The benefit of culture-independent methods to detect bacteria and fungi in re-infected root filled teeth: a pilot study.

AIM: To identify dominant microorganisms in root filled teeth with apical periodontitis by Pan-PCRs in comparison with a culture-dependent approach, focusing on fungal species profiling. METHODOLOGY: The root filling material (gutta-percha) removed from 42 teeth with periapical radiolucencies undergoing root canal retreatments was analysed by molecular genetics techniques. Real-Time Pan-PCRs were conducted for the diagnosis of predominant bacteria (targeting 16S rDNA) and fungi (targeting ITS1-2 region). Identification of microorganisms was performed by Sanger sequencing of the PCR products and BLAST analysis. Additionally, subgingival plaque samples were collected and cultured to review the composition of the microbial flora. The McNemar test and the repeated measures anova were used for statistical analyses (significance level was set at P < 0.05). RESULTS: Overall, 42/42 plaque samples had bacterial growth, whereas 32/42 gutta-percha samples had bacterial growth with a dominance of Streptococcus spp. (12/42) and Enterococcus faecalis (9/42). The mean number of bacterial taxa per gutta-percha sample was 1.6 cultivatable taxa, significantly lower than in the plaque sample that had six taxa/sample (P < 0.001). Fungus-specific cultures were negative for gutta-percha samples, and only one plaque sample had growth of a fungus. In total, 36/42 plaque samples were positive in bacterial Pan-PCRs. In bacterial Pan-PCRs of 31/42 gutta-percha samples, dominant microorganisms were identified including Streptococcus spp. (5/42) and E. faecalis (4/42). Moreover, in 7/42 gutta-percha samples, DNA of bacteria which are difficult-to-cultivate in microbiology routine culture (Acinetobacter,Pyramidobacter,Bacteroidetes,Synergistes,Atopobium and Pseudoramibacter) was found. DNA of Candida spp. was detected in 5/42 root canals by fungal Pan-PCR (1/5) and genus-specific Candida-PCR (5/5). CONCLUSIONS: Pan-PCR assays remain appropriate as a broad-range approach for the detection of a dominant pathogen in gutta-percha samples which have less diverse microbial composition. The molecular genetic Pan-PCR approach has the advantage of detecting microorganisms that are as-yet-uncultivable or difficult-to-cultivate and should be therefore complement conventional microbiological diagnostics.

Ocular Syphilis: Experience over 11 Years at a German Ophthalmology Reference Centre.

BACKGROUND: In accordance with worldwide data, the Robert Koch Institute (RKI) has reported a constant increase of syphilis cases in Germany over the past decade. METHODS: We analysed the data of all patients, referred to a Department of Ophthalmology in a tertiary referral centre in Düsseldorf, Germany between 2008 and 2019, who were tested for syphilis. The epidemiologic, demographic, clinical, diagnostic and therapeutic data were retrieved from the records and evaluated in a retrospective, descriptive, non-comparative study. RESULTS: Syphilis serology was positive in 32/1840 (1.7%) patients, and was evenly distributed over this period. 26 (81.3%) were male, 19 (59.4%) belonged to a risk group. Ocular syphilis was the primary diagnosis for 29 patients (90.6%). The most frequent manifestation was uveitis (n = 20, 62.5%). By the end of therapy, 19 patients (59.4%) had an improved visual acuity. CONCLUSION: The incidence of ocular syphilis cases has remained stable over the last decade.

Development and Assessment of a Simulator for in Vivo Confocal Microscopy in Fungal and Acanthamoeba Keratitis.

BACKGROUND AND PURPOSE: In vivo confocal microscopy (IVCM) is a non-invasive imaging technique that allows morphological analysis as a diagnostic approach of the cornea in real time, thus providing a suspected diagnosis of fungal or amoebic keratitis immediately, whereas culture or PCR require several days or even weeks. Since these infections are rare, it is difficult for ophthalmologists to gain the experience necessary to differentiate infection from normal findings or artefacts. The purpose of this project was to establish a simulator, on which physicians could practice as well as acquiring a database of IVCM images of fungal or amoebic keratitis and respective analyses. PATIENTS AND METHODS: An IVCM simulator was set up with cadaver human corneas, infected with either acanthamoeba, candida or aspergillus. Twenty-one ophthalmologists were trained in IVC microscopy first in a Dry Lab, then practically on the simulator. For evaluation, the participants were asked to fill out a standardized questionnaire, with a pre- and post-course self-assessment. RESULTS: The self-assessed theoretical and practical skills in differentiating infectious from non-infectious keratitis in IVCM significantly increased (p = 0.0001, p = 0.0002, respectively). The barrier to use this technique decreased (p = 0.0474). CONCLUSION: A very simple protocol based on a model of ex vivo corneal mycotic and amoebic infections can be used to train novices in the structured approach and diagnostic use of IVCM for corneal infections.

[Preliminary clinical study on the management of 26-34 week pregnancy with premature rupture of the membranes (PRM)]. / Etude clinique préliminaire sur la conduite à tenir dans la grossesse de 26-34 semaines avec rupture prématurée des membranes (RPM).

UNLABELLED: The aim of this study was to determine the optimal solution to the dilemma of "wait-and-see" or "evacuation of uterine contents" and to identify objective criteria in support of one or other of these management choices, in particular in 26-34 week pregnancies with PRM, which is the most difficult situation. On the basis of a clinical study of 54 cases, week pregnancies with PRM did not enable temporization for any considerable time because of the onset of amniotic infection, and a large number of premature newborn with a gestational age of 31-32 weeks, below 1500-1700 g respectively, died a short time after birth. CONCLUSIONS: In 26-34 week pregnancies with PRM, a "wait-and-see" attitude with its sometimes very considerable septic risks is justified by a high chance of fetal survival score (CFSS)--a concept suggested by the authors--, i.e. with a gestational age between 30-34 weeks and respective fetal weights (determined by ultrasonography) between 1400-2100 g, absence of intrauterine fetal pathology, favorable fetal vitality parameters, good prognosis foe vaginal delivery, low C-reactive protein levels, low-grade pathogenicity of cervicovaginal flora, etc. In their preliminary study, the authors suggest determination of a chance of fetal survival score (CFSS) which is of prognostic value and can be useful in determining optimal management in the dilemma raised by PRM in 26-34 week pregnancies.