Exercise Increases Glucose Transporter-4 Levels on Peripheral Blood Mononuclear Cells.

PURPOSE: Glucose transporter 4 (GLUT4) plays a key role in the pathophysiology of type 2 diabetes. Glucose transporter 4 is upregulated in response to exercise, enhancing cellular glucose transport in skeletal muscle tissue. This mechanism appears to remain intact in individuals with insulin resistance. Details of the mechanism are poorly understood and are challenging to study due to the invasive nature of muscle biopsy. Peripheral blood mononuclear cells (PBMC) have documented insulin-sensitive GLUT4 activity and may serve as a proxy tissue for studying skeletal muscle GLUT4. The purpose of this study was to investigate whether GLUT4 in PBMC is affected by conditioning. METHODS: We recruited 16 student athletes from the cross-country running and skiing teams and fifteen sedentary students matched for age and sex from the University of Alaska Fairbanks. Peripheral blood mononuclear cells were collected with mononuclear cell separation tubes. The GLUT4 concentrations were measured using a commercially available enzyme linked immunosorbent assay. Additionally, correlations between PBMC GLUT4 and common indicators of insulin resistance were examined. RESULTS: Results indicate significantly higher PBMC GLUT4 levels in conditioned athletes than in their sedentary counterparts, similar to what has been documented in myocytes. Females were observed to have higher PBMC GLUT4 levels than males. Correlations were not detected between PBMC GLUT4 and hemoglobin A1c, glucose, insulin, homeostatic model assessment of insulin resistance, body mass index, or body fat. CONCLUSIONS: This study provides evidence to support exploration of PBMC as a proxy tissue for studying GLUT4 response to exercise or other noninsulin factors.

25(OH)D levels in trained versus sedentary university students at 64° north.

PURPOSE: 25-hydroxyvitamin D (25[OH]D) deficiency is associated with compromised bone mineralisation, fatigue, suppressed immune function and unsatisfactory skeletal muscle recovery. We investigated the risk of 25(OH)D insufficiency or deficiency in endurance athletes compared to sedentary non-athletes living at 64° north. METHODS: University student-athletes (TS) and sedentary students (SS) volunteered to participate in this study. TS engaged in regular exercise while SS exercised no more than 20 minutes/week. Metabolic Equivalent of Task (MET) scores for participants were determined. Vitamin D intake was assessed using the National Cancer Institute's 24-hour food recall (ASA24). Fasting plasma 25(OH)D levels were quantified via enzyme-linked immunosorbent assay. RESULTS: TS reported higher activity levels than SS as assessed with MET-minutes/week and ranking of physical activity levels (p < 0.05). The reported mean daily intake of vitamin D was higher in TS compared to SS (p < 0.05) while 25(OH)D plasma levels were lower in TS than in SS (p < 0.05). In total, 43.8% of the TS were either insufficient (31.3%) or deficient (12.5%) in 25(OH)D, while none of the SS were insufficient and 13.3% were deficient. CONCLUSION: TS are at increased risk of 25(OH)D insufficiency or deficiency compared to their sedentary counterparts residing at the same latitude, despite higher vitamin D intake.

BUILDing BLaST: promoting rural students' biomedical research careers using a culturally responsive, one health approach.

BACKGROUND AND PURPOSE: Most postsecondary institutions in the state of Alaska (USA) have a broad mission to serve diverse students, many of whom come from schools in rural villages that are accessible only by plane, boat, or snowmobile. The major research university, the University of Alaska in Fairbanks (UAF), serves a population whereby 40% are from groups recognized as underrepresented in the biomedical workforce. The purpose of this article is to describe the Building Infrastructure Leading to Diversity (BUILD)-supported program in the state of Alaska that seeks to engage students from rural areas with a culturally relevant approach that is centered on the One Health paradigm, integrating human, animal, and environmental health. PROGRAM AND KEY HIGHLIGHTS: The Biomedical Learning and Student Training (BLaST) program distinguished by broad themes that address recruitment, retention, and success of students in biomedical programs, especially for students from rural backgrounds. Targeted rural outreach emphasizes that biomedical research includes research on the integration of human, animal, and environmental health. This One Health perspective gives personal relevance and connection to biomedical research. This outreach is expected to benefit student recruitment, as well as foster family and community support for pursuit of college degrees. BLaST promotes integration of research into undergraduate curricula through curriculum development, and by creating a new class of instructors, laboratory research and teaching technicians, who provide research mentorship, course instruction, and comprehensive advising. Finally, BLaST facilitates early and sustained undergraduate research experiences in collaborations with graduate students and faculty. IMPLICATIONS: BLaST's approach is highly adapted to the Alaskan educational and physical environment, but components and concepts could be adapted to other rural areas as a means to engage students from rural backgrounds, who often have a closer relationship with the natural environment than urban students.

The effect of acute exercise on GLUT4 levels in peripheral blood mononuclear cells of sled dogs.

Using sled dogs as exercise model, our objectives of this study were to 1) assess the effects of one acute bout of high-intensity exercise on surface GLUT4 concentrations on easily accessible peripheral blood mononuclear cells (PBMC) and 2) compare our findings with published research on exercise induced GLUT4 in skeletal muscle. During the exercise bout, dogs ran 5 miles at approximately 90% of VO2 max. PMBC were collected before exercise (baseline), immediately after exercise and after 24h recovery.GLUT4 was measured via ELISA. Acute exercise resulted in a significant increase on surface GLUT4 content on PBMC. GLUT4 was increased significantly immediately after exercise (~ 50%; p<0.05) and reduced slightly by 24h post-exercise as compared to baseline (~ 22%; p>0.05). An effect of acute exercise on GLUT4 levels translocated to the cell membrane was observed, with GLUT4 levels not yet returned to baseline after 24h post-exercise. In conclusion, the present investigation demonstrated that acute high-intensity exercise increased GLUT4 content at the surface of PBMC of sled dogs as it has been reported in skeletal muscle in other species. Our findings underline the potential use of peripheral blood mononuclear cell GLUT4 protein content as minimally invasive proxy to investigate relationships between insulin sensitivity, insulin resistance, GLUT4 expression and glucose metabolism.

Effects of postexercise feeding of a supplemental carbohydrate and protein bar with or without astaxanthin from Haematococcus pluvialis to exercise-conditioned dogs.

OBJECTIVE: To characterize the postprandial nutrient profiles of exercise-conditioned dogs fed a supplemental carbohydrate and protein bar with or without astaxanthin from Haematococcus pluvialis immediately after exercise. ANIMALS: 34 exercise-conditioned adult Husky-Pointer dogs. PROCEDURES: The study had 2 phases. During phase 1, postprandial plasma glucose concentration was determined for dogs fed a bar containing 25% protein and 18.5% or 37.4% maltodextrin plus dextrin (rapidly digestible carbohydrate; RDC), or dry kibble (30% protein and 0% RDC) immediately after exercise. During phase 2, dogs were exercised for 3 days and fed a bar (25% protein and 37.4% RDC) with (CPA; n = 8) or without (CP; 8) astaxanthin or no bar (control; 8) immediately after exercise. Pre- and postexercise concentrations of plasma biochemical analytes and serum amino acids were determined on days 1 and 3. RESULTS: Phase 1 postexercise glucose concentration was increased when dogs were provided the 37.4% RDC bar, but not 0% or 18.5% RDC. On day 3 of phase 2, the CPA group had the highest pre-exercise triglyceride concentration and significantly less decline in postexercise glucose concentration than did the CP and control groups. Mean glucose concentration for the CP and CPA groups was significantly higher than that for the control group between 15 and 60 minutes after bar consumption. Compared to immediately after exercise, branched-chain amino acid, tryptophan, leucine, and threonine concentrations 15 minutes after exercise were significantly higher for the CP and CPA groups, but were lower for the control group. CONCLUSIONS AND CLINICAL RELEVANCE: Dogs fed a bar with 37.4% RDCs and 25% protein immediately after exercise had increased blood nutrient concentrations for glycogen and protein synthesis, compared with control dogs.

Glucose transporter-4 in white blood cells of young and old sled dogs: a model for human biomarker development.

The insulin responsive glucose transporter, GLUT4 is found predominantly in muscle and adipose cells. Maratou and others (2007) reported that there is GLUT4 in white blood cells (WBC) collected from human subjects in response to insulin activation. This study was designed to validate the presence of GLUT4 in white blood cells of sled dogs and furthermore to investigate whether changes in levels of the GLUT4 protein might be associated with aging. Additionally, we examined the blood insulin concentration of two populations of dogs, young and old, before and after a meal to observe their insulin response. It is documented in skeletal muscle that GLUT4 expression is increased as a result of conditioning, making sled dogs an excellent model in the circumpolar north for studying the effects of exercise, nutrition and diabetes (Felsburg 2002; Kararli 2006). Blood was withdrawn from 11 healthy sled dogs: 6 young (1-5 years) and physically fit, conditioned for racing and 5 old (7-13 years), retired from racing. The insulin response was determined using blood plasma and ELISA. The buffy coat (containing WBC) was collected with a glass pipette after centrifugation and washed and suspended in 1x phosphate buffer. GLUT4 was measured using ELISA kits (USCN Life Sciences). The results validate that GLUT4 is present in white blood cells in sled dogs. Age had no significant effect in the concentration of GLUT4 between the populations of old and young dogs. A significant difference in insulin levels pre and post meal in young (0.13 ± 0.03 ng/mL (pre), 0.22 ± 0.04 ng/mL (post), p < 0.05) and old (0.13 ± 0.02 ng/mL (pre), 0.22 ± 0.03 ng/mL (post), p < 0.05) dogs was observed, displaying the typical postprandial insulin spike. No significant difference was found in insulin concentration comparing old versus young dogs. Our data shows that white blood cells in young (40.4 ± 2.4 ng/mL) and old (35.3 ± 8.8 ng/mL) sled dogs have quantifiable but non-significant different GLUT4 levels (p > 0.05). Detecting GLUT4 via an ELISA in white blood cells, opens up minimally invasive avenues for studying the underlying molecular mechanisms associated with insulin resistance in more complex, dynamic and physiological systems. This project was the first step in developing a protocol for this simple, technique with a potential clinical application for diagnosing insulin resistance.

Conditioning causes an increase in glucose transporter-4 levels in mononuclear cells in sled dogs.

This study was designed to investigate the effects of physical conditioning on the expression of the insulin sensitive glucose transporter-4 protein (GLUT4) on mononuclear cells and HOMA-IR levels in dogs and compared to results reported in human skeletal muscle and the skeletal muscle of rodent models. Blood was sampled from conditioned dogs (n = 8) and sedentary dogs (n = 8). The conditioned dogs were exercised four months prior the experiment and were following a uniform training protocol, whereas the sedentary dogs were not. GLUT4 expression in mononuclear cells and plasma insulin levels were measured using commercially available enzyme-linked immunosorbent assay (ELISA). Blood glucose levels were determined using blood plasma. HOMA-IR was calculated using plasma insulin and blood glucose levels using the linear approximation formula. Our results indicate that the state of conditioning had a significant effect on the GLUT4 expression at the surface of mononuclear cells. HOMA-IR was also affected by conditioning in dogs. GLUT4 levels in mononuclear cells of sled dogs were inversely correlated with the homeostasis model assessment of insulin sensitivity. This study demonstrates that conditioning increases GLUT4 levels in mononuclear cells of sled dogs as it has been previously reported in skeletal muscle. Our results support the potential of white blood cells as a proxy tissue for studying insulin signaling and may lead to development of a minimally invasive and direct marker of insulin resistance. This may be the first report of GLUT4 in mononuclear cells in response to exercise and measured with ELISA.

Mercury interferes with endogenous antioxidant levels in Yukon River subsistence-fed sled dogs.

Before adopting modern corn-and-grain-based western processed diets, circumpolar people had a high fat and protein subsistence diet and exhibited a low incidence of obesity, diabetes and cardiovascular disease. Some health benefits are attributable to a subsistence diet that is rich in omega-3 fatty acids and antioxidants. Pollution, both global and local, is a threat to wild foods, as it introduces contaminants into the food system. Northern indigenous people and their sled dogs are exposed to a variety of contaminants, including mercury, that accumulate in the fish and game that they consume. The sled dogs in Alaskan villages are maintained on the same subsistence foods as their human counterparts, primarily salmon, and therefore they can be used as a food systems model for researching the impact of changes in dietary components. In this study, the antioxidant status and mercury levels were measured for village sled dogs along the Yukon River. A reference kennel, maintained on a nutritionally balanced commercial diet, was also measured for comparison. Total antioxidant status was inversely correlated with the external stressor mercury.

Hair analysis in sled dogs (Canis lupus familiaris) illustrates a linkage of mercury exposure along the Yukon River with human subsistence food systems.

The dog has been an important biomedical research model and hair samples from sled dogs could be used as a biomarker of exposure to metals. Hair samples were used as a non-invasive indicator of mercury exposure in sled dogs fed commercial food and traditional village diets. Sled dog populations living in rural New York and Alaska were sampled in 2005 and 2006. Total mercury (THg) content was determined on the entire hair sample in sled dogs from reference sites in North Creek, New York and Salcha Alaska. Both sites fed a commercial feed for high performance dogs and had mean THg levels of 36.6 ng/g for New York sled dogs while Alaskan sled dogs, occasionally supplemented with fish oil showed THg mean of 58.2 ng/g. These THg levels are below levels that are suggested to cause adverse effects and should be considered baseline levels. Yukon River sled dogs had higher THg, ranging from 139 to 15,800 ng/g and showed decreasing mean levels from the delta area to upriver. There were significant differences between THg in the dogs from Russian Mission (10,908.3+/-3028 ng/g), the farthest west village, and Ft. Yukon (1822.4+/-1747 ng/g), the farthest east village. All village dogs along the Yukon had higher THg levels than the THg mean level (657+/-273 ng/g) of hair from ancient dogs of the Seward Peninsula.

Seasonal and diurnal melatonin production in exercising sled dogs.

Melatonin is a hormone that is released from the pineal gland into the blood stream and is controlled by nerve impulses from the suprachiasmatic nuclei. Melatonin synthesis, which is inhibited by light on the mammalian retina, peaks in plasma concentrations during the night. Though still a subject of intense research, melatonin in mammals is known to effect the reproductive system, thyroid function, and adaptations to seasonal changes. Sled dogs in Fairbanks, Alaska (65 degrees N) can be exposed to anywhere from 21 h of daylight in the summer to 4 h in the winter. While light may be the primary factor influencing melatonin production, we hypothesized that exercise may also affect melatonin production. In the current study, sled dogs were used to study seasonal and diurnal variation in melatonin production. Sled dogs by nature are elite athletes and therefore exercise was a focus in the study. Both exercise and non exercise dogs from 2 distinct latitudes were used. The peak in melatonin production was prolonged in high latitude dogs (65 degrees N), compared with lower latitude dogs (45 degrees N). Dogs at both latitudes show a reduction in peak melatonin levels with exercise, and winter melatonin levels in both locations were higher than the summer. Surprisingly, sled dogs in Alaska had lower melatonin levels than sled dogs in New York.

Total antioxidant power in sled dogs supplemented with blueberries and the comparison of blood parameters associated with exercise.

Oxidative damage from free radicals plays an important role in several diseases such as cancer, Alzheimer's disease, and heart disease. Research indicates that exercise contributes to oxidative stress. Fruits, such as blueberries, are good antioxidants because they contain phenolics that preferentially react with free radicals. Maintaining antioxidant levels by supplementing the diet with blueberries may prevent exercise-induced oxidative damage. The goal of our study was to compare antioxidant levels in sled dogs supplemented with blueberries on blood parameters within 48 h post-exercise. Though the exercise protocol did not cause unusual muscle damage as reflected in plasma creatine kinase and isoprostane levels, blueberry supplementation did elicit significantly elevated antioxidant status in sled dogs post exercise. This suggests that dogs fed blueberries while exercising as compared to dogs fed a control diet while exercising, may be better protected against oxidative damage.

Biochemical and metabolic changes due to exercise in sprint-racing sled dogs: implications for postexercise carbohydrate supplements and hydration management.

Evaluations of biochemical changes associated with spring-style sled dog racing indicate that differences in cortisol, lactate, and serum glucose levels suggest exercise of moderate duration (but high intensity) has metabolic demands that dif-fer from those for typical endurance sled dog racing. Additionally, hematocrit, albumin, sodium, chloride, and blood urea nitrogen levels decreased in one team of dogs, whereas there were mild increases in sodium, chloride, and blood urea nitrogen in the other team. These opposing biochemical findings suggest physiologic changes associated with differences in hydration status, likely attributed to different dietary and hydration strategies used by the respective kennels.

Effects of post-exercise supplements on glycogen repletion in skeletal muscle.

Post-exercise carbohydrate supplementation has been routinely used to enhance glycogen concentrations in skeletal muscle, particularly during multiple-day athletic events. Consumption of protein hydrolysates mixed with carbohydrate supplements during the post-exercise period may increase insulin response and cause glycogen repletion in skeletal muscle. A group of Alaskan sled dogs were selected to examine post-exercise supplementation in a paired crossover study design. The dogs were subjected to the same exercise regimen and provided one of three treatments-water, glucose polymers, or glucose polymers with protein hydrolysates-over a 2-month period. Parameters tested at various post-exercise time points included plasma insulin, glucagon and glucose concentrations, and skeletal muscle glycogen content to gain a better understanding of glucose metabolism and glycogen repletion. The results showed an enhanced insulin, glucose, and glucagon response immediately after supplementation and significantly increased glycogen concentrations in skeletal muscle within 24 hours when dogs received either of the glucose-containing supplements compared with water alone. There were no differences in the plasma parameters or skeletal muscle glycogen stores in dogs provided the glucose polymers alone or the glucose polymers plus protein hydrolysates. Thus, post-exercise carbohydrate supplementation increased muscle glycogen repletion, but inclusion of protein hydrolysates in the supplements provided no additional benefits.