Heat-labile Escherichia coli toxin enhances the induction of allergen-specific IgG antibodies in epicutaneous patch vaccination.

Epicutaneous allergen-specific immunotherapy (EPIT) is proposed as an alternative route for allergen-specific immunotherapy (AIT). The induction of allergen-specific blocking IgG antibodies represents an important mechanism underlying AIT, but has not been investigated for EPIT. Here, we compared the induction of allergen-specific blocking IgG in outbred guinea pigs which had been immunized with recombinant birch pollen allergen Bet v 1 using patch delivery system (PDS) with or without heat-labile toxin (LT) from Escherichia coli or subcutaneously with aluminum hydroxide (Alum)-adsorbed rBet v 1. Only subcutaneous immunization with Alum-adsorbed rBet v 1 and epicutaneous administration of rBet v 1 with PDS in combination with LT from E. coli induced allergen-specific IgG antibodies blocking allergic patients' IgE, but not immunization with rBet v 1 via PDS alone. Our results suggest that patch vaccination with rBet v 1 in combination with LT may be a promising strategy for allergen-specific immunotherapy against birch pollen allergy.

Heart rate variability in fibromyalgia patients and healthy controls during non-REM and REM sleep: a case-control study.

OBJECTIVES: To investigate heart rate variability (HRV) in fibromyalgia (FM) patients and healthy controls (HCs) during different sleep stages, and to examine the association of HRV with pain and sleep quality. METHOD: Polysomnography was recorded from 23 female FM patients and 22 age- and sex-matched HCs. HRV was recorded from bedtime until awakening including the standard deviation of normal-to-normal intervals (SDNN), the root mean square successive difference (RMSSD), and the low (LF; 0.04-0.15 Hz) and high (HF; 0.15-0.4 Hz) frequency power. Subjective scores of neck/shoulder pain and sleep quality were obtained at bedtime and awakening. RESULTS: Both patients and HCs showed high incidence of arousals per hour (FM: 16 ± 9.7; HCs: 17 ± 11). RMSSD was lower in patients than HCs during non-rapid eye movement (non-REM) stage 2 (N2) sleep (mean ± SD; 30 ± 12 ms vs. 42 ± 13 ms, p < 0.002) and during REM sleep (23 ± 11 ms vs. 37 ± 16 ms, p < 0.003). HRV did not differ between groups during N3 sleep (p > 0.19 for all comparisons). In patients, SDNN, RMSSD, and HF power showed modest positive correlations with sleep quality (HF power during N3 sleep showed the highest correlation; Spearman's ρ = 0.54) and modest negative correlations with neck/shoulder pain (RMSSD during N3 sleep showed the highest correlation with pain at bedtime; Spearman's ρ = -0.51). CONCLUSIONS: RMSSD, indicative of parasympathetic predominance, is attenuated in FM patients compared to HCs during N2 sleep and REM sleep. This difference was not present for the HF component. HRV during sleep in FM patients is moderately and positively associated with sleep quality and moderately and negatively associated with neck/shoulder pain.

Gender differences in work-home interplay and symptom perception among Swedish white-collar employees.

OBJECTIVE: To analyse gender differences in paid and unpaid workload and symptoms in matched groups of Swedish white-collar workers with children. DESIGN AND SETTING: Paid and unpaid workload and perceived stress from paid work, conflict between demands and control over household work were measured by a total workload (TWL) questionnaire. Some symptoms were rated with regard to frequency and severity as a measure of health. Cross-sectional analyses were performed. PARTICIPANTS: Matched groups of male (n = 440) and female (n = 529) well-educated white-collar workers in full-time employment, aged 32-58 years and living with children in the home. RESULTS: Women in higher positions in Sweden are healthier than the average population of women, but report more symptoms than men in the same position as well as more stress from paid work, more conflict between demands and a greater TWL. However, women also reported more control over household duties and TWL was not associated with more symptoms. The men were mainly focused on their paid work role and perhaps even more so than men in the general population as they were fairly resistant to feelings of conflicting demands. CONCLUSION: Even among matched groups of full-time employed, well-educated men and women, traditional gender differences in division of responsibilities and time allocation were found. Even though the women were healthy at this stage, they might risk future ill health, owing to high workload, stress and feelings of conflicting demands.

Co-existing structures of an mRNA stability determinant. The 5' region of the Escherichia coli and Serratia marcescens ompA mRNA.

The structure of untranslated regions of mRNA is thought to play a key role in the degradation of mRNAs by specific RNases. As a model system, in vitro transcripts of the stability determining 5' non-coding region of bacterial ompA mRNA were investigated by calculation of secondary structure models and by experiments applying the temperature-gradient gel electrophoresis (TGGE). For the theoretical prediction of secondary structures an algorithm was used, which yields the structure of lowest free energy as well as a large set of suboptimal structures. Three structures were predicted to co-exist in similar concentrations under native conditions. They denature in a low temperature transition leading to a unique structure which denatures in a high temperature transition. The prediction of three structures and two transitions could be confirmed experimentally by TGGE. Due to the use of transcripts of different length the conformational transitions could be attributed to distinct parts of the molecules. A pseudoknot structural motif was predicted theoretically, but could not be confirmed experimentally. Comparing ompA transcripts of E. coli and S. marcescens, a conservation of structural features could be shown in spite of a sequence homology of only 63%. Regarding the sequential folding of the transcript after synthesis, a metastable structure is formed first and is converted slowly into structures of lower free energy. The biological implications for in vivo degradation are discussed.

Cardiovascular reactivity to the cold pressor test as a predictor of hypertension.

Cardiovascular reactivity to stress is hypothesized to be a marker for subsequent neurogenic cardiovascular disease, but few prospective studies of this hypothesis are available. We studied 910 white male medical students who had their blood pressure and pulse rate measured before and during a cold pressor test in the years 1948-1964. Hypertensive status (requiring drug treatment) was ascertained by annual questionnaires in the 20- to 36-year follow-up period. An association was observed between maximum change in systolic blood pressure and later hypertension, with a cumulative incidence of hypertension by age 44 of 6.7%, 3.0%, and 2.4% for a change in systolic blood pressure in the upper, middle two, and lowest quartiles, respectively (Kaplan-Meier, p less than 0.02). After adjustment for study entry age, Quetelet Index, cigarette smoking, pretest systolic blood pressure, and paternal or maternal history of hypertension in a Cox model, the association persisted. The excess risk associated with systolic blood pressure reactivity was not apparent until the population aged some 20 years and was most apparent among those in whom hypertension developed before age 45 (relative risk = 2.5, 95% confidence intervals = 1.47, 4.71 for a 20 mm Hg change). Diastolic blood pressure and heart rate changes were not associated with later hypertension. These data suggest that persons prone to later hypertension manifest an altered physiology at a young age.

The differential stability of the Escherichia coli ompA and bla mRNA at various growth rates is not correlated to the efficiency of translation.

Using two monocistronic gene transcripts, bla and ompA, we have studied the relationship between mRNA stability and translational efficiency. It was found that changes in the ompA mRNA stability are not correlated with an alteration in translational efficiency. In addition, at slow bacterial growth rates, the ompA transcript is translated ten times more efficiently than the bla messenger although the stability of the two transcripts is about equal. At rapid bacterial growth rate, chloramphenicol slightly stabilises both the bla and ompA transcripts without affecting their characteristic difference in half-life. Thus, control of mRNA stability seems not necessarily to be mediated either by the efficiency of loading ribosomes on a transcript, or by the arrest or slowing down of translating ribosomes.

Characterization of RNA with unusual electrophoretic mobility from tissues of patients with Crohn's disease.

RNAs from tissues of patients with Crohn's disease that migrate off the diagonal in a two-dimensional gel electrophoresis system were partially characterized. One of the RNA species was a discrete cleavage product of region V2-9 of 28S rRNA; another is a conformer or variant of 5.8S rRNA; and a third is a mixture of unidentified fragments with mobility similar to that of 7S RNA. The yield of these species from resected tissue and their visualization by silver staining was very sensitive to the details of the preparative procedure. No evidence of viroid-like RNA was found within the range of molecular sizes (< 7S) that we examined.

Appearance of an arachidonic acid 15-lipoxygenase pathway upon differentiation of the human promyelocytic cell-line HL-60.

The metabolism of arachidonic acid and 15-HPETE was studied in a human promyelocytic cell line (HL-60). Upon exposure to DMSO, HL-60 cells undergo differentiation and acquire a 15-lipoxygenase activity while undifferentiated cells challenged with either arachidonic acid or 15-HPETE did not enzymatically transform these precursors. Products of the arachidonic acid 15-lipoxygenase pathway were identified by HPLC. UV-absorption and gas chromatography-mass spectrometry. Results indicate that upon differentiation HL-60 cells express a 15-lipoxygenase activity as well as the ability to transform 15-HPETE to 8,15-DHETEs and 14,15-DHETE. Moreover, these findings suggest that products of the 15-lipoxygenase cascade may be generated by a single cell system.

Formation of leukotriene C4 by human leukocytes exposed to monosodium urate crystals.

Monosodium urate (MSU) crystals stimulate the metabolism of arachidonic acid in mixed populations of human leukocytes. Leukocytes exposed to MSU crystals released leukotriene C4. Leukotriene C4 (LTC4) was characterized and detected by high-performance liquid chromatography (HPLC), UV absorption, bioassay with guinea pig ileum, and radioimmunoassay. Results indicate that MSU crystals stimulate the transformation of arachidonic acid and the formation of leukotriene C4 in human leukocytes; an effect inhibited by colchicine. Moreover, they suggest that LTC4 may serve as a mediator of inflammation in crystal-associated diseases.

Fibrinogen Caracas V, an abnormal fibrinogen with an Aalpha 532 Ser-->Cys substitution associated with thrombosis.

A new dysfibrinogenemia associated with thrombophilia has been identified in a Venezuelan kindred. Thrombin and Reptilase times were prolonged and the accelerating capacity of the patient's fibrin on the t-PA-induced plasminogen activation was decreased. In addition the affinity of fibrinogen for plasminogen was diminished. Permeability and electron microscopy studies revealed that the abnormal clot was made up of thin and densely packed fibres giving rise to a reduced fibrin gel porosity. This was confirmed by turbidity studies showing a decreased fibre mass/length ratio. Affected members were heterozygous for an Aalpha 532 Ser-->Cys mutation as demonstrated by genetic analyses. This abnormal fibrinogen has been designated as Fibrinogen Caracas V. The family study showed a convincing association between the mutation and thrombotic manifestations. The thrombotic tendency may be ascribed to lack of accelerating capacity of fibrin to induce fibrinolysis caused by an abnormal clot structure with thin fibres and reduced porosity.

Sex differences in sympathetic-adrenal medullary reactions induced by different stressors.

Male and female university students were exposed to two different stressors in each of two 110-min sessions, i.e., a cognitive task (color-word conflict) and repeated venipuncture. Catecholamine excretion, heart rate, and subjective reactions were measured. Control values were obtained under conditions of relaxation in the laboratory. Subjects of both sexes responded to both stressors by increased heart rate and feelings of unpleasantness and distress. The pattern of adrenaline excretion, however, differed between sexes: in males both stressors induced a significant increase, whereas in females adrenaline excretion remained on the same level under the two stress conditions as during relaxation; Noradrenaline excretion was not systematically affected by either stressor in either sex group.

Interaction of alcohol and reward in an achievement situation.

Effects of psychological stress on acute alcohol intoxication following intake of 2 ml whisky/kg body weight were examined in 12 healthy subjects. Each subject performed a 45-min psychomotor task under four conditions, two of which increased achievement stress by promising the subject a monetary reward for rapid and accurate performance. The conditions were (a) task only, (b) task + reward, (c) task + alcohol, and (d) task + reward + alcohol. In condition (b) reward improved performance and heightened both physiological and subjective arousal, and in condition (c) alcohol impaired performance, lowered subjective arousal, and increased physiological arousal. In condition (d) the reward counteracted the depressant effects of alcohol on performance and mood, whereas an additive, arousing effect was obtained in all physiological variables, i.e., catecholamine and cortisol excretion and heart rate.

Stress responses in low-status jobs and their relationship to health risks: musculoskeletal disorders.

Conditions typical of many low-status jobs are known to induce elevated stress. In keeping with this, blue-collar workers show elevated psychophysiological stress levels both during and after work compared with workers in more stimulating and flexible jobs. Health-related behaviors, such as cigarette smoking and drug abuse, that are known to contribute to the social gradient in health, can be seen as ways of coping with a stressful work situation in order to get short-term relief. Negative emotional states associated with low-status jobs, combined with a lack of economic resources, are also likely to reduce the individual's motivation to seek proper medical treatment and, thus, increase the risk that transient symptoms develop into chronic illness. With regard to musculoskeletal disorders, it is well documented that physically monotonous or repetitive work is associated with an increase in neck, shoulder, and low back pain problems. However, recent studies also report an association between psychosocial factors and muscle pain syndromes. Possible mechanisms explaining these findings involve the assumption that psychological stress may induce sustained activation of small, low-threshold motor units that may lead to degenerative processes, damage, and pain. Analysis of short periods of very low muscular electrical activity (EMG gaps) shows that female workers with a high frequency of EMG gaps seem to have less risk of developing myalgia problems than do workers with fewer gaps. Stress induced by psychosocial conditions at work, which is usually more lasting than that resulting from physical demands, may prevent the individual from shutting off their physiological activation and reduces the time for rest and recovery. In the modern work environment, with strong emphasis on a high work pace, competitiveness, and efficiency, it is possible that lack of relaxation is an even more important health problem than is the absolute level of contraction or the frequency of muscular activation.

Experimental thrombosis I: relation with fibrinogen and other haemostatic parameters.

Epidemiological studies have shown that the haemostatic parameters Fibrinogen (Fg), Factor VII (F VII), Factor VIII (F VIII), von Willebrand factor (vWF), Tissue Plasminogen Activator (t-PA), Plasminogen Activator Inhibitors (PAI) are risk factors/markers of ischemic cardiovascular disease. Ferritin (sFER) and Leukocytosis have also been implicated. In the present study we have followed the levels of fibrinogen, von Willebrand factor and thrombomodulin in relation to lipids, iron and the appearance of atherosclerotic lesions in New Zealand rabbits fed with a cholesterol enriched diet for a two-month period compared with a group of control rabbits. Hematocrit and white blood cell count (WBC) were measured in parallel. In hyperchlesterolemic rabbits the levels of fibrinogen and von Willebrand factor increased progressively, showing a positive correlation with the increasing cholesterol levels. There was an increase in soluble thrombomodulin beginning at the eighth week of study. In addition, these animals showed gross intimal atherosclerotic lesions in the whole extension of their aortas. Immunohistochemical studies showed the presence of fibrin(ogen) related antigen throughout the arterial wall and in the central portions of the atheromas. In the control group there was no formation of atherosclerotic plaques and all haemostatic, haematological and biochemical parameters were within the normal range. WBC and sFER levels were unaffected in both groups. Our results show that increased levels of fibrinogen and von Willebrand factor, known coronary risk factors, are strongly associated with the formation of atherosclerotic plaques in rabbits. The plaques contain a considerable amount of fibrinogen related antigen.

Fibrinogen Guarenas I: partial characterization of a new dysfibrinogenemia with an altered rate of fibrinopeptide release and an impaired polymerization.

A congenitally abnormal fibrinogen was isolated from the blood of a young woman with a severe bleeding diathesis. Coagulation tests showed a prolonged Thrombin and Reptilase time partially corrected by Ca2+. Polymerization of thrombin induced preformed fibrin monomers was severely impaired. Thrombin caused the release of fibrinopeptides with normal retention times on HPLC. However, the rate of release was abnormally slow and the total amount of fibrinopeptide A (FpA) released reached only approximately 50% of the theoretical maximum. The rate and quantity of FpA release was normal when Reptilase was used. Transmission Electron Microscopy (TEM) of Thrombin induced clots showed an altered clot structure characterized by a reduced mean fiber diameter. The mother has a polymerization defect similar to the propositus, her fibrinopeptide release is unaffected however. The father has a minor fibrinopeptide release defect suggesting the presence of two populations of fibrinogen. This study supports the idea that the fibrinogen isolated from the propositus has two defects inherited as separate genetic traits. This fibrinogen has been named Fibrinogen Guarenas I.

Sex differences in behaviour pattern and catecholamine and cortisol excretion in 3-6 year old day-care children.

Fifteen boys and eleven girls between three and six years of age were examined at a day-care center and at home as regards their catecholamine and cortisol excretion. Behaviour pattern was assessed by the MYTH-scale, which measures the competitiveness, impatience-anger and aggression components of the type A behaviour pattern in children (Matthews and Angulo, 1980). The boys obtained higher type A scores and excreted more adrenaline and noradrenaline than the girls, while cortisol excretion did not differ between the sexes. In view of previous findings, the results suggest that sex differences in catecholamine excretion in children are induced by sex related differences in behaviour. This relationship in childhood could be of relevance for sex differences in catecholamine responses observed in adulthood. In both sexes, adrenaline excretion was significantly elevated at the day-care center compared with the at-home levels, indicating that mental arousal was greater at the center. In a separate part of the study, eleven new children were tested while they were adjusting to the day-care situation; it was found that only noradrenaline levels during the first week at the center were significantly elevated.

Adrenal-medullary and adrenal-cortical responses to understimulation and overstimulation: comparison between type A and type B persons.

Male and female university students, classified as Type A and Type B persons with the aid of a questionnaire measuring coronary-prone behavior, were studied while doing mental work under laboratory conditions, characterized by understimulation and overstimulation, and while unoccupied (baseline session). The urinary excretion of adrenaline, noradrenaline and cortisol, heart rate (in the understimulation and baseline conditions) and self-reports were used as indices of arousal. The overall picture showed that both experimental conditions increased arousal, the rise being particularly pronounced for cortisol excretion during understimulation and for adrenaline excretion during overstimulation. Comparison between groups showed that cortisol excretion was higher in Type A than in Type B persons of both sexes during understimulation. Adrenaline excretion was about the same in both sexes (Type A and Type B) during understimulation, but was higher in males than in females during overstimulation. There were no group differences in performance.

Psychophysiological reactions to noise as modified by personal control over noise intensity.

Effects of personal control over noise intensity were studied in experimental situations where subjects performed mental arithmetic under noise exposure. Every other subject was offered a choice between noise intensities, and the next subject, serving as his yoked partner, had to submit to the same noise. Mean measures of catecholamine and cortisol excretion, and of heart rate and subjective effort and discomfort showed that subjects were more aroused in the yoked situation. There were, however, considerable interindividual differences, subjects classified as 'internals' or 'externals' on the basis of the Internal-External Locus of Control Scale responding to the two experimental conditions in congruence with their general beliefs and attitudes with regard to control.

Dissociation between sympathetic-adrenal and pituitary-adrenal responses to an achievement situation characterized by high controllability: comparison between type A and type B males and females.

Type A and Type B university students of both sexes performed a choice-reaction task that was designed to allow each subject at his/her preferred pace and to maintain control over the stimulus rate throughout a period of sustained work. Type As selected faster stimulus rates, and coped more effectively with their total work load than did Type Bs with their lesser load. The superior achievement of the Type A group was not reflected in self-reports of subjective arousal, not in physiological arousal indices, e.g. heart rate, and catecholamine and cortisol excretion. Comparison between sexes (Type A and Type B persons) showed that males reported exerting significantly more effort than females and, accordingly, secreted significantly more adrenaline. A common charcteristic of all groups was the dissociation between sympathetic-adrenal and pituitary-adrenal arousal, catecholamine output increasing and cortisol output decreasing in response to the achievement demands. The rise in catecholamine output was interpreted in terms of the mobilization of effort induced by the task, and the decrease of cortisol in terms of the high level of personal control in performing the task.

Note on Type A behavior and cardiovascular responses to challenge in 3-6-yr old children.

Using the MYTH-scale [1], the antecedents of Type A behavior were studied in fifteen healthy 3-6-yr old boys. Boys with scores above ('Type A') and below ('Type B') the median were compared in terms of systolic and diastolic blood pressure and heart rate during rest, and during 'emotional' and 'physical' challenge. It was found that 'Type A boys' responded to challenge with a greater increase in systolic blood pressure than 'Type B boys' (significant during physical challenge), while there were no differences during the rest condition. Diastolic blood pressure and heart rate did not differ markedly between the groups.