responsible for the functional inequivalence. Bone cells coordinately modulated mineralization as well as the cortical porosity associated with internal bone multicellular units (BMU)‐based remodeling to adjust tissue modulus to compensate for robustness. Although anecdotal evidence suggests that functional inequivalence is tolerated under normal loading conditions, our concern is that the functional deficit of slender tibias may contribute to fracture susceptibility under extreme loading conditions, such as intense exercise during military training or falls in the elderly. Thus, we show the natural variation in bone robustness was associated with predictable functional deficits that were attributable to cellular constraints limiting the amount of compensation permissible in human long bone. Whether these cellular constraints can be circumvented prophylactically to better equilibrate function among individuals remains to be determined.

Sex Differences in Parameters of Bone Strength in New Recruits : Beyond Bone Density

BACKGROUND Stress fracture (SF) injuries in new recruits have long been attributed to low bone mineral density (BMD). Low areal BMD assessed using two-dimensional dual-energy x-ray absorptiometry imaging, however, reflects structural density and is affected by smaller measures of bone geometry. Recent studies support a relationship between bone size and SF and indicate that slender bones are more susceptible to damage under identical loading conditions. Peripheral quantitative computed tomography (pQCT) is a three-dimensional imaging tool that provides measures of tissue density and geometry parameters of the tibia, a common site of SF. PURPOSE To evaluate sex differences in parameters of volumetric BMD (vBMD), geometry, and strength of the tibia in new recruits using a novel pQCT image analysis procedure. METHODS pQCT images were obtained from 128 healthy men and women (20 male, 108 female, aged 18-21 yr) entering a 4-month gender-integrated combat training program in the Israeli Defense Forces. Tibial scans taken at sites 4% (trabecular bone), 38%, and 66% (cortical bone) from the distal end plate were analyzed using MATLAB to assess whole-bone and regional parameters. Measures included vBMD, geometry (diameter, area, cortical thickness, and canal radius), and strength (moments of inertia and bone strength and slenderness indices). RESULTS With the exception of normalized canal radius, which did not differ between sexes, all measures of bone geometry (P < 0.0001) and strength (P < 0.0001 to P = 0.07) were greater in men. Women exhibited 2.7% to 3.0% greater cortical vBMD than men, whereas trabecular vBMD was 8.4% lower in women (P < 0.001). These differences remained significant after adjusting for body size. CONCLUSION Sex differences in bone geometry and mineralization of the tibia may contribute to a decreased ability to withstand the demands imposed by novel, repetitive exercise in untrained individuals entering recruit training.

Stress fracture and military medical readiness: bridging basic and applied research.

PURPOSE Military recruits and distance runners share a special risk of stress fracture injury. Recent efforts by US and Israeli military-sponsored researchers have uncovered important mechanisms and practical low-cost interventions. This article summarizes key findings relevant to prevention of stress fracture, including simple strategies to identify and to mitigate risk. METHODS Published research supported through the Bone Health and Military Medical Readiness research program and related military bone research was analyzed for contributions to preventing stress fracture in military recruits and optimizing bone health. RESULTS Thousands of military recruits helped test hypotheses about predictors of risk, safer exercise regimens, and rest, nutrition, gait training, and technology interventions to reduce stress fracture risk. Concurrent cellular, animal, and human laboratory studies were used to systematically investigate mechanisms of mechanical forces acting on bone and interactions through muscle, hormonal and genetic influences, and metabolism. The iterative and sometimes simultaneous process of basic discovery and field testing produced new knowledge that will provide safer science-based physical training. DISCUSSION Human training studies evaluating effects on bone require special commitment from investigators and funders due to volunteer compliance and attrition challenges. The findings from multiple studies indicate that measures of bone elasticity, fragility, and geometry are as important as bone mineral density in predicting fracture risk, with applications for new measurement technologies. Risk may be reduced by high intakes of calcium, vitamin D, and possibly protein (e.g., milk products). Prostaglandin E2, insulin-like growth factor 1, and estrogens are important mediators of osteogenesis, indicating reasons to limit the use of certain drugs (e.g., ibuprofen), to avoid excessive food restriction, and to treat hypogonadism. Abnormal gait may be a correctable risk factor. Brief daily vibration may stimulate bone mineral accretion similar to weight-bearing exercise. Genetic factors contribute importantly to bone quality, affecting fracture susceptibility and providing new insights into fracture healing and tissue reengineering.

Dietary intake and stress fractures among elite male combat recruits

BackgroundAppropriate and sufficient dietary intake is one of the main requirements for maintaining fitness and health. Inadequate energy intake may have a negative impact on physical performance which may result in injuries among physically active populations. The purpose of this research was to evaluate a possible relationship between dietary intake and stress fracture occurrence among combat recruits during basic training (BT).MethodsData was collected from 74 combat recruits (18.2 ± 0.6 yrs) in the Israeli Defense Forces. Data analyses included changes in anthropometric measures, dietary intake, blood iron and calcium levels. Measurements were taken on entry to 4-month BT and at the end of BT. The occurrence of stress reaction injury was followed prospectively during the entire 6-month training period.ResultsTwelve recruits were diagnosed with stress fracture in the tibia or femur (SF group). Sixty two recruits completed BT without stress fractures (NSF). Calcium and vitamin D intakes reported on induction day were lower in the SF group compared to the NSF group-38.9% for calcium (589 ± 92 and 964 ± 373 mg·d-1, respectively, p < 0.001), and-25.1% for vitamin D (117.9 ± 34.3 and 157.4 ± 93.3 IU·d-1, respectively, p < 0.001). During BT calcium and vitamin D intake continued to be at the same low values for the SF group but decreased for the NSF group and no significant differences were found between these two groups.ConclusionsThe development of stress fractures in young recruits during combat BT was associated with dietary deficiency before induction and during BT of mainly vitamin D and calcium. For the purpose of intervention, the fact that the main deficiency is before induction will need special consideration.