Marian L. Kohut

Aerobic exercise, but not flexibility/resistance exercise, reduces serum IL-18, CRP, and IL-6 independent of β-blockers, BMI, and psychosocial factors in older adults.

Increased serum levels of inflammatory mediators have been associated with numerous disease states including atherosclerosis, Type II diabetes, hypertension, depression, and overall mortality. We hypothesized that a long-term exercise intervention among older adults would reduce serum inflammatory cytokines, and this reduction would be mediated, in part, by improvements in psychosocial factors and/or by beta-adrenergic receptor mechanisms. Adults age 64 were randomly assigned to either an aerobic exercise treatment (CARDIO) or a flexibility/strength exercise treatment (FLEX) 3 days/week, 45 min/day for 10 months. A subgroup of subjects treated with non-selective beta(1)beta(2) adrenergic antagonists were included to evaluate the potential role of beta-adrenergic receptor adaptations as mediators of an exercise-induced change in inflammation. The inflammatory mediators [C-reactive protein (CRP), IL-6, tumor necrosis factor (TNF)-alpha, and IL-18] and the psychosocial factors (depression, perceived stress, optimism, sense of coherence, and social support) were measured pre- and post-intervention. The CARDIO treatment resulted in significant reductions in serum CRP, IL-6, and IL-18 compared to the FLEX treatment (significant treatment x time interaction, p<.05), whereas TNFalpha declined in both groups (main effect of time, p=.001). However, several psychosocial factors (depression, optimism, and sense of coherence) improved in both groups suggesting that the reduction of CRP, IL-6, and IL-18 in the CARDIO group was not mediated by improvements in psychosocial scores. With respect to the potential role of beta-adrenergic receptors, both CARDIO subjects treated with beta-adrenergic antagonists and those who were not treated with those medications demonstrated similar reductions in serum CRP, IL-6, IL-18, and TNFalpha. In summary, we have observed that an aerobic exercise intervention can significantly reduce serum inflammatory mediators, but beta-adrenergic receptors and psychosocial factors do not appear to be involved.

Endocrine and Lipid Responses to Chronic Androstenediol-Herbal Supplementation in 30 to 58 Year Old Men

Objective: The effectiveness of an androgenic nutritional supplement designed to enhance serum testosterone concentrations and prevent the formation of dihydrotestosterone and estrogen was investigated in healthy 30 to 58 year old men. Design: Subjects were randomly assigned to consume a nutritional supplement (AND-HB) containing 300-mg androstenediol, 480-mg saw palmetto, 450-mg indole-3-carbinol, 300-mg chrysin, 1,500 mg gamma-linolenic acid and 1,350-mg Tribulus terrestris per day (n = 28), or placebo (n = 27) for 28 days. Subjects were stratified into age groups to represent the fourth (30 year olds, n = 20), fifth (40 year olds, n = 20) and sixth (50 year olds, n = 16) decades of life. Measurements: Serum free testosterone, total testosterone, androstenedione, dihydrotestosterone, estradiol, prostate specific antigen and lipid concentrations were measured before supplementation and weekly for four weeks. Results: Basal serum total testosterone, estradiol, and prostate specific antigen (PSA) concentrations were not different between age groups. Basal serum free testosterone concentrations were higher (p < 0.05) in the 30- (70.5 ± 3.6 pmol/L) than in the 50 year olds (50.8 ± 4.5 pmol/L). Basal serum androstenedione and dihydrotestosterone (DHT) concentrations were significantly higher in the 30- (for androstenedione and DHT, respectively, 10.4 ± 0.6 nmol/L and 2198.2 ± 166.5 pmol/L) than in the 40- (6.8 ± 0.5 nmol/L and 1736.8 ± 152.0 pmol/L) or 50 year olds (6.0 ± 0.7 nmol/L and 1983.7 ± 147.8 pmol/L). Basal serum hormone concentrations did not differ between the treatment groups. Serum concentrations of total testosterone and PSA were unchanged by supplementation. Ingestion of AND-HB resulted in increased (p < 0.05) serum androstenedione (174%), free testosterone (37%), DHT (57%) and estradiol (86%) throughout the four weeks. There was no relationship between the increases in serum free testosterone, androstenedione, DHT, or estradiol and age (r2 = 0.08, 0.03, 0.05 and 0.02, respectively). Serum HDL-C concentrations were reduced (p < 0.05) by 0.14 mmol/L in AND-HB. Conclusions: These data indicate that ingestion of androstenediol combined with herbal products does not prevent the formation of estradiol and dihydrotestosterone.

Age effects on macrophage function vary by tissue site, nature of stimulant, and exercise behavior.

We explored the effects of aging on macrophage function in male BALB/c mice from three age groups: young (2 months), middle-aged (12 months), and old (21 months). Macrophages were collected from alveoli, peritonea, and spleens of each age group. Cells were cultured in vitro with LPS or LPS+IFN-gamma and assayed for production of IL-1, IL-12, NO, and TNF-alpha. Using herpes simplex virus-1, age-related changes in intrinsic antiviral resistance (plaque assay) and extrinsic antiviral resistance (NO and TNF-alpha production) were determined in alveolar and/or peritoneal macrophages. Effects of chronic exercise on age-related macrophage changes were examined. In vitro, macrophages from the alveoli and spleen of older mice generally produced more cytokine and NO compared to younger counterparts. Conversely, macrophages from the peritonea of older mice generally produced less cytokine and NO in vitro compared to younger counterparts. Alveolar macrophages from both old and young mice showed higher intrinsic antiviral resistance to HSV-1 compared to middle-aged mice, while peritoneal macrophages from young mice showed reduced intrinsic resistance compared to those from both middle-aged and old mice. When challenged with HSV-1, a trend towards decreased peritoneal macrophage production of TNF-alpha and decreased alveolar macrophage production of IL-12 with advancing age was found. Chronic moderate exercise tended to reverse age-associated changes in macrophage function in old mice.

Effects of androstenedione-herbal supplementation on serum sex hormone concentrations in 30- to 59-year-old men.

The effectiveness of a nutritional supplement designed to enhance serum testosterone concentrations and prevent the formation of dihydrotestosterone and estrogens from the ingested androgens was investigated in healthy 30- to 59-year old men. Subjects were randomly assigned to consume DION (300 mg androstenedione, 150 mg dehydroepiandrosterone, 540 mg saw palmetto, 300 mg indole-3-carbinol, 625 mg chrysin, and 750 mg Tribulus terrestris per day; n = 28) or placebo (n = 27) for 28 days. Serum free testosterone, total testosterone, androstenedione, dihydrotestosterone, estradiol, prostate-specific antigen (PSA), and lipid concentrations were measured before and throughout the 4-week supplementation period. Serum concentrations of total testosterone and PSA were unchanged by supplementation. DION increased (p < 0.05) serum androstenedione (342%), free testosterone (38%), dihydrotestosterone (71%), and estradiol (103%) concentrations. Serum HDL-C concentrations were reduced by 5.0 mg/dL in DION (p < 0.05). Increases in serum free testosterone (r2 = 0.01), androstenedione (r2 = 0.01), dihydrotestosterone (r2 = 0.03), or estradiol (r2 = 0.07) concentrations in DION were not related to age. While the ingestion of androstenedione combined with herbal products increased serum free testosterone concentrations in older men, these herbal products did not prevent the conversion of ingested androstenedione to estradiol and dihydrotestosterone.

Chronic Exercise Reduces Illness Severity, Decreases Viral Load, and Results in Greater Anti-Inflammatory Effects than Acute Exercise during Influenza Infection

BACKGROUND It is assumed that moderate exercise may improve resistance to infection and reduce inflammation, but there are limited data to support this assumption in an infection model. METHODS BALB/cJ mice were assigned to the following groups: no exercise (NON-EX), 1 session of acute exercise (A-EX), or chronic exercise for approximately 3.5 months (C-EX). Mice were infected with influenza (C-EX mice infected at rest; A-EX mice infected 15 min after exercise). RESULTS C-EX mice demonstrated the lowest severity of infection, assessed by body weight loss and food intake. There was less virus in the lungs at day 5 after infection in C-EX and A-EX mice compared with NON-EX mice (P = .02) and less virus at day 2 after infection only in C-EX mice (P = .07). Soon after infection (day 2), interleukin 6 (IL-6), monocyte chemoattractant protein 1 (MCP-1), macrophage inflammatory protein 1beta, and tumor necrosis factor alpha in the bronchoalveolar lavage (BAL) fluid were lower in C-EX and A-EX than in NON-EX mice. At day 5 after infection, the BAL fluid from C-EX (but not A-EX) mice had less IL-6, interleukin 12p40, granulocyte colony-stimulating factor, keratinococyte-derived chemokine, and MCP-1 than that from NON-EX mice. A trend toward reduced immunopathologic response was found in C-EX mice. CONCLUSIONS Chronic exercise resulted in reduced symptoms, virus load, and levels of inflammatory cytokine and chemokines. Acute exercise also showed some benefit, which was limited to the early phase of infection.

Echinacea increases arginase activity and has anti-inflammatory properties in RAW 264.7 macrophage cells, indicative of alternative macrophage activation

ETHNOPHARMACOLOGICAL RELEVANCE The genus Echinacea is a popular herbal immunomodulator. Recent reports indicate that Echinacea products inhibit nitric oxide (NO) production in activated macrophages. AIM OF THE STUDY In the present study we determined the inhibitory effects of alcohol extracts and individual fractions of alcohol extracts of Echinacea on NO production, and explored the mechanism underlying the pharmacological anti-inflammatory activity. MATERIALS AND METHODS Alcohol extracts of three medicinal Echinacea species, Echinacea angustifolia, Echinacea pallida and Echinacea purpurea, were prepared using Soxhlet apparatus and fractionated using HPLC. NO production by LPS activated RAW 264.7 macrophage cells was measured using a Griess reagent and iNOS detected using immunoblotting. In addition, effects on arginase activity were measured in RAW 264.7 cells stimulated with 8-bromo-cAMP +/- LPS. RESULTS Alcohol extracts of all three Echinacea species significantly inhibited NO production by lipopolysaccharide (LPS)-activated the RAW 264.7 macrophage cell line; among them Echinacea pallida was the most active. The Echinacea-mediated decrease in NO production was unlikely due to a direct scavenging of NO because the extracts did not directly inhibit NO released from an NO donor, sodium nitroprusside. An immunoblotting assay demonstrated that the extract of Echinacea pallida inhibited inducible nitric oxide synthase (iNOS) protein expression in LPS-treated macrophages. The enzymes iNOS and arginase metabolize a common substrate, l-arginine, but produce distinct biological effects. While iNOS is involved in inflammatory response and host defense, arginase participates actively in anti-inflammatory activation. Arginase activity of RAW 264.7 cells stimulated with 8-bromo-cAMP was significantly increased by alcohol extracts of all three Echinacea species. The polar fraction containing caffeic acid derivatives enhanced arginase activity, while the lipophilic fraction containing alkamides exhibited a potential of inhibiting NO production and iNOS expression. CONCLUSIONS These results suggest that the anti-inflammatory activity of Echinacea might be due to multiple active metabolites, which work together to switch macrophage activation from classical activation towards alternative activation.

Centrally located body fat is related to inflammatory markers in healthy postmenopausal women

Objective:C-reactive protein and fibrinogen are established atherosclerotic cardiovascular disease risk factors. These acute-phase proteins and the proinflammatory cytokines tumor necrosis factor &agr;, interleukin-6, and interleukin-1&bgr; may be elevated in obesity and with menopause. The purpose of this multicenter study was to identify whether centrally located fat and/or overall adiposity were related to these inflammatory markers in healthy postmenopausal women. Design:We used dual-energy x-ray absorptiometry to assess overall and regional body composition (fat mass in particular) in 242 postmenopausal women in relation to plasma fibrinogen, serum C-reactive protein, and these proinflammatory cytokines. Results:Multiple regression analyses revealed that 36% of the variability in C-reactive protein (F = 32.4, P ≤ 0.0001) was accounted for by androidal fat mass (16.1%, P ≤ 0.0001), white blood cells (5.6%, P ≤ 0.0001), and age (2.3%, P = 0.0045). Regression analyses revealed that 30% of the variability in fibrinogen (F = 24.5, P ≤ 0.0001) was accounted for by white blood cells (3.1%, P = 0.0015), hip fat mass (2.2%, P = 0.0081), years since menopause (0.9%, P = 0.082), and geographic site (P ≤ 0.0001). Our results indicated that androidal fat mass and hip fat mass contributed to C-reactive protein and fibrinogen, respectively, whereas we found no association between whole-body or regional fat measures and cytokines. Conclusion:Further study is warranted to determine the responsiveness of these acute-phase proteins and cytokines to loss of body fat through exercise and dietary intervention in postmenopausal women.

Contribution of Serum Inflammatory Markers to Changes in Bone Mineral Content and Density in Postmenopausal Women: A 1-Year Investigation

Bone formation and resorption are influenced by inflammatory processes. We examined the relationships among inflammatory markers and bone mineral content (BMC) and density (BMD) and determined the contribution of inflammatory markers to 1-yr changes in BMC and BMD in healthy postmenopausal women. This analysis included 242 women at baseline from our parent Soy Isoflavones for Reducing Bone Loss project who were randomly assigned to 1 of 3 treatment groups: placebo, 80 mg/d soy isoflavones, or 120 mg/d soy isoflavones. BMD and BMC from the lumbar spine (LS), total proximal femur (hip), and whole body were measured by dual energy X-ray absorptiometry and the 4% distal tibia by peripheral quantitative computed tomography. Serum inflammatory markers (C-reactive protein, interleukin [IL]-1 beta, IL-6, tumor necrosis factor-alpha [TNF-alpha], and white blood cell count [WBC]) were measured at baseline, 6, and 12 mo. Because of attrition or missing values, data analysis at 12 mo includes only 235 women. Significant associations among IL-6, TNF-alpha, and WBC were observed with percent change in LS, hip, and whole body BMC and BMD. Multiple regression analysis indicated that in combination inflammatory markers accounted for 1.1-6.1% of the variance to the observed 12-mo changes in BMC and BMD. Our results suggest that modifying inflammatory markers, even in healthy postmenopausal women, may possibly reduce bone loss.

Alcohol extract of Echinacea pallida reverses stress-delayed wound healing in mice

Healing of open skin wounds begins with an inflammatory response. Restraint stress has been well documented to delay wound closure, partially via glucocorticoid (GC)-mediated immunosuppression of inflammation. Echinacea, a popular herbal immunomodulator, is purported to be beneficial for wound healing. To test the hypothesis, an alcohol extract of E. pallida was administrated orally to mice for 3 days prior to, and 4 days post wounding with a dermal biopsy on the dorsum. Concomitantly, mice were exposed to 3 cycles of daily restraint stress prior to, and 4 cycles post wounding. Echinacea accelerated wound closure in the stressed mice, but had no apparent wound healing effect for the non-stressed mice when compared to their respective controls. To test if the positive healing effect is through modulation of GC release, plasma corticosterone concentrations were measured in unwounded mice treated with restraint stress and the herbal extract for 4 days. Plasma GC in restraint stressed mice gavaged with Echinacea was not different from mice treated with restraint only, but was increased compared to the vehicle control. This data suggests that the improved wound healing effect of Echinacea in stressed mice is not mediated through modulation of GC signaling.

Is job-related stress the link between cardiovascular disease and the law enforcement profession?

Objective: To determine whether job-related stress is associated with alterations in pro- and anti-atherogenic inflammatory mediators among law enforcement officers. Methods: Markers of vascular inflammation and the self-reported stress measures of perceived stress, vital exhaustion, job strain, effort-reward imbalance, and social support were compared between officers (N = 444) and non-officers (N = 166). Results: Officers had higher levels of IL-1&bgr;, IL-6, IL-10, and TNF-&agr; and lower levels of C-reactive protein and fibrinogen. No more than 4% of the variability in any of the inflammatory mediators was explained by any stress measure for either the two groups or the entire sample. Conclusions: Law enforcement officers may be at an increased risk for cardiovascular disease due to a relatively greater pro-inflammatory vascular environment. However, this increased risk cannot be attributed to either chronic stress or the work-related stress measures assessed here.