Tspan33 is Expressed in Transitional and Memory B Cells, but is not Responsible for High ADAM10 Expression.

Tetraspanins are a family of transmembrane proteins that form membrane microdomains. They play important roles in migration, adhesion and other cellular processes. TspanC8, a subfamily of tetraspanins, was found to associate and promote ADAM10 trafficking and cell surface localization. One of its members, Tspan33, is expressed in activated B cells. Using RT-PCR and flow cytometry, we analysed the pattern of expression of Tspan33 in B cells from healthy donors. We found Tspan33 expression in early and late stages of B cell development. However, Tspan33 expression did not correlate with ADAM10 surface expression. We also found expression of Tspan33 early in the activation process. Given its predominant expression in activated B cells and in several lymphomas, but not in naive B cells, we hypothesize that Tspan33 could be a potential target for therapeutic purposes.

[Evaluation of in vitro activity of ofloxacin against 73 strains of Chlamydia trachomatis isolated from gynecologic infections]. / Evaluation de l'activité in vitro de l'ofloxacine vis-à-vis de 73 souches de Chlamydia trachomatis isolées d'infections gynécologiques.

The in vitro activity of ofloxacin, levofloxacin, ciprofloxacin, doxycyclin, erythromycin and roxithromycin was determined against 73 recent clinical strains of Chlamydia trachomatis isolated from genital infections. The MICs 90% were: 0.4; 0.1; 1.6; 0.2; 1.6 and 0.1 mg/l respectively. 100% of strains were susceptible to ofloxacin, roxithromycin and doxycyclin. Erythromycin and ciprofloxacin had a lower in vitro activity against C. trachomatis.

[Tuberculosis en pediatric patients in the island of Gran Canaria]. / Tuberculosis en pacientes pediátricos en la isla de Gran Canaria.

OBJECTIVE: Tuberculosis is still an important sanitary problem in our country, which has a high rate of endemia. The pediatric population is highly susceptible to being exposed and has a high risk of developing the disease, as well as of suffering serious forms of it. The aim of this study was to analyze the clinical aspects and characteristics of pediatric patients who were diagnosed with tuberculosis in Gran Canaria during a nine years period. PATIENTS AND METHODS: The clinical charts of 49 children between 0 and 14 years of age who were diagnosed of having tuberculosis between 1986 and 1994 were revised. Tuberculosis was diagnosed when M. tuberculosis was isolated from at least one clinical sample. RESULTS: Of the children diagnosed, 69.4% were less than 5 years of age. Only one child had antibodies against HIV. Clinical forms were; pulmonary (33 cases), disseminated (13 cases), genitourinary (1 case), osteoarticular (1 case), and lymphatic (1 case). No association was found between any clinical form and any age group. The disseminated form presented more frequently with a negative Mantoux test (p < 0.01). In all cases lung disease was present and three patients also suffered meningitis. In most of the cases (77.8%), it was necessary to wait for the result of the culture to reach the diagnosis. Two children (4.1%) died and six (12.2%) had sequeale. CONCLUSIONS: Disseminated tuberculosis is relatively frequent (26.5%) in this population and is usually associated with a negative Mantoux test. Therefore, it appears that in this serious form of the disease, a high degree of clinical suspicion is needed and mycobacteria cultures must be done in clinical samples of different origins.

[Bacteria isolated in 1994-1995 in female upper genitalia infections and in male urethritis. Distribution and sensitivity to antibiotics]. / Bactéries isolées en 1994-1995 au cours des infections gynécologiques hautes et des uréthrites masculines. Distribution et sensibilité aux antibiotiques.

OBJECTIVE: Classify antibiotics according to their individual activity so as to identify those suitable for empiric therapy. METHODS: We studied bacterial strains isolated from patients with urethritis (n = 189) and upper genital tract infections (n = 163) between June 1994 and February 1995 in 3 hospital and 4 community laboratories. Upper genital tract infections were divided into two groups: proven infection on laparoscopy specimen (n = 79) and suspected infection with isolation of pathogen in cervical samples (n = 84). Pathogens isolated were: Chlamydia trachomatis in 36/12/15 cases respectively, Mycoplasma hominis in 12/20/13, Ureaplasma urealyticum in 55/30/15, Neisseria gonorrhoeae in 40/2/0, Haemophilus spp in 20/2/1, group B streptococci in 7/1/8, E. coli in 8/1/17 and miscellaneous in 11/8/15. The minimal inhibitory concentrations for all strains were determined in 4 laboratories for ofloxacin, erythromycin and doxycyclin against C. trachomatis, M. hominis and U. urealyticum, and for ofloxacin, erythromycin, doxycyclin, amoxicillin+clavulanate, cefotaxime and gentamicin against the other strains. The activity score (% susceptibility to each antibiotic weighted by the frequencies of each isolate in urethritis and upper genital tract infection based on recent French epidemiologic data) was calculated for each antibiotic. CONCLUSION: The antibiotics with the best empiric activity scores in urethritis were, in decreasing order: doxycyclin (90.4%), ofloxacin (88.1%), and erythromycin (50.2%). The most active combinations in upper genital tract infections were ofloxacin+amoxicillin (100%), doxycyclin+cefotaxime+metronidazole (95.9%) and doxycyclin+amoxicillin (95.3%).

[New vaccine against trachoma]. / Nouveau vaccin contre le trachome.

The new vaccine against trachoma is established upon a stimulation of MALT system by oral route. The amount of IgA produced using chlamydia trachomatis serovar C is sufficient to allow a protection against infections of the same species of chlamydiae and/or of others varieties. There is a cross immunity between different serovars and different species. The vaccine could bring a protection against trachoma, chlamydial infection of human and animals.

Genital human papillomavirus infections: correlation of cytological, colposcopic and histological features with viral types in women and their male partners.

To determine the incidence of anogenital papillomavirus infections and to assess the value of available diagnostic methods, we compared the cytological, colposcopic and histological features of anogenital papillomavirus-related lesions with their associated human papillomavirus types (HPV) in 300 women and in their male partners. HPV-type deoxyribonucleic acid was detected by blot hybridization in 398 out of 624 subclinical and clinically defined anogenital lesions. Whatever the site of the lesion, condylomas and low-grade intraepithelial neoplasia (IEN) were found in 84% of lesions associated with HPV 6-11, compared with 32% of lesions containing HPV 16-18 (P < 0.001). Among the HPV 16-18 associated lesions, high-grade cervical, vaginal, vulvar and anal intraepithelial neoplasias represented 45% (P < 0.001) of the lesions. In 65% of 23 cases of squamous anogenital cancer, HPV 16-18 and mixed types were present (P < 0.001). In 54% (161/300) of cases, the lesions were multicentric (161/300). On cytological examination, 27% of the samples gave false negative results. In cervical lesions, there was a good correlation between virological and colposcopic findings, but this was not true for extracervical mucous epithelia in the vagina or on the vulva. With peniscopy in the male partners 220 out of 410 had penile condylomatous lesions and more than half of the 350 male specimens examined by molecular hybridization contained HPV DNA. A correlation was found between the virus types in penile lesions or in cells of the distal urethra and in the cervical lesions of the sexual partner.(ABSTRACT TRUNCATED AT 250 WORDS)

Incidence of community-acquired pneumonia and Chlamydia pneumoniae infection: a prospective multicentre study.

This one year prospective multicentre study was designed to determine the incidence of community-acquired pneumonia in adults. It was carried out in primary health care centres and three reference hospitals, located in the 'Maresme' region (Barcelona, Spain) serving a population of 39,733 subjects over 13 years of age. Patients suspected of having contracted community-acquired pneumonia were visited by their family doctors and referred to the three reference hospitals for confirmation of the diagnosis. Patients attending the emergency services of these hospitals were also included. Urine and blood samples were obtained for culture, antigen detection, blood count, serological tests, blood gases and biochemical profile. The diagnosis of community-acquired pneumonia was made in 105 patients. Forty-six patients had an identifiable microbial etiology. Chlamydia pneumoniae was the most common pathogen (16 cases) followed by Streptococcus pneumoniae (13 cases) and Mycoplasma pneumoniae (8 cases). In conclusion; the annual incidence rate of community-acquired pneumonia in adults in this area was 2.6 cases per 1,000 inhabitants and Chlamydia pneumoniae was the most frequent causative pathogen.

[Epidemiology of Chlamydia trachomatis using analysis of gene encoding of the major outer membrane protein]. / Epidémiologie de Chlamydia trachomatis par l'analyse du gène de la protéine majeure de membrane externe.

One hundred and eight clinical strains and 24 reference strains of C. trachomatis were typed using differential restriction mapping of omp1, the gene which encodes the major outer membrane protein. The gene was obtained by polymerase chain reaction (PCR). This molecular typing method correlated well with serological typing. Eighty-four per cent of clinical strains were typed using the enzyme AluI alone. Heterogeneity was looked for among the most common serovars (E, F, and D; 62%, 17%, and 9%, respectively). Analysis of the PCR-amplified fourth variable domain of omp1 using denaturing gradient gel electrophoresis followed by direct sequencing of the variants disclosed substantial heterogeneity within the D serovar. Conversely, serovars E and F were homogeneous, with however a single variant strain of serovar E.

Human papillomavirus infection in esophageal squamous-cell carcinoma in western countries.

Recent studies have suggested that esophageal HPV infection could be a risk factor for esophageal squamous-cell carcinoma. The aims of our study were to assess the presence of HPV esophageal infection among French patients with esophageal squamous-cell carcinoma and to compare the prevalence of this infection among control patients exposed to similar known risk factors (alcohol and tobacco) and among non-exposed control patients. All patients had the following investigations: serum immunoglobulin level, T-lymphocyte subsets, cutaneous anergy test and endoscopy with biopsies from tumoral and normal areas. Three different methods were used for HPV-infection diagnosis: histological score, in situ hybridization intended for detection of HPV types 6, 11, 16, 18, 31 and 33, and dot blot intended for detection of HPV types 6/11 and 16/18. Five out of 12 patients with esophageal carcinoma had HPV esophageal infection. This infection did not result from impaired immune status. The most frequently observed types are HPV 16/18. None out of 17 exposed controls and only 1 out of 7 nonexposed controls had HPV esophageal infection (p less than 0.01). HPV infection may be implicated in the development of esophageal squamous-cell carcinoma in association with known risk factors.

Rapid genotyping of the Chlamydia trachomatis major outer membrane protein by the polymerase chain reaction.

Twenty one Chlamydia trachomatis reference strains and 40 clinical isolates belonging to the lymphogranuloma venerum (LGV) and trachoma biovars were genotyped by differential restriction mapping of the major-outer-membrane-protein gene (MOMP) obtained by the polymerase-chain reaction (PCR). AluI digestion of the PCR product distinguishes eight MOMP-genotypes corresponding to 8 serovars. Six additional enzymes (NlaIII, CfoI, EcoRI, HinfI, DdeI and FokI) further permit the discrimination of 10 MOMP-genotypes corresponding to the 10 remaining serovars of the species. AluI alone allows direct typing of 78% of the clinical isolates. AluI digestion patterns of mouse C. trachomatis biovar, a C. pneumoniae and two C. psittaci strains, studied for comparison, were clearly distinguishable from one another and from the C. trachomatis LGV and trachoma strains. These results indicate that MOMP genotyping by PCR is a valuable molecular tool for studying C. trachomatis epidemiology.

Estrogen and progesterone receptors in cervical human papillomavirus related lesions.

According to recent studies showing that human papillomavirus (HPV) infections can be influenced by sex steroid hormones, we performed estrogen (ER) and progesterone (PgR) receptor assays in fresh frozen biopsies of genital-HPV-related lesions. Seventy-three women with normal cervix, condyloma, low- and high-grade CIN and squamous carcinoma were evaluated in comparison with 15 persons with vulvar and 9 with penile papillomavirus-associated lesions. HPV genotypes were determined by dot-blot hybridization. Non-cervical lesions did not express HR. Condyloma on squamous metaplasia of the cervix and high-grade CIN expressed high levels of HR, particularly PgR (mean 4,086 and 4,518 fmoles/g tissue, respectively). Cervical squamous carcinoma expressed very low concentrations of PgR in a limited number of cases. High levels of PgR were correlated with high-grade CIN (p less than 0.05), HPV16-18-associated lesions (p less than 0.01) and ER were correlated to HPV6-11-related lesions (p less than 0.01). The levels were independent of age, cycle stage and oral contraception. Morphological localization of PgR, using an immunocytochemical method using a monoclonal antibody (MAb) (PR-ICA), showed intense homogeneous staining in the nuclei of the stromal fibroblasts underlying dysplastic epithelium and condyloma on squamous metaplasia. These results suggest that, under in vivo conditions, sex steroid hormones, particularly progesterone, may act indirectly on HPV-infected epithelial cells and be implicated as co-factors in HPV-related cervical neoplasia. They could explain the relative predisposition to malignant transformation of the cervix as compared with vulvar and penile mucosa.

A method for conforming the pH dependence of the michaelis parameters of nonallosteric enzymes to four kinetic schemes.

A nonlinear regression program for the analysis of the effect of pH on enzyme activity has been developed for the IBM micro range and compatible machines. The program conforms the V and Km pH profiles to one of four commonly occurring kinetic schemes. By using multiple linear regression the program computes initial estimates for the nonlinear search which are thus not required from the user. Two nonlinear optimization methods are included in order to effectively handle the nonnegativity constraints on the parameters. The program is user-friendly and provides plots of the fit and residual plots to help the user decide on the goodness of fit obtained. The program was coded in Turbo Pascal version 4.0 and runs on IBM micros or close compatibles with four types of graphics cards.

A program to estimate insulin sensitivity and pancreatic responsivity from an IVGTT using the minimal modeling technique.

A user-friendly program coded in PASCAL for the IBM PC has been developed to determine the etiology of impaired glucose tolerance using an intravenous glucose tolerance test (IVGTT). It makes use of the "minimal modeling technique," a method that has been shown to be adequate for the quantitative determination of insulin sensitivity and insulin resistance. Two models are used, the minimal model of glucose disappearance and the minimal model of insulin kinetics. The first model is described by two nonlinear ordinary differential equations (ODEs) which are solved numerically, and which yield the insulin sensitivity index SI. The second model is described by an ODE for which an explicit solution was obtained, and which yields the pancreatic responsivity parameters phi 1 and phi 2. The product SI.phi 2 can be used to segregate subjects into "good" and "low" tolerance types. The program provides best-fit plots along with numerical values of the parameters and their uncertainties, and requires little intervention from the user. The fact that it requires a noninvasive IVGTT as input and that it has been written for the ubiquitous IBM PC are added advantages.

[Statement: The evaluation of the Serodia-HIV and Serodia-ATLA FUJIREBIO-MILES kits]. / Communiqué: étude des trousses Sérodia-HIV et Sérodia-ATLA FUJIREBIO-MILES.

A new test, based on agglutination of gelatin particles (PA), sensitized with viral antigens of HIV, was applied on detection of the human immunodeficiency virus (HIV 1) antibody. Sensitivity compared to the ELISA tests (Organon, Dupond de Nemours and/or Elavia 2) was the same during the screening test (97.7 p. cent). Specificity was also acceptable when compared to the same tests (94 p. cent). This specificity remains acceptable with African sera (96 p. cent). During screening, 11.8 p. cent of tested sera were declared falsely positive by the Elisa classical techniques, against only 1.96 p. cent with the PA assay. The six sera remained positive with ELISA (false positives), whereas this positivity was not confirmed with the PA assay. Moreover, all the positive sera were confirmed with the Western blot HIV 1 assay (55.84 p. cent), HIV 1 + 2 (31.17 p. cent) or HIV 2 (13.0 p. cent). Amongst the five false positives pointed out in the european sera, all of them have shown in the Western blot the presence of one or two bands of the GAG protein. In this case, the assay whose easy use is attractive, can be adopted in screening serology and could be useful in African regions, as no further equipment is needed.

Phase II study of D.651, an oral vaccine designed to prevent recurrences of vulvovaginal candidiasis.

A vaccine has been prepared with ribosomes of Candida albicans serotypes a and b plus, as adjuvant, membrane proteoglycan from a nonencapsulated Klebsiella pneumoniae. A preliminary phase II trial without placebo control was conducted in 22 women with a history of frequent recurrences of vulvovaginal candidiasis (VVC). Initially, all the patients were treated locally with an antimycotic cream. Beginning at the same time, vaccine was taken orally in capsules containing 0.55 microgram active components. It was administered intermittently over six months, groups of women taking doses of two, three, six or nine capsules. Tolerance was excellent except for mild nausea, probably due to the excipients, in two patients taking nine capsules. Twenty patients completed the study. Only seven of them had a documented recurrence of VVC during the 6 months on vaccine. No recurrence occurred in the eight women taking six or nine capsules per day. Before the study, these 20 patients had had an average of 3.59 attacks of VVC per 6 months. On vaccine, the average rate of recurrence was only 0.55 attacks per 6 months. A multicentre placebo-controlled study of the efficacy of this vaccine is in progress to validate these encouraging preliminary results.

Temperature-jump studies on hemoglobin. Kinetic evidence for a non-quaternary isomerization process in deoxy- and carbonmonoxyhemoglobin.

Temperature jumps on mixtures of hemoglobin and pH indicators give rise to relaxation signals in the microsecond range. The pH and concentration dependences of the reciprocal relaxation time, 1/tau, may be rationalized on the basis of a reaction scheme in which a slow isomerization process in the protein moiety is coupled to a rapid co-operative ionization of two protons. At 11 degrees C the rate constants of the isomerization are kr = 4.2(+/- 1.8) x 10(4) s-1 and kf = 1.3(+/- 0.1) x 10(4) s-1 in deoxyhemoglobin; in carbonmonoxyhemoglobin they are kr = 3.9(+/- 1.3) x 10(4) s-1 and kf = 5.3(+/- 1.8) x 10(3) s-1. The pKa values of the coupled ionizing groups are 5.3 in deoxy- and 6.0 in carbonmonoxyhemoglobin. Modification of the CysF9(93) beta sulfhydryl group with iodoacetamide abolishes the pH dependence of 1/tau, suggesting that this sulfhydryl is involved in the isomerization process. Consideration of the X-ray structure of oxyhemoglobin allows a structural interpretation of the results. It is concluded that the isomerization may be important for the physiological function of hemoglobin, but that it is not identical with the quaternary structure transition.

[Results of detection and early treatment of Chlamydia trachomatis infections in pregnancy]. / Résultats du dépistage et traitement précoce des infections à Chlamydia trachomatis au cours de la grossesse.

The authors report and comment on the results of a prospective study of the usefulness of early treatment of Chlamydia trachomatis infections in pregnant women. The diagnosis is based on a serological examination carried out in the first trimester of pregnancy. 60 women were treated by antibiotics and they were compared with 51 women who were not treated. The two groups were statistically comparable. There was a significant drop in perinatal complications due to Chlamydia trachomatis. The difference was particularly significant as far as low birth weight and the incidence of conjunctivitis in the newborn was concerned. These results show how useful it is to carry out serological screening and early treatment of Chlamydia trachomatis infections in pregnancy.