Stimulus-specific regulation of CD63 and CD203c membrane expression in human basophils by the flavonoid quercetin.

BACKGROUND: Flavonoids, such as quercetin, were reported to inhibit histamine release and cytokine production by basophils, but there is no evidence describing their action on membrane markers and intracellular biochemical pathways. OBJECTIVE: The aim of the study was to examine the effect of several quercetin doses on an in vitro human basophil activation system that evaluates up-regulation of membrane markers in response to agonists. METHODS: Leukocyte buffy coats from K(2)-EDTA anti-coagulated blood were treated with different concentrations of quercetin and triggered with anti-IgE ("allergy model") and with N-formyl-Met-Leu-Phe (fMLP) ("inflammation model"). Basophils were captured as CD123(bright)/HLA-DR(non-expressing) cells in a flow cytometry analysis and fluorescence values of CD63-FITC, CD203c-PE and CD123-PECy5 were used to produce dose response curves. RESULTS: Quercetin at a dose of 10 microg/ml strongly inhibited CD63 and CD203c membrane up-regulation triggered by both agonists, but it neither affected cell viability nor changed the expression of the phenotypic marker CD123. The anti-IgE model appeared highly sensitive to the effect of quercetin: a dose as low as 0.01 microg/ml was able to significantly decrease CD63 and CD203c membrane expression. In the fMLP model the dose response was different: quercetin doses from 0.01 to 0.1 microg/ml significantly increased up-regulation of membrane markers, achieving the highest effect with CD63. CONCLUSION: Very low doses of quercetin, within the pharmacological range, inhibit IgE-mediated membrane marker's up-regulation but prime the response to the chemotactic peptide fMLP; this stimulus specificity may have implications on the possible therapeutic action of the flavonoid in different pathologies.

Double-blind, placebo-controlled homeopathic pathogenetic trials: symptom collection and analysis.

BACKGROUND: Homeopathic pathogenetic trials (provings) are fundamental to homeopathy. Since most of the data from available provings have not been statistically evaluated, it is unclear how specific reported symptoms are and how they differ from those reported by people taking placebo. METHOD: We combine and analyse data from two different homeopathic pathogenic trials--including 10 and 11 provers, respectively, and both including 30% placebo-to test the null hypothesis that there is no significant difference between the number of symptoms in placebo and verum groups. RESULTS: The principal results were: Placebo reported less symptoms than verum groups. Symptom distribution according to predefined classes (common symptoms increased in intensity and/or duration-, cured, old, new and exceptional) was statistically different between placebo and verum group at a high level of significance (P<0.001). Compared to verum, placebo provers reported less new and old but more common (increased in duration or intensity) symptoms. Within repertory categories, other differences were detected. The two groups differ in terms of the duration of each symptom and kinetics of symptoms: most symptoms were more persistent in verum than in placebo groups and verum provers recorded a decreasing number of symptoms with time. Placebo provers did not show such a temporal pattern. CONCLUSIONS: If confirmed by other studies these results would demonstrate the non-equivalence between homeopathic medicines in high dilution and placebo and contribute to the improvement of proving methodology and evaluation.

Complexity science and homeopathy: a synthetic overview.

Homeopathy is founded on 'holistic' and 'vitalistic' paradigms, which may be interpreted--at least in part--in terms of a framework provided by the theory of dynamic systems and of complexity. The conceptual models and some experimental findings from complexity science may support the paradoxical claims of similia principle and of dilution/dynamization effects. It is argued that better appreciation of three main properties of complex systems: non-linearity, self-organization, and dynamicity, will not only add to our basic understanding of homeopathic phenomena but also illuminate new directions for experimental investigations and therapeutic settings.

A double-blind, placebo-controlled study on the diagnostic accuracy of an electrodermal test in allergic subjects.

BACKGROUND: Many unconventional diagnostic procedures based on bioelectrical skin responses are presently widely used for allergic diseases, but rigorous experimental evaluations of their accuracy are still lacking. AIM: We assessed whether an electrodermal device can correctly diagnose respiratory allergy. METHODS: The diagnostic accuracy of the electrodermal device was assessed in double-blind fashion in 72 allergic patients and 28 healthy volunteers. A random sequence of substances in sealed vials, including histamine, allergens, immunoglobulins at various dilutions and physiological saline, were tested in duplicate in each subject. RESULTS: A wide variability of the measurements was found in most patients irrespective of their allergy status and of the substance tested. Allergic patients showed more negative skin electrical response at the second trial, compared to normal controls, independent of the tested substance. No significant difference in skin electrical response between allergens and negative controls could be detected. CONCLUSION: We conclude that the studied bioelectrical method, under blind testing, cannot correctly detect respiratory allergy.

Observational study of quality of life in patients with headache, receiving homeopathic treatment.

This study describes the results obtained from a prospective observational research of homeopathic treatment for patients suffering from headache (migraine with- and without aura and tension-type headache). Fifty-three patients were asked to complete the SF-36 questionnaire at the beginning of the treatment and after 4-6 months. The homeopathic medicine and potency were not pre-defined, but were adapted to each single patient according to individualised homeopathic prescription. Most patients (73.6%) completed the study. There was heterogeneity in the answers (patients in very poor health as well as those with only slight disorders). Analysis of the data according to the concept of 'intention-to-treat' showed that after therapy, the mean and median scores of all life quality dimensions rose. More than 60% of the cases experienced an improvement in pain and the limitations caused by pain, as well as in limitations in social activities and health in general. All the differences between pre/post post treatment were statistically highly significant, with the strongest results in the 'bodily pain' and 'vitality' parameters (P < 0.0001).

Cytokine and nitric oxide levels in a rat model of immunologic protection from adjuvant-induced arthritis.

In the present study we investigated the correlation between the progression of adjuvant arthritis induced by Mycobacterium butyricum and the production of nitric oxide and some pro- and anti-inflammatory cytokines in arthritic rats and in rats treated with low intra-peritoneal doses of Mycobacterium 3 and 10 days after arthritis induction. The intra-peritoneal administration of Mycobacterium antigen significantly inhibited disease development. Compared to healthy rats, a rise in serum and peritoneal pro-inflammatory cytokines was observed in all arthritic rats already from the 14 day. The treatment with intra-peritoneal Mycobacterium was associated with a significant reduction in IL-6 serum concentrations and a slight decrease of IFN-gamma production by peritoneal macrophages. Nitrite/nitrate plasma and peritoneal levels were significantly higher in all arthritic rats. Intra-peritoneal administration of Mycobacterium caused a further increase in nitrite/nitrate plasma concentrations, while no differences were evident in nitric oxide production by peritoneal macrophages. From our data it is evident that among the variables here investigated, IL-6 seems to be the more representative marker of the disease and of the treatment effect. A possible role of nitric oxide as a modulator rather than a direct mediator in this model of inflammation is discussed.

Defective platelet response to arachidonic acid and thromboxane A(2) in subjects with Pl(A2) polymorphism of beta(3) subunit (glycoprotein IIIa).

The membrane complex alpha(IIb)beta(3) is the major receptor for fibrinogen and is involved in platelet adhesion and aggregation. Evidence has been presented that the Pl(A2) allele of the beta(3) Pl(A1/A2) gene polymorphism might be an independent risk factor for coronary thrombosis, but the matter is still controversial. We investigated the relationship between this polymorphism and possible alterations of platelet functions in vitro. The platelet adhesion to fibrinogen-coated microplate wells and the aggregation induced by several different agonists were tested in 63 healthy volunteers, among them, 49 subjects with Pl(A1/A1) polymorphism, 12 subjects with Pl(A1/A2) polymorphism and two subjects with (PlA2/A2) polymorphism. Subjects with PlA1/A2 polymorphism or with Pl(A2/A2) polymorphism showed significantly lower platelet responses as compared with Pl(A1/A1) subjects when either arachidonic acid or the thromboxane A(2) analogue, U46619, were used as agonists. In resting condition and after thrombin or ADP stimulation, platelet function was normal in all the subjects. An increased sensitivity to the anti-aggregatory effect of acetylsalicylic acid was observed in platelets from subjects with the Pl(A2) allele. Finally, using a flow-cytometric evaluation and determining the beta-thromboglobulin plasma levels, we did not find any evidence of a Pl(A2) platelet hyper-reactivity ex vivo. Our findings are not consistent with the hypothesis that the purported increase of cardiovascular risk in these subjects may be as a result of platelet hyperactivation. On the contrary, the Pl(A2) allele is associated with a platelet functional deficiency, specifically linked to the activation of the fibrinogen receptor by thromboxane A(2).

Influence of an acute exercise on neutrophil and platelet adhesion, nitric oxide plasma metabolites in inactive and active subjects.

In this work we studied the influence of an acute exercise either on nitrite/nitrate plasma levels or on neutrophil and platelet adhesion in inactive and active subjects. Twelve healthy subjects (6 inactives and 6 actives) exercised on a racing cycle ergometer performing stepwise increases in intensity until reaching, within 5 min, a heart rate of 150 beats x min(-1) which represents an oxygen consumption of about 75 % of the individual maximum rate of oxygen uptake. From peripheral venous blood samples (drawn from all subjects before, immediately after the end of exercise, and 1 hour later) neutrophils and platelets were isolated to test plate adhesion, and nitrite/nitrate concentrations were measured in the plasma. Immediately after the acute exercise, in active subjects we observed a significant decrease in the percentage of neutrophil adhesion (7.96+/-2.38 vs. 14.10+/-3.14), associated with an increase in nitrite/nitrate plasma levels (81.38+/-10.76 vs. 41.08+/-8.13 micromol x l(-1)), restored by a 40 min pre-incubation with NG-nitro-L-arginine methyl ester (L-NAME). In unstimulated platelets we observed a significant lower percentage of platelet adhesion in active subjects compared to inactives after exercise. With thrombin or adenosine 5'-diphosphate as agonists platelet adhesion did not result significantly different in active subjects compared to inactives. In conclusion, our data show that physical exercise can induce changes in some cell activities, even if transient, and favour the generation of nitric oxide. The lower adhesion of neutrophils and platelets induced by regular exercise could be an important goal in the prevention of vascular and inflammatory diseases.

Differential effects of dietary supplementation with fish oil or soy lecithin on human platelet adhesion.

To investigate the possible regulating role of omega-6 and of omega-3 fatty acids on platelet adhesiveness, we randomised 60 volunteers into three groups to take 20 ml (equivalent to 0.3 g omega-6, 3.6 g omega-3; omega-6/omega-3 ratio 0.1) per day of a fish oil supplement, or to take 25 g (equivalent to 1.5 g omega-6, 0.5 g omega-3; omega-6/omega-3 ratio 3) per day of a soy lecithin supplement, or to continue on their usual diet without any supplement (control group) for a period of 15 days. Platelet adhesion on fibrinogen-coated 96-well microtitre plates was evaluated in the resting condition and after stimulation with 2 microM ADP or 0.02 U/ml thrombin. Compared to the values before the experimental period, the fish oil group showed a significant reduction in stimulated adhesion (with ADP: from 18.8% to 15.6%, p<0.01; with thrombin: from 24.4% to 20.8%, p<0.005), whereas no difference was noted in the resting condition (from 3.6% to 3.5%, NS). In the soy lecithin group, platelet adhesion was increased in all test conditions (with ADP: from 18.7% to 23.2%, p<0.001; with thrombin: from 24.0% to 29.9% p<0.001; resting: from 3.5% to 6.6%, p<0.001). No significant changes were observed in the control group. A good correlation was found between platelet adhesion data and the changes in the platelet fatty acid omega-6/omega-3 ratio caused by the different supplementations. Our results indicate an inhibitory effect of fish oil rich in omega-3 fatty acids on stimulated human platelet adhesiveness and a stimulatory effect of soy lecithin rich in omega-6 fatty acids on resting and stimulated adhesion. They suggest moreover that the omega-6/omega-3 ratio is a determinant of platelet adhesion.

Neutrophil migration, oxidative metabolism and adhesion in early onset periodontitis.

The aim of this study was to evaluate neutrophil function in patients suffering from the generalized form of early onset periodontitis (EOP). We investigated neutrophil migration in vivo and neutrophil superoxide production and adhesion in response to a variety of compounds; neutrophils were isolated both from blood and a skin experimental exudate of 15 patients with EOP and of 15 sex- and age-matched normal control subjects. No difference was found in neutrophil migration in vivo (71.2+/-16.4x10(6) and 68.8+/-10.7x10(6) PMN/cm2/24 h in patients affected by early onset periodontitis and normal subjects respectively) and in adhesion. The superoxide production in response to STZ and PMA was similar between the 2 groups, while superoxide production in response to fMLP was markedly lower in patients than in control subjects both in circulating neutrophils (5.6+/-2.2 versus 10.4+/-2.3 nmoles O2-/10(6) cells, p<0.0001) and in exudate neutrophils (16.3+/-4.3 versus 22.3+/-4.7 nmoles O2-/10(6) cells, p<0.005). In general, neutrophil function in patients suffering from early onset periodontitis does not differ from control subjects, suggesting that the overall defence function of these cells is normal. The only parameter that we have found to be different between the 2 groups is the low superoxide production after fMLP stimulation. The stimulus- and function-specificity of this defect in neutrophils from patients indicates the existence of a dysregulation of the signal transduction pathway distal to fMLP receptor and proximal to NADPH oxidase activation.

Expression of FCepsilonII/CD23 on human neutrophils isolated from rheumatoid arthritis patients.

CD23, the low affinity receptor for IgE, is a 45 kilodalton molecule belonging to the C-type lectin family, some members of which have been identified as adhesion molecules. Since it has been described upregulated in different cells in chronic inflammatory diseases and in rheumatoid arthritis in particular, where neutrophils are directly involved in tissue damage, our interest, in this work, has been focused on the expression and regulation of this antigen on neutrophil membrane. We studied 22 patients suffering from rheumatoid arthritis and 22 healthy control subjects. CD23 expression on neutrophil membrane was analyzed by immunofluorescence. Neutrophils of 9 out of 22 patients expressed CD23 molecules, neutrophils of 11 out of 22 patients expressed CD23 only after 24 h of incubation in RPMI; only 2 out of 22 patients did not express the CD23 antigen on neutrophil membrane either after isolation or after a 24 h incubation. On the contrary neutrophils isolated from healthy subjects did not express CD23 molecules upon isolation. Only in 7/22 control subjects neutrophils resulted positive after 24 h of incubation in RPMI. Moreover, we found that in our experimental conditions the presence of IFN-g or GM-CSF alone or in combination with IL-4 inhibited CD23 expression during the 24 h incubation. Our results show that there is a strong association between neutrophil ability to express CD23 and rheumatoid arthritis, and that such expression may be regulated by GM-CSF, IFN-gamma and IL-4.

Dual effects of a homeopathic mineral complex on carrageenan-induced oedema in rats.

Carrageenan oedema, a classical experimental model commonly used to test activity of anti-inflammatory drugs, was used to evaluate the therapeutic activity of a low-potency mineral complex (MC). The MC was administered in the right plantar surface of albino rats 60 min before, simultaneously and 30 min after injection of carrageenan, an irritant which causes a local, transitory increase of fluid volume. The administration of the MC 60 min before the injection of carrageenan primed the animal to enhanced inflammatory response to the irritant. The administration of MC contemporarily to carrageenan did not modify the kinetic and the extent of the oedema, while the administration of the MC 30 min after the induction of the oedema significantly reduced the early phase of the inflammatory reaction. This indicated that the therapeutic action of this MC is not due to conventional anti-inflammatory effect but to activation of endogenous regulatory mechanisms, a phenomenon which may be regarded as a simple application of the 'similia rule'.

Homeopathic terminology: a consensus quest.

The aim of this project is to develop a common homeopathic terminology to improve communication. A questionnaire was sent by email to an international group of experts. As a result of an iterative process we propose that a number of terms which are inaccurate, unclear or have become outdated should be replaced by new terms. The main areas in which terminology should be updated are: concepts relating to: homeopathic pharmacology, research, homeopathic medicine, the principle of similarity, homeostasis and disease imitation, miasms, experimental homeopathy, provings and pathogenic trials.

Dual effects of a homeopathic mineral complex on carrageenan-induced oedema in rats

Carrageenan oedema, a classical experimental model commonly used to test activity of anti-inflammatory drugs, was used to evaluate the therapeutic activity of a low-potency mineral complex (MC). The MC was administered in... (AU)

Homeopathic terminology: a consensus quest

The aim of this project is to develop a common homeopathic terminology to improve communication. A questionnaire was sent by email to an international group of experts.As a result of an iterative process we propose that a number of terms which are... (AU)

Invasive aspergillosis in neutropenic patients: rapid neutrophil recovery is a risk factor for severe pulmonary complications.

BACKGROUND: In invasive aspergillosis, the duration of neutropenia is an accepted risk factor, and recovery from neutropenia is generally associated with a favourable outcome. However, the rapidity of granulocyte recovery may rarely be associated with adverse sequelae. The purpose of this study was to define the relationship between neutrophil (polymorphonuclear, PMN) recovery after chemotherapy-induced bone marrow aplasia and the occurrence of severe pulmonary complications (haemoptysis, pneumothorax and death) in patients with haematological malignancies who developed invasive fungal pneumonias. METHODS: Twenty consecutive patients were retrospectively studied; eight of them had developed pulmonary events between 5 and 11 days after neutrophil recovery that followed deep neutropenia (PMN < 100 microL-1). RESULTS: Five patients had haemoptysis (one of these also had pneumothorax) and three had pneumothorax. According to the multiplicative logistic model, the odds of occurrence of a pulmonary event increased significantly with increasing PMN count on the fifth day (P < 0.001). Five of the eight patients who had pulmonary complications died. Also, the risk of death was larger in the presence of rapid neutrophil recovery, although the difference was not statistically significant (P = 0.111). Analysis of clinical and laboratory data showed that the risk of pulmonary complications significantly increased when the neutrophil concentration was > 4500 microL-1 on day 5 after deep granulocyte neutropenia (PMN < 100 microL-1). There was no correlation between pulmonary complications, dosage of amphotericin B and deaths. CONCLUSION: The occurrence of life-threatening complications in patients with invasive fungal pneumonia is closely related to rapid PMN recovery.

A microplate-based colorimetric assay of the total peroxyl radical trapping capability of human plasma.

We developed a colorimetric assay estimating the radical-scavenging activity of human plasma. The test is based on a measure, in 96-well microplates at 450 nm, of the bleaching of carotenoid crocin by peroxyl radicals generated during thermal decomposition of 2, 2'-azobis-(2-amidinopopane) dihydrochloride (ABAP). The inhibition of this bleaching is a function of the antioxidant power of substances added to incubation mixture. We determined the optimal conditions for a sensitive, rapid, and reproducible assay of 50% inhibitory capacity (IC50) of a range of antioxidant substances and of plasma. Only a total of 200 microl of plasma is required in a complete dose-inhibition curve. The IC50 of normal human plasma resulted of 2.70 microl of plasma/250 microl assay volume. The total antioxidant capability (TAC) of plasma was defined as the reciprocal of IC50 and its value in a group of 19 healthy adults resulted in 0. 369 +/- 0.06. Intraassay and interassay coefficients of variation of plasma TAC were 6.13 and 4.80%, respectively. Measurement of samples with different uric acid concentration showed that antioxidant activity of uric acid accounts for approximately two-thirds of TAC.

[Homeopathy in the perspective of scientific research]. / L'omeopatia nella prospettiva della ricerca scientifica.

A significant portion of the traditional concepts of homeopathy (similia principle, experimentation on healthy humans, the cure of whole person, the use of minimum doses or high dilution/potency of medicines) are amenable to investigations conducted according to criteria accepted by biomedical science. Even though many randomized and controlled clinical studies seem to demonstrate the efficacy of some homeopathic medicines and their superiority to placebo, other trials have given negative results. A definite clear answer is not yet possible due to the scarce quality of some published reports, to lack of reproduction by independent investigators and to the uncertainty regarding the methodologies to be used for testing the claims of homeopathy. As regards the possible physiopathological, biophysical and pharmacological explanations for the action of homeopathic remedies, there are models which tend to set the similia principle as a general expression of the action-reaction principle, within the context of dynamic systems theory. The clarification of the more controversial aspects regarding dilution/potency of medicines remains tied to several promising developments in physics of condensed matter, in chaos theory and in biophysics.

Effect of Traumeel S, a homeopathic formulation, on blood-induced inflammation in rats.

OBJECTIVE: To evaluate the activity of Traumeel S (TRS), a homeopathic formulation containing Arnica montana and other plant extracts and minerals on an animal model of traumatic inflammation. DESIGN: TRS and individual components thereof were administered locally to rats 1 h before hind-paw injection with 0.1 ml of homologous blood and the development of oedema was measured over five hours. In each experiment, a control group was treated with saline. MAIN OUTCOME MEASURES: Paw volume of each rat was measured before oedema and 1, 3, and 5 h after oedema induction. Serum levels of IL-6 were determined at hour 5. RESULTS: The decrease of paw oedema, associated with the process of healing, was more rapid in rats treated with TRS (P < 0.05 after 3 h and P < 0.01 after 5 h). Similar effects were also induced by separate injection of most, but not all, TRS ingredients. The efficacy of complete mixture of TRS was higher than the combination of a selection of active components. TRS also reduced oedema development when administered after the oedema induction. The therapeutic effect of TRS was associated with a significant decrease of systemic interleukin-6 production. CONCLUSION: TRS seems to act by speeding up the healing process instead of blocking the development of oedema from the beginning. Moreover, its effect cannot be considered as the 'sum' of its active components and probably a synergistic interaction occurs to determine the final effect.