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Humic Substances (HSs) of the Chubut River (Patagonia‐Argentina) have been investigated, during two consecutive years and different climatic seasons, in order to obtain a first approach at their spatial and temporal distribution, as well as their origin and relation with environmental characteristics. Absorbance at 250 nm and limnological variables were measured and the data processed by different statistical tools. We found that the processes developed in the lower sections of Chubut River are dominated by those produced in the Florentino Ameghino Reservoir. The riverine HSs are present in very low concentrations, have mainly autochthonous origin, exhibit spatial homogeneity and temporal variability similar to nutrients (nitrates and soluble reactive phosphorus). Copyright © 2010 John Wiley & Sons, Ltd.

Objective Chlamydia trachomatis is a common sexually transmitted micro-organism. The impact of chlamydial infection on semen parameters and male fertility is controversial. Our purpose was to determine the prevalence of C. trachomatis in the male partners of infertile couples in Tunisia and to assess the relationship between chlamydial infection markers and male infertility. Methods Chlamydial DNA in urethral and in semen specimens was determined by using the Cobas Amplicor polymerase chain reaction (PCR) assay and chlamydial immunoglobulin G (IgG) antibodies were measured by micro-immunofluorescence in serum samples in 92 male partners, with or without pathological standard semen parameters, according to the guidelines of the World Health Organization (sperm count, progressive sperm motility, sperm morphology and sperm viability). In parallel, chlamydial infection markers in endocervical material were determined by PCR and chlamydial IgG antibodies were measured by micro-immunofluorescence in serum samples from the female partners of the patients. Results C. trachomatis was found in 35.9% (33/92) of the male partners of the infertile couples and in 38% (35/92) of their female partners. There was a significant correlation between the detection of C. trachomatis in both partners (p = 0.004). C. trachomatis DNA was detected in 18.5% (17/92) of urethral specimens and in 16.3% (15/92) of semen specimens. Chlamydial IgG antibodies were present in 9.8% (9/92) of the serum samples. A standard semen analysis showed that 88% (81/92) were pathological. Sperm viability, progressive sperm motility, morphology and sperm concentration were abnormal in 73.8%, 70.2%, 34.5% and 13%, respectively, of the 92 evaluated semen specimens. Comparison of the parameters of the standard semen analysis between the male partners of the infertile couples with or without chlamydial infection markers showed that only the presence of C. trachomatis DNA in semen samples can affect sperm motility. Parameters of the standard semen analysis were not significantly related either to the detection of chlamydial DNA in urethral samples or to the presence of serum chlamydial antibodies. Conclusion Our results show that C. trachomatis seems to be widespread among the male partners of infertile couples in Tunisia and show that this organism can affect sperm motility and, thus, can play an important role in the etiology of male infertility.

The sequelae to infection with Chlamydia trachomatis in women are an established cause of tubal infertility. However, little is known about chlamydial infection and male infertility. The main objective of this study was to evaluate the presence of asymptomatic C. trachomatis infections in urethral and semen specimens from the male members of infertile couples by means of four different methods: the direct fluorescence antibodies assay, cell culture, the Roche Cobas Amplicor polymerase chain reaction, and the presence of chlamydial local IgA antibodies by the recombinant antibody-enzyme-linked immunosorbent assay. One or more chlamydial infection markers were detected in 42 (45.7%) of the 92 examined urethral and semen specimens from the male partners of infertile couples. C. trachomatis was detected in 23.9% (22/92) of urethral specimens and in 35.9% (33/92) of semen specimens. Although there was a significant correlation between the detection of one or more chlamydial infection markers in urethral and semen specimens (p = 0.01), no significant correlation was found between the detection of C. trachomatis DNA in these samples. Furthermore, no significant association was found between the presence of chlamydial local IgA antibodies and the detection of C. trachomatis. The discrepancies in positive results found between some techniques for the detection of C. trachomatis in urethral and semen specimens might be explained by variations in the sensitivities and specificities of the tests carried out and the use of specimens from different anatomical locations. Our findings suggest that C. trachomatis seems to be widespread among the male partners of infertile couples in Tunisia. The detection of C. trachomatis in urethral or semen specimens can serve as a marker for the presence of this organism in the genital tract, which is not necessarily the cause of male infertility. The study of the correlation between the detection of chlamydial infection markers and the parameters of male fertility seems to be necessary in order to determine the direct link between chlamydial infection and male infertility and to choose the most efficient technique and most suitable specimen with which to diagnose C. trachomatis-associated male infertility.

The humoral immune response to Chlamydia trachomatis 10-kDa heat shock protein (Chsp10) in populations of Russian and French origin was studied by using a recombinant Chsp10 enzyme-linked immunosorbent assay. A physiological but not a serological correlation of Chsp10 exposure with Chsp60 exposure was observed in the Russian population. In the French population studied, there was a significant association between detection of anti-r-Chsp10 immunoglobulin G (IgG) antibodies and chronic genital tract infections. Chsp10 residues 50 to 67 were found to contain an immunodominant although not universal B epitope. Cross-reactions with Chlamydia pneumoniae or Escherichia coli GroES protein are limited but may occur. Our study suggests that detection of anti-Chsp10 IgG antibodies is associated with chronicity of C. trachomatis genital tract infection and does not parallel that of anti-Chsp60 IgG antibodies.

Chlamydia trachomatis(CT) est le microorganisme sexuellement transmissible le plus fréquemment responsable d’infection génitale. Si la fréquence de l’infection à CT est connue chez la femme enceinte et chez la femme à risque de maladies sexuellement transmissibles, peu de données existent chez l’adolescente non exposée en milieu africain. Nous avons étudié la prévalence des anticorps anti-CT dans une population de 295 adolescents (154 filles, 141 garçons) scolarisés âgés de 13 à 19 ans, parallèlement à celle d’un groupe témoin constitué de 190 enfants âgés de 6 à 12 ans. Les anticorps anti-CT apparaissent vers l’âge de 15 ans. La prévalence est de 6,1% dans cette population et elle prédomine davantage chez la fille (10,4%) que chez le garçon (1,4%). Les données suggèrent que l’adolescent apparemment asymptomatique est concerné par l’infection à CT et que les sujets doivent être dépistés et traités. La fréquence chez l’adolescente mérite toute l’attention car le jeune organisme est davantage prédisposé à des infections récidivantes et à leurs complications.

AimsTo examine the species specificity of the microimmunofluorescence test (MIF) and assess a time resolved fluoroscopic immunoassay (TRIA) for measuring IgG antibodies to C pneumoniae.MethodsSera from 1020 subjects were tested by MIF for IgG, IgM, and IgA antibodies to C pneumoniae, C trachomatis, and C psittaci; 501 serum samples were also tested by TRIA for IgG antibodies to C pneumoniae.ResultsC pneumoniae antibody titres as measured by MIF were correlated with those for C psittaci and C trachomatis. It was estimated that on average, one third of the twofold dilution steps that make up the final C pneumoniae antibody titre may be due to cross reacting genus specific antibody. The results of TRIA correlated well with those of MIF. In 75% of cases, the TRIA result predicted a three titre range within which the actual MIF result would fall.ConclusionsMIF does not appear to be as species specific as claimed. TRIA is unlikely to be as specific but as it is completely objective, easier to perform, amenable to automation, and gives reproducible results, it is a rapid and useful method for comparing populations.

C. TRACHOMATIS(CT) infections of the upper genital tract in women are either acute, sub acute or chronic. CT infection has a tendency to be chronic, latent and persistent as a consequence of the host immune reaction to CT major outer membrane protein, 57 Kd heat shock protein and lipopolysaccharide. Chlamydial persistence can be induced as a result of inflammatory and/or immune regulated cytokines, Interferon γ depletion of tryptophan causes a stress response involving development of abnormal forms with increased levels of stress response proteins which maintain host immune responses with continuous fibrin exudate. The main clinical consequences are acute and chronic pelvic inflammatory disease, with infertility, ectopic pregnancy and, less frequently, chronic pelvic pain as late sequelae. PID, when acute, is marked by bilateral pelvic pain, plus other infectious signs in typical cases: fever, leucorrhea, red and purulent cervix. In 50% cases, infectious signs are slight or absent or there is an atypical clinical situation. Laparoscopy is the key for diagnosis. It allows the surgeon to have a direct look at the pelvic organs and perform microbiologic and histologic sampling. In severe cases, laparoscopy allows the surgeon to aspirate the purulent discharge and successfully treat pelvic abscesses. Chronic PID usually is clinically silent. It is in most cases discovered some years after the onset of CT infection, in women operated on for tubal infertility or ectopic pregnancy. Further studies, to evaluate treatments efficiency in chronic cases and factors leading to ectopic pregnancy or to recurrence, are indicated. © 1996 Wiley‐Liss, Inc.

We have compared in two separate studies the kinetics of ceftriaxone and cefotaxime in 8 cirrhotic patients with ascites and 8 control subjects after a single 20 min intravenous infusion of 1 g of each drug.

Abstract: An enzyme-linked immunosorbent assay (ELISA) using a soluble antigen prepared from the D strain ofChlamydia trachomatis was used for titration of IgG antibodies toChlamydia trachomatis in 153 sera from 126 patients with non-specific genital infection and from 27 healthy subjects. The results were compared to those obtained with the micro-immunofluorescence method and were in complete agreement in 143 of the 153 sera. The ELISA proved to be reproducible and as sensitive as the micro-immunofluorescence method.

The ability of an enzyme-linked immunosorbent assay (ELISA) to detect antibodies toChlamydia trachomatis was evaluated in 100 sera using three different antigen preparations as substrates (sonicated organisms, Triton X solubilized antigen and SDS solubilized antigen). The results were compared to those obtained by a standard microimmunofluorescence assay. The results obtained by the three ELISA techniques and the microimmunofluorescence method were in relatively good agreement (76%); some discrepant results were observed in sera with a low antibody titer. There was good agreement of results obtained by the three ELISA techniques (84%). The microimmunofluorescence method showed the greatest sensitivity. Assuming the microimmunofluorescence method accurately demonstrates antibodies, the ELISA using Triton X solubilized antigen showed the highest degree of specificity (97%), and the ELISA with sonicated organisms the greatest sensitivity (82%) and accuracy (86%).

We used a mouse model of acute chlamydial salpingitis to evaluate the efficacy of roxithromycin in preventing irreversible inflammatory damage leading to tubal infertility. Female C3H/He mice were genitally inoculated with a human strain of Chlamydia trachomatis and then treated with roxithromycin glutamate subcutaneously. Treatment was initiated either 7 or 10 days postinfection (p.i.) and continued for 7 days at a dosage of 50 or 100 mg/kg of body weight per 24 h. The course of the disease was monitored serologically, bacteriologically, and histologically. At the end of the treatment, the mice were encaged with males and their reproductive capacity was recorded over a 19-week period. The protective effect of roxithromycin was assessed in terms of fertility parameters in comparison with values for noninfected control mice. When treatment was initiated on day 7 p.i. and given in twice-daily 25-mg/kg doses, all the mice remained fertile and the total number of offspring was similar to that of sham-infected mice (17.3 +/- 3.3 versus 17.2 +/- 2.3). When treatment was initiated on day 10 p.i. and given in a single daily dose of 50 or 100 mg/kg, 90 and 70% of the mice, respectively, remained fertile; however, in terms of total offspring, fertility was lower in the group treated with the lower dose (5.6 +/- 1.4 versus 13.0 +/- 3.8). Roxithromycin was found to be effective against C. trachomatis in the mouse genital tract, but fertility was only partially preserved when the time between infection and treatment was prolonged.

Commensal de l'oropharynx de l'homme, Branhamella catarrhalisa longtemps été considérée comme non pathogène. Ces dernières années ce germe a été reconnu comme un agent opportuniste responsable de surinfections bronchiques. Les cas de pneumonies alvéolaires sont eux très rarement rapportés. De novembre 1985 à février 1987 nous avons isolé 89 souches de Branhamella catarrhalisdont 64 à partir de sécrétions bronchiques de l'adulte. Dans 8 cas les malades présentaient une pneumonie alvéolaire. Les tableaux cliniques étaient graves et la ventilation assistée a été nécessaire quatre fois. Les 8 patients ont guéri. Des circonstances favorisantes de l'infection ont été retrouvées dans tous les cas. Nous avons été frappés par la rapidité d'évolution de ces pneumonies alvéolaires. Six des huit souches de Branhamella catarrhalissécrétaient une β-lactamase et étaient résistantes à la Pénicilline.

The Western blot (immunoblot) patterns of 56 serum specimens, all examined previously by the microimmunofluorescence (MIF) test for species-specific Chlamydia antibodies, were analyzed. Predominant specific-antibody activity was directed to the 170-, 155-, 145-, 120-, 115-, 100-, 57-, and 38-kDa proteins of Chlamydia trachomatis and to the 175-, 130-, 110-, 98-, and 30-kDa proteins of Chlamydia pneumoniae. All of these antigens appeared to be species specific. The reactivity with 90-, 80-, 75-, 62- or 60-, and 55-kDa proteins and the major outer membrane protein appeared to be genus specific. Fourteen serum samples which had identical titers of immunoglobulin G as determined by the MIF test were investigated by Western blotting. We found that nine serum samples had anti-C. trachomatis protein profiles and two had anti-C. pneumoniae protein profiles. The double seropositivity observed by MIF corresponded with cross-reactivity to genus-specific antibodies. As for the three remaining serum specimens, we observed identical protein profiles for C. trachomatis and C. pneumoniae, confirming the double seropositivity experienced with the MIF test. Western blotting can differentiate between specific reactions and interfering noise from other, partly cross-reacting chlamydial species.

A mouse model of salpingitis and subsequent tubal infertility induced by a human strain of Chlamydia trachomatis has been studied. C3H/He female mice were inoculated into the ovarian bursa. Some of the mice (six infected, five controls) were killed on days 15 and 23 and the remaining animals (10 infected, 10 controls) were mated on day 15. On day 15, the infection was maximal with intratubal inflammation, elevated antichlamydial antibody titre and positive cultures in 12 cases out of 16. After 19 weeks of housing with the male, the proportion of fertilized females was significantly lower in the infected group (20% versus 100% in the control group P less than 0.01). In the killed mice, hydrosalpinx and or tubal occlusion were noted at this time in nine cases out of 10, despite an apparent bacteriological healing.

We identified, by two-dimensional electrophoretic analysis and microsequencing, a protein of Chlamydia trachomatis elementary bodies which corresponds to the polypeptide (pgp3) encoded by open reading frame 3 (ORF3). Amino acid analysis showed that the first residue (Gly) found in the native protein is the one encoded by the second ORF3 codon, implying a typical bacterial removal of the first Met residue. Relatively large amounts of recombinant pgp3 (r-pgp3) in a stable, water-soluble form were obtained by overexpressing ORF3 in Escherichia coli and purifying the product from periplasmic extracts under nondenaturing conditions. These r-pgp3 preparations allowed specific detection of anti-pgp3 antibodies by enzyme-linked immunosorbent assay. Analysis of a group of 170 sera from healthy blood donors and from patients who were seropositive or -negative for C. trachomatis and Chlamydia pneumoniae showed that an immune response to pgp3 occurs in the majority (ca. 81%) of patients with sexually transmitted diseases who are seropositive for C. trachomatis and generally correlates with the response to cell surface antigens. No reaction between r-pgp3 and C. pneumoniae-positive sera was detected.

Examination of complementary replicas obtained by freeze-fracture of Chlamydia psittaci revealed, at the level of the plasma membrane, a progressive differentiation of "crate-like formations," which likely correspond to transmembranal pores. Recognition of "early" and "late" stages observed in the intermediate bodies permitted detailed study of the developmental cycle of this chlamydia.

Résumé L'étude dynamique intracellulaire du cycle de Chlamydia psittaci a été réalisée en microcinématographie à contraste de phase.

Résumé L'étude ultrastructurale des membranes de Chlamydia psittaci au cours de son développement intracellulaire dépend des fixateurs utilisés.

Summary.: Stress fracture of the clavicle is extremely rare, even in sports medicine. The authors have treated a young man who, for several years practised a traditional physical activity in which groups of people make human towers by standing on each others shoulders. For the last three years the patient experienced pain in the medial aspect of the right clavicle when he increased the intensity of his training. Bone scintigraphy revealed endosteal thickening which was indicative of a stress reaction. The mechanism of injury, the diagnosis of stress fracture, and a review of the literature are presented.

A microparticle enzyme immunoassay (LEIA) that uses passive latex agglutination was used to detect cytomegalovirus immunoglobulin G antibodies in 495 serum samples. LEIA was in excellent concordance with latex agglutination (97%) and an enzyme-linked immunosorbent assay (98.3%). The sensitivity and specificity of LEIA compared with those of the enzyme-linked immunosorbent assay were 100 and 92.5%, respectively.

THE indirect fluorescent antibody technique was used to investigate the possible presence of antibodies in the lacrimal fluid or, to be more exact, the fluid of the conjunctival sac of 21 patients with active trachoma (Stage II–III), nearly all of them children, 9–14 years old. In most instances, the small quantity of fluid normally present in the conjunctival sac, or produced by slight mechanical irritation rather than freely flowing tears, was examined. The fluids, obtained with the aid of sterile pipettes used for white blood-cell count, were diluted 1 : 10 in phosphate buffered saline pH 7.4. Trachoma elementary body antigens were prepared and examinations were carried out by techniques previously described1–3. Commercial fluorescein isothiocyanate labelled anti-human γ-globulin (Microbiological Associates, Bethesda) and rhodamine were used for the purpose of staining. Fluorescence with trachoma antigen was observed with the conjunctival fluids of ten from the twenty-one cases of trachoma. The conjunctival fluids from twenty-two trachoma-free children gave negative results.

C. TRACHOMATIS (CT) infections of the upper genital tract in women are either acute, sub acute or chronic. CT infection has a tendency to be chronic, latent and persistent as a consequence of the host immune reaction to CT major outer membrane protein, 57 Kd heat shock protein and lipopolysaccharide. Chlamydial persistence can be induced as a result of inflammatory and/or immune regulated cytokines, Interferon γ depletion of tryptophan causes a stress response involving development of abnormal forms with increased levels of stress response proteins which maintain host immune responses with continuous fibrin exudate.

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.