Comparison of culture and real-time polymerase chain reaction methods for detection of Mycoplasma hominis in amniotic fluids samples.

BACKGROUND: Mycoplasma hominis is often present in the amniotic cavity with microbial invasion associated with spontaneous preterm labor. Conventional culture method is the gold standard for detection of Mycoplasmas, but real-time polymerase chain reaction (real-time PCR) has revolutionized the diagnosis of M. hominis. OBJECTIVE: The purpose of this study is the comparison of the culture methodology with real-time PCR for the detection of M. hominis in amniotic fluid samples. METHODS: Amniotic fluid samples were collected from 65 pregnant women (age range: 25-45 years) previously followed at an infertility clinic. They were collected by transabdominal genetic amniocentesis during 16-21 weeks of gestation. Amniotic fluids were inoculated in SP4 broth for 48-72 h, and after becoming alkaline, culture suspension was spread on A7 agar plate for 1 week till the typical colonies seen in "fried-egg" morphology under stereomicroscope. DNA was extracted using a QIAGEN Mini DNA kit. The real-time-PCR was performed using Rotor-Gene Q Real-time PCR instrument. A melting-curve analysis was also performed. Sensitivity, specificity, positive predictive value (PPV), and negative predictive value (NPV) were measured by real-time PCR by taking culture as gold standard. RESULTS: Sixty-five women in 16-21 weeks of gestation, with a mean age of 33 ± 5.06 years, were enrolled into this study. M. hominis detected by culture and real-time PCR assay was 72% (47/65) and 69% (45/65), respectively. 66% (43/65) specimens were positive by both methods. Real-time PCR sensitivity was 91.5%, specificity 88.9%, PPV 95.6%, and NPV 80%. CONCLUSION: Rapid detection of Mycoplasmas causing maternal complications such as neonatal infections and preterm labor in pregnancy by real-time PCR may be important and necessary. The high sensitivity and shorter time requirement of real-time PCR support its further development for diagnosis of Mycoplasma infections.

Clinical effects of probiotics containing Bacillus species on gingivitis: a pilot randomized controlled trial.

BACKGROUND AND OBJECTIVE: Lactobacillus spp. and bifidobacteria are the most frequently used probiotics in oral health research. However, although probiotic effects have been suggested for other genera, such as bacilli, no trials are available to describe the effect of bacilli probiotics on gingivitis in humans. The aim of the present study was to evaluate the clinical effects of a bacilli-containing toothpaste, a mouthrinse and a toothbrush cleaner versus a placebo in patients with generalized gingivitis. MATERIAL AND METHODS: In this double-blind placebo-controlled randomized clinical trial, nonsmoking, systemically healthy patients with generalized gingivitis were included. They used a placebo or an experimental probiotic Bacillus subtilis-, Bacillus megaterium- and Bacillus pumulus-containing toothpaste, mouthrinse and toothbrush cleaner for 8 wk. Primary outcome measures of interest were plaque and gingivitis index, and the secondary outcome measures were pocket probing depth and bleeding on probing. RESULTS: Twenty male and 20 female patients were randomized over the two groups. All participants could be included in the final analysis. Although plaque and gingivitis indices were significantly reduced after 8 wk, no intergroup differences could be found at any time point. Also, for the secondary outcome measure, intragroup but no intergroup differences could be detected. No harm or unintended effects were reported by the patients after using the study products. CONCLUSIONS: This study did not show any statistically significant differences between a placebo and a bacilli-containing toothpaste, mouthrinse and toothbrush cleaner on gingivitis parameters.

Effects of prior sustained tactile stimulation on the somatosensory response to the sudden change of intensity in humans: an magnetoencephalography study.

The rapid detection of sensory changes is important to survival. The change-detection system should relate closely to memory since it requires the brain to separate a new stimulus from past sensory status. To clarify effects of past sensory status on processing in the human somatosensory cortex, brain responses to an abrupt change of intensity in a train of electrical pulses applied to the hand were recorded by magnetoencephalography (MEG). In Experiment 1, effects of the magnitude of deviance (1.0, 0.5, 0.3, 0.2, and 0.1 mA) between conditioning and test stimuli were examined. In Experiment 2, effects of the duration of the conditioning stimulus (3, 1.5, 1.0, and 0.5 s) were examined. The abrupt change in stimulus intensity activated the contralateral primary (cSI) and secondary somatosensory cortex (cSII). The amplitude of the cSI and cSII activity was dependent on not only the magnitude of the change in intensity but also the length of the conditioning stimulus prior to the change, suggesting that storage of prior tactile information was involved in generating these responses. The possibility that an activity of onset (with no conditioning stimulus) would be involved in the change-related activity was also discussed.

Case report: peritonitis by Penicillium spp. in a patient undergoing continuous ambulatory peritoneal dialysis.

Even though prominent technical improvements in continuous ambulatory peritoneal dialysis (CAPD) treatments during the last decade, peritonitis keeps its place as an important cause of morbidity and mortality in these patients. Among them fungal peritonitis is happened to be the most difficult one to deal with and comes out serious clinical presentation. It is presented here a case of CAPD related fungal peritonitis caused by Penicillium spp. This case experienced recently relapsing bacterial episodes of peritonitis and received long term antibiotics intraperitoneally and systemically. Eventually, Penicillium spp. was detected in several cultures of peritoneal effluent and also tip of Tenckhoff catheter, therefore it was considered as a causative agent. Then, the catheter was removed and amphotericin B therapy was performed. But the general condition of the patient did not improve till surgically drainage of peritoneal collection which was determined by MR (Magnetic Resonance) examination of abdomen after antifungal treatment was completed and Penicillium spp. in the drainage samples was not determined anymore.

Differential inhibition of mollicute growth: an approach to development of selective media for specific mollicutes.

The energy-generating pathways of Mycoplasma spp. are diverse. Thus, it was predicted that the ability of inhibitors of these pathways to block growth would vary among species. This prediction was tested with 14 Mycoplasma species and potential inhibitors. The greatest differentiation among test species was obtained using fluoride, iodoacetate (IAA), beta-fluoropyruvate (FP), cibacron blue (CB), L-citrulline, and carbonyl cyanide m-chlorophenylhydrazone. A range of other potential inhibitors, including L-arginine analogues, had little inhibitory effect on growth, and D-arginine was shown to be a growth substrate for arginine-hydrolyzing species. Fluoride selectively inhibited the growth of mycoplasmas that were able only to ferment sugars to lactate and/or to hydrolyze arginine. In contrast, IAA was most effective against organic acid-oxidizing species, and L-citrulline inhibited arginine-hydrolyzing species. Mycoplasma verecundum, a species for which energy sources have not been identified, was relatively resistant to FP. Similarly, Acholeplasma laidlawii was distinguished by its CB resistance.