Utility of opium seed extract tests in preventing hypersensitivity reactions during surgery.

BACKGROUND: Anaphylaxis during anaesthesia is fatal in 3-9% of patients and analgesics, including opioids, and is the second most common medicament-related cause, although the prevalence is underestimated. We recently found that patients may generate IgE antibodies to opium seeds. OBJECTIVES: To determine the diagnostic accuracy of specific antibodies to morphine, codeine, rocuronium and oil body and aqueous fractions of Papaver somniferum seeds in the diagnosis and prevention of allergy to opioids. METHODS: Patients with hypersensitivity reactions during surgery, and severe clinical allergy (pollen, tobacco), and illicit heroin users were selected. The sensitivity, specificity and predictive values of in vivo and in vitro diagnostic techniques including oil body and aqueous fractions of P. somniferum seeds were measured. RESULTS: We studied 203 patients, with mean age 35.1±17.1 and 200 healthy controls. Patients sensitised to heroin or with hypersensitivity reactions during surgery responded to P. somniferum seed tests. Of patients not known to be sensitised to opioids, the highest positivity was in patients sensitised to tobacco (p<0.001). Opium seed skin tests and IgE, especially the oil body fraction, were more sensitive (64.2%) and specific (98.4%) than morphine, codeine and rocuronium tests for opioid sensitivity. Pollen allergy was not a risk factor for sensitisation to morphine. CONCLUSIONS: Sensitivity to opioids and intraoperative anaphylaxis can be diagnosed by routine tests. IgE and skin tests for the oil body fraction of P. somniferum had the highest sensitivity for sensitisation to opioids.

Clinical value of morphine, pholcodine and poppy seed IgE assays in drug-abusers and allergic people.

BACKGROUND: The diagnosis of anaphylactic reactions due to opiates during anaesthesia can be difficult, since in most cases various drugs may have been administered. Detection of specific IgE to poppy seed might be a marker for sensitisation to opiates in allergic people and heroin-abusers. This study assessed the clinical value of morphine, pholcodine and poppy seed skin-prick and IgE determination in people suffering hypersensitivity reactions during anaesthesia or analgesia and drug-abusers with allergic symptoms. METHODS: We selected heroin abusers and patients who suffered severe reactions during anaesthesia and analgesia from a database of 23,873 patients. The diagnostic yield (sensitivity, specificity and predictive value) of prick and IgE tests in determining opiate allergy was analysed. RESULTS: Overall, 149 patients and 200 controls, mean age 32.9 ± 14.7 years, were included. All patients with positive prick to opiates showed positive prick and IgE to poppy seeds, but not to morphine or pholcodine IgE. Among drug-abusers, 13/42 patients (31%) presented opium hypersensitivity confirmed by challenge tests. Among non-drug abusers, sensitisation to opiates was higher in people allergic to tobacco (25%), P<.001. Prick tests and IgE against poppy seed had a good sensitivity (95.6% and 82.6%, respectively) and specificity (98.5% and 100%, respectively) in the diagnosis of opiate allergy. CONCLUSIONS: Opiates may be significant allergens. Drug-abusers and people sensitised to tobacco are at risk. Both the prick and specific IgE tests efficiently detected sensitisation to opiates. The highest levels were related to more-severe clinical profiles.

Clinical significance of cross-reactivity between tobacco and latex.

BACKGROUND: Allergen cross-reactivity between tobacco and other species of Solanaceae family (tomato, potato, aubergine and egg plant) have been reported. We have recently studied IgE response to tobacco in asthmatic patients sensitised to Lolium perenne (Perennial rye grass pollen) and have found that 30% of the tobacco responsive patients also have latex sensitisation. OBJECTIVE: The aim of our study was to investigate the possibility of cross-reactivity between tobacco and latex in asthmatic patients with IgE response to latex. METHODS: A study was performed on tobacco and latex exposure in 15 patients who suffered from asthma and latex sensitisation and who were randomly chosen from our database of latex-sensitive patients. To identify tobacco and latex as possible allergens that might cause clinical specific responses, all these patients were tested with prick-tests, specific IgE to tobacco, latex and related allergens, bronchial challenge, and patch tests with tobacco, latex and nicotine. Immunological response was evaluated with immunoblotting, immunoblotting-inhibition and EAST-inhibition tests. RESULTS: Positive prick and bronchial challenge with specific IgE>0.35 kU/L to tobacco was demonstrated in 11 asthmatics who were also sensitised to rye grass. Tobacco IgE level was related with sensitisation to latex (p<0.002), but not to other vegetables belonging to the Solanaceae family. EAST-inhibition and immunoblotting-inhibition showed the existence of cross-reactivity between tobacco and latex. CONCLUSIONS: Cross-reactivity exists between latex and tobacco allergens. Smoker patients with IgE response to tobacco may be a risk population for latex sensitisation.

Anisakis simplex allergy after eating chicken meat.

BACKGROUND: Allergic reactions to food can be produced by contaminants that induce sensitization. Among these, Anisakis simplex can cause seafood infestation, and allergic symptoms (urticaria-angioedema, anaphylaxis, and asthma) can follow the eating or handling of affected fish. Although seafood is the principal source of human infections by this parasite, we have found allergic symptoms in 8 patients previously diagnosed as having A simplex sensitization after they ate chicken meat. Chicken feed usually has a high proportion of fishmeal, which might possibly be contaminated by this nematode. OBJECTIVE: The aim of our study was to determine whether parasite proteins present in chicken meat could be responsible for the symptoms reported by these subjects. METHODS: We carried out in vivo tests (prick, bronchial challenge, and double-blind placebo-controlled challenge with meat chicken) in these 8 patients. We performed immunoblotting using the sera from the 8 patients and controls in order to detect A simplex sensitization. We also investigated the presence of A simplex proteins in sera from chickens fed with fishmeal and in other sera from chickens fed only with cereals. We excluded sensitization to other chicken nematodes by serologic methods. RESULTS: All 8 patients presented positive prick and challenges to A simplex. When we used serum from chickens fed with fishmeal as the antigen in blotting, patients 3, 4, 5, 6, 7, and 8 recognized a band of 16 kd, also obtained when using pools of fish-shellfish and A simplex larva. No detection was observed with sera from chickens fed with only cereals. CONCLUSION: We provide evidence, based on in vivo and in vitro tests, that subjects highly sensitized to A simplex can detect the presence of Anisakis species allergens in chicken meat.

Differential scanning calorimetry investigations on LPS and free lipids A of the bacterial cell wall.

Differential scanning calorimetry (DSC) was used to investigate the thermal stability and behaviour of the lipopolysaccharides (LPS) of the outer membrane of Gram-negative bacteria and their lipid portion. DSC curves of LPS show thermal features between 200 and 129 degrees C (depolymerization) and between -13 and -36 degrees C (cooling phase transition). Both effects were related to the relative strength of the linkage types in the O-chain structure and to their capacity for intermolecular hydrogen bonding. DSC curves of lipids A show endotherm peaks between 40 and 24 degrees C, around 15 degrees C and between -23 and -4 degrees C. Based on these effects, strong differences in thermal behaviour can be observed between that of Brucella and Vibrio cholerae on the one hand, and that of Escherichia coli and Shigella flexneri on the other. Fluidity of the acyl chains and lyotropism, which are important parameters in expression of biological activities, are discussed using the above data. To explain some properties, fluidity could be related to the temperature of the gel<==>liquid crystalline (beta<==>alpha) phase transition, which occurs at physiological temperature. Nevertheless, fluidity could be related to temperature of the previous thermal effect (between 6 and 20 degrees C), for which a softening or partial melting of the sample has been evidenced. The thermal effect measured between -23 degrees C and -4 degrees C indicates a greatly reduced water concentration of lipid A from Brucella, thus explaining its early fusion process and its activity by means of hydrophobic interactions.

Reference values and methods comparison of a new testosterone assay on the AxSYM system.

OBJECTIVES: We have contributed to the Design Validation Protocol of the new Abbott AxSYM Testosterone assay with a study on reference values and methods comparison. DESIGN AND METHODS: For reference values a population of 45 women and 30 men was tested. In methods comparison, 132 samples for the AxSYM vs. ACS-180, and 30 for the AxSYM vs. Elecsys were used. Pearson and intraclass concordance coefficients and Passing-Bablock test were performed for overall group, men and women. RESULTS: Reference values were 0.9-3.1 (females) and 1.0-30.2 nmol/L (males). Globally, a good agreement between methods in both the AxSYM vs. ACS-180 (slope: 0.88, y-intercept: 0.67, r = 0.961) and the AxSYM vs. Elecsys (slope: 1.08, y-intercept: 0.31, r = 0.935) studies was found. Slightly worse results were observed for women. CONCLUSIONS: The reference range of testosterone by the AxSYM system matches with those published for other methods. An acceptable agreement between the AxSYM Testosterone assay and both a classical (ACS-180) and a more recent (Elecsys) methodology was observed.

Identification of species of Brucella using Fourier transform infrared spectroscopy.

Fourier transform infrared spectroscopy (FTIR) is a technique that has been used over the years in chemical analysis for the identification of substances and is one that may be applied to the characterisation of microorganisms. The marked tendency of Brucella towards variation in the smooth rough phase, together with the laboriousness and risk involved in the methods used in their identification, make their classification difficult. We studied the type strains of the different species and biovars of Brucella and 11 isolates of human origin of Brucella melitensis, six corresponding to biovar 1, one to biovar 2 and five to biovar 3. The results of linear discriminant analysis performed using the data provide an above 95% likelihood of correct classification, over half of which are in fact above 99% for the vast majority of Brucella strains. Only one case of B. melitensis biovar 1 has been incorrectly classified. The rest of the microorganisms studied (Staphylococcus aureus, Strteptococcus pyogenes, Enterococcus faecalis, Corynebacterium pseudodiphtheriticum, Clostridium perfringens, Escherichia coli, Acinetobacter calcoaceticus and Pseudomonas aeruginosa) have been classified correctly in all cases to a likelihood of over 80%. In the graphic representation of the analysis, a grouping of these can be seen in clusters, which include the different species. One of these comprises B. melitensis, another Brucella abortus, and another wider one is made up of Brucella suis. The Brucella canis, Brucella ovis and Brucella neotomae strains appear separate from the previously described groups.

Thermolytical techniques to characterize fungal polysaccharides and bacterial lipopolysaccharides.

The present work constitutes an entirely novel contribution in the scope of microbiology and especially in taxonomy, introducing thermolysis curves as a rapid method of characterization of fungal polysaccharides and bacterial lipopolysaccharides. The thermal analysis techniques applied were thermogravimetry and derivative thermogravimetry (TG-DTG), differential thermal analysis (DTA), and differential scanning calorimetry (DSC). Each thermogram of a sample is represented by one or a few temperatures and, in DSC, by complementary enthalpy data. The temperatures of the thermograms from structurally unknown polysaccharides are compared with those used as references, and thus, information on their composition, linkage types, and anomeric configuration can be deduced. The situation is more complicated for bacterial lipopolysaccharides, but in whatever mode, a structural estimation is always possible. In the course of the development and validation of the thermal method, structural findings on relative stabilities of linkage types (valuable in carbohydrate research) have been recognized and are therefore also described in this work.

Scanning electron microscopy images and energy-dispersive X-ray microanalysis of the stapes in otosclerosis and Van der Hoeve syndrome.

OBJECTIVE/HYPOTHESIS: The objective of this study was to evaluate the morphological and microchemical changes that affect sclerotic stapes in otospongiosis and van der Hoeve syndrome. METHODS: A scanning electron microscope equipped with an energy-dispersive x-ray analyzer was used in the experiments. RESULTS: In otosclerosis, focal lesions are poorly mineralized, with low calcium salt and reduced calcium-to-phosphorus (Ca/P) ratio (1.9:1). This finding correlates with a spongiotic type of lesion and indicates unstable mineralization with possible change from hydroxyapatite to calcium triphosphate. In van der Hoeve syndrome the presence of magnesium in stapes suggests osteoclastic function stimulation. The osteoclasts secrete many protons, causing an acidified microenvironment. Brushite is formed, and Ca/P ratio decreases in comparison with that of control patients (2.0:1 vs. 2.6:1).

[Otosclerosis and Van der Hoeven's syndrome: a contribution]. / Aportación al estudio de la otosclerosis y del síndrome de Van der Hoeve.

Morphological and microchemical changes that effect to the otosclerotic stape in the Van der Hoeve's syndrome were examined with a scanning electron microscope equipped with an energy dispersive X-ray fluorescence. Using the Ca/P ratio as criterion--measured by the characteristic x-ray fluorescence--it was shown that the Van der Hoeve stape had a higher Ca/P ratio (2.6:1) as compared to the normal stape (2:1). The Van der Hoeve's syndrome lesions as poorly mineralized, with low calcium salt and apparent increase of phosphates. This finding indicates a possible change from hydroxyapatite (or apatite) to brushite, which imply an acidification of bone.

[Soluble CD40 ligand: a potential marker of cardiovascular risk]. / El ligando CD40 soluble: marcador potencial de riesgo cardiovascular.

Recombinant human soluble CD40 ligand, also named CD145 or gp 39, is a 16.3 kD glycoprotein containing 149 aa residues comprising the receptor binding TNF-like domain of CD40 ligand. It is expressed on antigen-presenting cells such as B cells, macrophages, dendritic cells and thymic epithelial cells and it constitutes the nexus between the inflammatory system and the vascular thrombotic processes. Its gene is located in the long arm of the human X chromosome. Prognostic evaluation of the residual fixed atherosclerotic plaque is insufficient to predict clinical course. Currently, studies have been done that demonstrate the participation of the immunoinflammatory system in the genesis and complications of the atherosclerotic condition. In the future, the most specific biomarkers of vulnerability will be very useful in the daily practice (interleukins, CD40, etc.). The soluble CD40 ligand together with its CD40 receptor are overexpressed in experimental and human atherosclerotic lesions. This leads to an increase of mediators for the development of atherosclerosis. Both significantly contribute to the inflammatory processes that leads to atherosclerosis and thrombosis.

Enhancement of tomato allergenicity after treatment with plant hormones.

BACKGROUND: Practical applications to enhance the productivity of agriculture by using plants with improved resistance to pathogens are expected to increase in the near future. Although tomato has been widely investigated for breeding purposes, there have been no studies on tomato allergenicity after plant hormones treatments. METHODS: Prick by prick tests were carried out with different tomato samples (fruits grown under biological conditions without addition of chemical products, and treated with ethylene and salicylic acid) in eight patients with ages between 12 and 27 years who suffered from anaphylaxis episodes after eating raw tomatoes. An immunoblot experiment with the different tomato extracts was performed using sera from these eight patients and controls. RESULTS: The wheals obtained in prick tests were significantly higher with the extracts of tomato treated with ethylene and SAA (chi(2) = 31.3, p < 0.0001) and the patients who presented higher wheal diameters in skin tests were those who had more severe episodes of anaphylaxis. Neither the protein stain nor the IgE immunodetection patterns clearly varied between the untreated and the hormone-treated samples. CONCLUSIONS: In the case of anaphylaxis induced by tomato, the treatment with plant hormones induced a higher cutaneous response than with non-treated tomato, but the "in vitro" response was similar.

Comparison of two homogeneous assays with a precipitation method and an ultracentrifugation method for the measurement of HDL-cholesterol.

We report on the performance of four HDL-cholesterol assays: a homogeneous method based on polyethylene glycol-modified enzymes/alpha-cyclodextrin sulfate (PEGME; Kyowa); a homogeneous method based on polyanion-polymer/detergent (PPD; Daiichi); the usual precipitation method with phosphotungstic acid/MgCl2 (PTA); and an ultracentrifugation (UC) procedure. The homogeneous HDL-cholesterol assays (performed with automated analyzers) were precise and correlated well with the PTA and UC assays. The specificity and accuracy of the PEGME method were better than those of the PPD method.

Shewanella putrefaciens in a fuel-in-water emulsion from the Prestige oil spill.

Microorganisms that colonize the fuel-in-water emulsion from the Prestige spill have been compared with those from Exxon-Valdez. Both emulsions contained non-fermentative gram-negative rods but unlike Exxon-Valdez's, the Prestige's spill contained anaerobic bacteria and no fungi. Our main finding has been the identification of Shewanella putrefaciens , a bacterium promising for bioremediation.

The first known use of vermillion.

Vermillion has been shown to be useful in preserving human bones from 5000 years ago. Remarkably well-preserved human bones have been found in the dolmenic burial 'La Velilla' in Osorno (Palencia, Spain), carefully covered by pulverized cinnabar (vermillion) which ensured their preservation even in non-favorable climatic conditions. We believe the red powder was deliberately deposited for preservative use because no cinnabar mine is to be found within 160 km, because of the large amount (hundreds of kilograms) used, and because its composition, red mercuric sulphide, is similar to that of preparations used in technical embalming. This finding pushes back the data of the use of mercury ore for preservation by four millennia in South America, and by at least one millennium in the Old World. Chemical and thermal analyses of vermillion in La Velilla have demonstrated its great purity and shown that the cinnabar was pulverized and washed (but not heated), producing a bright red-orange tone.

Living in towers as risk factor of pollen allergy.

BACKGROUND: The effects of fluctuations in pollen counts have important implications for health services. Little research on the clinical implications of the vertical distribution of pollen in allergy symptoms has been carried out. METHODS: We have investigated the allergic symptoms of a population of 17 171 patients coming from our health area (Valladolid, Spain) and living in the city and villages. We compared the prevalence of sensitization to different pollens: Graminae, trees and shrub in relation with the floor of the building where they were living. RESULTS: Relative risk of pollen sensitization (confidence index, CI 95%) was higher in patients who were living at high floors than in those patients who were living in lower floors or at street level, independently of rural or urban conditions The chi-square showed a lineal trend in this relationship in the case of sensitization to grass pollen: chi(2): 1794, P > 0.00001 CONCLUSIONS: On the basis of our local investigations, natural pollen sensitization appears to increase with height where the patient lives. This paper reports clinical results on the influence of vertical pollen distribution in pollen allergy.

Anaphylaxis associated with antiphospholipid syndrome.

BACKGROUND: To our knowledge, no previously published reports have described food-induced anaphylaxis associated with the antiphospholipid syndrome. OBJECTIVE: We undertook a study of four patients with thrombosis associated with the antiphospholipid syndrome after each patient experienced anaphylaxis attributable to ingestion of vegetal foods. METHODS: IgE antibody levels to various foods were determined in serum specimens from the study patients, and skin prick tests with the same allergens were conducted to determine their in vivo responses. Hematologic, cardiopulmonary, vascular, and rheumatologic studies were also performed. IgG anticardiolipin antibody levels were determined by ELISA. RESULTS: All four patients fulfilled the criteria for antiphospholipid syndrome and had high levels of specific IgE antibodies for certain food allergens. By immunoblot analysis, the presence of serum IgE specific for a 45-kD protein band in an almond extract was detected in these four patients who experienced food-related anaphylaxis. No specific IgE was detected in sera from normal subjects. No IgE antibodies specific for the food panallergen lipid transfer proteins were detected. CONCLUSIONS: This is the first report of severe food-precipitated anaphylaxis associated with the antiphospholipid syndrome and the first description of a patient with allergy to blackberry. The possible involvement of food panallergens distinct from lipid transfer proteins is also discussed.