In Vivo Evolution of Butane Oxidation by Terminal Alkane Hydroxylases AlkB and CYP153A6

Enzymes of the AlkB and CYP153 families catalyze the first step in the catabolism of medium-chain-length alkanes, selective oxidation of the alkane to the 1-alkanol, and enable their host organisms to utilize alkanes as carbon sources. Small, gaseous alkanes, however, are converted to alkanols by evolutionarily unrelated methane monooxygenases. Propane and butane can be oxidized by CYP enzymes engineered in the laboratory, but these produce predominantly the 2-alkanols. Here we report the in vivo-directed evolution of two medium-chain-length terminal alkane hydroxylases, the integral membrane di-iron enzyme AlkB from Pseudomonas putida GPo1 and the class II-type soluble CYP153A6 from Mycobacterium sp. strain HXN-1500, to enhance their activity on small alkanes. We established a P. putida evolution system that enables selection for terminal alkane hydroxylase activity and used it to select propane- and butane-oxidizing enzymes based on enhanced growth complementation of an adapted P. putida GPo12(pGEc47{Delta}B) strain. The resulting enzymes exhibited higher rates of 1-butanol production from butane and maintained their preference for terminal hydroxylation. This in vivo evolution system could be useful for directed evolution of enzymes that function efficiently to hydroxylate small alkanes in engineered hosts

Biocatalysis. Biological systems for the production of chemicals

Biocatalysis harnesses the catalytic potential of enzymes to produce building blocks and end-products for the pharmaceutical and chemical industry. Located at the interface between fermentation processes and petrol-based chemistry, biotransformation processes broaden the toolbox for bioconversion of organic compounds to functionalized product

Subjective and objective assessment of executive functions in Parkinson's disease

AbstractImpairments in executive functions (EF) in Parkinson's disease (PD) will have a negative influence on daily life. For the assessment objective and subjective measurement approaches are used. It is however unknown whether these approaches contribute in a different way to the assessment of EF in PD. Thirty-nine PD patients and 24 healthy participants completed the Dysexecutive questionnaire (DEX; subjective measure) and the Frontal Assessment Battery (FAB; objective measure). PD patients showed impaired EF (FAB) and reported more problems with EF in daily life (DEX) than healthy participants. The performance on the FAB could however not be explained by the problems with EF that were reported by PD patients (DEX) and vice versa. In conclusion, not all PD patients who show impairments in EF report them and not all PD patients who report problems with EF in daily life show impairments according to objective measurement. Both measures thus contribute in a different way to the assessment of EF in PD patients. However, it has to be considered that the FAB is not a critical test to assess cognition in PD, since these patients also suffer from posterior abnormalities including memory and visuo-spatial deficits which are strong predictors for PD dementia

Abnormal Parietal Function in Conversion Paresis

The etiology of medically unexplained symptoms such as conversion disorder is poorly understood. This is partly because the interpretation of neuroimaging results in conversion paresis has been complicated by the use of different control groups, tasks and statistical comparisons. The present study includes these different aspects in a single data set. In our study we included both normal controls and feigners to control for conversion paresis. We studied both movement execution and imagery, and we contrasted both within-group and between-group activation. Moreover, to reveal hemisphere-specific effects that have not been reported before, we performed these analyses using both flipped and unflipped data. This approach resulted in the identification of abnormal parietal activation which was specific for conversion paresis patients. Patients also showed reduced activity in the prefrontal cortex, supramarginal gyrus and precuneus, including hemisphere-specific activation that is lateralized in the same hemisphere, regardless of right- or left-sided paresis. We propose that these regions are candidates for an interface between psychological mechanisms and disturbed higher-order motor control. Our study presents an integrative neurophysiological view of the mechanisms that contribute to the etiology of this puzzling psychological disorder, which can be further investigated with other types of conversion symptoms

Functional (psychogenic) movement disorders associated with normal scores in psychological questionnaires:A case control study

AbstractObjectiveFunctional movement disorders (FMDs) fall within the broader category called functional neurological symptom disorder (FNSD). New DSM-5 criteria for FNSD no longer require the presence of a ‘psychological conflict’ suggesting that some patients with FMD may not have obvious psychological comorbidity. We studied patients with FMD in comparison to patients with a neurological movement disorder (MD) and healthy controls (HC) to identify whether there is a subgroup of patients with FMD who have normal psychological test scores.MethodsWe assessed self-rated measures of depression/anxiety (SCL-90), dissociation and personality disorder (PDQ-4) in patients attending neurological clinics and healthy controls. The proportion of patients scoring within normal ranges was determined, and the levels of somatic and psychological symptoms were compared between the three groups.ResultsAmong the FMD group, 39% (20/51) scored within the normal range for all measures compared to 38% (13/34) of MD subjects and 89% (47/53) of healthy controls. There were no differences in overall scores in the SCL-90 and PDQ-4 between FMD and MD patients. FMD patients also did not differ from controls on a self-rated measure of personality pathology.ConclusionOur data show that a substantial proportion of patients with FMD score within the normal range in psychological questionnaires, lending some support to the new DSM-5 criteria

Virtual reality facial emotion recognition in social environments:An eye-tracking study

BACKGROUND: Virtual reality (VR) enables the administration of realistic and dynamic stimuli within a social context for the assessment and training of emotion recognition. We tested a novel VR emotion recognition task by comparing emotion recognition across a VR, video and photo task, investigating covariates of recognition and exploring visual attention in VR. METHODS: Healthy individuals (n = 100) completed three emotion recognition tasks; a photo, video and VR task. During the VR task, emotions of virtual characters (avatars) in a VR street environment were rated, and eye-tracking was recorded in VR. RESULTS: Recognition accuracy in VR (overall 75%) was comparable to the photo and video task. However, there were some differences; disgust and happiness had lower accuracy rates in VR, and better accuracy was achieved for surprise and anger in VR compared to the video task. Participants spent more time identifying disgust, fear and sadness than surprise and happiness. In general, attention was directed longer to the eye and nose areas than the mouth. DISCUSSION: Immersive VR tasks can be used for training and assessment of emotion recognition. VR enables easily controllable avatars within environments relevant for daily life. Validated emotional expressions and tasks will be of relevance for clinical applications

Virtual reality based cognitive behavioral therapy for paranoia:Effects on mental states and the dynamics among them

Background: Negative affective processes may contribute to maintenance of paranoia in patients with psychosis, and vice versa. Successful treatment may break these pathological symptom networks. This study examined whether treatment with virtual reality based cognitive behavioral therapy (VR-CBT) for paranoia influences momentary affective states, and whether VR-CBT changes the adverse interplay between affective states and paranoia. Methods: Patients with a psychotic disorder (n = 91) were randomized to 16-session VR-CBT or treatment as usual (TAU). With the experience sampling method (structured diary technique) mental states were assessed for 6–10 days at baseline, posttreatment and 6-month follow-up. Multilevel analysis were performed to establish treatment effects and time-lagged associations between mental states, that were visualized with networks of mental states. Results: Average levels of paranoia (feeling suspicious [b = −032., p = .04], disliked [b = −49., p < .01] and hurt [b = −0.52, p < .01]) and negative affect (anxious [b = −0.37, p = .01], down [b = −0.33, p = .04] and insecure [b = −0.17, p = .03) improved more after VR-CBT than TAU, but positive affect did not. Baseline mental state networks had few significant connections, with most stable connections being autocorrelations of mental states. The interplay between affective states and paranoia did not change in response to treatment. A trend reduction in average intranode connections (autocorrelations) was found after VR-CBT (b = −0.07, p = .08), indicating that mental states reinforce themselves less after treatment. Conclusions: VR-CBT reduced paranoid symptoms and lowered levels of negative affect in daily life, but did not affect the extent to which mental states influenced each other. Findings do suggest that as a result of treatment mental states regain flexibility

Executive Functioning in Daily Life in Parkinson's Disease: Initiative, Planning and Multi-Task Performance

Impairments in executive functioning are frequently observed in Parkinson's disease (PD). However, executive functioning needed in daily life is difficult to measure. Considering this difficulty the Cognitive Effort Test (CET) was recently developed. In this multi-task test the goals are specified but participants are free in their approach. This study applies the CET in PD patients and investigates whether initiative, planning and multi-tasking are associated with aspects of executive functions and psychomotor speed. Thirty-six PD patients with a mild to moderate disease severity and thirty-four healthy participants were included in this study. PD patients planned and demonstrated more sequential task execution, which was associated with a decreased psychomotor speed. Furthermore, patients with a moderate PD planned to execute fewer tasks at the same time than patients with a mild PD. No differences were found between these groups for multi-tasking. In conclusion, PD patients planned and executed the tasks of the CET sequentially rather than in parallel presumably reflecting a compensation strategy for a decreased psychomotor speed. Furthermore, patients with moderate PD appeared to take their impairments into consideration when planning how to engage the tasks of the test. This compensation could not be detected in patients with mild PD

Profiling Mechanisms of Alkane Hydroxylase Activity In Vivo Using the Diagnostic Substrate Norcarane

SummaryMechanistically informative chemical probes are used to characterize the activity of functional alkane hydroxylases in whole cells. Norcarane is a substrate used to reveal the lifetime of radical intermediates formed during alkane oxidation. Results from oxidations of this probe with organisms that contain the two most prevalent medium-chain-length alkane-oxidizing metalloenzymes, alkane ω-monooxygenase (AlkB) and cytochrome P450 (CYP), are reported. The results—radical lifetimes of 1–7 ns for AlkB and less than 100 ps for CYP—indicate that these two classes of enzymes are mechanistically distinguishable and that whole-cell mechanistic assays can identify the active hydroxylase. The oxidation of norcarane by several recently isolated strains (Hydrocarboniphaga effusa AP103, rJ4, and rJ5, whose alkane-oxidizing enzymes have not yet been identified) is also reported. Radical lifetimes of 1–3 ns are observed, consistent with these organisms containing an AlkB-like enzyme and inconsistent with their employing a CYP-like enzyme for growth on hydrocarbons

Harnessing plant biomass for biofuels and biomaterials

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/72519/1/j.1365-313X.2008.03512.x.pd