Impact of polyunsaturated vegetable oils on adiponectin levels, glycaemia and blood lipids in individuals with type 2 diabetes: a randomised, double-blind intervention study.

BACKGROUND: Low adiponectin levels are discussed as risk factor for cardiovascular events. This is of special importance in individuals with type 2 diabetes (T2DM) because they are at higher risk for cardiovascular diseases. The present study aimed to investigate the effect of two plant oils rich in polyunsaturated fatty acids (PUFA), with different content of omega-3 fatty acids, on adiponectin levels, glucose and lipid metabolism in T2DM individuals treated either with insulin or oral anti-diabetics (OAD). METHODS: Ninety-two subjects with T2DM [34 treated with insulin (T2DM-Ins) and 58 treated with OAD (T2DM-OAD)] participated in this randomised, double-blind, parallel intervention study. Individuals received either 9 g of nut oil (n-3:n-6 ratio: 1.3 : 6.1) or mixed oil (n-3:n-6 ratio: 0.6 : 5.7) per day for 10 weeks. The fatty acid profile, tocopherol, adiponectin levels and parameters regarding glucose and lipid metabolism were assessed at baseline, during and after the intervention. RESULTS: Compliance was confirmed by significant increases in γ-tocopherol and PUFA in both oil groups. An increase in adiponectin levels in T2DM-Ins participants (+6.84% in nut oil and +4.47% in mixed oil group after 10 weeks compared to baseline) was observed, albeit not significantly different from T2DM-OAD individuals (P = 0.051). Lipid and glucose metabolism were not affected by the intervention. CONCLUSIONS: The present study provides evidence that a small and easy change in dietary behaviour towards better fat quality moderately increases adiponectin levels in T2DM-Ins subjects, independently of the administered plant oil.

Intake of a resveratrol-containing dietary supplement has no impact on DNA stability in healthy subjects.

It has been postulated that the beneficial health effects of dietary supplements and of red wines which contain resveratrol (RES) are due to the anti-oxidative properties of this phenolic compound, but evidence for protection against reactive oxygen species is mainly based on results of in vitro experiments and high-dose animal experiments. Aim of this study was to find out if intake of a RES-containing supplement protects healthy humans against oxidative DNA-damage and alters their redox status. Therefore, an intervention trial was conducted in which the participants (n=12) consumed a RES-containing supplement over a period of five days. At the start, after one day and after five days of consumption, and after a washout period DNA stability was measured in single cell gel electrophoresis (SCGE) assays with peripheral blood lymphocytes. These tests were conducted (a) under standard conditions, which reflect single- and double-strand DNA breaks, (b) after treatment of the cells with hydrogen peroxide, which enables detection of alterations of the ROS sensitivity, and (c) by use of formamidopyrimidine DNA-glycosylase (FPG), which provides information on formation of oxidatively damaged bases (pyrimidines). Furthermore, the biochemical parameters TAC (total antioxidant capacity) and oxLDL (oxidized low-density lipoprotein), which reflect the redox status, and C-reactive protein (CRP), a marker of inflammation, were monitored. The intake of the supplement had no significant impact on the DNA stability parameters and on the different biomarkers of the redox status. Our results indicate that intake of 6mg RES per day via the supplement does not cause DNA-protective or antioxidant effects. This amount is equivalent to or lower than that reached after intake of many (ca. 50%) of the RES-containing preparations which are currently on the market in Middle Europe, and is contained in 0.3-2L red wine.

Recombinant allergen-based monitoring of antibody responses during injection grass pollen immunotherapy and after 5 years of discontinuation.

BACKGROUND: Subcutaneous injection immunotherapy (SCIT) is considered as antigen-specific and disease-modifying treatment with long-lasting effect. METHODS: We used a panel of recombinant grass pollen allergens for analyzing allergen-specific IgE, IgG(1) -IgG(4) , IgM, IgA, and light-chain (kappa, lambda) responses in grass pollen-allergic patients who had received one course of injection immunotherapy (SCIT) with an aluminum hydroxide-adsorbed grass pollen extract or only anti-inflammatory treatment. Serum samples were analyzed before and after 5 months of treatment as well as after 5 years. RESULTS: After 5 months of SCIT but not of anti-inflammatory treatment, IgG(1) > IgG(4) > IgG(2) > IgA antibody responses using both kappa and lambda light chains specific for major grass pollen allergens (Phl p 1, Phl p 5, Phl p 6, Phl p 2) increased significantly, whereas specific IgM or IgG(3) levels were unaltered. Allergen-dependent basophil degranulation was only inhibited with SCIT sera containing therapy-induced allergen-specific IgG antibodies. Likewise, decreases in Phl p 1- and Phl p 5-specific IgE levels and significant (P<0.001) reduction in symptom and medication scores were found only in the SCIT group but not in the group of patients receiving anti-inflammatory treatment. After 5 years, allergen-specific IgG antibody levels in the SCIT group had returned to baseline levels and there was no significant difference regarding symptoms between the SCIT and non-SCIT groups. CONCLUSION: The results from our observational study demonstrate that only SCIT but not anti-inflammatory treatment induces allergen-specific IgG and reduces boosts of allergen-specific IgE production but that one SCIT course was not sufficient to achieve long-term immunological and clinical effects.

Analysis of the antibody responses induced by subcutaneous injection immunotherapy with birch and Fagales pollen extracts adsorbed onto aluminum hydroxide.

BACKGROUND: Allergen-specific subcutaneous immunotherapy (SCIT) is an antigen-specific therapy of IgE-mediated allergies. In the present study, we analyze the epitope specificities of antibody responses induced by SCIT with allergen extracts from pollen of trees belonging to the order Fagales (birch, alder, hazel) adsorbed onto aluminum hydroxide. METHODS: The IgE, IgG1-4 and IgA responses to defined recombinant allergens (birch pollen: Bet v 1; alder pollen: Aln g 1; hazel pollen: Cor a 1; apple: Mal d 1) as well as to Bet v 1-derived recombinant fragments and synthetic peptides were analyzed in sera from patients who had undergone SCIT for different periods of time. RESULTS: Long-term SCIT (>1 year; cumulative dose >1,000,000 SQ units) induced more pronounced IgG1, IgG2 and IgG4 responses to Bet v 1 and Bet v 1-related allergens according to the degree of sequence homology (Bet v 1>Aln g 1>Cor a 1>Mal d 1) than short-term SCIT (<1 year; cumulative dose <1,000,000 SQ units). In contrast to patients treated for <1 year, patients treated for >1 year mounted distinct IgG1, IgG2 and IgG4 responses against sequential Bet v 1 epitopes. No relevant allergen-specific IgA or IgG3 responses were induced by short- or long-term SCIT. Using a competitive ELISA assay, it could be shown that serum IgG from patients undergoing long-term SCIT inhibited IgE reactivity to Bet v 1 better than IgG from patients undergoing short-term SCIT. CONCLUSION: SCIT with allergen extracts adsorbed onto aluminum hydroxide induces IgG responses against new epitopes that block IgE binding and cross-react with structurally related allergens depending, among other factors, on duration of treatment and cumulative injected dose.

Different allergenic activity of grass pollen allergens revealed by skin testing.

BACKGROUND: Grass pollen is one of the most important allergen sources. The aim of this study was to compare the in vivo allergenic activity of two recently characterized major grass pollen allergens, Phl p 4 and Phl p 13, with three established major grass pollen allergens, Phl p 1, Phl p 2 and Phl p 5 as a basis for the formulation of a grass pollen allergy vaccine based on purified allergens. MATERIAL AND METHODS: Eighty-two grass pollen allergic patients were skin prick tested with serial dilutions of approximately equimolar concentrations of the purified allergens in a double-blind study. RESULTS: Phl p 4 and Phl p 13 were identified as major grass pollen allergens according to IgE binding frequency (Phl p 4: 85%; Phl p 13: 56%), but exhibited a five to nine-fold lower allergenic skin reactivity compared to Phl p 1, Phl p 2 or Phl p 5. CONCLUSION: Our results indicate that Phl p 4 and Phl p 13 are not essential components for a therapeutic grass pollen vaccine and underpin the importance of evaluating the in vivo allergenic activity of individual allergens for the formulation of therapeutic vaccines based on purified allergens.

Coffee consumption protects human lymphocytes against oxidative and 3-amino-1-methyl-5H-pyrido[4,3-b]indole acetate (Trp-P-2) induced DNA-damage: results of an experimental study with human volunteers.

Aim of the study was to investigate the impact of coffee on DNA-stability in humans. DNA-damage was monitored in lymphocytes of eight individuals with single cell gel electrophoresis assays before and after consumption of 600 ml coffee (400 ml paper filtered and 200 ml metal filtered/d) for five days. Under standard conditions, no alteration of DNA-migration was seen, but a strong reduction of DNA-migration attributable to endogenous formation of oxidised purines and pyrimidines was detected with restriction enzymes; furthermore DNA-damage caused by reactive oxygen radicals (H2O2 treatment) and by the heterocyclic aromatic amine 3-amino-1-methyl-5H-pyrido[4,3-b]indole-acetate was significantly reduced after coffee consumption by 17% and 35%, respectively. Also in in vitro experiments, inhibition of H2O2 induced DNA-damage was observed with coffee at low concentrations (

The role of Foxp3+ T cells in long-term efficacy of prophylactic and therapeutic mucosal tolerance induction in mice.

BACKGROUND: Mucosal tolerance induction is suggested as treatment strategy for allergic diseases. Using a murine model of birch pollen (BP) allergy we investigated the long-term efficacy and the underlying mechanisms of mucosal tolerance induction with two structurally different molecules in a prophylactic and in a therapeutic set-up. METHODS: The three-dimensional major BP allergen Bet v 1 or a nonconformational hypoallergenic fragment thereof was intranasally applied before (prophylaxis) or after sensitization (therapy). RESULTS: In the prophylactic application both the Bet v 1 allergen and the fragment prevented allergic sensitization, and this effect lasted for 1 year. In the therapeutic approach established allergic immune responses were also suppressed after treatment with either of the molecules. However, a long-lasting curative effect (6 months) was only achieved with the Bet v 1 allergen but not with the Bet v 1 fragment. Real-time reverse transcriptase polymerase chain reaction (RT-PCR) analysis of splenocytes revealed that tolerance induction with the Bet v 1 allergen was associated with enhanced expression of transforming growth factor (TGF)-beta, interleukin (IL)-10, and Foxp3 mRNA in CD4+ T cells, whereas treatment with the fragment led to the induction of either Foxp3 (prophylaxis) or IL-10 (therapy) alone. CONCLUSION: From these data we concluded (i) that the mechanisms underlying peripheral tolerance are linked to the conformation of the antigen, (ii) that mucosal tolerance is mediated by separate regulatory cell subsets, and (iii) that the long-term efficacy of immunosuppression is associated with the presence of Foxp3+ T cells.

Moulds in housing: visual inspection and spore counts comparison--implications for future strategies in the public health setting.

OBJECTIVES: Among the problems of health related to living conditions presented to a public consulting centre for environmental medicine in Vienna, the main point at issue was damp housing and mould growth. On answering this demand indoor exposure to fungal spores was identified by visual semi-quantitative assessment of fungal growth on the one hand and quantitative measurement of viable spores on the other hand. The validity and practicability of this approach was investigated. METHOD: We applied these two simple methods in a standardised form and compared the results in a field study, which has been conducted between 1995 and 1999. In 197 rooms in Viennese flats (183 of which were claimed to be "mouldy") fungal spore concentrations were measured by RCS-impactor and Rose-Bengal-agar. Outdoor measurements were taken simultaneously outside the buildings and were used for reference. RESULTS: Independent classification of visible growth of mould in flats correlated significantly (r = 0.635; p < 0.0001) with the ratio of indoor/ outdoor concentration of fungal spores. CONCLUSIONS: It is usually possible to classify the severity of the problem just by local inspection. Visible plaques should always be an indication for sanitary measures. However, spore counting is indicated for quantifying health relevant exposures, for scientific comparisons and in documentation and follow up such as for litigation purposes. Quantitative data bear more credibility and so help to prove the need of sanitation.

Double-blind, randomised, placebo controlled low level laser therapy study in patients with primary Raynaud's phenomenon.

BACKGROUND: No causal treatment of primary Raynaud's phenomenon is available due to its unclear aetiology. Low level laser therapy (LLLT) is applied in a multitude of medical conditions often without sufficient evidence of efficacy and established mechanisms. To asses the effect of this therapy in patients with primary Raynaud's phenomenon a randomised, double blind, placebo controlled cross over study was designed. PATIENTS AND METHODS: Absolute and relative frequency and intensity of vasospastic attacks during three weeks of either LLLT or placebo therapy and results of infrared thermography before onset and at the end of both therapy sequences were evaluated in 15 patients with primary Raynaud's phenomenon. RESULTS: Frequency of Raynaud's attacks was not significantly affected by low level laser therapy. Compared to placebo a significantly lower intensity of attacks during laser irradiation was observed, but no transfer effect occurred. Additionally the mean temperature gradient after cold exposure was reduced after laser irradiation, while the number of fingers showing prolonged rewarming was unaffected. CONCLUSION: Though further studies are necessary to confirm these results we could demonstrate for the first time in a double blind placebo controlled clinical trial that low laser therapy is a potential candidate for an effective therapy of Raynaud's phenomenon, although effects seem to be of short duration.

Khat (Catha edulis) consumption causes genotoxic effects in humans.

We used the micronucleus (MN) test to determine the genetic damage caused by khat, a widely consumed psychostimulant plant, in exfoliated cells of volunteers who chewed the drug on a regular basis. In the first study in which we compared the frequency of MN in buccal and bladder mucosa cells in 20 khat consumers (10-160 g/day) and 10 controls, a pronounced (8-fold) increase in micronucleated buccal mucosa cells was seen among khat consumers; khat consumption did not lead to a detectable elevation of micronucleated bladder mucosa cells. Among heavy khat chewers, 81% of the MN had a centromere signal indicating that khat is aneuploidogenic. To investigate the effect of simultaneous consumption of tobacco and alcoholic beverages, we compared the MN frequency in buccal cells of 25 khat consumers (20-85 g/day) who smoked cigarettes (15-60/day) and drank alcoholic beverages (15-80 g of pure ethanol/day) with a control group (control group I) of 25 individuals matched for age, body weight, tobacco and alcohol consumption and with another control group of 25 individuals (control group II) not consuming any of the drugs. The frequency of buccal mucosa cells with MN was higher in control group I than in group II and the effect of khat, tobacco and alcohol was found to be additive. A time-kinetics study on khat-induced MN showed that the highest frequency of MN was observed during the fourth week after consumption. In light of the large body of evidence on the close association between genetic damage and cancer, these results suggest that khat consumption, especially when accompanied by alcohol and tobacco consumption, might be a potential cause of oral malignancy.

Detection of genotoxic effects of heavy metal contaminated soils with plant bioassays.

Aim of the present study was the development of a bioassay which enables the detection of genotoxic effects of heavy metal contaminated soils. In the first part of the present study, the data base on metal effects in plant bioassays was extended. Four metal salts, namely Cr(VI)O3, Cr(III)Cl3, Ni(II)Cl2 and Sb(III)Cl3 were tested comparatively in MN tests with pollen tetrad cells of Tradescantia clone #4430 and in meristematic root tip cells of Vicia faba. With Cr6+ and Ni2+, clear-cut dose-effects were observed in a range between 0.75 and 10.0 mM, whereas this was not the case with Cr3+ (range tested 1.25-10 mM) and Sb3+ (range 0.30-5.25 mM). In Vicia, negative results were obtained with the four metal salts under all conditions of test. To compare the mutagenic potencies of the metals, the increases of the regression curves (k-values) were calculated, they indicate the number of MN induced per mM in 100 tetrad cells. The corresponding values for Cr6+ and Ni2+ are 0.87 and 1.05, respectively. It appears that the Tradescantia system is in particular sensitive towards those metal species which cause DNA damage in animals and man such as Cr6+, Cd2+, Ni2+, and Zn2+, whereas no clear positive results were obtained with less harmful metal ions such as Cu2+, Cr3+ or Sb3+. In the second part of the study, the mutagenic effects of four metal contaminated soils and two types of standardized leachates (pH 4.0 and pH 7.0) of these soils were tested in Tradescantia and in Vicia. In addition, chemical analyses were carried out to determine the metal concentrations in the soils and in the extracts. Two of the samples contained highly elevated levels of a number of metals (Zn, Pb, Cu, Cd, Sb, As), one soil came from the Central Austrian Alps and contained high As levels only. Direct exposure of the Tradescantia plants in the soils resulted in a drastic increase of the MN frequencies over the background. The lowest effect was seen with the Slovakian soil which contained in particular Sb and As (4.5-fold increase over the background), with the other soils, the induced frequencies were 11-15-fold over the control values. On the contrary, negative results were obtained upon exposure of Tradescantia cuttings in the leachates and upon implantation of germinated Vicia beans in the soils. The results of the present study indicate that Trad-MN assays with direct exposure of intact plants is an appropriate method which enables to detect genotoxic effects of metal contaminated soils in situ. This simple and fast biomonitoring assays might be a valuable supplement to analytical analyses of contaminated soils.

Evaluation of diarrhoea management of health professionals trained at the Diarrhoea Training Unit of Rawalpindi General Hospital.

To promote the use of oral rehydration therapy (ORT), a training programme was launched by The Pakistan National Programme for Control of Diarrhoeal Disease (CDD) by establishing the Diarrhoea Training Units (DTUs). Physicians trained at DTU were designated to establish functioning oral rehydration therapy (ORT) corners at their health centres and train health workers in delivery of facilities on standard diarrhoea case management. The study was designed to evaluate the functioning of ORT corners three years after their establishment. The study revealed that ORT corners have failed to achieve the main objectives of DTU programme. Twenty-four out of 49 ORT corners were non-functional after three years of their establishment, mainly due to frequent transfers of trained staff. In 22 ORT corners evaluated, performance of health professionals was far from satisfactory, 19 out of 22 doctors were found to have inadequate performance in diarrhoea management and only 3 out of 7 LHVs performed adequately. LHVs could not consistently deliver health education messages to mothers. There seems to be a lack of interest and willingness to participate actively, as more than 50% of both doctors and LHVs did not consider ORT work as their job. We conclude that the ultimate objectives of improved and appropriate diarrhoea case management through ORT corners have not been achieved.

Phorbol-12,13-dibutyrate improves the quality of cytogenetic preparation in lymphoid malignancies.

In cytogenetic preparation of lymphoid malignancies we investigated the quantitative and qualitative impact of phorbol-12,-13-dibutyrate (P) and of this tumor promoter in combination with the calcium ionophore A23187 (PA). Using parallel cultures of unstimulated and stimulated preparations, the effect was examined in 13 patients with malignant lymphomas and six patients with acute lymphoblastic leukemias (ALL). Focusing on high-quality analyzable metaphases, the best results were found in seven of 13 cases with lymphomas and five of six patients with ALL in the cultures supplemented with phorbol-12,13-dibutyrate. The yield of metaphases of good quality regarding length, spreading, and banding of chromosomes was regularly better in P-stimulated 24-hour culture (p < 0.05), followed by 48-hour cultures stimulated with P alone. Addition of the calcium-ionophore was of no further benefit. The yield of the unstimulated direct harvest was rather poor in nearly all patients investigated. Because no mutagenic effect of P was observed, the use of this mitogen may offer interesting perspectives in cytogenetic analysis of lymphoid malignancies and perhaps also in other tumors with low mitotic indexes.

Maternal perceptions of pneumonia and pneumonia signs in Pakistani children.

Fifty mothers of children attending a hospital outpatient clinic with non-severe pneumonia (fast breathing but no chest indrawing) were interviewed in depth. Maternal perceptions and practices with clinical significance were documented. Results showed that most mothers initially tried "heat-producing" home remedies designed to counter the "coldness" of the disease, allowed only 2 days for any particular allopathic medicine to work, and did not go to the same practitioner twice. When mothers were asked what had alarmed them enough to come to the hospital, the symptoms named most frequently were persistent severe cough and high fever, inability to sleep and excessive crying. Fast breathing was spontaneously mentioned by only a few, although when questioned, 32/50 said that they had noticed it. The mothers who had prior experience with child pneumonia were more likely to notice fast breathing and also came to the hospital earlier than those who were inexperienced. Relatively higher levels of maternal education and income were suggestively associated with bringing a female child rather than a male child for pneumonia treatment. Fewer than half of the mothers knew where air goes when a person breathes in and where the lungs are located. Most held treatment preferences at odds with the protocols proposed for the national ARI program currently being initiated in Pakistan, e.g. they said that a doctor should use a stethoscope, should prescribe suspensions rather than tablets and should give injections. This study provides baseline data on attitudes and behaviors that can either be built on in that program or addressed through public education campaigns.

The median rise of leucocytes predicts the optimal days for leukapheresis in children.

22 consecutive patients with a median age of 11 years (range 3-23) underwent 103 leucaphereses after a chemotherapy induced aplasia. They suffered from various solid tumors and hematological malignancies. The stem cell yield of 69 aphereses which were done during the 8 days following the first platelet rise, was median 1.02 x 10(4) myeloid committed stem cells (CFU-GM)/kg/apheresis (range 0.02-13.3), whereas only 0.27 x 10(4) CFU-GM/kg/apheresis (range 0.01-1.11) were obtained in 34 "random" collections (p less than .05). The CFU-GM yield of 69 "well timed" collections depended on the absolute number of circulating leucocytes (p = .002) and mononuclear cells (p less than .001). The median daily increment of leucocytes was a reliable predictor of the stem cell yield (p = .002) and could be used to select the optimal days for leucaphereses during the well timed period. By careful timing alone high stem cell numbers could be collected in various malignancies without using exogenous growth factors.

A destabilization theory on health impairments by night- and shift work--some tests about its predictive value.

270 blue collar workers (181 shift and 89 day workers) of an Austrian oil refinery were investigated in 1976/77. 125 of them (60% of those still employed) were reinvestigated in 1981/82 (91 shift and 34 day workers). Subjects were interviewed about family situation, working conditions, sleep quality, risk factors and health problems. These informations were used for a partial test of our destabilization hypothesis, which states that a dynamic equilibrium between the degree of adaptation to the working sphere, the social sphere and the recreation sphere is a necessary condition for the preservation of health and that shift work, by its direct impact on all these activity spheres, tends to disturb the equilibrium, thus leading either directly or by increasing risk factors to diminished wellbeing and health impairment. It could be demonstrated that the causal structures of the formation of health problems differ considerably between shift and day workers in the predicted way. Furthermore, it was shown that shift and day workers differ with respect to the amount of destabilization and that within shift workers the degree of destabilization is a useful predictor of health impairment.

[Algesimetric measurement of the effect of cerebral electrotherapy on the pain threshold (author's transl)]. / Agesimetrische Messungen von Veränderungen der Schmerzschwelle unter zerebraler Elektrotherapie (CET, Elektroanästhesie)

The effect of cerebral electrotherapy (CET) on the pain threshold was investigated in 3 test persons, on 5 points of the neck of these persons. Altogether 1200 algesimetric measurements were performed. The statistical evaluation showed a significant tedency (p less than 0.01) towards an elevation of the pain threshold by CET.