Diagnosis of bacterial vaginosis by a new multiplex peptide nucleic acid fluorescence in situ hybridization method

Bacterial vaginosis (BV) is one of most common vaginal infections. However, its diagnosis by classical methods reveals low specificity. Our goal was to evaluate the accuracy diagnosis of 150 vaginal samples with research gold standard methods and our Peptide Nucleic Acid (PNA) probes by Fluorescence in situHybridization (FISH) methodology. Also, we described the first PNA-FISH methodology for BV diagnosis, which provides results in approximately 3 h. The results showed a sensitivity of 84.6% (95% confidence interval (CI), from 64.3 to 95.0%) and a specificity of 97.6% (95% CI [92.699.4%]), demonstrating the higher specificity of the PNA-FISH method and showing false positive results in BV diagnosis commonly obtained by the classical methods. This methodology combines the specificity of PNA probes for Lactobacillus species and G. vaginalis visualization and the calculation of the microscopic field by Nugent score, allowing a trustful evaluation of the bacteria present in vaginal microflora and avoiding the occurrence of misleading diagnostics. Therefore, the PNA-FISH methodology represents a valuable alternative for BV diagnosis.This work was supported by European Union funds (FEDER/COMPETE) and by national funds (FCT) under the project FCOMP-01-0124-FEDER-008991 (PTDC/BIA-MIC/098228/2008). Antonio Machado was funded by the FCT individual fellowship SFRH/BD/62375/2009. Other fundings came from the FCT Strategic Project PEst-OE/EQB/LA0023/2013, the Project NORTE-07-0124-FEDER-000027, co-funded by the Programa Operacional Regional do Norte (ON.2-O Novo Norte), QREN, FEDER, and the project FCOMP-01-0124-FEDER-008991 (RECI/BBB-EBI/0179/2012). Nuno Cerca is an Investigador FCT (Portuguese national funds). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript

Transabdominal Ultrasonography of the Small Bowel

In the era of double balloon enteroscopy, capsule endoscopy, CT, and MRI enterography is transabdominal ultrasonography (TUS) underestimated method for evaluation of small bowel pathology. As often initial imagine method in abdominal complaints, nowadays has TUS much better diagnostic potential than two decades ago. High-resolution ultrasound probes with harmonic imaging significantly improve resolution of bowel wall in real time, with possibility to asses bowel peristalsis. Color flow doppler enables evaluation of intramural bowel vascularisation, pulse wave doppler helps to quantificate flow in coeliac and superior mesenteric arteries. Small intestine contrast ultrasonography with oral contrast fluid, as well as contrast enhanced ultrasonography with intravenous microbubble contrast also improves small bowel imaging. We present a review of small intestine pathology that should be detected during ultrasound examinations, discuss technical requirements, advantages and limitations of TUS, typical ultrasound signs of Crohn's disease, ileus, celiac disease, intussusception, infectious enteritis, tumours, ischemic and haemorrhagic conditions of small bowel. In the hands of experienced investigator, despite some significant limitations(obesity, meteorism), is transabdominal ultrasonography reliable, noninvasive and inexpensive alternative method to computerised tomography (CT) and magnetic resonance imaging (MRI) in small bowel examination

Antagonistic effects of ocean acidification and rising sea surface temperature on the dissolution of coral reef carbonate sediments

Increasing atmospheric CO2 is raising sea surface temperature (SST) and increasing seawater CO2 concentrations, resulting in a lower oceanic pH (ocean acidification; OA), which is expected to reduce the accretion of coral reef ecosystems. Although sediments comprise most of the calcium carbonate (CaCO3) within coral reefs, no in situ studies have looked at the combined effects of increased SST and OA on the dissolution of coral reef CaCO3 sediments. In situ benthic chamber incubations were used to measure dissolution rates in permeable CaCO3 sands under future OA and SST scenarios in a coral reef lagoon on Australia's Great Barrier Reef (Heron Island). End of century (2100) simulations (temperature +2.7°C and pH -0.3) shifted carbonate sediments from net precipitating to net dissolving. Warming increased the rate of benthic respiration (R) by 29% per 1°C and lowered the ratio of productivity to respiration (P/R; delta P/R = -0.23), which increased the rate of CaCO3 sediment dissolution (average net increase of 18.9 mmol CaCO3/m**2/d for business as usual scenarios). This is most likely due to the influence of warming on benthic P/R which, in turn, was an important control on sediment dissolution through the respiratory production of CO2. The effect of increasing CO2 on CaCO3 sediment dissolution (average net increase of 6.5 mmol CaCO3/m**2/d for business as usual scenarios) was significantly less than the effect of warming. However, the combined effect of increasing both SST and pCO2 on CaCO3 sediment dissolution was non-additive (average net increase of 5.6 mmol CaCO3/m**2/d) due to the different responses of the benthic community. This study highlights that benthic biogeochemical processes such as metabolism and associated CaCO3 sediment dissolution respond rapidly to changes in SST and OA, and that the response to multiple environmental changes are not necessarily additive

Rapid and Accurate Identification of Candida albicans Isolates by Use of PNA FISHFlow▿

We developed the simple, rapid (1 h), and accurate PNA FISHFlow method for the identification of Candida albicans. The method exploits unique in solution in situ hybridization conditions under which the cells are simultaneously fixed and hybridized. This method facilitates the accurate identification of clinical yeast isolates using two scoring techniques: flow cytometry and fluorescence microscopy