Aerobic, resistance, or combined exercise training and cardiovascular risk profile in overweight or obese adults: the CardioRACE trial.

BACKGROUND AND AIMS: To determine the comparative efficacy of resistance, aerobic, and combined resistance plus aerobic exercise on cardiovascular disease (CVD) risk profile. METHODS: This randomized controlled trial enrolled 406 adults aged 35-70 years with overweight or obesity and elevated blood pressure. Participants were randomly assigned to resistance (n = 102), aerobic (n = 101), combined resistance plus aerobic exercise (n = 101), or no-exercise control (n = 102). All exercise participants were prescribed 1 h of time-matched supervised exercise (the combination group with 30 min of each resistance and aerobic exercise) three times per week for 1 year. The primary outcome was the change from baseline to 1 year in the standardized composite Z-score of four well-established CVD risk factors: systolic blood pressure, low-density lipoprotein (LDL) cholesterol, fasting glucose, and per cent body fat. RESULTS: Among 406 participants (53% women), 381 (94%) completed 1-year follow-up. Compared with the control group, the composite Z-score decreased at 1 year, which indicates improved CVD risk profile, in the aerobic {mean difference, -0.15 [95% confidence interval (CI): -0.27 to -0.04]; P = .01} and combination [mean difference, -0.16 (95% CI: -0.27 to -0.04); P = .009] groups, but not in the resistance [mean difference, -0.02 (95% CI: -0.14 to 0.09); P = .69] group. Both aerobic and combination groups had greater reductions in the composite Z-score compared with the resistance group (both P = .03), and there was no difference between the aerobic and combination groups (P = .96). Regarding the four individual CVD risk factors, only per cent body fat decreased in all three exercise groups at 1 year, but systolic blood pressure, LDL cholesterol, and fasting glucose did not decrease in any exercise groups, compared with the control group. CONCLUSIONS: In adults with overweight or obesity, aerobic exercise alone or combined resistance plus aerobic exercise, but not resistance exercise alone, improved composite CVD risk profile compared with the control.

Novel nanoadjuvants balance immune activation with modest inflammation: implications for older adult vaccines.

BACKGROUND: Age-associated impairments of immune response and inflammaging likely contribute to poor vaccine efficacy. An appropriate balance between activation of immune memory and inflammatory response may be more effective in vaccines for older adults; attempts to overcome reduced efficacy have included the addition of adjuvants or increased antigenic dose. Next generation vaccine formulations may also use biomaterials to both deliver and adjuvant vaccine antigens. In the context of aging, it is important to determine the degree to which new biomaterials may enhance antigen-presenting cell (APC) functions without inducing potent inflammatory responses of APCs or other immune cell types (e.g., T cells). However, the effect of newer biomaterials on these cell types from young and older adults remains unknown. RESULTS: In this pilot study, cells from young and older adults were used to evaluate the effect of novel biomaterials such as polyanhydride nanoparticles (NP) and pentablock copolymer micelles (Mi) and cyclic dinucleotides (CDN; a STING agonist) on cytokine and chemokine secretion in comparison to standard immune activators such as lipopolysaccharide (LPS) and PMA/ionomycin. The NP treatment showed adjuvant-like activity with induction of inflammatory cytokines, growth factors, and select chemokines in peripheral blood mononuclear cells (PBMCs) of both young (n = 6) and older adults (n = 4), yet the degree of activation was generally less than LPS. Treatment with Mi or CDN resulted in minimal induction of cytokines and chemokine secretion with the exception of increased IFN-α and IL-12p70 by CDN. Age-related decreases were observed across multiple cytokines and chemokines, yet IFN-α, IL-12, and IL-7 production by NP or CDN stimulation was equal to or greater than in cells from younger adults. Consistent with these results in aged humans, a combination nanovaccine composed of NP, Mi, and CDN administered to aged mice resulted in a greater percentage of antigen-specific CD4+ T cells and greater effector memory cells in draining lymph nodes compared to an imiquimod-adjuvanted vaccine. CONCLUSIONS: Overall, our novel biomaterials demonstrated a modest induction of cytokine secretion with a minimal inflammatory profile. These findings suggest a unique role for biomaterial nanoadjuvants in the development of next generation vaccines for older adults.

"Just right" combinations of adjuvants with nanoscale carriers activate aged dendritic cells without overt inflammation.

BACKGROUND: The loss in age-related immunological markers, known as immunosenescence, is caused by a combination of factors, one of which is inflammaging. Inflammaging is associated with the continuous basal generation of proinflammatory cytokines. Studies have demonstrated that inflammaging reduces the effectiveness of vaccines. Strategies aimed at modifying baseline inflammation are being developed to improve vaccination responses in older adults. Dendritic cells have attracted attention as an age-specific target because of their significance in immunization as antigen presenting cells that stimulate T lymphocytes. RESULTS: In this study, bone marrow derived dendritic cells (BMDCs) were generated from aged mice and used to investigate the effects of combinations of adjuvants, including Toll-like receptor, NOD2, and STING agonists with polyanhydride nanoparticles and pentablock copolymer micelles under in vitro conditions. Cellular stimulation was characterized via expression of costimulatory molecules, T cell-activating cytokines, proinflammatory cytokines, and chemokines. Our results indicate that multiple TLR agonists substantially increase costimulatory molecule expression and cytokines associated with T cell activation and inflammation in culture. In contrast, NOD2 and STING agonists had only a moderate effect on BMDC activation, while nanoparticles and micelles had no effect by themselves. However, when nanoparticles and micelles were combined with a TLR9 agonist, a reduction in the production of proinflammatory cytokines was observed while maintaining increased production of T cell activating cytokines and enhancing cell surface marker expression. Additionally, combining nanoparticles and micelles with a STING agonist resulted in a synergistic impact on the upregulation of costimulatory molecules and an increase in cytokine secretion from BMDCs linked with T cell activation without excessive secretion of proinflammatory cytokines. CONCLUSIONS: These studies provide new insights into rational adjuvant selection for vaccines for older adults. Combining appropriate adjuvants with nanoparticles and micelles may lead to balanced immune activation characterized by low inflammation, setting the stage for designing next generation vaccines that can induce mucosal immunity in older adults.

Exercise after influenza or COVID-19 vaccination increases serum antibody without an increase in side effects.

Vaccination is an effective public health measure, yet vaccine efficacy varies across different populations. Adjuvants improve vaccine efficacy but often increase reactogenicity. An unconventional behavioral "adjuvant" is physical exercise at the time of vaccination. Here, in separate experiments, we examined the effect of 90-minute light- to moderate-intensity cycle ergometer or outdoor walk/jog aerobic exercise performed once after immunization on serum antibody response to three different vaccines (2009 pandemic influenza H1N1, seasonal influenza, and COVID-19). Exercise took place after influenza vaccination or after the first dose of Pfizer-BioNTech COVID-19 vaccine. A mouse model of influenza A immunization was used to examine the effect of exercise on antibody response and the role of IFNα as a potential mechanism by treating mice with anti-IFNα antibody. The results show that 90 min of exercise consistently increased serum antibody to each vaccine four weeks post-immunization, and IFNα may partially contribute to the exercise-related benefit. Exercise did not increase side effects after the COVID-19 vaccination. These findings suggest that adults who exercise regularly may increase antibody response to influenza or COVID-19 vaccine by performing a single session of light- to moderate-intensity exercise post-immunization.

Racial/ethnic variations in inflammatory markers: exploring the role of sleep duration and sleep efficiency.

Individuals from minoritized racial/ethnic groups have higher levels of circulating inflammatory markers. However, the mechanisms underlying these differences remain understudied. The objective of this study was to examine racial/ethnic variations in multiple markers of inflammation and whether impaired sleep contributes to these racial/ethnic differences. Nurses from two regional hospitals in Texas (n = 377; 71.62% White; 6.90% Black; 11.14% Hispanic, 10.34% Asian; mean age = 39.46; 91.78% female) completed seven days of sleep diaries and actigraphy to assess mean and variability in total sleep time (TST) and sleep efficiency (SE). On day 7, blood was drawn to assess 4 inflammatory markers: C-reactive protein (CRP), Interleukin-6 (IL-6), Interleukin-1 beta (IL-1ß), and tumor necrosis factor-alpha (TNF-α). Results from regression models showed differences in inflammatory markers by race/ethnicity, adjusting for age and gender. The associations between sleep parameters and inflammatory markers also varied by race/ethnicity. Among White nurses, lower mean and greater variability in actigraphy-determined TST and greater variability in diary-determined TST were associated with higher levels of IL-6. Among Black nurses, lower mean diary-determined SE was associated with higher levels of IL-6 and IL-1ß. Among Hispanic nurses, greater diary-determined mean TST was associated with higher CRP. Among Asian nurses, greater intraindividual variability in actigraphy-determined SE was associated with lower CRP. Among nurses, we did not find racial/ethnic disparities in levels of inflammation. However, analyses revealed differential relationships between sleep and inflammatory markers by race/ethnicity. Results highlight the importance of using a within-group approach to understand predictors of inflammatory markers.

Characterizing Patterns of Nurses' Daily Sleep Health: a Latent Profile Analysis.

BACKGROUND: Nursing is a demanding occupation characterized by dramatic sleep disruptions. Yet most studies on nurses' sleep treat sleep disturbances as a homogenous construct and do not use daily measures to address recall biases. Using person-centered analyses, we examined heterogeneity in nurses' daily sleep patterns in relation to psychological and physical health. METHODS: Nurses (N = 392; 92% female, mean age = 39.54 years) completed 14 daily sleep diaries to assess sleep duration, efficiency, quality, and nightmare severity, as well as measures of psychological functioning and a blood draw to assess inflammatory markers interleukin-6 (IL-6) and C-reactive protein (CRP). Using recommended fit indices and a 3-step approach, latent profile analysis was used to identify the best-fitting class solution. RESULTS: The best-fitting solution suggested three classes: (1) "Poor Overall Sleep" (11.2%), (2) "Nightmares Only" (8.4%), (3) "Good Overall Sleep" (80.4%). Compared to nurses in the Good Overall Sleep class, nurses in the Poor Overall Sleep or Nightmares Only classes were more likely to be shift workers and had greater stress, PTSD symptoms, depression, anxiety, and insomnia severity. In multivariate models, every one-unit increase in insomnia severity and IL-6 was associated with a 33% and a 21% increase in the odds of being in the Poor Overall Sleep compared to the Good Overall Sleep class, respectively. CONCLUSION: Nurses with more severe and diverse sleep disturbances experience worse health and may be in greatest need of sleep-related and other clinical interventions.

Intraindividual Variability in Sleep and Levels of Systemic Inflammation in Nurses.

OBJECTIVE: Disturbed sleep is common among nurses and is associated with morbidity and mortality. Inflammation may be one mechanism linking sleep and disease. However, most studies rely on retrospective questionnaires to assess sleep, which fail to account for night-to-night fluctuations in sleep across time (i.e., intraindividual variability [IIV]). We examined prospective associations between mean and IIV in sleep with inflammation markers in nurses. METHODS: Participants were 392 nurses (mean age = 39.54 years, 92% female, 23% night-shift working) who completed 7 days of sleep diaries and actigraphy to assess mean and IIV in total sleep time and sleep efficiency. Blood was drawn on day 7 to assess inflammation markers C-reactive protein, interleukin (IL)-6, tumor necrosis factor α, and IL-1ß. RESULTS: Greater IIV in total sleep time-measured via both actigraphy and sleep diary-was associated with higher IL-6 (actigraphy: b = 0.05, p = .046, sr = 0.01; diary: b = 0.04, p = .030, sr = 0.01) and IL-1ß (actigraphy: b = 0.12, p = .008, sr = 0.02; diary: b = 0.09, p = .025, sr = 0.01), but not C-reactive protein or tumor necrosis factor α. IIV in actigraphy- and sleep diary-determined sleep efficiency was not associated with inflammation biomarkers, nor were any mean sleep variables. Shift work did not moderate any associations. CONCLUSIONS: Nurses with more variable sleep durations had elevated levels of inflammation, which may increase risk for development of inflammatory-related diseases. Research should investigate how sleep regularization may change levels of inflammation and improve health.

Comparison of the Cardiovascular Benefits of Resistance, Aerobic, and Combined Exercise (CardioRACE): Rationale, design, and methods.

BACKGROUND: The benefits of aerobic exercise (AE) for cardiovascular disease (CVD) have been well documented. Resistance exercise (RE) has been traditionally examined for its effects on bone density, physical function, or metabolic health, yet few data exist regarding the benefits of RE, independent of and combined with AE, for CVD prevention. This randomized controlled trial, "Comparison of the Cardiovascular Benefits of Resistance, Aerobic, and Combined Exercise (CardioRACE)," is designed to determine the relative benefits of RE, AE, or combined RE plus AE training on CVD risk factors. METHODS: Participants are 406 inactive men and women (35-70 years) with a body mass index of 25-40 kg/m2 and blood pressure (BP) of 120-139/80-89 mm Hg without taking antihypertensive medications. Participants are randomly assigned to RE only, AE only, combined RE and AE (CE), or a no exercise control group. Participants perform supervised exercise at 50%-80% of their relative maximum intensity for both AE and RE, 3 times a week for 60 minutes per session, for 1 year (all 3 groups are time matched). RESULTS: The primary outcome is a composite z score including resting BP, low-density lipoprotein cholesterol (LDL-C), fasting glucose, and percent body fat, which is assessed at baseline, 6 months, and 12 months. Diet and outside physical activity are measured throughout the intervention for 1 year. CONCLUSION: CardioRACE (ClinicalTrials.govNCT03069092) will fill an important knowledge gap regarding the effects of RE, alone or in addition to the well-documented effects of AE. CardioRACE will help generate more comprehensive and synergistic clinical and public health strategies to prevent CVD.

Is Insomnia a Risk Factor for Decreased Influenza Vaccine Response?

Healthy young adult college students (N = 133) with Insomnia (n = 65) or No Insomnia (n = 68) were compared on influenza serum antibody levels pre- and four weeks postvaccination. Volunteers underwent structured clinical interviews for sleep disorders to ensure insomnia diagnoses, as well as psychiatric interviews, physical examinations, and drug testing to ensure comorbid health problems were not potential confounds. There were significant time (both groups had increases in antibody levels pre- to postvaccination) and group (Insomnia group had lower HI antibody levels overall) main effects, but the time × group interaction was nonsignificant. Exploratory analyses did find significant PSQI x Time (p < .001) and Insomnia Status × Time (p = .002) interaction effects. Results indicate insomnia may be a risk factor for lowered immunity to the influenza virus.

Alkaloids and athlete immune function: caffeine, theophylline, gingerol, ephedrine, and their congeners.

Plant alkaloids are found in foods, beverages, and supplements consumed by athletes for daily nutrition, performance enhancement, and immune function improvement. This paper examined possible immunomodulatory roles of alkaloids in exercise contexts, with a focus on human studies. Four representative groups were scrutinized: (a) caffeine (guaranine, mateine); (b) theophylline and its isomers, theobromine and paraxanthine; (c) ginger alkaloids including gingerols and shogaol; and (d) ephedra alkaloids such as ephedrine and pseudoephedrine. Emerging or prospective alkaloid sources (Goji berry, Noni berry, and bloodroot) were also considered. Human in vitro and in vivo studies on alkaloids and immune function were often conflicting. Caffeine may be immunomodulatory in vivo depending on subject characteristics, exercise characteristics, and immune parameters measured. Caffeine may exhibit antioxidant capacities. Ginger may exert in vivo anti-inflammatory effects in certain populations, but it is unclear whether these effects are due to alkaloids or other biochemicals. Evidence for an immunomodulatory role of alkaloids in energy drinks, cocoa, or ephedra products in vivo is weak to nonexistent. For alkaloid sources derived from plants, variability in the reviewed studies may be due to the presence of unrecognized alkaloids or non-alkaloid compounds (which may themselves be immunomodulatory), and pre-experimental factors such as agricultural or manufacturing differences. Athletes should not look to alkaloids or alkaloid-rich sources as a means of improving immune function given their inconsistent activities, safety concerns, and lack of commercial regulation.

Effects of aerobic exercise training on cerebral pulsatile hemodynamics in middle-aged adults with elevated blood pressure/stage 1 hypertension.

Mechanisms behind the protective effects of aerobic exercise on brain health remain elusive but may be vascular in origin and relate to cerebral pulsatility. This pilot study investigated the effects of 12-wk aerobic exercise training on cerebral pulsatility and its vascular contributors (large artery stiffness, characteristic impedance) in at-risk middle-aged adults. Twenty-eight inactive middle-aged adults with elevated blood pressure or stage 1 hypertension were assigned to either moderate/vigorous aerobic exercise training (AET) for 3 days/wk or no-exercise control (CON) group. Middle cerebral artery (MCA) pulsatility index (PI), large artery (i.e., aorta, carotid) stiffness, and characteristic impedance were assessed via Doppler and tonometry at baseline, 6, and 12 wk, whereas cardiorespiratory fitness (V̇o2peak) was assessed via incremental exercise test and cognitive function via computerized battery at baseline and 12 wk. V̇o2peak increased 6% in AET and decreased 4% in CON (P < 0.05). Proximal aortic compliance increased (P = 0.04, partial η2 = 0.14) and aortic characteristic impedance decreased (P = 0.02, partial η2 = 0.17) with AET but not CON. Cerebral pulsatility showed a medium-to-large effect size increase with AET, although not statistically significant (P = 0.07, partial η2 = 0.11) compared with CON. Working memory reaction time improved with AET but not CON (P = 0.02, partial η2 = 0.20). Our data suggest 12-wk AET elicited improvements in central vascular hemodynamics (e.g., proximal aortic compliance and characteristic impedance) along with apparent, paradoxical increases in cerebral pulsatile hemodynamics.NEW & NOTEWORTHY We identify differential central versus cerebrovascular responses to 12 wk of aerobic exercise training in middle-aged adults. Although proximal aortic compliance and characteristic impedance improved after 12 wk of exercise, cerebral pulsatility tended to unexpectedly increase. These data suggest short-term aerobic exercise training may lead to more immediate benefits in the central vasculature, whereas longer duration exercise training may be required for beneficial changes in pulsatility within the cerebrovasculature.

Severe mitral valve insufficiency caused by standard surgical aortic valve implantation and its reparation using suture-less prosthesis.

BACKGROUND: Aortic valve stenosis is the most frequent cardiac valve pathology in the western world. Surgical aortic valve replacement is the gold standard for the treatment of significant degenerative aortic valve diseases. CASE PRESENTATION: This case report highlights an unexpected abnormal iatrogenic shortening of the aorto-mitral continuity and its deformity, during traditional AVR using sutured stented aortic prosthesis as the first choice, which caused significant mitral valve regurgitation. The suture-less prosthesis was a rescue choice to restore the geometry and eliminate the deformation of the aorto-mitral continuity. CONCLUSIONS: Aortic valve replacement using suture-less prosthesis could be a valuable optional choice for lowering the risk of deformation of the aortic annulus and aorto-mitral continuity. It might provide better outcomes in combined procedures.

Mitochondrial Mass of Naïve T Cells Is Associated with Aerobic Fitness and Energy Expenditure of Active and Inactive Adults.

PURPOSE: Chronic exercise training is known to induce metabolic changes, but whether these adaptations extend to lymphocytes and how this may affect immune function remains largely unknown. This study was conducted to determine the extent to which mitochondrial characteristics of naïve T cells differ according to fitness status and to further examine the energy production pathways of cells from aerobically trained and inactive participants. METHODS: Blood was collected from 30 aerobically active (>6 h·wk -1 ) or inactive (<90 min·wk -1 ) men and women. Naïve T cell mitochondrial mass, membrane potential, and biogenesis were assessed with flow cytometry. Participants completed a treadmill maximal oxygen consumption (V̇O 2peak ) test and wore a physical activity monitor for 1 wk. In a subset of participants, naïve CD8 + T cell activation-induced glycolytic and mitochondrial ATP production was measured. RESULTS: Active participants exhibited 16.7% more naïve CD8 + T cell mitochondrial mass ( P = 0.046), 34% greater daily energy expenditure ( P < 0.001), and 39.6% higher relative V̇O 2peak ( P < 0.001), along with 33.9% lower relative body fatness ( P < 0.001). Among all participants, naïve CD8 + T cell mitochondrial mass was correlated with estimated energy expenditure ( r = 0.36, P = 0.048) and V̇O 2peak ( r = 0.47, P = 0.009). There were no significant differences in ATP production, mitochondrial biogenesis, or mitochondrial membrane potential between active and inactive groups. CONCLUSIONS: This is the first study to examine the effects of aerobic exercise training status on metabolic parameters within human naïve T cells. Findings suggest that mitochondrial adaptations in certain immune cell types are positively associated with aerobic fitness and energy expenditure. This study provides a foundation for future development of prophylactic and therapeutic interventions targeting specific immune cell subsets to improve the immune response and overall health.

Exercise duration modulates upper and lower respiratory fluid cellularity, antiviral activity, and lung gene expression.

Exercise has substantial health benefits, but the effects of exercise on immune status and susceptibility to respiratory infections are less clear. Furthermore, there is limited research examining the effects of prolonged exercise on local respiratory immunity and antiviral activity. To assess the upper respiratory tract in response to exercise, we collected nasal lavage fluid (NALF) from human subjects (1) at rest, (2) after 45 min of moderate-intensity exercise, and (3) after 180 min of moderate-intensity exercise. To assess immune responses of the lower respiratory tract, we utilized a murine model to examine the effect of exercise duration on bronchoalveolar lavage (BAL) fluid immune cell content and lung gene expression. NALF cell counts did not change after 45 min of exercise, whereas 180 min significantly increased total cells and leukocytes in NALF. Importantly, fold change in NALF leukocytes correlated with the post-exercise fatigue rating in the 180-min exercise condition. The acellular portion of NALF contained strong antiviral activity against Influenza A in both resting and exercise paradigms. In mice undergoing moderate-intensity exercise, BAL total cells and neutrophils decreased in response to 45 or 90 min of exercise. In lung lobes, increased expression of heat shock proteins suggested that cellular stress occurred in response to exercise. However, a broad upregulation of inflammatory genes was not observed, even at 180 min of exercise. This work demonstrates that exercise duration differentially alters the cellularity of respiratory tract fluids, antiviral activity, and gene expression. These changes in local mucosal immunity may influence resistance to respiratory viruses, including influenza or possibly other pathogens in which nasal mucosa plays a protective role, such as rhinovirus or SARS-CoV-2.

Age-dependent associations among insomnia, depression, and inflammation in nurses.

OBJECTIVE: Insomnia and depression have been inconsistently associated with inflammation. Age may be one important moderator of these associations. This study examined associations between insomnia and depression with inflammatory biomarkers in nurses and how these associations varied by age. Design: Participants were 392 nurses ages 18-65 (Mage = 39.54 years ± 11.15, 92% female) recruited from two hospitals. Main outcome measures: Participants completed surveys to assess insomnia and depression symptoms. Serum samples were obtained and analysed for inflammatory biomarkers interleukin-6 (IL-6), C-reactive protein (CRP), interleukin-1 beta (IL-1ß), and tumour necrosis factor alpha (TNF-α). Results: Neither insomnia nor depression symptoms were associated with inflammatory biomarkers. Older age was associated with higher IL-1ß, and age moderated the effects of depression symptoms on CRP and TNF-α: Greater depression symptoms were associated with higher CRP (b = .14, p = .017) and TNF-α (b = .008, p = .165) among older nurses only. Conclusion: Results suggest older nurses with higher depression symptoms may be at increased risk for elevated inflammation. Interventions should consider the role of age-related processes in modifying health and well-being. Given relatively low levels of depression in the current sample, future studies should replicate results in clinical and non-nurse samples.

Echinacea-induced cytosolic Ca2+ elevation in HEK293.

BACKGROUND: With a traditional medical use for treatment of various ailments, herbal preparations of Echinacea are now popularly used to improve immune responses. One likely mode of action is that alkamides from Echinacea bind to cannabinoid type 2 (CB2) receptors and induce a transient increase in intracellular Ca2+. Here, we show that unidentified compounds from Echinacea purpurea induce cytosolic Ca2+ elevation in non-immune-related cells, which lack CB2 receptors and that the Ca2+ elevation is not influenced by alkamides. METHODS: A non-immune human cell line, HEK293, was chosen to evaluate E. purpurea root extracts and constituents as potential regulators of intracellular Ca2+ levels. Changes in cytosolic Ca2+ levels were monitored and visualized by intracellular calcium imaging. U73122, a phospholipase C inhibitor, and 2-aminoethoxydiphenyl borate (2-APB), an antagonist of inositol-1,4,5-trisphosphate (IP3) receptor, were tested to determine the mechanism of this Ca2+ signaling pathway. E. purpurea root ethanol extracts were fractionated by preparative HPLC, screened for bioactivity on HEK293 cells and by GC-MS for potential constituent(s) responsible for this bioactivity. RESULTS: A rapid transient increase in cytosolic Ca2+ levels occurs when E. purpurea extracts are applied to HEK293 cells. These stimulatory effects are phospholipase C and IP3 receptor dependent. Echinacea-evoked responses could not be blocked by SR 144528, a specific CB2 receptor antagonist, indicating that CB2 is not involved. Ca2+ elevation is sustained after the Echinacea-induced Ca2+ release from intracellular Ca2+ stores; this longer-term effect is abolished by 2-APB, indicating a possible store operated calcium entry involvement. Of 28 HPLC fractions from E. purpurea root extracts, six induce cytosolic Ca2+ increase. Interestingly, GC-MS analysis of these fractions, as well as treatment of HEK293 cells with known individual and combined chemicals, indicates the components thought to be responsible for the major immunomodulatory bioactivity of Echinacea do not explain the observed Ca2+ response. Rather, lipophilic constituents of unknown structures are associated with this bioactivity. CONCLUSIONS: Our data indicate that as yet unidentified constituents from Echinacea stimulate an IP3 receptor and phospholipase C mediation of cytosolic Ca2+ levels in non-immune mammalian cells. This pathway is distinct from that induced in immune associated cells via the CB2 receptor.

Effects of Echinacea extracts on macrophage antiviral activities.

Type I interferons are a class of cytokines synthesized by leukocytes such as macrophages that limit viral replication. We hypothesized that one mechanism whereby Echinacea spp. extracts may enhance immunity is through modulating interferon-associated macrophage pathways. We used herpes simplex viral infection in the murine macrophage cell line RAW264.7 and monitored virus-induced cell death, interferon secretion, and two intracellular proteins that indicate activation of interferon pathways. Cells were incubated with control media or extracts from four different species (E. angustifolia, E. purpurea, E. tennesseensis, E. pallida). Cells incubated with extracts prior to infection showed very modest enhancement of viability, and no increase in the secretion of interferons alpha or beta as compared to control cells. Virus-infected macrophages treated with extracts from E. purpurea showed a small (<2-fold) induction of guanylate binding protein (GBP) production, but no effect of extracts from other species was observed. In virus-infected cells, all the extracts increased the amount of inducible nitric oxide synthase (iNOS) protein, and this effect varied by type of extraction preparation. Together, these results suggest that any potential antiviral activities of Echinacea spp. extracts are likely not mediated through large inductions of Type I interferon, but may involve iNOS.

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 tennesseensis ethanol tinctures harbor cytokine- and proliferation-enhancing capacities.

BACKGROUND: Members of the genus Echinacea are used medicinally to treat upper respiratory infections such as colds and influenza. The aim of the present investigation was to characterize the phytomedicinal properties of the American federally endangered species Echinacea tennesseensis. METHODS: Fifty-percent ethanol tinctures were prepared from roots, stems, leaves, and flowers and tested separately for their ability to influence production of IL-1beta, IL-2, IL-10, and TNF-alpha as well as proliferation by young human adult peripheral blood mononuclear cells (PMBC) in vitro. Tincture aliquots were stored at three different temperatures (4, -20, and -80 degrees C) for 21h before testing. At 1-month post-extraction, tinctures stored at -20 degrees C were tested again for cytokine modulation. Phytochemical analyses were performed using HPLC. RESULTS: Fresh root, leaf, and flower tinctures stimulated PBMC proliferation. Fresh root tinctures alone stimulated IL-1beta, IL-10, and TNF-alpha production. No tinctures modulated IL-2 production. Stem tinctures showed no activity. Storage temperature did not influence any outcomes. Root tinctures maintained their ability to modulate IL-1beta, IL-10, and TNF-alpha production after 1month of storage at -20 degrees C. CONCLUSIONS: These results suggest E. tennesseensis harbors phytomedicinal properties that vary by plant organ, with roots demonstrating the strongest activities.

A direct synthesis of 5,6-dihydroindolo[2,1-a]isoquinolines that exhibit immunosuppressive activity.

Dihydroindolo[2,1-a]isoquinolines were synthesized from tetrahydroisoquinolines and alpha-fluoroaldehydes by a novel two-step procedure. These compounds exhibited significant immunosuppressive activity against IL-2, IL-10 and IFN-gamma.