The study was performed to investigate the prevalence of Chlamydia trachomatis and Mycoplasma genitalium in a population of infertile couples from Iran and how this relates to tubal factor infertility, pregnancy rate and outcome of pregnancy. Blood, semen and first-void urine samples were obtained from 250 infertile couples and 250 fertile women as a control. Infertile couples were followed up after 24 months to determine diagnosis, referral for assisted conception, any pregnancy and pregnancy outcome. Data were analysed with regard to the results of (i) serological analysis for specific antibodies to C. trachomatis in serum; (ii) the presence of C. trachomatis and M. genitaliumDNA in first-void urine; and (iii) in a semen sample of the male partner. Prevalence of C. trachomatis in our study population was comparable to other studies using similar methods and test specimens. No evidence of M. genitalium infection was found. Detection of C. trachomatis in one partner rarely correlated with infection in the other. The risk of tubal factor infertility and the probability of pregnancy and pregnancy outcome were unrelated to the results of serological tests for C. trachomatis antibodies or the presence of C. trachomatisDNA in first-void urine of both partners and in a semen sample provided by the male.

Objectives The aim of the study was to qualitatively and semiquantitatively characterize the expression of the principal HIV co-receptors chemokine (C-C motif) receptor 5 (CCR5) and chemokine (C-X-C motif) receptor 4 (CXCR4) on susceptible CD4 T-helper cell, monocyte/macrophage and Langerhans dendritic cell populations within the cervical epithelia of asymptomatic women attending a genitourinary medicine clinic. Methods Of 77 asymptomatic women recruited, 35 were excluded: 21 because they were found to have bacterial vaginosis, eight because they were found to have candida and six for other reasons. Cervical cytobrush samples from 11 women with Chlamydia trachomatis infection and 31 women without any detectable genital infection were stained with fluorescently labelled antibodies specific for cell surface CCR5, CXCR4, CD4, CD3, CD1a and CD19 expression, then analysed by flow cytometry. Results CD4/CD3 T-helper cells (84%), CD1a Langerhans dendritic cells (75%) and CD4/CD14 monocytes/macrophages (59%) were detected in the samples. CCR5 and CXCR4 HIV co-receptor expression was observed on 4686% of the above subsets. CD1a cells exhibited significantly higher CCR5 and CXCR4 positivity and median fluorescence than CD4 cells and higher CXCR4 positivity and median fluorescence than CD14 cells (P?<?0.05 or less). Increased detection of CCR5 over CXCR4 was seen in CD14 cells (P?<?0.05). No significant differences in CCR5 or CXCR4 expression were found in samples from asymptomatic women with or without chlamydial infection. Conclusions Co-receptor expression confirms the potential for CD1a Langerhans cells, monocytes/macrophages and T-helper cells in the cervix as primary targets for HIV infection. Previously observed selective transmission of CCR5-tropic isolates cannot be accounted for by a lack of CXCR4-expressing CD4 cervical immune cells. We were unable to identify any specific impact of chlamydial infection on co-receptor expression in this study.

Chlamydia trachomatis is an important bacterial cause of infertility. In men, the recommended specimen for diagnosing chlamydial infection is urine, with little or no emphasis placed on testing semen. A systematic review of the literature was conducted to search for studies in which men undergoing investigations for infertility produced both samples of urine and semen that were tested concurrently for C. trachomatis. An analysis of these studies was then performed. From 322 papers identified from the US National Library of Medicine PubMed database, 14 were selected for a detailed review and 11 were analysed further. Overall, the size of the study groups was only modest and differences between the studies included variation in geography and test methodology, especially whether commercial testing systems had been used. There was also a lack of consistency with regard to including men with azoospermia. Patients were typically 30-35 years old and the median duration of infertility was about 4 years. Of those patients positive for C. trachomatis in the 11 studies, 56% could be detected in both semen and urine, 20% only in urine and 23% only in semen. Deficiencies in existing studies would not allow for a meta-analysis, emphasizing the need for further research in this area for which a number of recommendations are made.

The aim of this study was to determine whether interleukin (IL)-6 and IL-8 concentrations, as well as numbers of seminal leukocytes in a population of infertile men, some of whom were Chlamydia trachomatis positive, were related to chlamydial infection. Our patient group included 255 men attending for diagnostic semen analysis as part of infertility investigations. Significantly raised levels of IL-8, but not IL-6, were found in C trachomatis-infected patients but not in uninfected patients. Raised IL-8 levels in semen were also associated with an increase in semen volume. There was a relationship between C trachomatis infection and lower progressive motile sperm, as well as an increase in seminal leukocytes. The overall prevalence rate for C trachomatis was 6.2%, and more infections were detected in semen than in first void urine. This study supports the suggestion that IL-8 might be used as a marker for male genital tract infection, especially when due to C trachomatis. In this study, there was a relationship between the presence of C trachomatis in semen and alterations of some semen parameters. Further investigations should be performed to understand the disparities of first void urine and semen testing for detection of C trachomatis in males.

BACKGROUND: Previous work has shown that co-incubation of human sperm with Chlamydia trachomatis serovars E and LGV leads to premature sperm death and that this is due primarily to chlamydial lipopolysaccharide (LPS).Here, we investigated the possible involvement of apoptosis in this premature sperm death.METHODS: Highly motile preparations of sperm from normozoospermic patients were co-incubated for 6 h with extracted LPS from C. trachomatis serovars E and LGV.Three different methods were used to determine if LPS-treated sperm underwent apoptosis, including: (i) flow cytometry; (ii) measurement of ADP:ATP ratios; and (iii) measurement of mono-and oligonucleosomal DNA fragments.Caspase activity was also investigated by fluorimetry and by use of a pan-caspase inhibitor and caspase-3 inhibitor.RESULTS: All three methods used for detection indicated that C. trachomatis LPS induced some apoptosis in sperm after 6 h when compared with a staurosporine (apoptosispositive) control.Moreover, a greater degree of apoptosis was seen with C. trachomatis serovar E than with serovar LGV.It was also shown that C. trachomatis LPS-induced apoptosis of sperm could be blocked with a pan-caspase inhibitor and a caspase-3 inhibitor.Moreover, by using a fluorogenic substrate, apoptosis was shown to be caspasemediated.CONCLUSIONS: In general it is believed that apoptosis does not occur in C. trachomatis-infected host cells.However, using three different methods, our findings clearly indicate that co-incubation of sperm with C. trachomatis LPS results in cellular death which is in part due to apoptosis and is caspase-mediated.These findings provide an explanation as to how C. trachomatis can mediate premature death in human sperm.

There is increasing evidence that the function of human spermatozoa can be significantly affected by direct exposure to the bacterium Chlamydia trachomatis. This may contribute to sub-fertility in infected individuals by a route that is independent of any damage to the reproductive epithelium. In addition, if a C. trachomatis infection is undiagnosed it could contribute to poor outcomes in assisted conception techniques such as in vitro fertilization. The antibiotics routinely used in IVF culture systems are largely ineffective against chlamydia, emphasizing the importance of screening patients prior to treatment. Moreover, given the many thousands of semen samples provided for analysis by men in primary care (many of which will never undergo assisted conception treatment), it is suggested that this may represent a wasted opportunity to provide screening (and treatment) for the infection using an appropriate test specimen and without the need for additional hospital visits.

Sulphated glycosaminoglycans, such as heparan sulphate, have been shown to be essential for the infectivity of many organisms. The aims of this study were to verify the role of sulphated glycosaminoglycans in chlamydial infection and to investigate whether they are present on chlamydia or chlamydial host cells. The effect of undersulphation of host cells and chlamydial elementary bodies was examined using sodium chlorate. Also studied was whether any inhibitory effect was reversible. The results strongly suggest that Chlamydia trachomatis does not produce heparan sulphate and that heparan sulphate of the host cell is necessary and sufficient to mediate chlamydial infection. The essential role played by the sulphate constituents of the host-cell glycosaminoglycan in the infectivity of LGV serovars, and to a lesser extent of serovar E, was also confirmed.

Recognition of bacterial lipopolysaccharide (LPS) is critical in the host defence against Gramnegative infection. While enterobacterial LPS signals via Toll-like receptor 4 (TLR4), it has recently been reported that the LPS of Leptospira interrogans, Legionella pneumophila, Rhizobium species Sin-1 and at least one strain of Porphyromonas gingivalis are capable of signalling via TLR2. Using a TLR transfection assay and measurement of an NF-kappaB-sensitive promoter region, the results show that the LPS of Bacteroides fragilis NCTC-9343, Chlamydia trachomatis LGV-1 and Pseudomonas aeruginosa PAC-611 also signal via TLR2 and it is pointed out that all TLR2-signalling LPS discovered to date demonstrate relatively weak endotoxicity in some models and structural features distinct from those LPS shown to signal via TLR4.

There is increasing evidence that the function of human spermatozoa can be significantly affected by direct exposure to the bacterium Chlamydia trachomatis. This may contribute to sub-fertility in infected individuals by a route that is independent of any damage to the reproductive epithelium. In addition, if a C. trachomatis infection is undiagnosed it could contribute to poor outcomes in assisted conception techniques such as in vitro fertilization. The antibiotics routinely used in IVF culture systems are largely ineffective against chlamydia, emphasizing the importance of screening patients prior to treatment. Moreover, given the many thousands of semen samples provided for analysis by men in primary care (many of which will never undergo assisted conception treatment), it is suggested that this may represent a wasted opportunity to provide screening (and treatment) for the infection using an appropriate test specimen and without the need for additional hospital visits.

Elementary bodies (EBs) of Chlamydia trachomatis serovar E are more toxic to sperm than those from serovar LGV. In this study, lipopolysaccharide (LPS) was prepared from the EBs of both serovars and incubated with human spermatozoa at concentrations that matched the LPS concentration of EBs. The effects of EBs and LPS on sperm motility, viability and acrosomal status were then determined. Sperm motility was measured by computer-assisted sperm analysis and the hypo-osmotic swelling test was used to determine the proportion of dead cells. Acrosomal status was examined using a standard mAb assay. Over a 6 h incubation, LPS from both serovars resulted in a marked reduction in sperm motility (and a concomitant increase in the proportion of dead spermatozoa) in a manner similar to that seen in response to EBs of serovar E. In addition, when sperm were incubated with a range of doses of EBs and LPS, probit analysis revealed that the greater spermicidal effects of EBs from serovar E (when compared with serovar LGV) were not observed when sperm were incubated with LPS from the two serovars. This suggests that the more potent effect of EBs of serovar E cannot be explained entirely by differences in the composition of LPS. Interestingly, Escherichia coli LPS was required in doses 500 times more concentrated than chlamydial LPS in order to kill a similar proportion of sperm, suggesting that bacterial LPSs may differ in their spermicidal properties. However, that chlamydial LPS was spermicidal was demonstrated by the use of polymyxin B (a polycationic antibiotic known to neutralize LPS effects), confirming that the effects observed were primarily a result of LPS activity.

The ability of heparan sulfate, heparin, and other glycosaminoglycans to inhibit the infectivity of Chlamydia trachomatis serovars E and LGV was examined using a simple competitive inhibition assay with three cell types from the human female reproductive tract, including primary human endosalpingeal cells. With the majority of the glycosaminoglycans tested, LGV was more significantly inhibited than serovar E. We have compared chlamydial infectivity between a wild-type Chinese hamster ovary cell line and two glycosaminoglycan-deficient cell lines. LGV was shown to be unable to infect heparan sulfate-deficient and GAG-deficient Chinese hamster ovary cell lines, whereas the E serovar infected these cells as efficiently as the control (nondeficient) cells. These two sets of experiments confirmed that serovar LGV is more dependent on a heparan sulfate-related mechanism of infectivity than is serovar E. This is further supported by the fact that attempts to purify a heparan sulfate-like molecule from either serovar cultured in glycosaminoglycan-deficient cell lines were nonproductive. Previous reports have suggested that chlamydia are able to produce a heparan sulfate-like molecule that is important for attachment and infectivity. We have attempted to detect possible binding of a specific heparan sulfate antibody to C. trachomatis by flow cytometry. Results showed no binding of the heparan sulfate antibody to C. trachomatis serovar LGV or E. Our results strongly indicate that chlamydiae do not produce a heparan sulfate-like molecule but rather use host cell heparan sulfate in order to infect cells.

The aim of this work was to investigate the effect of elementary bodies (EB) of Chlamydia trachomatis serovars E and LGV on sperm motility, viability and acrosomal status. Highly motile preparations of spermatozoa from normozoospermic patients were co-incubated for 6 h with 0.54x10(6) EB per ml. At 1, 3 and 6 h of incubation, sperm motility was determined by computer-assisted semen analysis (CASA) and the proportion of dead cells determined by the hypo-osmotic swelling (HOS) test. Acrosomal status was also examined using a standard monoclonal antibody assay. In the absence of EB, the percentage of motile spermatozoa remained >69% over the 6h incubation and the proportion of dead spermatozoa at <12%. However, during the incubation with EB of serovar E there was a significant decline in the percentage of motile spermatozoa (P < 0.05), and a corresponding increase in the proportion of dead spermatozoa (P < 0.05) at all time-points. However, following incubation with serovar LGV, only the percentage of dead spermatozoa after 6 h incubation was significantly different from the control (P < 0.05). The amount of acrosome-reacted spermatozoa remained unchanged (<16%) in all incubations at all time-points. Dose-response experiments indicated that increasing the concentration of EB to 2.5x10(6) per ml did not significantly alter the results. Furthermore, co-incubation of spermatozoa with dead EB (killed by heat treatment) abolished the chlamydia-mediated response, indicating that the effect is a result of the live organism and not soluble components or membrane elements. These data suggest that a detrimental effect on sperm function by some serovars may be an as yet unrecognized component of infertility problems.

The prevalence of chlamydial DNA determined by PCR and in-situ hybridisation (ISH) in fresh tissue specimens (endometrium, fallopian tube and ovary) was investigated in 33 women presenting with ectopic pregnancy (EP), 14 women with tubal factor infertility (TFI) and 50 control patients from the UK and the West Indies. In the UK EP group, chlamydial DNA was detected by PCR in 56% of patients; similar results were found in the Trinidad EP group (67%). In the TFI group, chlamydial DNA was detected in (71%) of patients by PCR. The detection of Chlamydia trachomatis DNA by ISH was highest in the TFI group (43%). Women presenting with EP and TFI showed evidence of previous or current genital C. trachomatis infection, underlining the importance of this microorganism in the development of these conditions. Importantly, chlamydial DNA could be detected in DNA preparations from the endometrium, fallopian tube and ovary of EP and TFI patients at the time of surgery.

Editor,—A recent study in Jamaica by Dowe et al using cell culture and a direct fluorescence assay (DFA) showed a prevalence of Chlamydia trachomatis infection in 47% of gynaecological patients.1 Unfortunately, there are no comparable data for cell culture and DFA in Trinidad. Moreover, we cannot find any reports on serological studies for C trachomatis IgG antibody in the West Indies. In an attempt to shed more light on prevalence of C trachomatis IgG antibody in pregnant women in Trinidad, we collected 56 serum specimens (mean age of patients 27 years) with ethics committee approval from one clinic at the general hospital, Port of Spain. As well as testing these sera …

ABSTRACT Elementary bodies (EBs) of the obligate intracellular bacteriumChlamydia trachomatis are responsible for the first step of attachment to host cells. We have studied the effects of EBs on human sperm protein tyrosine phosphorylation, which is important to sperm function. Indirect immunofluorescence using antiphosphotyrosine antibodies showed that serovar E, but not LGV, caused increased tyrosine phosphorylation which was localized to the sperm tail region. Immunoblotting revealed that serovar E caused a marked increase in tyrosine phosphorylation of 80- and 95-kDa sperm proteins, whereas serovar LGV caused increased phosphorylation of only the 80-kDa moiety. Considering the importance of tyrosine phosphorylation for sperm capacitation and other aspects of sperm function, we conclude that EBs may affect these events.

It has been postulated that certain organic acids produced by the anaerobes associated with bacterial vaginosis (BV) could prevent a purulent response in this infection. Varying concentrations of pure succinic, acetic and lactic acids were incubated in vitro with a monocytic cell line (MonoMac 6). High inhibition of chemotaxis was produced by succinic acid; lower inhibition and no inhibition was shown by acetic acid and lactic acid respectively. Succinic and acetic acids were detected in high concentrations in the vaginal fluid of women with BV and in culture supernates of Prevotella and Mobiluncus spp.; these acids impaired chemotaxis of MonoMac 6 cells in vitro. The vaginal fluids of normal women and the culture supernates of Lactobacillus spp. had no effect on chemotaxis.

ABSTRACT The conventional method for antimicrobial susceptibility testing of Chlamydia trachomatis is subjective and potentially misleading. We have developed a reverse transcriptase PCR (RT-PCR)-based method which is more sensitive and less subjective than the conventional method. Using 16 strains of C. trachomatis in triplicate assays, we found the RT-PCR method consistently more sensitive than the conventional technique for all eight antimicrobials tested, with resultant MICs determined by RT-PCR ranging from 1.6-fold higher (erythromycin) to ≥195-fold higher (amoxicillin).

Objective: Pain is a common reason for patients to present to a doctor. Many patients with tester have seen their doctor with pain during the days before the rash and tester sine herpete is well described. If early varicella tester virus (VZV) reactivation could be identified confidently it could provide an opportunity for early antiviral intervention, This prospective study was performed to assess how often patients presenting to their general practitioner with unilateral pain of no obvious clinical cause proved to have evidence of VZV reactivation,Methods: Fifty-seven patients were recruited and followed for 28 days; laboratory testing included VZV polymerase chain reaction (PCR) from peripheral blood mononuclear cells. VZV IgG, IgA and IgM. The control group consisted of 81 blood donors.Results: Only two study patients developed the rash of tester. There was no significant difference in PCR or serological responses between the study group and control group. Clinical characteristics did not enable identification of patients presenting to their doctor with unilateral pain who had prodromal zoster.Conclusion: There was no evidence on clinical or laboratory tests used in this study to support the view that reactivation of VZV is a common cause of unexplained unilateral pain. (C) 1999 The British Infection Society.

The aim of this study was to analyse individual polar lipid analogues, within each lipid family present, of fusobacteria using fast atom bombardment mass spectrometry (FAB‐MS). Polar lipid extracts were prepared, washed and dried. Samples, dispersed in a matrix ofm‐nitrobenzyl alcohol, were analysed by negative ion FAB‐MS using xenon as the reagent gas. Major anion peaks observed in the low mass region of mass/charge (m/z), 211, 221, 225, 227, 239, 241, 249, 251, 253, 255, 273, 277, 279, 281, 289 and 291, were consistent with the presence of C13:1, C14:3, C14:1, C14:0, C15:1, C15:0, C16:3, C16:2, C16:1, C16:0, unknown, C18:3, C18:2, C18:1, unknown and C19:3 carboxylate anions. In the high mass region, major anion peaks observed with m/z 644, 646, 648, 660, 662, 672, 673, 674, 686, 688, 689, 690, 698, 700, 701, 703, 714, 716, 717 and 719 were consistent with the presence of phosphatidylethanolamine (PE) (29:2), PE (29:1), PE (29:0), PE (30:1), PE (30:0), PE (31:2), first isotope of PE (31:2), PE (31:1), PE (32:2), PE (32:1), first isotope peak of PE (32:1), PE (30:0), PE (33:3), PE (33:2), phosphatidylglycerol (PG) (31:3), PG (31:2), PE (34:2), PE (34:1), PG (32:2) and PG (32:1). We conclude that FAB‐MS can provide data on individual analogues of PE and PG from Fusobacterium spp. not readily obtained by other means. Furthermore, the phospholipid profile is diagnostic for the genus.

Members of the genus Aeromonas are responsible for a significant number of intestinal and extra-intestinal infections in humans as well as other animals. However, their true role in enteric disease has been hindered by an inadequate human volunteer study, lack of an appropriate animal model and deficiencies in diagnostic procedures. The latter are probably responsible in part for the small number of outbreaks that have been reported. Despite these difficulties, several recent outbreaks and epidemiological studies have indicated strong association between isolation of Aeromonas and enteric disease. This has further stimulated research into potential pathogenic properties of Aeromonas, and much progress has been made over the last few years with regard to colonization, protection from the host and extracellular factors. It is hoped that these advances in elucidating the role of potential pathogenicity factors will be matched by interest and diagnostic studies, so that together the true nature of Aeromonas in disease will be more closely understood.