[Environmental factors for asthma severity and allergy: results from the EGEA study]. / Facteurs environnementaux de l'asthme sévère et de l'allergie: résultats de l'étude EGEA.

INTRODUCTION: EGEA (Epidemiological study on the genetics and environment of asthma, bronchial hyperresponsiveness and atopy), a case control and family study including 2048 individuals, was initiated to look for environmental and genetic risk factors for asthma. A synthesis of the results obtained since 2002 on phenotypic and environmental aspects of asthma severity and allergy are presented in this article. METHODS AND RESULTS: The results support a role for hormonal factors in asthma severity and in various allergic markers of asthma. A greater body mass index was related to a more severe asthma in women with early menarche. Associations between markers of allergy (eosinophils, IgE and atopy) and hormonal dependent events in women (premenstrual asthma, menopause and oral contraceptive use) have been found. In asthmatics, exposure to agents known to be associated with occupational asthma, active and passive smoking were associated with an increased clinical asthma severity score. The study underlines the protective role of country living and exposure to pets in early life on allergy markers in adulthood, supporting the hygiene hypothesis. CONCLUSIONS: New hypothesis will be tested in the near future from the second stage of this survey.

The protective role of country living on skin prick tests, immunoglobulin E and asthma in adults from the Epidemiological study on the Genetics and Environment of Asthma, bronchial hyper-responsiveness and atopy.

BACKGROUND: Farming environment and traditional lifestyle seem to protect from childhood allergy. OBJECTIVE: The aim is to analyse the relationships of living in the country to asthma, positive skin prick tests and IgE among adults considering various windows of exposure over the life-span. METHODS: The study concerns 805 adults drawn from the Epidemiological study on the Genetics and Environment of Asthma, bronchial hyper-responsiveness and atopy (EGEA) (asthmatic cases, non-asthmatic controls, and parents of cases with and without asthma). Ever living in the country concerned 55% of the subjects. Early (beginning < 1 years), childhood (beginning < or = 16 years), prolonged (duration > or = 10 years) and current life in the country were studied. RESULTS: The results based on the case control and family components of the study show that IgE levels were significantly lower in those who ever lived in the country and in particular in those who lived for > or = 10 years. Positive skin prick tests (SPT) were significantly less prevalent in those who ever lived in the country and in particular in those with childhood (< or = 16 years) exposure. These associations remained independent of age, sex, smoking or asthma with IgE levels of 64 vs. 88 IU/mL; P = 0.004 for those ever living in the country vs. others and odds ratio for SPT positivity of 0.72 (95% CI [0.53-0.98]). In the more specific group with traditional mode of heating in childhood (use of wood) associations were stronger. The association with asthma, studied in parents of asthmatic probands showed that fathers, but not mothers, of asthmatics were significantly less often asthmatic themselves in relation to country living. CONCLUSION: Country life protects from asthma and adulthood allergy. The protective effect is not restricted to exposure in early childhood.

Degenerated pIX-IVa2 adenoviral vector sequences lowers reacquisition of the E1 genes during virus amplification in 293 cells.

A critical issue for E1-deleted adenoviral vectors manufactured from 293 cells is the emergence of replication-competent adenovirus (RCA). These contaminants arise through homologous recombination between identical sequences framing the E1 locus displayed by 293 cells, and the vector backbones. Modified recombinogenic sequences (syngen) were thus introduced within the vector backbone, and virus viability and RCA emergence were assessed. Syngen#1 is a synthetic sequence displaying silent point mutations in the pIX and IVa2 coding regions. A side by side comparison of Ad5CMV/p53 (E1-deleted adenovirus expressing the p53 tumor suppressor gene) and AVdeltaE1#1CMV/p53 (with syngen#1 in place of wild-type sequences) demonstrated a normal productivity for the modified construct. The altered sequences did not impair p53-mediated apoptosis in a model tumor cell line. Most importantly, a statistically significant decrease in terms of RCA occurrence could also be demonstrated. Degenerescence of the recombinogenic sequences could be further accentuated by modifying noncoding pIX region (syngen #2), with no effect on virus productivity and stability. We concluded that these vector modifications constitute a feasible strategy to reduce RCA emergence during amplification in 293 cells. This approach could also be applied to decrease reincorporation of the E1 genes during amplification of deltaE1deltaE4 vectors in 293/E4-trans-complementing cells.

Relationships of active and passive smoking to total IgE in adults of the Epidemiological Study of the Genetics and Environment of Asthma, Bronchial Hyperresponsiveness, and Atopy (EGEA).

The increase of total IgE in relation to active smoking has been shown in the general population, but little is known about subjects with a personal or family history of asthma. The objective of this report is to analyze the relationships of active and passive smoking to total IgE in the Epidemiological Study of the Genetics and Environment of Asthma, Bronchial Hyperresponsiveness, and Atopy (EGEA). The sample studied includes 122 asthmatic probands, 430 first-degree relatives, and 190 control subjects, age 25 to 54 yr. As expected, first-degree relatives had total IgE intermediate between cases and control subjects and men had higher values than women. Current smokers had significantly higher IgE than never smokers. The relationship was statistically significant restricting the analysis in asthmatic probands. In a model taking into account gender, personal and familial history of asthma, socio- occupational class, and the nonindependence of subjects from the same family, IgE were in current smokers, ex-smokers, and never smokers 128, 61, and 76 IU/ml and 77, 41, and 55 IU/ml in men (p = 0.01) and women (p = 0. 05), respectively. The relation was independent of skin test response. Some increase in IgE was observed in both men and women first-degree relatives in relation to passive smoking. That relation was statistically significant in women only (adjusted for asthma values: 103 IU/ml versus 48 IU/ ml, p = 0.02). Results show that an increase in total IgE in relation to active smoking may be evidenced even in asthmatics despite the healthy smoker effect. Susceptible subjects, such as women who are first-degree relatives of asthmatics, may increase total IgE in relation to passive smoking.

Segregation analysis of IgE levels in 335 French families (EGEA) using different strategies to correct for the ascertainment through a correlated trait (asthma).

The main objective of this study was to search for a major gene controlling total serum immunoglobulin E (IgE) levels, an intermediate phenotype for asthma and allergy. We studied 335 French nuclear families of the EGEA study (Epidemiological study of the Genetics and Environment of Asthma), ascertained through asthmatic probands (123 are parents in the family, 212 children). Segregation analyses were performed by regressive models, which can take into account a major gene effect, various sources of familial covariation (genetic and/or environmental) as well as measured risk factors (i.e. , age, sex, smoking habits). Different strategies were considered to account for the mode of ascertainment of the families through a correlated trait (asthma): the ascertainment mode was either ignored (strategy A) or taken into account by adjusting IgE levels for the position in the family, i.e., probands, blood relatives, spouses (strategy B) or excluding the asthmatic children-probands and computing the likelihood of each family conditionally on parents' IgE levels (strategy C). Whereas a major gene effect could not be detected with strategy A, strategies B and C showed evidence for the transmission of a dominant major gene for high IgE levels, which was more significant with strategy B. This gene does not interact with any of the covariates and is responsible for approximately 15% of IgE variation (the allele frequency is 0.65).

A regression survival model for testing the proportional hazards hypothesis.

A semi-parametric generalization of the proportional hazards regression model is defined, whereby the hazard functions can cross for different values of the covariates. In the two-sample comparison, it includes in particular the case of two Weibull distributions differing in scale and shape parameters. A global test of the proportional hazards assumption is proposed against such defined alternatives. Its power in the two-sample case is compared to that of previously described tests by using simulation experiments. Survival data of patients with breast carcinoma, including several prognostic factors, are presented as an illustration.

Bronchial hyperresponsiveness as a predictor of wheezing in a follow-up study of healthy men.

We assessed the relationship between bronchial hyperresponsiveness (BHR) and the onset of wheezing 5 years later, by epidemiological analysis of 194 working men without asthma or wheezing at the first examination. In 1985/ 1986 and 1990/1991, subjects answered a British Medical Research Council questionnaire and performed lung function measurements and methacholine challenge tests (total dose 6 mg). BHR was measured in three ways: (1) FEV1 fall > or = 20% (PD20+); (2) the two-point response slope expressed as percentage decline of FEV1/dose, and (3) a four-parameter model: FEV1 at dose (d)/ prechallenge FEV1 = ONE-k(d-delta)+a, where 'k' is the slope of the relative variation of FEV1 with the dose, 'delta' the threshold dose, and 'alpha' a shape factor. In the 13 new wheezers, the mean values of the two-point slope and of k were significantly increased, and the proportion of reactors was almost threefold (the latter was not statistically significant). Among nonsmokers, delta was significantly lower in new wheezers than in the others, whereas the slope and k had similar mean values. Among smokers, new wheezers had increased mean values for the slope and k, and an increased proportion of reactors, whereas delta was not decreased. Thus, BHR was a significant predictor of wheezing, independent of the method of analysis. Moreover, the model distinguished between two components of bronchial response: wheezing was predicted by sensitivity (delta) in nonsmokers, and by reactivity (k) in smokers.

Statistical evidence for two major proteins in freeze-fractured gastric parietal cell tubulovesicles and canaliculus.

In a previous work, resting and acid-secreting rabbit gastric mucosa were freeze-fractured and shadowed at 45 degrees with Pt-C. The shadow widths of proteic particles of tubulovesicle and canaliculus membranes were measured and compared. It was concluded that the frequency distributions of widths are significantly different in resting and secreting membranes and that each distribution accounts for several subpopulations of homogeneous particles. In the present study, an attempt is made to describe the experimental distributions as a mixture of those of two major proteins, say A and B and their aggregates (AA, AB and BB). The modelling, although simple, gave a very satisfactory statistical fit between observed and computed distributions. The comparison of parameters calculated from histamine and ranitidine experimental data further improves the fits and finally, component A accounts for 69% of the particles. Most replica of A particles are heart-shaped and the median shadow widths are 6.1 and 6.8 nm in canaliculus and tubulovesicles respectively. The component B accounts for 31% of the particles. They mainly appear as small barrels and the median shadow widths are 8.8 and 10.3 nm in canaliculus and tubulovesicles respectively. According to calculated parameters and observed particle replica, the onset of secretion does not change the relative ratio of proteins but changes their shapes. Component A should be the (H+,K+)-ATPase whereas debate on the identity of B is wide open.

[A new model for the study of filarial physiopathology: Molinema dessetae in its definitive natural host, Proechimys oris. 1]. / Un nouveau modèle d'étude de la physiopathologie filarienne: Molinema dessetae chez son hôte définitif naturel Proechimys oris. Première partie.

The filarial rodent model Molinema dessetae/Proechimys oris, recently adapted in the laboratory allows studies on the host/parasite relationships. Development of filarial worm was described elsewhere, and the model standardized. Biological, parasitological data and antifilarial drug response were determined. Natural models can seldom be used in laboratory; in non natural models many informations on the host-parasite relationships are lost and pathological studies can only be fragmentary and punctual. With inbred Proechimys oris we have the possibility to investigate with a natural filarial model. Three rodent groups have been studied for over a period of one year. The first group comprises eleven parasite free animals; the second one, eight rodents infected once by L3, and the last with eight rodents regularly re-infected during the period of experiment. Several biological parameters were considered: weight, total red and white blood cell counts, haemoglobinemia, mean cell volume, differential leucocyte count, blood urea nitrogen, total serum proteins and electrophoretic pattern, endogenous creatinine clearance, circulating immune complexes. On the whole, more than 19,700 biological data were obtained and several multivariate statistical analysis were necessary for a valid interpretation. The Principal Component Analysis (ACP) shows in each infected rodent distinct trajectories related either with the age of the host or with the filarial infection itself. Three graphic methods were used: individual graphics of each rodent, graphics of each rodent group and, individual and group statistical center of gravity. At the end of the experiments, the rodents were autopsied and a detailed histological study of the whole body was made. Different lesions and stages of inflammatory processes found were analysed and classified as follows: granulomatous nodule with eosinophilic and neutrophilic polymorphs; foreign-body granulomatous nodule; sclerocicatricial nodule; mononucleate cell infiltrate. Secondary pathological alterations, such as irritative lesions and blocage of lymph and blood vessels, were found. A comprehensive study of all our qualitative histological results was performed by a second multivariate analysis: the Correspondence Analysis (AFC). This analysis confirmed the heterogeneity of each Proechimys infected and multi-infected groups. The cross checking between the ACP and AFC allows the separation of two physiopathological filarial groups; the first group (A) is characterized by moderate biological variations and the absence of hepatosplenic lesions.(ABSTRACT TRUNCATED AT 400 WORDS)

Evidence that cycloleucine affects the high-affinity systems of amino acid uptake in cultured human fibroblasts.

The influence of cycloleucine on kinetic parameters of uptake of L-alanine, L-proline and L-leucine into cultured human fibroblasts was examined under initial-rate conditions with substrate concentrations of 0.05-10 mM and 5 mM-cycloleucine. Kinetic data obtained by computer analysis showed that, in the absence of cycloleucine, cell uptake was heterogeneous for each amino acid. L-Alanine and L-leucine entered by two transport systems with different affinities; L-proline was taken up by one saturable transport system plus a diffusion-like process. This heterogeneity disappeared in the presence of cycloleucine, since the high-affinity systems were no longer detectable. The remaining process had the same kinetic constants as the low-affinity system for alanine and leucine and a KD similar to the diffusion constant for proline. The influence of cycloleucine on the amino acid uptake was not specific either to the amino acid concerned or to a particular transport system, since the three neutral amino acid-transport systems, A, ASC and L, were involved in these experiments. This influence was shown to be unaffected by the absence of Na+ (for leucine uptake). ATP content of the cells was identical in the presence or in the absence of cycloleucine.