The influence of a residual group in low-molecular-weight allergoids of Artemisia vulgaris pollen on their allergenicity, IgE- and IgG-binding properties.

BACKGROUND: Reaction of epsilon-amino groups of lysine with potassium cyanate, maleic, or succinic anhydride leads to allergoids of low molecular weight. No study has been performed to compare their properties and investigate the influence of a residual group on allergenicity and human IgE- and IgG-binding of these derivatives. METHODS: Allergoids of a pollen extract of Artemisia vulgaris were obtained by means of potassium cyanate, and succinic and maleic anhydride. Biochemical properties were investigated by determination of amino groups, enzyme activity, isoelectric focusing IEF and SDS-PAGE. IgE- and IgG-binding was determined using immunoblots and ELISA inhibition. Allergenicity was investigated by skin prick tests (SPT) on a group of 52 patients, of which 6 were control subjects, 30 were patients with no previous immunotherapy (IT), and 16 were patients undergoing immunotherapy. RESULTS: The same degree of amino-group modification (more than 85%), residual enzyme activity (less then 15%), IEF, and SDS-PAGE pattern were noted. In the immunoblots of IgE-binding, there was more pronounced reduction in the succinyl and maleyl derivatives than in the carbamyl one. IgG-binding was less affected by carbamylation than by acid anhydride modification. The SPT showed that the succinylated derivative had the most reduced allergenicity (98% showed a reduced wheal diameter when tested with the succinyl derivative, 87% with the maleyl allergoid, and 83% with the carbamyl allergoid). The most significant difference among allergoids could be seen in the group of patients with high skin reactivity (83% of patients showed no reaction to the succinyl derivative when compared to the value of 28% for the carbamyl derivative or 22% for the maleyl derivative). CONCLUSIONS: According to our results, all three modification procedures yielded allergoids with a similar extent of modification. No single biochemical parameter investigated in the study could predict the degree of reduced allergenicity in vivo. The most reduced allergenicity was seen in the succinyl derivative while the preservation of IgG binding epitopes was of the highest degree for the carbamyl derivative.

[Skin reactivity in the histamine-equivalent skin test in hypersensitivity to grass pollen]. / Reaktivnost koze i sistem "prik-test jednak histaminskom" kod preosetljivih na polene trava.

INTRODUCTION: Skin prick-test has been accepted as one of diagnostic criteria for atopic diseases. In accordance with the recommendations of the EAACI (European Academy of Allergy and Clinical Immunology), the Subcommittee for skin testing, allergen specific skin prick-test should be estimated in relation to positive control (histamine hydrochloride in the HEP system/histamine equivalent prick/) [1-4]. The purpose of this study was to establish a recording system of skin sensitivity to allergen by using HEP method in our patients. MATERIAL AND METHODS: The study was performed in 75 persons (49 females, 26 males, mean age 34.6 years) suffering from atopic diseases (hay fever, allergic asthma or rhinoconjunctivitis) and susceptible to one of pollen allergens. Skin prick tests were performed and recorded as said before [2-6]. We used a house standardized (Torlak Institute) allergen solution of 5000 AU/mL (pollen of Dactylis glomerata, Lolium perennae, Phleum pratense), 7500 AU/mL (Phleum pratense) and 10,000 AU/mL (Phleum pratense) and histamine-hydrochloride solution (1 mg/mL and 10 mg/mL). Total and allergen specific serum IgE was made before the study by ELISA immunoassay (Pharmacia, Uppsala, EIA RAST Phadesim) in all selected persons and results were recorded in Phadebas RAST unit given by the manufacturer. According to the serum concentration of specific IgE, patients were classified into groups (low susceptibility, intermediate sensitivity and highly susceptible persons). Skin reactions were recorded according to histamine skin sensitivity (as reaction equal or larger in diameter than histamine papule). Additionally, measured allergenic and histamine wheal were sorted and frequency of positive tests was calculated. Skin testing was performed with the approval of the Ethic Committee of our Centre and with the written patients' consent. For statistical analysis we used chi 2 test, regression analysis and Kolmogar-Smirnov nonparametric test. Results for the confidence of 95% (p < 0.05), were considered significant. RESULTS: In a group of 30 persons, susceptible to Phleum pratense pollen (Table 1, Graph 1 and 2), it was found that the number of positive tests was equally distributed among groups; it was dependent on the allergen concentrations, and was higher when using H1 (p < 0.01). When H10 was used as reference solution, only allergens of the highest concentration provided a significant number of positive tests (p < 0.01). There was no difference among groups of patients depending on the concentration of used histamine solution. In all 75 persons, we estimated skin-prick test performed by allergen solution of 5000 AU/mL and compared it with HI reaction. It was calculated that the most frequent histamine papule diameter was 3 mm and this value was farther used as the end-diameter in HEP system. Patients were classified in groups on the basis of serum IgE concentrations (RAST). The estimation of criterion positivity revealed the highest significance by using criterion Ap > Hp (p < 0.001). At the same time, if the criterion Ap > 2Hp was used we were sure that 99% of our patients were highly sensitive to allergen we tested. DISCUSSION: In the study it was established that allergen concentration of 5000 AU/mL and histamine solution of 1 mg/mL are sufficient to estimate skin-prick test in the HEP system. We found that the end diameter of histamine-papula was 3 mm. This finding suggests the good utility of HEP system. In addition, it has been proven that absolute value of papula diameter is not a critical parameter for the estimation of positivity or discrimation among patients regarding their susceptibility to allergen. By this system we found that criterion Ap > Hp, provides a significantly higher number of positive tests (p < 0.001). The recommended criterion Ap > Hp gives no discrimination among tested groups of patients, while using the criterion Ap > 2Hp we diagnosed highly susceptible persons (p < 0.01). (ABSTRACT TRUNCATED

Protein sgp 120 as a marker of an acquired angioedema.

BACKGROUND: Angioedema is not an event peculiar to an acute-phase response. We registered the appearance of angioedema in four nonatopic patients suffering from diverse inflammatory stimuli. MATERIALS AND METHODS: Serum concentrations of C-reactive protein (CRP), haptoglobin, alpha1-antitrypsin, alpha2-macroglobulin, C3, C4, C1INH and protein sgp 120 (classified as a regulatory protein of the complement system) were assessed over 7 consecutive days, beginning with the onset of angioedema in these four patients. RESULTS: Instead of returning to the normal level, CRP peaked again on the 4th day (A-test, P<0.05) and declined significantly by the 7th day (A-test, P<0.05). Serum levels of sgp 120, C3c and C1INH increased gradually up to the 5th day but only sgp 120 was significantly raised in relation to the initial value (P< 0.05). CONCLUSION: In our patients, we registered four diverse profiles of the acute-phase proteins (Kruskal-Wallis test, P<0.05) probably due to the diverse nature of the pre-existing inflammatory stimuli. According to these results, the unexpected appearance of angioedema implies the contribution of sgp 120 as the main source of fragments with kinin-like properties.

[Interleukin 4, total and allergen-specific immunoglobulin E antibodies in the blood of individuals with an atopic constitution]. / Interleukin-cetiri, ukupna i alergen-specificna antitela imunoglobulina E u serumu osoba s atopijskom konstitucijom.

INTRODUCTION: The recently published data suggest that atopic patients have an enhanced ability to produce IL-4, even in response to antigens other than common environmental allergens or helminthic components. This aberrant IL-4 production by Th cells may be one of the immune alterations encoded by nonMHC genes in the control of basal IgE level [1-7]. The existence of atopen-CD4+ T cell clones that have diverse repertoires might be relevant for aetiopathogenesis of atopic diseases [8-10]. Being subjected continuously to the environmental antigens (allergens), the immune system, especially T cells, is permanently activated and stimulated in response to that challenge. Due to that, substantial amount of diverse cytokines (and their soluble receptors) could be found in the serum. Accordingly, it could be expected to find raised serum IL-4 concentration in atopic persons. In this study we investigated concentrations of IL-4, total IgE and allergen-specific IgE in sera of atopic persons susceptible to pollen allergens. The main goal was to estimate whether the immunologic profile of atopics is defined by increased serum concentration of IL-4 apart from total and allergen-specific serum IgE. METHODS: Patients. We selected 9 atopic patients in the range of 19-35 years, susceptible to grass or weed pollens and suffering from allergic rhinoconjuctivitis. There were 6 females and 3 males (mean age 31.2 years). Atopic allergy was proven by clinical history, skin prick test and by means of in vitro test (positive serum allergen-specific IgE antibodies, RAST/radioallergosorbent tests [11]. As controls we randomly selected B blood donors (6 females, 2 males, mean age 31.3). Study design. The study was carried out from August till September 1995. None of selected patients had previously accomplished allergen-specific immunotherapy nor had been medicated by corticosteroids (topic or systemic) [12-15]. Selected persons were not allowed to take medications such as antihistamines, beta-2 agonists or tricyclic antidepressants at least 10 days prior to the study. We performed in vitro tests for determining concentrations of IL-4, total and pollen-specific serum IgE. Sera were sampled in the morning and were stored frozen at -20 degrees C until analysis. Determination of IL-4, total and allergen specific IgE in serum. Interleukin-4 measurement was carried out in blind fashion with an ELISA kit (Intertest-4 ELISA, Genzyme, Cambridge) according to the manufacturer's instructions. A reference curve was obtained by plotting the IL-4 concentration of several standard dilutions versus an absorbency. The determination limit was 0.045 pg/ml. For determining total serum IgE we used commercially available enzyme immunoassay (EIA Phadezym IgE PRIST, Pharmacia, Uppsala). Normal values was up to 120 IU/L (international unit per ml). Allergen specific IgE was determined by semiquantitative EIA method (RAST Phadezym, Pharmacia, Uppsala) and expressed in Phadebas RAST Unit (PRU) according to standard curve done by manufacturer. Study was carried out after approval of the Ethic Committee of our Hospital and with the obtained patients consents. For statistical analysis we used nonparametric tests (U-test and the Spearman rank correlation). For all comparisons, statistical significance was considered to be present if p < 0.05. RESULTS: Among selected patients, 4 persons had high concentration of serum IgE antibodies (RAST class 4) against grass pollens, two persons had IgE-specific (RAST, class 4) to weed pollens and 2 patients had IgE antibodies (RAST class 4) against weed and grass pollens simultaneously. RAST of class 3 was registered in the serum of one person. Registered serum total IgE, allergen-specifc IgE and IL-4 are shown in Table 1. Serum IL-4 was significantly higher in atopics (U-test, SR 43.53, p.05) as well as total IgE (ER 493, p.05). In atopics, IL-4 was not in correlation with either total or allergen-specific IgE (IL-4 versus total IgE, p = +0.190;

[Specific skin reactions induced by individual pollen preparations in hypersensitivity persons]. / Specificna reaktivnost koze izazvana pojedinacnim pripravcima polenakod preosetljivih osoba.

INTRODUCTION: Skin tests are the most useful single modality for demonstrating an IgE-mediated mechanism underlying clinical symptoms. Skin reactivity to an allergen depends on person's exposure and genetic factors influencing the IgE response. However, the reproducibility of allergy skin tests has been shown to be variable depending, among other things, on the allergen extract employed and the kind of technique used. These variations have led to controversy regarding clinical relevancy of allergy skin testing [1-7]. We examined the relationship of skin-prick test and serum allergen-specific IgE in pollen susceptible adult persons. We estimated changes in the immediate skin reactivity to pollen-allergens by testing patients on more than one occasion: a) during and out of pollen season; b) before and after completing specific immunotherapy, and c) who had been tested with diverse concentrations of the same pollen-allergen. We correlated the degree of skin reactivity at the initial and subsequent testings with the aim of estimate diagnostic relevancy of such performed allergy skin testings. MATERIAL AND METHODS: Study was performed in 64 persons (aged from 17 to 43 years) suffering from rhinitis or rhinoconjunctivitis or asthma. They were diagnosed for the first time [8, 9]. None of selected persons had proven dermographism [1, 9]. Study protocol Skin prick tests were performed according to recommended protocol reviewed elsewhere [9]. Blood samples for total and allergen-specific IgE were taken in the morning on the test-day and sera were stored until analyses. Testing was performed with in house standardized extract of grass pollens (each of 5000 AU/allergy unit per milliliter): Dactylis glomerata and Phleum pratense and Lolium perenne (produced by Torlak Institute, Belgrade). Testing was performed in selected groups of patients: a) during and out of pollen season; b) before and after allergen-specific immunotherapy, and c) with three diverse concentrations of Phleum pratense allergenic extract (5000 AU, 7500 AU and 10,000 AU/ml). During the study a medication was not allowed at least 7 days prior to the skin testing (14 days for hydroxyzine). Alergen-specific IgE in serum (RAST) Determination of serum allergen-specific IgE was performed by using commercially available Pharmacia kit (EIA RAST Phadesim). For allergen-specific IgE results were expressed in classes (1 minimal to 4-maximal concentrations) determined by manufacturer. Study was performed under approval of the Ethic Committee of our hospital. For statistical analysis we used t-test and test of variance (ANOVA). Results were considered significant if p < 0.05. RESULTS: Influence of allergen-specific serum IgE concentration on local skin reactivity was estimated in 23 persons (17 females, 6 males), and is shown in Table 1. The largest papule diameter was registered in high susceptible persons (RAST class 4). Influences of season when testing was performed and specific immunotherapy were estimated in 26 persons (19 females, 7 males) (Figures 1 and 2). The effect of diverse allergenic extract concentrations was estimated in 15 adults (9 females, 6 males, mean age 26.4 years) and results are shown in Figure 3. DISCUSSION: In the study it has been shown that serum concentration of allergen-specific IgE positively correlated with the skin reactivity to pollen-extract. In addition, allergen specific immunotherapy as well as the concentrations of the used allergenic extract influenced significantly local skin reactivity. On the contrary, season when testing had no influence. According to our results, it seems that skin reactions (papule), equal or larger than 6 mm in diameter, could be of diagnostic value concerning pollen-monotest application (Table 1). Meanwhile, with significance of 95% we could postulate that in persons tested with the lowest concentration of allergen-solution (5000 AU/ml), skin prick reactivity defined by the papule of 4 mm or larger can replace in vitro

Lack of cross-reactivity between casein and gliadin in sera from coeliac disease patients.

BACKGROUND: While searching for antigliadin and anticasein antibodies in sera from coeliac patients we investigated more than 20 persons suffering from this enteropathy. In some of sera, we registered antibodies against gliadin and in parallel the appearance of anticasein antibodies. A high percentage of glutamic acid has been proven by amino acid analysis in both these proteins. Therefore we try to demonstrate the existence of cross-reactivity between them in sera from coeliac patients. METHODS: We analyzed sera from 5 adult patients by using Western blot (SDS-PAGE and immunoblot assay). All of them had previously proven serum antibodies against casein and gliadin. In the procedure, antigliadin-antibodies were removed from sera by adsorption with gliadin. Adsorbed antibodies were then eluted from the adsorbent. RESULTS: Adsorbed samples retained anticasein activity. Eluates had a strong reactivity to gliadin but not to casein. CONCLUSION: No cross-reactivity restricted to any of the immunoglobulin isotypes (IgG, IgM, IgA) had been proven between these two proteins. Before the final conclusion, we believe the study has to be completed by using partly of totally disassembled proteins by intestinal enzymes. It might be that enzymatic digestion could open some epitopes shared by both these proteins.

[Total allergen-specific IgE antibodies in the serum and skin reactivity induced by a mixture of pollens in allergic individuals]. / Ukupna alergen-specificna IgE antitela u serumu i reakcija koze izazvana mesavinama polena kod preosetljivih osoba.

Skin puncture test is fast and precise method for revealing IgE-mediated allergen hypersensitivity. In the diagnosis of pollinosis and other atrophic diseases, expensive tests for revealing the allergen-specific IgE antibodies in the serum are frequently inaccessible. The subject of our investigation was how and if the skin test with mixture of pollen could replace the expensive in-vitro method in diagnostic procedure. In 41 patients with pollinosis were performed skin tests by the mixture of pollen of grass, weeds and trees (produced by Torlak Institute, Belgrade) and determined the concentration of overall and allergen-specific IgE serum (EIA, enzyme immuno-assay, Pharmacia Uppsala; RAST Phadesim, multidisk, Pharmacia). Obtained results pointed out that the sex of investigated subjects did not influence the size of skin reaction in case when the pollen mixture (grass, weed or trees) was used. With high significance rate was established that in skin puncture test, just the size of the reaction, defined by 5 mm papule or larger, indicated the hypersensitive person. We consider that this can be taken as the diagnostic criterion and replace in-vitro method for determination of allergen-specific IgE antibodies in serum. Also, overall IgE of serum positively correlated (p < 0.05) with the size of skin reaction only in the persons with extremely high concentrations (RAST class 4) allergen-specific IgE antibodies in serum.

Early effects of rush immunotherapy with Dermatophagoides pteronyssinus in asthmatics.

In this study we assessed the effects of Dermatophagoides pteronyssinus (Dpt) rush immunotherapy in comparison with placebo treatment in our asthmatic patients. Fourteen highly Dpt-susceptible adults were randomized in two groups (immunotherapy and placebo) and treated in single-blind manner. Patients were selected according to the recommendation of the immunotherapy position paper (1993). To minimize side effects we modified the protocol by adjusting allergen doses for each patient separately. Immunologic (total and Dpt-specific serum IgE and IgG antibodies/EIA, Pharmacia) and clinical parameters (spirometry, medication score and skin testing) were recorded before treatment, after 2 weeks, at the second month and after 4 months of immunotherapy onset. None of the patients had life-threatening side effects in the course of the treatment. The results obtained demonstrated significant influence of immunotherapy on Dpt-specific serum IgG synthesis (Kruskal Wallis test, p < 0.05) and on the late phase skin reaction with Dpt (U-test, p < 0.05) at the end of the second month of immunotherapy onset. In the immunotherapy group, we also registered a negative correlation between concentrations of Dpt-specific serum IgE and IgG antibodies (p = -0.83; p < 0.05), at the end of the second month. In addition, diversities among patients, expressed by immunologic parameters, were related to the amount of delivered allergen. There were no significant differences between groups concerning medication score from opposite to better FEV, PEFR and dPEFR results (Kruskal Wallis, p < 0.05) in the placebo group. In conclusion, Dpt-specific serum IgG concentration, immunologic score and late phase skin reactivity to allergen appeared to be the valid parameters of rush-immunotherapy achievement, while delivered allergen dose also seemed to be an influencing factor.

[Significance of monitoring histamine serum levels, cryoglobulins and complement titers during cold tests in patients with cold allergy and Raynaud's phenomenon]. / Znacaj pracenja vrednosti histaminemije, krioglobulina i titra komplementa u toku testa hladjenja u pacijenata sa alergijom na hladnocu i Raynaud fenomenom.

The aim of our investigations is to evaluate blood histamine, complement titer and cryoglobulins before and after cold stimulation (extremity water immersion test) in patients with cold urticaria and Raynaud phenomenon. In our investigations patients were put into five groups according to the following diagnoses: cold urticaria (40 patients), chronic urticaria (15 patients), Urticaria-Vasculitis Syndrome (15 patients), Raynauds Disease (16 patients), and Raynauds phenomenon (15 patients). The investigations demonstrated that the test was always positive in cold urticaria and that a high percentage of patients had a rise in blood histamine. This was found in 50% of the patients while cryoglobulins were found in one third of the patients. Consequently they can be grouped in secondary cold urticarias. A number of patients with chronic urticaria and Urticaria-Vasculitis Syndrome gave a positive history of cold sensitivity. Positive tests were accompanied by a rise in histamine, although (less frequently) there was a decrease in the complement titer and the occurence of cryoglobulins. Raynauds Disease was accompanied by a positive test in 94% of the patients, while patients with Raynauds Phenomenon (whose etiology was Thoracic Outlet Syndrome) had a negative test (60% of the patients). Significant changes were not found in the remaining groups. Positive tests were accompanied by abnormal values of cryoglobulins and of complement titer as well as an increase in histamine. The rise in histamine was not significant in Raynauds Phenomenon, while one patient had a decrease in the complement titer and the appearance of cryoglobulins.

[Atopy and drug allergy]. / Odnos atopije i alergije na lekove.

UNLABELLED: The aim of our study is to show possible correlation between atopy and drug allergy, and the clinical manifestations of drug allergy that may have predictive value. There were 150 patients with positive history of drug allergy, ages 18-73 years. Cutaneous anr/or dose provocative testing was done, depending on the drug. Drug allergy was found in 58 patients (40%). There were 18 patients (12%) with atopic diathesis of which 9 (50%) were found to have drug allergy. Of 17 patients diagnosed as anaphylaxis, 13 (76%) had drug allergy, penicillin being the most common offending drug (7 parients). Of 75 patients diagnosed as urticaria, drug allergy was prouded in 36 (47%) patients. Positive drug allergy was found as follows (percentage wise): trimetoprim-sulfometoksazol 46% (13 tested, 6 positive); penicillin 35,8% (54 tested, 19 positive); noraminophenazone 31% (22 tested, 6 positive); muscle relaxants and general anaesthetics 26% (23 tested, 6 single positive). CONCLUSIONS: Athophy is not predisposing factor for drug allergy; 60% of patients did not have drug allergy although stated in medical documents; anaphylaxis has the highest predictive value, followed by urticardia and fixed drug eruption (when trimetoprim-sulfometoxasol is concerned).

[Histologic manifestations in the skin in alpha-lactoalbumin intradermal tests in patients with chronic urticaria]. / Histoloske pojave u kozi u toku intradermnog testa alfa-laktoalbuminom kod bolesnika s hronicnom urtikarijom.

Intradermal injection with alpha-lactoalbumine, 2000 PNU/ml was carried out in seven patients (1 male, 6 females, mean age 32.4 years) suffering from milk intolerance. None of the patients had a positive allergen-specific IgE antibodies in the sera (RAST). Skin test was performed on the volar part of the forearms at the same time. The area of skin reaction was estimated and measured within 36 hours on the right arm. Skin biopsy was done with a 4 mm punch after 8 hours from site of antigen injection on the left arm. Various cells infiltrating the site of antigen instillation were present. Leukocytoclastic vasculitis was only found in patients with dual intradermal reaction (immediate and late onset of response; 8 12 hours). These results indicate that local skin reaction to alpha-lactoal-bumine is mediated by diverse immune mechanisms, and that it may be of clinical significance for the diagnosis of cow milk allergy in adults.

[Clinico-laboratory characteristics of physical urticaria]. / Klinicko-laboratorijske karakteristike fizickih urtikarija.

Physical urticarias represent a specific reaction of the skin, which is characterised by local erythema, edema and pruritus at the site where the physical stimulus has been applied. However, local changes can be associated with various systemic manifestations. Specific cutaneous reactions due to physical factors may be the first manifestation of a systemic and/or infectious disease, which should be taken into account as physical urticarias generally occur in young patients. In view of the above mentioned, as well as unpredictable course of the disease and relatively limited therapeutic effects, the etiology and pathogenesis of the physical urticarias are one of the most interesting fields of investigation in contemporary medicine.

[Effect of ranitidine on the pharmacokinetics of theophylline in patients with obstructive pulmonary disease]. / Moguci uticaj ranitidina na farmakokinetiku teofilina u bolesnika sa opstruktivnim bolestima pluca.

Investigations were performed on 20 patients with mild and moderately severe forms of bronchiale asthma or chronic obstructive bronchitis. Theophylline for oral use (sustained release form) and Ranitidine (second generation H2-blocker) were used in the first group of patients (7 females and 3 males, average age 37.9). Dosage of theophylline from patients sera was monitored at 3, 6 and 12 hours, beginning with the initial morning dose. The second group of patients (9 females and 1 male, average age 46.3) were on Ranitidine therapy, along with 250 mg intravenous theophylline-ethylene-diamine. Fractionated dosage of theophylline was done within 12 hours. Pharmacokinetic parameters were calculated according to the concentration of theophylline in the sera of patients in the second group. Significant differences were not found by statistical analysis (T-test) in the pharmacokinetic parameters of theophylline although significantly higher concentrations of theophylline were found in the sera of patients in the first group. This indicated that there are individual differences on the metabolism of theophylline drugs with concomitant use of Ranitidine.

[Proteinuria in dermal vasculitis (incidence, degree and possible mechanisms of onset)]. / Proteinurija u dermalnim vasculitisima (ucestalost, stepen i moguci mehanizmi nastanka).

The paper comprises 63 patients with diagnosed vasculitis of the skin who had abnormal findings in the urine. Thirteen of these patients had positive skin findings and abnormal findings in the urine, with concomitant attacks of dyspnea as seen in bronchial asthma. These patients account for 20% of the entire group, while there was an incidence of 37% of systemic manifestations. Skin findings: 45% had a maculopapular rash, 36% had urticarial findings, 13% had urticaria and angioedema and 6% had angioedema alone. Nonspecific biologic syndrome of evolution was statistically significant while no significant changes were found in the number of white blood cells and eosinophils. The degree of proteinuria ranged from 0.1 g/L to 1.16 g/L. Most patients with proteinuria above 0.25 g/L had microhematuria. Slightly over 50% of the patients had signs of complement activation by the alternative pathway, along with the presence of cryoglobulins. Increased histamine in the serum was found in over 50% of the patients although the values of histamine did not correlate with the degree of proteinuria. Proteinuria was not detected in patients with very high values of histamine (and without signs of vasculitis), which indicates that histamine itself responsible for changes in the glomeruli. There is a possibility that local tissue hyperhistaminemia is responsible for the increased permeability of the basal membrane of the glomeruli. According to the obtained results, the etiology of proteinuria and microhematuria should be pursued in the pathogenesis of vasculitis as signs of complement activation indicate. Other possible causes for proteinuria were excluded. The proteinuria was selective, benign according to its course and degree, occurred concomitantly with skin findings and was absent during remission of the disease.

[Laboratory study of patients with endemic nephropathy]. / Prilog laboratorijskim ispitivanjima bolesnika sa endemskom nefropatijom.

The introduction deals with the main characteristics of two representatives of high risk pregnancies: diabetes mellitus and EPH gestoses. Particular interest was shown in risks that may occur in pregnant diabetics, with which they must be acquainted. Some theories are given on the pathogenesis of EPH gestoses. The aim of our investigation was to determine the concentration at which immunoglobulins G, M and A pass in cord blood, amniotic fluid and urine, and whether there is a significant difference between control group and among pregnant diabetics or those with EPH gestoses. The investigation was performed by the nephelometric technique on the Immunochemistry Analyzer. Each group consisted of 20 women, with a total of 60. Immunoglobulins A, M and G were determined in the mothers' sera, cord blood or amniotic fluid. IgG was obtained in urine in measurable concentrations. A significant increase of IgG was found in the urine of pregnant diabetics. IgM was significantly increased in the sera of diabetic mothers. IgA was significantly increased in pregnant diabetics, while both high risk pregnancy groups had an increased IgA in cord blood. The values of IgA in amniotic fluid were decreased in the EPH gestoses group in comparison to the group of diabetics. The authors find these variations interesting and feel they should be followed in other high risk pregnancy groups.

[The effect of ranitidine on theophylline pharmacokinetics in patients with obstructive lung disease]. / Uticaj ranitidina na farmakokinetiku teofilina u bolesnika sa opstrukcionim bolestima pluca.

The study was performed in 20 patients with mild and moderately severe forms of bronchial asthma or chronic obstructive bronchitis. Theophylline for oral use (sustained release form) and ranitidine was used in the first group of patients (7 females and 3 males, mean years of age 37.9). Dosage of theophylline from patients' sera was monitored prior and at 3, 6, and 12 hours beginning with the initial morning dose. The second group of patients (9 females and 1 male, mean age 46.3 years) were on ranitidine therapy along with 250 mg intravenous theophylline-ethylene-diamine. Fractionated dosage of theophylline was made within 12 hours. Pharmacokinetic parameters were calculated according to the concentration of theophylline in the sera of patients in the second group. In the sera of patients in the first group significantly higher concentrations of theophylline were found at 3 and 6 hour of the initial morning dose (p less than 0.05). In the second group a significant difference was not registered for the assessed pharmacokinetic parameters. This investigation indicates that there are individual differences of the metabolism of theophylline drugs with concommitant use of ranitidine.

[The oral milk provocation test in patients with chronic urticaria]. / Oralni provokacioni test mlekom u bolesnika sa hronicnom urtikarijom.

Histamine and sera complement were followed-up in patients with chronic urticaria during oral provoking test to cow milk. Three patients with a positive history to cow milk sensitisation showed a significant decrease in C3 concentration within 180 minutes of the test. This was followed by increased histamine and skin eruption. According to our results (skin test reactivity and clinical evaluation of the provoking test) several mechanisms were involved in the sensitisation to cow milk in our patients.