Effects of a Spirulina-based dietary supplement on cytokine production from allergic rhinitis patients.

Spirulina represents a blue-green alga that is widely produced and commercialized as a dietary supplement for modulating immune functions, as well as ameliorating a variety of diseases. We have previously shown that the in vitro culture of Spirulina with human peripheral blood mononuclear cells (PBMCs) modulated the production of cytokines. In the present study, we evaluated the impact of a Spirulina-based dietary supplement (Earthrise Nutritionals, Inc., Irvine, CA) on patients with allergic rhinitis by assessing the production of cytokines [interleukin (IL)-4, interferon (IFN)-gamma, and IL-2] critical in regulating immunoglobulin E-mediated allergy. In a randomized double-blinded crossover study versus placebo, allergic individuals were fed daily with either placebo or Spirulina, at 1,000 mg or 2,000 mg, for 12 weeks. PBMCs isolated before and after the Spirulina feeding were stimulated with phytohemagglutinin (PHA) prior to determining the levels of cytokine from cell culture supernatants. Although Spirulina seemed to be ineffective at modulating the secretion of Th1 cytokines (IFN-gamma and IL-2), we discovered that Spirulina, administered at 2,000 mg/day, significantly reduced IL-4 levels by 32% from PHA-stimulated cells. These results indicate that Spirulina can modulate the Th profile in patients with allergic rhinitis by suppressing the differentiation of Th2 cells mediated, in part, by inhibiting the production of IL-4. To our knowledge, this is the first human feeding study that demonstrates the protective effects of Spirulina towards allergic rhinitis.

Cocoa flavonols and procyanidins promote transforming growth factor-beta1 homeostasis in peripheral blood mononuclear cells.

Evidence suggests that certain flavan-3-ols and procyanidins (FP) can have a positive influence on cardiovascular health. It has been previously reported that FP isolated from cocoa can potentially modulate the level and production of several signaling molecules associated with immune function and inflammation, including several cytokines and eicosanoids. In the present study, we examined whether FP fractions monomers through decamers modulate secretion of the cytokine transforming growth factor (TGF)-beta(1) from resting human peripheral blood mononuclear cells (PBMC). A total of 13 healthy subjects were studied and grouped according to their baseline production of TGF-beta(1). When cells from individuals with low baseline levels of TGF-beta(1) (n = 7) were stimulated by individual FP fractions (25 microg/ml), TGF-beta(1) release was enhanced in the range of 15%-66% over baseline (P < 0.05; monomer, dimer, and tetramer). The low-molecular-weight FP fractions (or=hexamer), with the monomer and dimer inducing the greatest increases (66% and 68%, respectively). In contrast to the above, TGF-beta(1) secretion from high TGF-beta(1) baseline subjects (n = 6) was inhibited by individual FP fractions (P < 0.05; trimer through decamer). The inhibition was most pronounced with trimeric through decameric fractions (28%-42%), and monomers and dimers moderately inhibited TGF-beta(1) release (17% and 23%, respectively). Given the vascular actions associated with TGF-beta(1), we suggest that in healthy individuals, homeostatic modulation of its production by FP offers an additional mechanism by which FP-rich foods can potentially benefit cardiovascular health.

Modulation of TNF-alpha secretion in peripheral blood mononuclear cells by cocoa flavanols and procyanidins.

Epidemiological reports have suggested that the consumption of foods rich in flavonoids is associated with a lower incidence of certain degenerative diseases, including cardiovascular disease. Flavanols and their related oligomers, the procyanidins CFP, isolated from cocoa can modulate the production and level of several signaling molecules associated with immune function and inflammation in vitro, including several cytokines and eicosanoids. To further elucidate the potential immuno-modulatory functions of flavanol-rich cocoa, the present investigation examined whether isolated CFP fractions (monomers through decamers) influence the secretion of tumor necrosis factor-alpha (TNF-alpha) from resting and phytohemagluttinin (PHA)-stimulated human peripheral blood mononuclear cells (PBMC). We used an in vitro culture system where PBMC from 14 healthy subjects were introduced to individual CFP fractions for 72 h prior to measuring the levels of TNF-alpha released. The intermediate-sized CFP fractions (tetramers through octamers) were the most active on resting cells, causing a 3-4 fold increase in TNF-alpha relative to media baseline. The monomers and dimers were the least stimulatory of the fractions tested, displaying a 42 and 31% increase, respectively, over media control, whereas the trimers, nonamers and decamers showed an intermediate stimulation of this cytokine. In the presence of PHA, the intermediate-sized CFP fractions again were the most active, enhancing TNF-alpha secretion in the range of 48-128% relative to the PHA control. The monomers and dimers were slightly inhibitory (-1.5 and -15%, respectively), while trimers, nonamers and decamers stimulated moderate increases in TNF-alpha levels (13, 19 and 15%, respectively). The above results lend support to the concept that CFP can be immunomodulatory. The stimulation of TNF-alpha secretion may contribute to the putative beneficial effects of dietary flavanoids against microbial infection and tumorigenesis.

Effect of cocoa flavanols and their related oligomers on the secretion of interleukin-5 in peripheral blood mononuclear cells.

We previously showed that flavanols and their related oligomers (FLO) isolated from cocoa can have immunomodulatory effects on production of the cytokines interleukin-1beta (IL-1beta), IL-2, and IL-4. In the present study, we examined whether selected FLO fractions isolated from cocoa (monomer through decamer) modulate IL-5 protein secretion from resting and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC). Although FLO fractions were unstimulatory for IL-5 secretion in resting cells, PHA-induced IL-5 release from PBMC was markedly affected by certain FLO fractions. The monomeric and small oligomeric (dimer and trimer) fractions enhanced PHA stimulation by 50%, 54%, and 43%, respectively. In contrast, the larger oligomeric fractions (hexamer through decamer) inhibited IL-5 release in the range of 18% to 39%; the tetramer and pentamer showed intermediate effects. The increment in IL-5 suggests that FLO may preferentially stimulate immunoglobulin A. We suggest that in the oral cavity this could result in reduction in the risk for dental caries and periodontal disease. This work offers additional data for consideration of the health benefits of dietary FLO from a variety of foods, including those benefits associated specifically with consumption of some cocoas and chocolates.

A histomorphological investigation of the effect of cyclosporin on trabecular bone of the rat mandibular condyle.

The effect of cyclosporin A (CSA) on the condylar trabecular bone was evaluated by microscopy. Twenty, 5-week-old male Sprague-Dawley rats were divided into a treated and a control group. Animals in the treated group received CSA, 15 mg/kg body weight, by gastric feeding daily for 4 weeks; controls received the vehicle only. Five animals from each group were killed at the end of weeks 2 and 4. After histological processing, 10 tissue sections from the mid-part of the mandibular condyle were examined. Generally, a histopathological osteopenia was observed around the condyle after CSA treatment, especially at the end of week 4. In the control animals, the trabecular bone volume steadily increased from weeks 2 to 4 (from 0.46+/-0.07 to 0.61+/-0.07 mm(3)/mm(3)). However, the bone volume was significantly less in the CSA group than in the control group at both times (0.33+/-0.02 vs 0.46+/-0.07 and 0.26+/-0.07 vs 0.61+/-0.07 mm(3)/mm(3) for CSA vs control group at the end of weeks 2 and 4, respectively). Conversely, an increased marrow volume was observed in the CSA group at both these times (0.60+/-0.02 vs 0.42+/-0.08 and 0.71+/-0.06 vs 0.31+/-0.06 mm(3)/mm(3) for CSA vs control group at the end of weeks 2 and 4, respectively). Decreases were also observed in trabecular thickness, osteoid seam width, osteoid volume and cortical bone width. Because trabecular bone mass, osteoid mass and cortical bone thickness all showed a decrease after CSA at both times, an inhibitory effect of CSA on trabecular bone formation in the mandibular condyle is proposed.

Cyclosporin-induced gingival overgrowth at the newly formed edentulous ridge in rats: a morphological and histometric evaluation.

BACKGROUND: Since cyclosporin A (CsA)-induced overgrowth seldom occurs at sites distant from teeth, the periodontal ligament has been considered significant. The aim of this study was to examine overgrowth occurrence at the edentulous ridge--the sites without the ligament--after CsA therapy in rats. METHODS: After extracting all right maxillary molars, 16 Sprague-Dawley rats underwent a 2-week healing period. The animals were separated into CsA and control groups. CsA rats received 15 mg/kg of CsA by gastric feeding for 4 weeks, while the control group received only mineral oil. At the end of study, all animals were sacrificed and stone models were immediately obtained by rubber-based impressions. The edentulous ridge morphology, including the bucco-lingual width and the vertical height, was measured on the models. For histometry, 10 sections were selected from the edentulous ridge of each animal after undecalcified tissue preparation. The soft tissue areas of the edentulous ridge and the trabecular bone morphology of the dental alveolus were measured. RESULTS: CsA therapy produced a significant increase of the ridge width and height, measured from the stone models, when compared to the control group. Under histometry, CsA resulted in a significant increase of the epithelium, connective tissue, and total soft tissue areas. The measured trabecular bone volume was affected by both examining factors: the drug therapy and the location of the dental alveolus. CsA therapy produced a significant loss of bone volume but a significant increase of the bone-specific surface area. Although the mean osteoid volume was similar between CsA and control groups, a significant decrease of the fractional formation surface in the CsA group was revealed. CONCLUSIONS: An enlarged edentulous ridge and an altered dental alveolar bone morphology were observed in CsA-treated animals at the end of the study; therefore, we suggest that CsA may induce not only a soft tissue overgrowth but also an alveolar bone alteration at the edentulous ridge. The hypothesis that tooth or periodontal ligament is an essential component for the overgrowth development is questioned.

The effect of cocoa procyanidins on the transcription and secretion of interleukin 1 beta in peripheral blood mononuclear cells.

Recent data has demonstrated that cacao liquor polyphenols (procyanidins) have antioxidant activity, inhibit mRNA expression of interleukin-2 and are potent inhibitors of acute inflammation. Given the widespread ingestion of cocoa in many cultures, we investigated whether cocoa, in its isolated procyanidin fractions (monomer through decamer), would modulate synthesis of the pro-inflammatory cytokine, interleukin-1 beta. Both resting and phytohemagglutinin (PHA)-stimulated peripheral blood mononuclear cells (PBMC) were investigated at the levels of transcription and protein secretion. Individual cocoa fractions were shown to augment constitutive IL-1 beta gene expression, although values varied between subjects. Interestingly, the smaller fractions of cocoa (monomer-tetramer) consistently reduced IL-1 beta expression of PHA-stimulated cells by 1-15%, while the larger oligomers (pentamer-decamer) increased expression by 4-52%. These data, observed at the transcription level, were reflected in protein levels in PHA-induced PBMC. The presence or absence of PHA did not alter the effects of the cocoa procyanidins with the exception of the pentamer. This study offers additional data for the consideration of the health-benefits of dietary polyphenols from a wide variety of foods, including those benefits associated specifically with cocoa and chocolate consumption.

Effect of spirulina on the secretion of cytokines from peripheral blood mononuclear cells.

ABSTRACT The purpose of this study was to evaluate the immunomodulatory activity of Spirulina, a bluegreen alga used as a food supplement. The effects of Spirulina on the secretion of three cytokines from unstimulated and stimulated human peripheral blood mononuclear cells (PBMC) were examined. In resting PBMC, Spirulina stimulated secretion of interleukin (IL)-1beta, IL-4, and interferon (IFN)-gamma to nearly 2.0, 3.3, and 13.6 times basal levels, respectively. Spirulina induced levels of IFN-gamma (229 +/- 104 pg/ml) that were comparable to those seen after phytohemagglutinin (PHA) stimulation (476 +/- 121 pg/ml). However, it was much less mitogenic than PHA (13.1 +/- 6.9 pg/ml) with respect to the induction of IL-4 secretion (0.34 +/- 0.1 pg/ml). In PHA-stimulated cells, Spirulina enhanced secretion of IL-1beta, IL-4, and IFN-beta by 2.9, 4.0., and 1.6 times, respectively. Although Spirulina stimulates several cytokines, it is clearly more effective in the generation of a Thl-type response. This in vitro study offers additional data for consideration of the potential therapeutic benefits of Spirulina.

Perceived need for local anesthesia in tooth drilling among Anglo-Americans, Chinese, and Scandinavians.

This study explored ethnic differences in perceptions of pain and the need for local anesthesia for tooth drilling among age- and gender-matched Anglo-American, Mandarin Chinese, and Scandinavian dentists (n = 129) and adult patients (n = 396) using a systematic qualitative research strategy. Semistructured qualitative interviews determined: (a) the relative frequency of use or nonuse of anesthetic for similarly specified tooth drilling, (b) the reasons for nonuse of anesthetic as reported by dentists about their patients, and (c) the distribution of reasons for not using anesthetic. American dentists (n = 51) reported that about 1% of their adult patients did not use anesthetic compared with 90% among Chinese (n = 31) and 37.5% among Scandinavian dentists (n = 40). Of patients, Americans (n = 112) reported 6% nonuse of anesthetic for tooth drilling compared with 90% of 159 Chinese and 54% of 125 Scandinavians. Reasons among Anglo-Americans and Scandinavians were similar (ranked): the sensation was tolerable, to avoid numb feelings afterwards, and fear of injections. Danish patients were an exception; the fact that they had paid extra and out-of-pocket for anesthetic ranked second. In contrast, Chinese dentists made their decisions not to use anesthetics because they explained drilling as only a suan or "sourish" sensation, whereas injections were described as "painful." It was concluded that ethnic pain beliefs and differences in health-care systems are powerful psychosocial variables that affect pain perception and the perceived need for anesthetic.

Acute pain and use of local anesthesia: tooth drilling and childbirth labor pain beliefs among Anglo-Americans, Chinese, and Scandinavians.

Differences in ethnic beliefs about the perceived need for local anesthesia for tooth drilling and childbirth labor were surveyed among Anglo-Americans, Mandarin Chinese, and Scandinavians (89 dentists and 251 patients) matched for age, gender, and occupation. Subjects matched survey questionnaire items selected from previously reported interview results to estimate (a) their beliefs about the possible use of anesthetic for tooth drilling and labor pain compared with other possible remedies and (b) the choice of pain descriptors associated with the use of nonuse of anesthetic, including descriptions of injection pain. Multidimensional scaling, Gamma, and Chi-square statistics as well as odds ratios and Spearman's correlations were employed in the analysis. Seventy-seven percent of American informants reported the use of anesthetics as possible remedies for drilling and 51% reported the use of anesthetics for labor pain compared with 34% that reported the use of anesthetics among Chinese for drilling and 5% for labor pain and 70% among Scandinavians for drilling and 35% for labor pain. Most Americans and Swedes described tooth-drilling sensations as sharp, most Chinese used descriptors such as sharp and "sourish" (suan), and most Danes used words like shooting (jagende). By rank, Americans described labor pain as cramping, sharp, and excruciating, Chinese used words like sharp, intermittent, and horrible, Danes used words like shooting, tiring, and sharp, and Swedes used words like tiring, "good," yet horrible. Preferred pain descriptors for drilling, birth, and injection pains varied significantly by ethnicity. Results corroborated conclusions of a qualitative study about pain beliefs in relation to perceived needs for anesthetic in tooth drilling. Samples used to obtain the results were estimated to approach qualitative representativity for these urban ethnic groups.

Consensus analysis: reliability, validity, and informant accuracy in use of American and Mandarin Chinese pain descriptors.

A quantitative method for validating qualitative interview results and checking sample parameters is described and illustrated using common pain descriptions among a sample of Anglo-American and mandarin Chinese patients and dentists matched by age and gender. Assumptions were that subjects were members of a sociocultural group (e.g. ethnic or professional/lay) and answered questions independently about a monotonic domain (e.g. pain). Subjects answered 18 true/false items derived from and selected to reflect pain perceptions consistent with published and unpublished interview data. Estimates of consistency in use of descriptors within groups, validity of description, accuracy of individuals compared with others in their group, and minimum required sample size were calculated using Cronbach's alpha, factor analysis, and Bayesian probability. Ethnic and professional differences within and across groups were also tested using multidimensional scaling (MDS) and hypothesis testing. Consensus (consistency of subject response by group) was .99 among Anglo-Americans and .97 among Chinese. Mean subject accuracy was .81 for Americans and .57 for Chinese, indicating the need for larger numbers of Chinese to supplement each others' statements. However, larger numbers of subjects were recruited than actually required for each ethnic group at .95 confidence limits. MDS showed similarities in use of descriptors within ethnic groups, while there were differences (p < .001) between Chinese and American groups. Use of covalidating questionnaires that reflect results of qualitative interviews are recommended in order to estimate sample parameters such as intersubject agreement, individual subject accuracy, and minimum required sample sizes.

Fear of injections and report of negative dentist behavior among Caucasian American and Taiwanese adults from dental school clinics.

Fear of injections and reports of negative dentist behavior and associations with dental anxiety and avoidance of treatment were explored among 951 adults from dental school clinics in Iowa City, Iowa and Taipei, Taiwan. Use and fear of anesthetic injections and negative dentist behavior were assessed by written questionnaire to test associations with demographics, overall dental anxiety (Dental Anxiety Scale or "DAS") and utilization behaviors. Frequency and logistic regression analyses showed that use of dental anesthetics for routine treatment was much greater overall among caucasian Americans than Taiwanese, as was fear of injections. Taiwanese and Americans with high dental anxiety (DAS 12) had similar high fear of injections, but inspite of similar fears about dental drilling, high anxiety Taiwanese reported using much less local anesthesia for routine treatments than did high anxiety Americans. Report of condescending remarks to patients ("put downs") by dentists was mainly an American phenomenon associated with high dental anxiety. Avoidance of appointment making was high for persons afraid of injections and for Americans reporting negative dentist behavior. Avoidance was highest in subjects with high dental anxiety. That predominant characteristics or etiologies of dental anxiety can differ by cultural differences in dental health care systems, dentist beliefs and/or expectations of patients within those systems was discussed in relation to the literature.