Cheryl A. Rowe

Standardized capsule of Camellia sinensis lowers cardiovascular risk factors in a randomized, double-blind, placebo-controlled study

OBJECTIVE Previous studies examining the effect of tea drinking on cardiovascular health have produced mixed results due to their observational nature and qualitatively and quantitatively imprecise definitions of active tea components. The objective of this study was to determine if a standardized and defined decaffeinated green tea (Camellia sinensis) product lowers blood pressure, serum lipids, oxidative stress, and markers of chronic inflammation. METHODS A randomized, double-blind, placebo-controlled, parallel study on 111 healthy adult volunteers 21-70 y old was performed. We administered a standardized capsule of Camellia sinensis compounds (CSC) twice a day. Before and after 3 wk, blood pressure, serum lipids, serum amyloid-alpha (a marker of chronic inflammation), and serum malondialdehyde (a marker of oxidative stress) were measured. RESULTS After 3 wk, CSC lowered systolic and diastolic blood pressures by 5 and 4 mmHg, respectively. After 3 mo, systolic blood pressure remained significantly lower. CSC lowered serum amyloid-alpha by 42% and lowered malondialdehyde by 11.9%. In men, there were 10- and 9-mg/dL reductions in total and low-density lipoprotein (LDL) cholesterol, respectively. In all subjects with a baseline LDL cholesterol level >99 mg/dL, there was 9 mg/dL lowering of total and LDL cholesterol. Adverse effects were mild and few and not different from placebo. CONCLUSION CSC was effective for decreasing, in as quickly as 3 wk, blood pressure, LDL cholesterol, oxidative stress, and a marker of chronic inflammation, all independent cardiovascular risk factors.

Specific Formulation of Camellia sinensis Prevents Cold and Flu Symptoms and Enhances γδ T Cell Function: A Randomized, Double-Blind, Placebo-Controlled Study

Objective: Determine if a specific formulation of Camellia sinensis (CSF) can prevent illness and symptoms due to cold and flu, and enhance γδ T cell function Methods: Design: Randomized, double-blind, placebo-controlled study. Subjects: Healthy adults 18–70 years old. Intervention: Proprietary formulation of Camellia sinensis (green tea) capsules, or a placebo, twice a day, for 3 months. Measures of Outcome: As assessed by daily symptom logs, percentage of subjects experiencing cold and flu symptoms, number of days subjects experienced symptoms, and percentage of subjects seeking medical treatment. Mean in vivo and ex vivo proliferative and interferon gamma responses of subjects’ peripheral blood mononuclear cells to γδ T cell antigen stimulation. Results: Among subjects taking CSF there were 32.1% fewer subjects with symptoms (P = 0.035), 22.9% fewer overall illnesses of at least 2 days duration (P = 0.092), and 35.6% fewer symptom days (P < 0.002), compared to subjects taking placebo. γδ T cells from subjects taking CSF proliferated 28% more (P = 0.017) and secreted 26% more IFN-γ (P = 0.046) in response to γδ T cell antigens, as compared to γδ T cells from subjects taking placebo. CSF was well-tolerated. Conclusions: This proprietary formulation of CSF is a safe and effective dietary supplement for preventing cold and flu symptoms, and for enhancing γδ T cell function.

Preparation, characterization, and induction of cell apoptosis of cocoa procyanidins–gelatin–chitosan nanoparticles

Cocoa procyanidins (CPs)-gelatin-chitosan nanoparticles were fabricated based on the procyanidin-protein and electrostatic interactions, with an objective to enhance the stability and bioactivity of CPs. The CPs were purified using chromatographic methods and analyzed using HPLC equipped with a fluorescence detector (FLD) and mass spectrometer (MS). The purified CPs had a purity of 53.1% (w/w) and contained procyanidin oligomers (from monomer to decamers) and polymers, with polymers being the predominant component (26.4%, w/w). Different CPs-gelatin-chitosan mass ratios were tested to investigate the effects of formulation on the nanoparticle fabrication. Using CPs-gelatin-chitosan mass ratio of 0.75:1:0.5, the resultant nanoparticles had a particle size of 344.7 nm, zeta-potential of +29.8 mV, particle yield of 51.4%, loading efficiency of 50.1%, and loading capacity of 20.5%. The CPs-gelatin-chitosan nanoparticles were spherical as observed by scanning electron microscopy (SEM). Fourier transform infrared spectroscopy (FTIR) suggested that the primary interaction between the CPs and gelatin was hydrogen bond and hydrophobic interaction, while electrostatic interaction was the main binding force between chitosan and CPs-gelatin nanoparticles. Nanoencapsulation of the CPs significantly improved the stability of the CPs at 60°C. The CPs-gelatin-chitosan nanoparticles showed the same apoptotic effects at lower concentrations in human acute monocytic leukemia THP-1 cells compared with the CPs in solution.

Regular Consumption of Concord Grape Juice Benefits Human Immunity

γδ T cells are important immune surveillance cells residing in epithelial layers lining the intestine, lung, and reproductive tract. The main objective of this study was to test the hypothesis that consumption of dietary compounds from grapes would modify γδ T-cell function. Other factors related to immune function after grape juice consumption were also tested. A randomized, double-blind, placebo-controlled, parallel intervention was conducted: 100% grape juice made from Concord grapes or a placebo beverage was consumed by 85 individuals daily for 9 weeks. Subjects were asked not to consume other red, blue, and purple fruits during the study. Blood samples, taken at the beginning and the end, were analyzed for γδ T-cell numbers and proliferation, vitamin C, antioxidant capacity, and the ability to protect DNA from strand breaks. Those consuming the grape juice had significantly greater numbers of circulating γδ T cells and higher serum vitamin C levels compared to the placebo by two-way repeated-measure analysis of variance (P < .05). Individuals consuming the placebo had lower serum antioxidant activity, less γδ T-cell proliferation, and increased DNA strand breaks when challenged with H(2)O(2). Analysis of the data by structural equation modeling confirmed that 61% of the variance in biological functions at 9 weeks was due to grape juice consumption. Based on conventional statistical analyses, as well as on sophisticated modeling techniques, regular consumption of purple grape juice in the absence of other red, blue, or purple fruits benefited immunity in healthy, middle-aged human subjects.

UNIT 19.11 Influenza Virus

This unit contains several methods for infecting mice with influenza virus. It also includes protocols needed to propagate influenza virus in hen eggs, quantitate virus titers (in tissue culture medium and in influenza‐infected mouse serum), and adopt human isolates of influenza for growth in mice. Methods for measuring the 50% mouse lethal dose are also included. Finally, protocols for generating anti‐influenza cytotoxic T lymphocytes (CTL) from splenocyte precursors and harvesting pulmonary CTL following respiratory tract challenge of mice with influenza virus are provided.

Consuming Lentinula edodes (Shiitake) Mushrooms Daily Improves Human Immunity: A Randomized Dietary Intervention in Healthy Young Adults

Background: Mushrooms are widely cited for their medicinal qualities, yet very few human intervention studies have been done using contemporary guidelines. Objective: The aim of this study was to determine whether consumption of whole, dried Lentinula edodes (shiitake) mushrooms could improve human immune function. Primary objectives were to ascertain whether L. edodes consumption would improve γδ-T cell proliferation and activation responses, quantify a dose response, and elicit cytokine secretion patterns. Secondary objectives included determining changes in natural killer T (NK-T) cell proliferation and activation, secretory immunoglobulin A (sIgA) in saliva, and C-reactive protein (CRP) in serum. Design: Fifty-two healthy males and females, aged 21–41 years, participated in a 4-week parallel group study, consuming either 5 or 10 g of mushrooms daily. Each subject had blood drawn before and after 4 weeks of daily L. edodes consumption. Saliva and serum were also collected. Peripheral blood mononuclear cells were cultured in autologous serum for 24 hours or 6 days, stained, and examined by flow cytometry. Results: Eating L. edodes for 4 weeks resulted in increased ex vivo proliferation of γδ-T (60% more, p < 0.0001) and NK-T (2-fold more, p < 0.0001) cells. Both cell types also demonstrated a greater ability to express activation receptors, suggesting that consuming mushrooms improved cell effector function. The increase in sIgA implied improved gut immunity. The reduction in CRP suggested lower inflammation. The pattern of cytokines secreted before and after mushroom consumption was significantly different; consumption resulted in increased interleukin (IL)-4, IL-10, tumor necrosis factor (TNF)-α, and IL-1α levels, a decreased macrophage inflammatory protein-1α/chemokine C-C ligand 3 (MIP-1α/CCL3) level, and no change to IL-6, IL-1β, MIP-1β, IL-17 and interferon (IFN)-γ levels. Conclusions: Regular L. edodes consumption resulted in improved immunity, as seen by improved cell proliferation and activation and increased sIgA production. The changes observed in cytokine and serum CRP levels suggest that these improvements occurred under conditions that were less inflammatory than those that existed before consumption.