Differential expression of groEL-1, incB, pyk-F, tal, hctA and omcB genes during Chlamydia trachomatis developmental cycle.

Chlamydia trachomatis infects squamous and columnar epithelia at the mucosal surface. Research on gene expression patterns of C. trachomatis has predominantly focused on non-native host cells, with limited data on growth kinetics and gene expression of chlamydia in keratinocytes. Here, we investigated whether early, mid, and late chlamydial genes observed in HeLa cell line studies were co-ordinately regulated at the transcriptional level even in the keratinized cell line model and whether the expression was stage-specific during the developmental cycle. HaCaT cell lines were infected with chlamydia clinical isolates (US151and serovar E) and reference strain (L2 434). Expression of groEL-1, incB, pyk-F, tal, hctA, and omcB genes was conducted with comparative real-time PCR and transcriptional events during the chlamydial developmental cycle using transmission electron microscopy. The relative expression level of each gene and fold difference were calculated using the 2-ΔΔCT method. The expression of groEL-1 and pyk-F genes was highest at 2 hours post-infection (hpi) in the L2 434 and serovar E. The expression of incB gene increased at 2 hpi in L2 434 and serovar E but peaked at 12 hpi in serovar E. L2 434 and US151 had similar tal expression profiles. Increased expression of hctA and omcB genes were found at 2 and 36 hpi in L2 434. Both clinical isolates and reference strains presented the normal chlamydial replication cycle comprising elementary bodies and reticulate bodies within 36 hpi. We show different gene expression patterns between clinical isolates and reference strain during in vitro infection of keratinocytes, with reference strain-inducing consistent expression of genes. These findings confirm that keratinocytes are appropriate cell lines to interrogate cell differentiation, growth kinetics, and gene expression of C. trachomatis infection. Furthermore, more studies with different clinical isolates and genes are needed to better understand the Chlamydial pathogenesis in keratinocytes.

Determinants of symptomatic vulvovaginal candidiasis among human immunodeficiency virus type 1 infected women in rural KwaZulu-Natal, South Africa.

INTRODUCTION: We sought to determine the association between HIV-induced immunosuppression, virologic correlates, and vulvovaginal candidiasis (VVC). METHODS: This is a retrospective cohort study, where HIV infected and uninfected women were studied with VVC being the primary outcome. Ninety-seven HIV-infected and 101 HIV-uninfected women were enrolled between June and December 2011. Cases of VVC were confirmed. HIV RNA load was determined by RT-PCR and CD4 counts were obtained from medical records. RESULTS: Fifty-two of 97 (53.6%) HIV-infected and 38/101 (37.6%) HIV-uninfected women were diagnosed with VVC (P = 0.032). The relative risk for VVC amongst HIV-infected patients was 1.53 (95% CI: 1.04-2 P = 0.024). Cases of VVC increased at CD4+ T cell count below 200 cells/mm(3) (P < 0.0001) and plasma HIV RNA load above 10 000 copies/mL (P < 0.0001). VVC was associated with increased genital shedding of HIV (P = 0.002), and there was a linear correlation between plasma HIV load and genital HIV shedding (r = 0.540; R (2) = 0.292; P < 0.0001). Women on HAART were 4-fold less likely (P = 0.029) to develop VVC. CONCLUSION: CD4 counts below 200 cells/mm(3) and plasma HIV loads ≥10 000 copies/mL were significantly associated with VVC.

Intensified specimen collection to improve tuberculosis diagnosis in children from Rural South Africa, an observational study.

BACKGROUND: In drug-resistant TB settings, specimen collection is critical for drug-susceptibility testing (DST). This observational study included multiple specimen types collected from pediatric TB suspects with the aim to determine diagnostic yield and inform clinical practice in children with drug-resistant and drug-susceptible TB. METHODS: From 03/2009-07/2010, TB suspects aged ≥6 months and ≤12 years were recruited among outpatient and inpatient settings. Subjects were new TB suspects or had persistent symptoms despite ≥2 months of TB treatment. The protocol included collection of a single blood and urine specimen, a single sputum induction and, if inpatients and <5 years of age, collection of 3 gastric aspirates (GA). Samples were cultured on solid and/or liquid media. DST was by 1% proportion method. RESULTS: Among 118 children with possible, probable or confirmed TB, the mean age was 4.9 years [SD 3.2] and 64 (62%) of those tested were HIV-positive. Eight (7%) subjects were culture-positive from at least one specimen; yield did not differ by HIV status or TB treatment history. Among those with positive cultures, 7/8 (88%) were from induced sputum, 5/6 (83%) from GA, 3/8 (38%) from blood, and 3/7 (43%) from urine. In subjects with both induced sputum and GA collection, sputum provided one additional case compared to GA. Multidrug resistant (MDR)-TB was detected by urine culture alone in one child >5 years old. Pan-resistant extensively drug resistant (XDR)-TB was identified by cultures from all sites in one subject. CONCLUSIONS: TB was cultured from HIV-positive and -negative children, and allowed for identification of MDR and XDR-TB cases. Urine and induced sputum each provided an additional TB diagnosis and, when compared to GA, may be considered a less invasive, same-day method of specimen collection for childhood TB suspects. This study illustrates the continued challenges and limitations of available strategies for pediatric TB diagnostics.

Aetiology of sexually transmitted infections in Maputo, Mozambique.

INTRODUCTION: The study sought to ascertain the prevalence of the aetiological agents of genital discharge and genital ulcer diseases in Maputo, Mozambique. METHODOLOGY: Consecutive consenting patients presenting to the Centro de Saúde do Porto in Maputo between March and April 2005 with genital discharge syndrome and/or genital ulcer diseases were recruited. Specimens were collected for the identification of STI pathogens. RESULTS: Of 346 recruited patients, 164 were male and 182 female. The prevalence of confirmed single aetiological agents for male urethritis was as follows: N. gonorrhoeae, 35%; C. trachomatis, 10%; and M. genitalium 4%. For vaginal discharge, N. gonorrhoeae was found in 11% of the women tested, followed by C. trachomatis (6.5%), bacterial vaginosis (34%), and T. vaginalis (2%). The prevalence of genital ulcers was as follows: Herpes simplex virus type 2, 62%; H. ducreyi 4 %; and C. trachomatis biovar LGV, 4%. Five percent of patients with genital ulcers had a positive syphilis serology (RPR ≥ 1:8 and confirmed by TPHA) and 35% of all tested patients were HIV-1/2 infected. Cases of mixed infections were present in 5%, 11% and 3% of patients with male urethritis, vaginal discharge, and genital ulcers respectively. CONCLUSION: The classic sexually transmitted infection aetiologies are still prevalent in Maputo. The study highlights the need for a periodic surveillance to inform syndromic management protocols.

Aetiology of Sexually Transmitted Infections in Maputo; Mozambique

Introduction: The study sought to ascertain the prevalence of the aetiological agents of genital discharge and genital ulcer diseases in Maputo; Mozambique. Methodology: Consecutive consenting patients presenting to the Centro de Saude do Porto in Maputo between March and April 2005 with genital discharge syndrome and/or genital ulcer diseases were recruited. Specimens were collected for the identification of STI pathogens. Results: Of 346 recruited patients; 164 were male and 182 female. The prevalence of confirmed single aetiological agents for male urethritis was as follows: N. gonorrhoeae; 35; C. trachomatis; 10; and M. genitalium 4. For vaginal discharge; N. gonorrhoeae was found in11of the women tested; followed by C. trachomatis (6.5); bacterial vaginosis (34); and T. vaginalis (2). The prevalence of genital ulcers was as follows: Herpes simplex virus type 2; 62; H. ducreyi 4; and C. trachomatis biovar LGV; 4. Five percent of patients with genital ulcers had a positive syphilis serology (RPR = 1:8 and confirmed by TPHA) and 35of all tested patients were HIV-1/2 infected. Cases of mixed infections were present in 5; 11and 3of patients with male urethritis; vaginal discharge; and genital ulcers respectively. Conclusion: The classic sexually transmitted infection aetiologies are still prevalent in Maputo. The study highlights the need for a periodic surveillance to inform syndromic management protocols

Antimicrobial susceptibility profile of Neisseria gonorrhoeae isolated from patients attending a STD facility in Maputo, Mozambique.

OBJECTIVE: To ascertain the effectiveness of kanamycin for the treatment of gonorrhoea in Maputo, Mozambique. METHODS & DESIGN: A cross-sectional study design was employed. Urethral and cervical specimens were collected for the isolation of Neisseria gonorrhoeae from patients attending Centro de Saúde do Porto. Antimicrobial susceptibilities were determined for kanamycin, spectinomycin, ciprofloxacin, ceftriaxone, cefixime, tetracycline and penicillin. RESULTS: Twenty-two (40%) Neisseria gonorrhoeae isolates were intermediate and 4(7%) were resistant to kanamycin; 42(77%) displayed high level resistance to tetracycline (MIC > or = 16 mg/L); 34 (65%) were penicillinase producers, and 52 (95%) had spectinomycin MICs of 64 mg/L. All isolates were susceptible to ciprofloxacin (MIC < or = 0.06 mg/L), ceftriaxone (MIC < or = 0.015 mg/L) and cefixime (MIC < or = 0.015 mg/L). CONCLUSION: The observations underscore the need for broader susceptibility surveillance studies to elucidate the pattern and extent of drug resistance in Mozambique. A review of the current treatment guidelines for genital discharge syndrome is warranted.