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Radioimmunosorbent assays for determination of serum content of the neutrophil proteins myeloperoxidase and lactoferrin are described. Serial studies were performed in patients with neutropenia. In 2 cases of cyclic neutropenia the myeloperoxidase level showed slight variations within the normal range during the cycle while lactoferrin displayed a clear correlation with neutrophil counts. In 1 case with persistent agranulocytosis myeloperoxidase was normal but lactoferrin was extremely low. During the regeneration phase of drug-induced neutropenia neutrophil counts and serum lactoferrin increased in a parallel fashion. Since serum myeloperoxidase was normal during profounded neutropenia it is suggested to derive primarily from myeloperoxidase-rich granulopoietic precursor cells of the marrow. Serum lactoferrin on the other hand seems to derive from leakage of more mature granulopoietic cells of blood and marrow. Studies of neutrophil proteins of serum may aid in evaluation of neutropenic patients.

Summary A descriptive case study was performed on 75 patients with NF1 (neurofibromatosis type 1) from the CNNF (Brazil) database. Serum IgE levels were determined using the IgE radioimmunosorbent test, with the reference values of 75–502 IU mL−1. The patients were divided into groups, with 25 patients presenting plexiform neurofibromas, 25 presenting neurofibromas and 25 presenting no neurofibromas. The purposes of this study were to determine the serum IgE levels of patients with NF1 presenting plexiform neurofibromas, neurofibromas and no neurofibromas, as well as to determine possible correlations between serum IgE levels and the size of the plexiform neurofibromas and neurofibromas presented by these patients. Elevated serum IgE levels were observed in all the patient groups. We did not observe a correlation between IgE levels and age in these patients; however, we did observe correlations between IgE levels and neurofibroma and plexiform neurofibroma size. We suggest further studies to confirm these results and to investigate in greater depth the possible role of IgE in the development and growth of neurofibromas and plexiform neurofibromas in NF1.

The repeated manipulation of feeding schedule has a marked influence on the chronopharmaological aspects of many drugs. In this study, we investigated the role of endogenous glucocorticoid in the mechanism by which restricting the feeding time modulates the analgesic effect of morphine. Male ICR mice were housed under a light-dark cycle (light on from 07:00 to 19:00) with food and water ad libitum or under repeated time-restricted feeding (feeding time from 09:00 to 17:00) for 2 weeks before the experiment. Under the ad libitum feeding, mRNA levels of µ-opioid receptor and its binding capacity in mouse brainstem increased around the early dark phase, following the 24-h variation in circulating glucocorticoid levels. As a consequence, potent analgesic effects of morphine were observed in mice injected with the drug during the dark phase. Daily restricted feeding modulated the time-dependency of µ-opioid receptor function, accompanied by the alteration of the rhythm in circulating glucocorticoid levels. Under the time-restricted feeding, potent analgesic effects of morphine were found in mice injected with the drug during the light phase. Because the manipulation of feeding schedule was unable to produce the food-entrainable rhythm in the expression of µ-opioid receptor in the brainstem of adrenalectomized mice, endogenous rhythm of glucocorticoid secretion seems to be involved in the mechanism by which the time-restricted feeding modulates the analgesic effects of morphine. Keywords:: food-entainale rhythm, feeding schedule, glucocorticoid, μ-opioid receptor, morphine

We evaluated a new, commercially developed radioimmunoprecipitation assay for measuring glutamic acid decarboxylase (GAD) antibodies by using recombinant human GAD65. The intra- and interassay CVs were 8.0% (n = 20) and 8.6% (n = 15), respectively. We found GAD antibodies in 74% (23 of 31; 95% confidence interval 55-88%), 70% (14 of 20; 46-88%), and 65% (28 of 43; 49-79%) of patients at, respectively, < or = 1 year, 1-2 years, and 2-4 years after the onset of insulin-dependent diabetes mellitus (IDDM) and in 30% (30 of 99; 21-40%) of patients with long-term diabetes (4-22 years). We also detected GAD antibodies in 8% (9 of 106; 4-16%) of patients with non-insulin-dependent diabetes mellitus (NIDDM). The frequency of GAD antibodies in the NIDDM group was markedly higher in the insulin-deficient patients [67% (6 of 9; 30-93%)], who initially were nonketotic and non-insulin-dependent for > or = 6 months but later became insulin dependent, than in the non-insulin-deficient patients [3% (3 of 97; 1-9%)]. This new commercial assay is easy to use and provides a specific and sensitive method for evaluating GAD antibodies in IDDM.

The association between heavy metals exposure and respiratory diseases or allergic sensitization showing high serum immunoglobulin E (IgE) has been suggested. However, previous findings have been inconsistent and the mechanisms responsible remain unclear. We evaluated heavy metal exposure and its association with coughing, itchy eyes in chronic cough patients with different IgE levels. Ninety outpatients in Kanazawa University Hospital were recruited between January–June 2011. Subjects whose total IgE measured by radioimmunosorbent test were asked to record their daily symptoms. We collected daily total suspended particles (TSP) from which concentrations of calcium (Ca), cadmium (Cd), chromium (Cr), iron (Fe), manganese (Mn), nickel (Ni), and lead (Pb) were determined then divided into high and low level groups. Generalized estimating equations were applied to compute the relationship between concentrations of these metals and symptoms. All metals at high levels were significantly associated with itchy eyes compared with low levels, with exception of Ca, the six others were significant in patients with IgE < 250 IU/mL. Cd, Fe, Mn had association with coughing (odds ratio-OR (95% confidence interval-CI): 1.13 (1.03, 1.24), 1.22 (1.05, 1.42), and 1.13 (1.01, 1.27), respectively), this relationship remained significant for Cd (OR (95% CI): 1.14 (1.03, 1.27)) and Mn (OR (95% CI): 1.15 (1.00, 1.31)) in patients with lower IgE. Our findings demonstrate the relationship between aerial heavy metals and itchy eyes, coughing in chronic cough patients, suggesting these symptoms may be due to a non-IgE mediated mechanism.

Hymenoptera venom allergy is a growing problem in Spain. This problem has a special relevance in areas where population is frequently exposed to hymenoptera stings, being paediatric patients a high risk population. Immunotherapy with hymenoptera venom is an effective and safe treatment for these patients. However, there is a lack of data on the role of this treatment on paediatric population. For this reason, from the data base of the Allergy Unit from Hospital Xeral (Lugo, Spain) which includes 560 patients, have been analyzed the 21 paediatric patients, all of them treated with venom immunotherapy. Eighteen patients completed the treatment. The maintenance dose administered was 100 microg. Two systemic reactions (both with an Apis extract) were registered. Cutaneous test and specific IgE shown a statistical significant reduction at the end of treatment (p =.0004 and p.0001 respectively). Seven patients (33 %) suffered a spontaneous re-stung during maintenance phase or after immunotherapy was completed. In 4 patients there was no allergic reaction and the other 3 children suffered a mild local reaction. In conclusion, venom immunotherapy is a safe and effective treatment in paediatric patients with hymenoptera venom allergy, being necessary to increase the experience on this specific segment of the allergic population.

Historical data were obtained by questionnaire and telephone survey on 20 of 24 women with reported allergic reactions to a psyllium-containing cereal, Heartwise. Protein fractions from this new cereal, as well as from psyllium mucilloid and a psyllium-containing laxative, Metamucil, were extracted, quantitated, and separated by sodium dodecyl sulfate-polyacrylamide gel electrophoresis. Patients' sera were collected, and specific IgE and IgG antibodies to these psyllium antigens were detected by immunoblotting techniques. Of the 20 women evaluated, all but six were nurses. Eighteen (90%) of the women had historical and/or laboratory evidence of atopy. Exposures included ingestion or dispensing of psyllium-containing products. Only three women denied prior exposure to psyllium. Symptoms developed shortly after small amounts of the cereal were ingested and most commonly included moderate to severe wheezing, throat and chest tightness, and urticaria. All the women required medical therapy, 11 (55%) in an emergency room. Specific IgE and IgG antibodies to various psyllium protein fractions were documented in all the subjects. It was concluded that individuals sensitized by occupational exposure to psyllium dust are at high risk for allergic reactions to ingested psyllium-containing products.

The presence of immunoreactive cells in periapical inflammatory lesions suggests that immune responses participate in the disease process. The purpose of this study was to determine the presence and concentration of immunoglobulins in the supernatant fluids of explant cultures of periapical lesions. Ninety periapical lesions that had been contiguous with the apex of a root were removed and maintained in explant cultures for 96 h. Tissue culture medium was replenished at 24, 48, 72, and 96 h. Double diffusion in agarose assays demonstrated the presence of IgG in 100% of the 24-h supernatant fluids and IgA in 65% of the 24-h supernatant fluids. However, IgM was not detected. Radial immunodiffusion assays were used to detect and quantitate IgG, IgA, and IgM in samples of 24-h supernatant fluids from 90 explant cultures. IgG was the predominant immunoglobulin followed by IgA. A radioimmunosorbent test was used to detect and quantitate IgE in samples of 24-h supernatant fluids from 90 explant cultures. Forty of the 90 supernatant fluids contained measurable IgE. All detected immunoglobulins decreased in concentration in daily supernatant fluids with time (24, 48, 72, and 96 h) in the culture.

The relative merits of antibody response and virus shedding as markers of cytomegalovirus (CMV) infection among cyclosporin-treated renal allograft recipients were analysed. CMV-specific antibody was assayed by IgG-specific radioimmunosorbent test (RIST) and by complement fixation test (CFT). CMV shedding was assayed by virus isolation and by the rapid test immediate early nuclear antigen detection (IENAD). RIST and CFT detected seroconversion in similar numbers of patients, but the former test was the more sensitive when CMV antibody was sought in pretransplant sera to differentiate primary from recurrent infection. IENAD detected or excluded CMV shedding for more urine specimens than virus isolation (462/515 [90%] vs. 366/515 [71%]), but the reverse applied to saliva specimens (33/57 [58%] vs. 54/57 [95%]). The high specificity of IENAD allowed positive results by IENAD to be accepted when virus isolation failed to provide a result. IENAD was, however, less sensitive than virus isolation even when specimens yielding CMV by IENAD, but no result by virus isolation, were included in the analysis (27/44 [61%] vs. 38/44 [86%]). Assays of both antibody response and virus shedding were required to maximise the diagnosis of recurrent CMV infections, but the former assay detected all primary CMV infections. The diagnostic implications of these results are discussed.

An immunoradiometric assay (radio-immunosorbent test; RIST) for the detection of IgG antibodies to human herpesvirus 4 [human cytomegalovirus (CMV)] has been developed. The technique utilizes CMV antigen passively adsorbed to a polyvinyl microtitration plate and a radiolabelled murine monoclonal anti-human IgG antibody to detect binding of human antibody to the ‘solid phase’ reagent. The assay was optimized, and its specificity confirmed by testing paired acute and convalescent sera from patients with acute CMV or other human herpesvirus infections. To determine the assay's sensitivity 1433 blood donor sera were examined. The RIST was more sensitive than a standard complement fixation (CFT), in that 53% of these sera were positive by RIST and 48% positive by CFT. There were 1303 concordant results, 88 sera positive only by RIST and 19 sera were only positive by CFT. These discrepant results remained after an attempt to exclude false positive reactivity; their significance is discussed. Use of a monoclonal anti-human IgG antibody in the RIST reduced non-specific binding to the control uninfected cell antigen such that blood donor sera could be tested in the assay using only a CMV antigen without generating an unacceptable false positive rate.

The study comprised 28 patients from the centre of Spain (Madrid and surroundings) who had suffered systemic reactions when stung by vespids. Specific IgE antibodies to Vespula spp. and Polistes spp. venoms were measured by RAST. All patients had positive RAST to Vespula venom and half of them also had positive RAST to Polistes venom. A patient can be sensitive to both venoms due to either a clinical sensitization to both venoms, or common antigenic determinants. To differentiate these states we used a RAST inhibition assay. We could inhibit Polistes RAST with either Polistes or Vespula venom to a similar degree. Inhibition of Vespula RAST was possible with Vespula venom, but only to a limited degree with Polistes venom. Direct RAST and RAST inhibition studies indicate that in our geographic region sensitization to Vespula venom is more common than to Polistes venom and Polistes might have cross-reactivity in our patients.

A study was performed in order to assess the value of the Phadiatop, a new in vitro allergy test, in general practice. A total of 248 patients were screened for bronchial allergy with PRIST, RAST and the Phadiatop. Information about the patient's history and the results of the PRIST, Phadiatop and RAST tests were presented consecutively to the general practitioner and his diagnoses based on and evolving from these were evaluated and compared with the diagnosis of a specialist. The general practitioner's diagnosis based on the results of the Phadiatop was the closest of all the tests to the specialist diagnosis. It is concluded that Phadiatop is of great use in diagnosing bronchial allergy in general practice.

To evaluate the prevalence of latex allergy in a population of children with spina bifida (SB) and to assess the role of early exposure to latex products and others risk factors.SB is related with an higher incidence of latex allergic reactions. These patients received repeated surgical procedures, implant of latex-containing materials and catheterization.Eighty consecutive subjects affected with SB besides answering a questionnaire, underwent a skin-prick test (SPT) to latex and the determination of the specific serum IgE (RAST CAP) to latex. 40% (32/80) of the patients showed a latex sensitization with specific IgE0.7 kU/I but only twelve of the 32 sensitized patients (40%) suffered from clinical reactions to latex (urticaria, conjunctivitis, angioedema, rhinitis, bronchial asthma). Number of surgical procedures, but particularly early exposure to latex and familiarity for allergy are correlated with latex allergy (p0.01).Latex allergy in SB children is multifactorial situation related with a disease-associated propensity for latex sensitization, early exposure and number of surgical procedures. Prophylactic measures to avoid the exposure, not only in the sanitary environment, through the institution of latex-safe routes and every day, prevent potentially serious allergic reactions.

BACKGROUND In clinical studies involving rapid-acting analogues (RAAs), insulin immunoreactivity is frequently measured, including endogenous, regular insulin (RI) and RAA immunoreactivities. Such a procedure implies equivalent cross-reactivities of all insulins present in serum. Commercially available human insulin immunoassays have been widely used, but their limitations (including hemolysis and anti-insulin antibodies) were not fully investigated. The aims of our study were to compare cross-reactivities of RI and RAAs in buffer and in serum and to investigate insulin immunoassay pitfalls. METHODS Cross-reactivities were assessed using Bi-insulin IRMA (Schering Cis-Bio International) in phosphate-buffered saline (PBS)-1% bovine serum albumin (BSA) and in pools of sera spiked with RI and RAAs (lispro and aspart). To investigate the influence of hemolysis, a pool of sera spiked with RAA was mixed with a concentrated hemolysate (final hemoglobin concentration 10 g/L) and incubated for 3 h at room temperature. To determine interference by anti-insulin antibodies, insulin was removed using charcoal from 18 sera with anti-insulin antibodies and from 17 sera without detectable anti-insulin antibodies. These insulin-free samples were then spiked with RI and RAAs and the immunoreactivity was determined. RESULTS Compared with buffer, cross-reactivity in serum for RI, lispro and aspart was lower (35%, 29% and 26% lower, respectively). Hemolysis degraded almost all RI and RAAs contained in the serum (>or=95%). Anti-insulin antibody interference was significant for RI and RAAs (p

Type I hypersensitivity is a major problem in public health, often requiring numerous investigations which aim to diagnose atopy and identify the causative allergen. Among these investigations, several blood tests, mostly using immunoenzymatic methods, can be performed to measure total and specific IgE levels. Type I hypersensitivity can also be investigated with assays which quantitate several mediators released after cellular activation induced by allergens.

This paper describes a case of eosinophilic granuloma of the soft tissue in the cheek. The patient was a 52-year-old man, who was a repairman of vintage cars. He had a history of inhaling large quantities of dust.Preoperative hematologic examination showed a white blood cell count of 5600 with 12% eosinophils. A radioimmunosorbent test (RIST) revealed an immunoglobulin E level of 5217.3 IU/dl. In antigen examinations, antibodies against house dust and ticks were abnormal on radioallergosorbent tests (RAST) of immunoglobulin E.The lesion was enucleated under GO-NLA. The patient was also provided guidance how to improve his work environment.There is no evidence of lesion recurrence as of 14 months after operation.

Monitoring beta2-microglobulin (beta2M) in biological fluids has gained considerable interest in pathologies such as haematologic malignancies, renal diseases, and chronic inflammatory diseases. Due to limitations of the RIA in the routine laboratory, we measure beta2M with non-isotopic methods. 189 patients suffering from myeloma (n=66), end stage renal failure (n=54) or inflammation (n=69) were included in this study. beta2M was determined in serum, urine and dialysate using an immunoenzymometric assay with chemiluminescence detection [Immulite Diagnostic Products Corporation (DPC), La Garenne Colombes, France] and an immunoturbidimetric assay (Olympus, Rungis, France). The data were compared with a radioimmunoassay (Immunotech, Marseille, France) taken as a reference. Using serum samples, the immunoenzymometric assay with chemiluminescence detection and the immunoturbidimetric assay have reliable analytical performances. Values obtained with serum samples are highly correlated with the radioimmunoassay (DPC/RIA r2=0.84; Olympus/RIA r2=0.94) whatever the type of pathology; however an over-estimation which could be related to cross reactivity with beta2M fragments was observed with the RIA method as suggested by crossover calibration and recovery studies. Values obtained with urinary samples (n=96) are closely related to those obtained with the RIA (DPC/RIA r2 = 0.98; Olympus/RIA: r2=0.99). Despite the low levels observed in dialysate (n=57) good correlations were observed between Olympus vs DPC (r2=0.85). By contrast, the two non-isotope methods are poorly related with the RIA method (DPC vs RIA r2=0.47 and Olympus vs RIA r2=0.54). In conclusion, the immunoenzymometric assay with chemiluminescence detection or the immunoturbidimetric assay could be used in the routine laboratory in order to determine beta2M in plasma, urine and dialysate.

To study renal functional reserve and 24-h beta 2-microglobulinuria (B2-MGU) in patients with different stages of essential hypertension (EH) and to evaluate acceptability of these parameters as early markers of preclinical renal impairment in EH.We examined 150 patients (mean age 42.1 +/- 3.4 years) with EH without overt clinical manifestations of hypertensive nephropathy. Renal functional reserve (RFR) was determined with the use of acute oral protein loading and was defined as an increase in endogenic creatinine clearance in percent. B2-MGU was evaluated by radioimmunoassay (RIO-BETA-2-MICRO kit). Radioimmunoassay detection of microalbuminuria (MAU) was performed with the use of the standard kit Albumin-RIA.RFR appeared to be significantly lower in patients with EH compared to the control group, and its level significantly decreased with aggravation of EH. Both B2-MGU and MAU were significantly higher in patients with EH and significantly increased with aggravation of EH.We suggest that, apart from MAU, RFR abnormalities and excessive B2-MGU should be used as markers of preclinical renal impairment in EH.

It is well established that malnutrition affects the immune response and increases the susceptibility to parasitic infection. In the present study we evaluated some aspects of the cellular and cytokine network that regulate the IgE responses which are important components of host defence mechanisms against helminthic parasites in children infected with the intestinal helminth Ascaris lumbricoides and with differing degrees of malnutrition. We found a defective T cell response in malnourished children as indicated by diminished levels of circulating total (CD3+) helper (CD4+) IL-2-receptor-bearing (CD4+CD25+) and memory helper T cell responses (CD4+CD45RO+) in keeping with the decreased specific IgE levels against Ascaris lumbricoides. In contrast the proportions of total B cells (CD20+) and those bearing the low-affinity IgE receptor (CD23+) were increased in the moderated malnourished children. Moreover serum IL-4 levels and total IgE were also increased in these children.We suggest that malnutrition can cause an imbalance in T cell subpopulations that may lead to a defective T cell maturation and a decreased specific anti-Ascaris IgE response thus increasing the susceptibility to such infections. The high levels of total IgE observed may be related to a non-specific stimulation of the proliferation of activated B cells probably caused by helminthic parasites and other infectious agents that are frequent in malnourished children. (authors)

The formation of electrostatic aggregates was studied by analysis of two types of virus-containing liquids: initial warm liquid collected at temperature 37 degrees and the same liquid stored over the night at temperature 4 degrees C. The formation of virus aggregates was revealed at 4 degrees C. The aggregates formed at temperature 4 degrees C had a relatively high HA/NP ratio in comparison with unassociated virus analyzed at 37 degrees. HA-enriched aggregates were found in the precipitate formed under short-term high-speed centrifugation as well as in "heavy arm" of the virus profile in the saccharose gradient. Aggregates formed at 4 degrees C dissociated at 37 degrees. The ability to form aggregates is reversible and correlates with the virus concentration. It is shown also that virus containing liquid contains heterogenic structures with molecular weight under 2000 kD having HA involved in the forming aggregates enriching HA. Possible nature of low-molecular HA-containing structures involved in the aggregates and nature of relations stabilizing aggregates are discussed.

Background A systemic reaction to mycobacteria biases the balance of T helper cell types 1 and 2 toward T helper cell type 1. BCG vaccination mimics some characteristics of mycobacterial infection. Children who have undergone tuberculin conversion after BCG vaccination seem to be more likely to lose their atopic symptoms. Inhibition of both allergic response and airway hyperreactivity after vaccination for mycobacteria has been observed in animal experiments. Objective To evaluate the effects that BCG vaccination has on the serological status of allergic people. Participants and Methods This study included 20 volunteers with a history of allergic rhinitis who were required to undergo BCG vaccination by Italian law. Epicutaneous allergy testing with a panel of common seasonal and perennial inhalational allergens and 2 blood withdrawals were performed. The serum total IgE levels and the serum allergen-specific IgE levels of each individual were measured just before BCG vaccination and again 4 months later. Total IgE levels were determined using the paper radioimmunosorbent test, and allergen-specific IgE levels were determined using the radioallergosorbent test. Results Total IgE and allergen-specific IgE levels were significantly decreased after BCG vaccination (P= .004 andP Conclusion BCG, an effective stimulus for cell-mediated immunity, deserves further study to evaluate its ability to modulate the immune response associated with allergic rhinitis.

OBJECTIVE: To analyze the correlation of risk factors to the occurrence of urinary tract infection in full-term newborn infants. PATIENTS AND METHODS: Retrospective study (1997) including full-term infants having a positive urine culture by bag specimen. Urine collection was based on: fever, weight loss > 10% of birth weight, nonspecific symptoms (feeding intolerance, failure to thrive, hypoactivity, debilitate suction, irritability), or renal and urinary tract malformations. In these cases, another urine culture by suprapubic bladder aspiration was collected to confirm the diagnosis. To compare and validate the risk factors in each group, the selected cases were divided into two groups: Group I - positive urine culture by bag specimen collection and negative urine culture by suprapubic aspiration, and Group II - positive urine culture by bag specimen collection and positive urine culture by suprapubic aspiration . RESULTS: Sixty one infants were studied, Group I, n = 42 (68.9%) and Group II, n = 19 (31.1%). The selected risk factors (associated infectious diseases, use of broad-spectrum antibiotics, renal and urinary tract malformations, mechanical ventilation, parenteral nutrition and intravascular catheter) were more frequent in Group II (p37,8ºC), perda de peso>10% do peso de nascimento, alterações do estado geral (recusa alimentar, ganho insuficiente de peso e hipoatividade) ou presença de malformações nefro-urológicas. Nesses recém-nascidos foi realizada punção suprapúbica para confirmação diagnóstica. Os recém-nascidos foram divididos em dois grupos, segundo o resultado das uroculturas: Grupo I (diagnóstico presuntivo de infecção urinária) e Grupo II (diagnóstico confirmado de infecção urinária), para avaliação dos fatores de risco pela análise do risco relativo. RESULTADOS: Foram estudadas 61 crianças (5,1% dos recém-nascidos de termo) - Grupo I n=42 (68,9%) e Grupo II n=19 (31,1%). Os fatores de risco avaliados (patologias infecciosas associadas, uso prévio de antibióticos, malformações nefro-urológicas, ventilação mecânica, nutrição parenteral e o uso de cateteres) foram mais freqüentes no Grupo II (p

Guidelines to the multitude of allergologic tests are defined. But, in practical experience, skin prick test are the first and the best test for allergy diagnosis. Positive skin prick test refer to sensitization in accordance with clinical history.

PURPOSE Tear IgE has been considered to play an important role in allergic conjunctivitis and the measurement of tear IgE concentrations can help to diagnose this condition. Locally produced IgE levels have been shown to be the largest contributor to the severity of allergic conjunctivitis. METHODS One hundred and thirteen allergic conjunctivitis patients (70 seasonal allergic conjunctivitis (SAC), 21 perennial allergic conjunctivitis (PAC), 22 vernal keratoconjunctivitis (VKC)), 14 bacterial conjunctivitis (BC) patients, 13 epidemic keratoconjunctivitis (EKC) patients and 18 normal controls were recruited. Tear samples were collected using the microcapillary method and tear IgE levels were measured using an immunoenzyme assay. RESULTS Tear IgE concentrations showed significant increases in the VKC (322.2 +/- 45.7 ng/ml), SAC (194.7 +/- 21.7 ng/ml) and PAC (134.8 +/- 23.1 ng/ml) groups when compared with controls (52.1 +/- 9.7 ng/ml, p < 0.01). No significant difference was found between EKC (97.2 +/- 11.7 ng/ml) and BC (92.6 +/- 13.8 ng/ml) groups and controls (p = 0.1). CONCLUSIONS Tear IgE concentrations showed a significant increase in allergic conjunctivitis patients when compared with controls. Our results suggest that measuring tear IgE concentrations can help to diagnose allergic conjunctivitis.

To investigate the new criteria for skin prick test (SPT) of seventy-four atopic infants (2-5 months of age at the first visit, Mean 3.8 months, M:F = 54:20) to diagnose for hypersensitivity to egg white. It was classified into three groups by reaction type of SPT in the first visit. Group A were the infants who seemed only late (6 hours) or delayed (48 hours) reaction (n = 26). Group B were seemed immediate (15 minutes) and late or delayed reactions (n = 26), Group C were seemed only immediate reaction (n = 23). Atopic infants and controlled infants without no symptom but have any atopic disease a relative in the third degree, agreed to undergo SPT in the first visit, the prior were undergo 9-12 months of age, too. Serum total IgE (RIST), serum specific IgE antibody of egg white (EWRAST) and peripheral eosinophil counts in the blood (Eo. counts) were determined at the same time of SPT in atopic infants. The best criterion for SPT was the longest diameter of a erythema were greater than 3 mm at late and/or delayed reaction (Sensitivity: 100%, Specificity: 60%) in group A. Two third of infants in group. A were seemed immediate reaction and EWRAST levels were increased to larger than gread two at 9-12 months (p0.001). RIST levels and Eo. counts at the first visit were increased compared with the normal levels in the all groups, the prior and EWRAST levels in group B were higher than group A or C (p0.05, p0.05). RIST and EWRAST levels in group A at 9-12 months were higher than the first visit (p0.05, p0.01). In conclusion, SPT in atopic early infants were seemed several reactions at the first visit, but all reactions were useful for diagnose for hypersensitivity to egg white.

Background. Eosinophil cationic protein (ECP) has been reported to reflect the eosinophil inflammatory activity in asthma. However, the relative impact of asthma symptoms and atopic eczema upon serum (s)-ECP in asthmatic children has not been established. Objectives. To examine s-ECP levels and s-myeloperoxidase (MPO) in relation to asthma symptoms and atopic eczema in asthmatic children. Methods. S-ECP and s-MPO were assessed in relation to symptom activity, lung function, exercise induced bronchoconstriction and bronchial responsiveness in 101 children; median age 9 years, range 1-16 years; with moderate to severe asthma, admitted to Voksentoppen Center. Result. S-ECP was significantly higher in children with persistent compared to episodic or no asthma symptoms in the past four weeks, S-ECP was also higher in children with atopic compared to non-atopic asthma, as well as in those with active compared to past history of no history of atopic eczema. SMPO was higher in children with persistent asthma symptoms, but did not differ in relation to atopy of eczema state. Persistent asthma symptoms had the greatest impact upon s-ECP levels, followed by atopy and active eczema. Conclusion. S-ECP may be used in assessing symptom activity in asthmatic children, but with the realisation that active eczema and the presence of atopy may also influence levels.

Three groups of HIV-positive men and a control group of healthy subjects were evaluated simultaneously by delayed-type skin tests with recall antigens detection of CD4 cell counts in peripheral blood and the IgE serum levels. Delayed-type skin test reactivity and CD4 cell counts in peripheral blood decreased while IgE serum levels increased as immune imbalance progressed with the worsening of HIV infection (p = 0.003 between controls and HIV-positive patients). The existence of atopy did not significantly influence IgE serum levels in the groups of HIV-positive patients (p0.2). Candidin appeared as a useful antigen in the delayed-type skin tests considering that it was the only antigen that remained positive with low values of CD4 cell counts (or = 250/mm3). The detection of serum IgE levels as well as the performance of delayed-type skin tests with recall antigens are useful tools to evaluate immunological status whereas the number of CD4 in peripheral blood is critical for determining the initiation of antiretroviral therapy.

This paper examines the familial aggregation of physician-diagnosed asthma in relation to the age- and sex-standardized total serum IgE levels of children and their parents in a sample of the general population in Tucson, Arizona, that has been followed in a longitudinal study for over 20 yr. There were 591 nuclear families containing 1,177 children who provided information about the presence or absence of a physician diagnosis of asthma. The serum IgE data were less complete: both parents and one or more of their children in 251 of the nuclear families, containing 468 children, had serum IgE levels measured. There was a very strong tendency for asthmatic patients to have asthmatic children, but only a small part of this appeared to be related to the familial aggregation of total serum IgE. In the absence of an asthmatic parent, there was a slight but significantly higher prevalence of asthma in children of whom both parents had IgE levels in the highest tertile. Very high rates of children's asthma depended on there being an asthmatic parent who also had at least moderate levels of serum IgE. It was also shown that asthmatic children have considerably higher total IgE levels than would be expected on the basis of their parents' IgE levels alone. The data appear compatible with several familial-aggregation hypotheses and a strong environmental influence determining which children are likely to develop asthma. We speculate that the inflammation in the airways of asthmatic patients itself tends to increase the serum IgE level, possibly secondary to mediators that it generates.

The first method used for detection of growth hormone-binding protein (GHBP) in biological fluids was based on the incubation of the sample with radiolabeled GH followed by separation of bound and free GH by gel exclusion chromatography. Recently, other methods have been developed which are faster and easier to use. These methods include variants of the original binding/column assay (e.g., separation of bound and free GH is obtained by immunoprecipitation, charcoal adsorption, ion exchange chromatography, or HPLC), and a ligand-mediated immunofunctional assay (LIFA), in which a monoclonal antibody is used to capture the GHBP on a microtiter plate; all binding sites are saturated with GH and an anti-GH antibody is used to detect the amount of GH (endogenous and exogenous) bound to the GHBP. To permit comparison of results obtained by different methods we have cross-validated the LIFA with two different binding assays: (i) the original long column assay (column assay), and (ii) an assay based on immunoprecipitation (RIPA) of the GH/GHBP complex with an anti-GHBP antibody.

This study, on 185 subjects, has given evaluation of total IgE by the IMX technique in comparison with CAP PRIST, and of the MATRIX* technique for the measurement of sIgE in comparison with the reference technique of CAP RAST**, skin tests and clinical history. The two techniques of CAP and MATRIX are judged to be comparable (Correlation percentage 88.2%). The correlation of MATRIX with skin tests and clinical history is satisfactory (respectively 85.9% and 95%). The results were obtained with four allergens (D. pteronyssinus, Timothy grass, cat and dog). Separate assays confirmed the reliability of the technique.

A panel of 421 cytomegalovirus (CMV) seronegative-platelet donors was followed using complement-fixation (CFT) and IgG-specific radioimmunosorbent tests (RIST) to detect seroconversion. During 4623 person-months of observation 2452 serum specimens were tested, and the annual rate of seroreactivity detected by RIST was 56%. However, most of the positive RIST results were followed by negative results with later serum samples. CFT seroreactivity was a poor predictor of RIST seroreactivity. The high rate of transient seroreactivity detected by RIST may have reflected the lack of absolute specificity of the test. Alternatively, the results could illustrate the difficulty of determining whether an individual harbours latent CMV infection using antibody tests when some seronegative individuals appear to be latently infected. In the light of our observations recommendations for CMV antibody testing of blood donors are proposed.

Determination of total and specific IgE using reactive test strips may afford a useful screening method in the in vitro diagnosis of allergens owing to easy use and possible correlation with established methods. T.D. LETI reactive strips for the semiquantitative determination of total IgE is a method combining classical fundamentals (sandwich anti-IgE-IgE-labeled anti-IgE) and the use of strip methods. It also permits determination of specific IgE levels in a wide range of Mediterranean and food (10) allergens. In a preliminary assessment, this method was correlated with isotopic PRIST for total IgE and with Phadebas RAST for specific IgE. The most important features are: a) no additional equipment besides serum or whole blood; b) cheaper and faster than PRIST for total IgE; c) correlation coefficient of 0.88 in the proposed range (n = 20) and 0.91 (n = 25) in the 10-500 U/ml range for PRIST/T.D. LETI for total IgE; d) specific IgE determination strips present sensitivity and specificity of 88 and 91%, respectively, in 180 allergens assayed in relation to Phadebas RAST. In conclusion, the in vitro diagnosis of allergens using reactive strips is currently a practical and simple method. By optimizing the top of the scale for total IgE and reducing the volume and/or time in specific IgE, it may prove to be an important screening method.

The diagnosis of allergic airway diseases, especially allergic rhinitis, is based on medical history, clinical examination and further tests. Since the detection of IgE, total IgE in serum has become a major tool in the diagnosis of immediate allergy. Although in-vivo-tests like skin prick test are performed in almost every allergologist's practice, the capability of determining IgE was restricted to special laboratories. As shown for semiquantitative glucose dipsticks, a rapid and easy method has advantages for both patient and doctor. A recently developed dipstick test (Visagnost, in U.S. Quidel allergy screen) for allergy screening now enables every practitioner to determine total IgE levels within less than one hour. For the evaluation of the diagnostic value of this dipstick test, we compared the total IgE levels in the sera of 56 patients with two modern methods (Magic Lite, Ciba-Corning, and Cap-FEIA, Pharmacia). Visagnost is an enzyme-immuno-assay with the solid phase on the dipstick. It can be performed in serum, plasma or whole blood, in this study the test was performed in serum. Magic lite is a combined solid-liquid-phase immuno-assay with a final chemiluminescent reaction, the incubation time is about two hours. Cap-FEIA is a solid phase sandwich ELISA with a fluorescent reaction and an incubation time of at least four hours. Patient's sera were shock-frozen in liquid nitrogen and stored at -80 degrees C until examination. Visagnost gave results in four ranges of minor 20 U/ml, 21-50 U/ml, 51-100 U/ml and more than 100 U/ml.(ABSTRACT TRUNCATED AT 250 WORDS)

The authors determined the content of total immunoglobulins of the E class in the blood plasma: native (IgE) and exhausted by a streptococcal allergen in the dynamics of the disease in 145 patients with erysipelas. The level of antistreptococcal reagins was evaluated. It was established that at the onset of erysipelas the concentration of IgE and a-IgE showed a short-term increase in 52 +/- 4% and 87 +/- 3% of patients correspondingly. During convalescence the level of these antibodies did not differ from that of the normal excluding high risk patients for recurrence. The participation of IgE-mediated type of allergic reactions in the pathogenesis of erysipelas is suggested. It is recommended to use the content of IgE and a-IgE for the prognosis of immediate and remote recurrences of erysipelas.

The allergenicity of cow's milk whey proteins, purified by high performance liquid chromatography (HPLC), was examined by the radio-allergosorbent test (RAST) against the sera of children immediately hypersensitive to milk. beta-lactoglobulin and alpha-lactalbumin bound specific IgE in the sera of 63% and 75% of these patients respectively. These allergens were tested for cross reactivity with each other by RAST inhibition. Both inhibited the binding of IgE, in the sera of allergic patients, to the other protein. Two possible determinant peptides, one from beta-lactoglobulin and one from alpha-lactalbumin, were selected by computer prediction of antigenic sites and synthesized by the fluorenylmethoxycarbonyl (FMOC)-polyamide method. The peptides were adsorbed to nitrocellulose discs and used in further RAST studies with sera from the allergic children. Both peptides bound specific IgE in the RAST assay.

PRIST was used to examine the time course of changes in the content of total blood IgE in 128 children aged 3 to 15 years suffering from chronic virus hepatitis and liver cirrhosis. A well-defined relationship was discovered between the above parameter and allergic background as well as the process activity in the liver, pointing to the capacity of that class immunoglobulins to reflect the intensity of not only allergic reactions but also of immunopathological processes lying at the basis of chronic active hepatic diseases.

A rapid enzyme immunoassay system was developed for the detection and quantitative measurement of Total IgE and specific IgE in human serum. The system utilizes a unique solid phase configuration called Acti Tip for the separation phase and an enzyme base detection system. Measurement and quantitation of Total IgE and allergen specific IgE is achieved through the use of a compact photometer pregrogrammed to print out IgE results in units and classes. Only 10 microliters of serum and 45 minutes reaction time is required to measure Total IgE. Allergen specific IgE antibodies are detected and quantitated in 4 hours. A very good agreement exists between test results obtained by the Acti Tip Allerg E and Allerg Ens test systems and the Pharmacia Phadebas PRIST and RAST tests. The correlation coefficients between the two systems is in excess of 90%. The Dexall Acti Tip system offers a number of significant advantages over the currently available in vitro techniques for the detection and quantitation of Total IgE and allergen specific IgE.

We specified three standard levels of IgE RIST (10, 20, 60 IU/ml) for the ninety-four (94) allergic clients of 0-year infants, who classified with three groups. Each group was furthermore divided in two groups such as a group under the standard level and a group exceeding the level respectively. Next, using the Multiple Factor Analysis Type II, we examined quantitative analysis that each of three classified groups was influenced on 12 allergic factors of sex, existence and inexistence of wheezing and atopic dermatitis, family history of allergic diseases, eosinophil counts. IgE RAST scores and antibody titers against egg white, milk and soybean, IgE RAST scores to Dermatophagoides farinae, while the we discussed how the standard levels were adequately reacted to the analysis. As a results, strong allergic factors which influenced the three groups commonly were two factors of IgE RAST scores to egg white and D.f. In particular, the factor which influenced the group of 10 IU/ml standard level was eosinophil counts and family history of allergic diseases. Furthermore, the strong factors which influenced the group classified on 60 IU/ml standard outstandingly appeared in IgE RAST scores to soybean, however, we did not find any difference between three groups. On the other hand, when we specified the standard level at 100/mm3, 200/mm3, 400/mm3 in regard to the eosinophil counts under the same method and examined results, we found strong allergic factors which influenced family history of allergic diseases. IgE RAST scores to egg white and soybean. However, the study scarcely recognize any difference between the three groups.

Food allergies may lead to local and systemic allergic reactions. Participation of respiration tract is rare and is only noticed in combination with additional systemic allergic reactions. Food allergies seem not to have influence on the origin of bronchial hyperreactivity or Asthma. Furthermore the question of genetic influence on the origin of children's allergies and possible preventive measures are discussed.

The efficiency of the new screening tests for atopy, Phadiatop® and CAP Phadiatop®, was studied by comparing their results with a clinical diagnosis of atopy in 100 consecutive adults with asthma and/or rhinitis. Further, the diagnostic efficiency of a combination of Phadiatop and a few standardized questions was studied. The Phadiatop was found to have a specificity of 0.98, and a sensitivity of 0.92 and the CAP Phadiatop a specificity of 0.94 and a sensitivity of 0.96. When the Phadiatop was combined with a few questions, a sensitivity of 1.00 was achieved. It is concluded that Phadiatop and CAP Phadiatop have a higher diagnostic precision than other hitherto used methods for screening of atopic allergy. The place of Phadiatop in a diagnostic flow chart is suggested.

Forty-six adult asthmatics allergic to D. pteronyssinus (Dp) participated in a 2-year study. Thirty-one underwent hyposensitization (HS-group). Fifteen were treated with Dp-extract (Dp-group), and 16 with a similar extract modified by monomethoxypolyethylene glycol with reduced allergenicity (mPEG-Dp-group). Fifteen patients served as controls. Dp-specific antibodies and histamine release from blood basophils were determined and compared with Dp-sensitivity in lungs and skin. In addition, IgG and IgE against the major allergen Der p I were followed in a subgroup. Dp-specific IgG, IgG1, and IgG4 increased significantly in both HS-treated groups after 1 and 2 years (median: 2.5- to 11.6-fold). IgG4 was not induced if maintenance dose during the first year was less than 20,000 BU. Median skin sensitivity decreased 4.4- to 8.2-fold after 1 year and 7.4- to 21.4-fold after 2 years. Der p I specific IgG response was unrelated to the occurrence or change in IgE with the same specificity. The mPEG-Dp-extract tended to have less effect on skin sensitivity and immunological parameters, differences reaching statistical significance for skin sensitivity only. In the HS-group, the decrease in bronchial sensitivity was significantly correlated to a decrease in IgE (r = 0.36), IgG1/IgG4 (r = 0.49), Dp-specific histamine release (r = 0.58), and to an increase in Dp-specific IgG4 (r = -0.36) and IgG4/IgE (r = -0.48). In patients improving clinically, Dp-specific IgG4/IgE increased, and median Dp-specific IgE was reduced to 80% compared with an increase to 150-160% seen in the unchanged or deteriorated group (P less than 0.05). Findings indicate an improvement of effect, if the allergen dose is sufficient to reduce specific IgE and/or induce an IgG and especially IgG4 response.