Clinical Manifestations and Genomic Evaluation of Melioidosis Outbreak among Children after Sporting Event, Australia.

Melioidosis, caused by the environmental gram-negative bacterium Burkholderia pseudomallei, usually develops in adults with predisposing conditions and in Australia more commonly occurs during the monsoonal wet season. We report an outbreak of 7 cases of melioidosis in immunocompetent children in Australia. All the children had participated in a single-day sporting event during the dry season in a tropical region of Australia, and all had limited cutaneous disease. All case-patients had an adverse reaction to oral trimethoprim/sulfamethoxazole treatment, necessitating its discontinuation. We describe the clinical features, environmental sampling, genomic epidemiologic investigation, and public health response to the outbreak. Management of this outbreak shows the potential benefits of making melioidosis a notifiable disease. The approach used could also be used as a framework for similar outbreaks in the future.

Workflow to facilitate the detection of new psychoactive substances and drugs of abuse in influent urban wastewater.

The complexity around the dynamic markets for new psychoactive substances (NPS) forces researchers to develop and apply innovative analytical strategies to detect and identify them in influent urban wastewater. In this work a comprehensive suspect screening workflow following liquid chromatography - high resolution mass spectrometry analysis was established utilising the open-source InSpectra data processing platform and the HighResNPS library. In total, 278 urban influent wastewater samples from 47 sites in 16 countries were collected to investigate the presence of NPS and other drugs of abuse. A total of 50 compounds were detected in samples from at least one site. Most compounds found were prescription drugs such as gabapentin (detection frequency 79%), codeine (40%) and pregabalin (15%). However, cocaine was the most found illicit drug (83%), in all countries where samples were collected apart from the Republic of Korea and China. Eight NPS were also identified with this protocol: 3-methylmethcathinone 11%), eutylone (6%), etizolam (2%), 3-chloromethcathinone (4%), mitragynine (6%), phenibut (2%), 25I-NBOH (2%) and trimethoxyamphetamine (2%). The latter three have not previously been reported in municipal wastewater samples. The workflow employed allowed the prioritisation of features to be further investigated, reducing processing time and gaining in confidence in their identification.

Investigation and Resolution of Interference in the LC-QTOF-MS Detection of 4-MePPP.

An interference resulting in the false-positive detection of the synthetic cathinone 4-MePPP in urine was suspected following the recent addition of 4-MePPP spectral data to an LC-QTOF-MS drug library. Although positive detection criteria were achieved, it was noted that all urine samples suspected of containing 4-MePPP also concurrently contained high levels of tramadol and its associated metabolites. Using QTOF-MS software elucidation tools, candidate compounds for the suspected interference were proposed. To provide further confidence in the identity of the interference, in silico fragmentation tools were used to match product ions generated in the analysis with product ions predicted from the theoretical fragmentation of candidate compounds. The ability of the suspected interference to subsequently produce the required product ions for spectral library identification of 4-MePPP was also tested. This information was used to provide a high preliminary confidence in the compound identity prior to purchase and subsequent confirmation with certified reference material. A co-eluting isobaric interference was identified and confirmed as an in-source fragment of the tramadol metabolite, N,N-bisdesmethyltramadol. Proposed resolutions for this interference are also described and subsequently validated by retrospective interrogation of previous cases of suspected interference.

Quadrupole Time-of-Flight Mass Spectrometry: A Paradigm Shift in Toxicology Screening Applications.

The screening of biological samples for the presence of illicit or legal substances is an important frontline tool in both clinical and forensic toxicology. In the clinical setting, drug screening is a useful tool for the clinician in improving patient care and guiding treatment. Analytical approaches for the screening of drugs in biological samples are extensive and well documented, though many rapid screening techniques often lack appropriate sensitivity and specificity, requiring careful clinical interpretation. The continuous emergence of new psychoactive substances presents a considerable analytical challenge in maintaining up-to-date methods for the detection of relevant drugs. Adapting and validating methods for the detection of new substances can be a complicated and costly undertaking. There is also a considerable lag time between the emergence of new drugs and the release of commercial assays for detection. Quadrupole time-of-flight mass spectrometry (Q-TOF-MS) has gained considerable attention over the last decade as an analytical technique that is capable of meeting the challenges of a rapidly changing drug landscape. Exhibiting both high sensitivity and specificity in drug detection, Q-TOF-MS also allows methods to be rapidly updated for newly emerging psychoactive agents. The coupling of Q-TOF-MS with techniques such as liquid or gas chromatography can provide both rapid and comprehensive screening solutions that are gaining popularity in the clinical laboratory setting.

An electrophysiological signal that precisely tracks the emergence of error awareness.

Recent electrophysiological research has sought to elucidate the neural mechanisms necessary for the conscious awareness of action errors. Much of this work has focused on the error positivity (Pe), a neural signal that is specifically elicited by errors that have been consciously perceived. While awareness appears to be an essential prerequisite for eliciting the Pe, the precise functional role of this component has not been identified. Twenty-nine participants performed a novel variant of the Go/No-go Error Awareness Task (EAT) in which awareness of commission errors was indicated via a separate speeded manual response. Independent component analysis (ICA) was used to isolate the Pe from other stimulus- and response-evoked signals. Single-trial analysis revealed that Pe peak latency was highly correlated with the latency at which awareness was indicated. Furthermore, the Pe was more closely related to the timing of awareness than it was to the initial erroneous response. This finding was confirmed in a separate study which derived IC weights from a control condition in which no indication of awareness was required, thus ruling out motor confounds. A receiver-operating-characteristic (ROC) curve analysis showed that the Pe could reliably predict whether an error would be consciously perceived up to 400 ms before the average awareness response. Finally, Pe latency and amplitude were found to be significantly correlated with overall error awareness levels between subjects. Our data show for the first time that the temporal dynamics of the Pe trace the emergence of error awareness. These findings have important implications for interpreting the results of clinical EEG studies of error processing.