Systemic Metabolomic Changes in Blood Samples of Lung Cancer Patients Identified by Gas Chromatography Time-of-Flight Mass Spectrometry.

Lung cancer is a leading cause of cancer deaths worldwide. Metabolic alterations in tumor cells coupled with systemic indicators of the host response to tumor development have the potential to yield blood profiles with clinical utility for diagnosis and monitoring of treatment. We report results from two separate studies using gas chromatography time-of-flight mass spectrometry (GC-TOF MS) to profile metabolites in human blood samples that significantly differ from non-small cell lung cancer (NSCLC) adenocarcinoma and other lung cancer cases. Metabolomic analysis of blood samples from the two studies yielded a total of 437 metabolites, of which 148 were identified as known compounds and 289 identified as unknown compounds. Differential analysis identified 15 known metabolites in one study and 18 in a second study that were statistically different (p-values <0.05). Levels of maltose, palmitic acid, glycerol, ethanolamine, glutamic acid, and lactic acid were increased in cancer samples while amino acids tryptophan, lysine and histidine decreased. Many of the metabolites were found to be significantly different in both studies, suggesting that metabolomics appears to be robust enough to find systemic changes from lung cancer, thus showing the potential of this type of analysis for lung cancer detection

Conscious coupling: The challenges and opportunities of cascading enzymatic microreactors

The continuous production of high value or difficult to synthesize products is of increasing interest to the pharmaceutical industry. Cascading reaction systems have already been employed for chemical synthesis with great success, allowing a quick change in reaction conditions and addition of new reactants as well as removal of side products. A cascading system can remove the need for isolating unstable intermediates, increasing the yield of a synthetic pathway. Based on the success for chemical synthesis, the question arises how cascading systems could be beneficial to chemo-enzymatic or biocatalytic synthesis. Microreactors, with their rapid mass and heat transfer, small reaction volumes and short diffusion pathways, are promising tools for the development of such processes. In this mini-review, the authors provide an overview of recent examples of cascaded microreactors. Special attention will be paid to how microreactors are combined and the challenges as well as opportunities that arise from such combinations. Selected chemical reaction cascades will be used to illustrate this concept, before the discussion is widened to include chemo-enzymatic and multi-enzyme cascades. The authors also present the state of the art of online and at-line monitoring for enzymatic microreactor cascades. Finally, the authors review work-up and purification steps and their integration with microreactor cascades, highlighting the potential and the challenges of integrated cascades

Attention and the motion after effect

We measured the effects of attentional distraction on the time course and asymptote of motion adaptation strength, using visual search performance (percent correct and reaction time). In the first two experiments, participantsadapted to a spatial array of moving Gaborpatches, either all vertically oriented (Experiment 1) or randomly oriented (Experiment 2). On each trial the adaptingarray was followed by a test array in which all of the test patchesexcept one were identical in orientation and movement direction to their retinotopically corresponding adaptors, but thetarget moved in the opposite direction to its adaptor. Participantswere required to identify the location of the changed targetwith a mouse click. The ability to do so increased with the number of adapting trials. Neither search speed nor accuracy was affected by an attentionallydemanding conjunction task at the fixation point during adaptation, suggesting low-level (pre-attentive) sites in the visual pathway for the adaptation. In Experiment 3 the same participants were required to identify the one element in the test array that was slowly moving. Reaction times in this case were elevated following adaptation, but once again there was nosignificant effect of the distracting task upon performance.In Experiment 4 participants were required to make eye movements, so that retinotopically corresponding adaptors could be distinguished from spatiotopically corresponding adaptors.Performance in Experiments 1 and 2 correlated positively with reaction times in Experiment 3, suggesting a general trait for adaptation strength

Molecular and Engineering Perspectives of the Biocatalysis Interface to Chemical Synthesis

The sustainable use of limited resources by nature to provide target molecules with biocatalytic reactions continues to be a role model for chemical synthesis. The application of biocatalysts to functional group transformations is shaped by the various parallel influences like e.g. the search for selectivity, the shift from fossil-based to biobased raw materials and the economy of molecular transformations like atom economy and step economy. As safety, health and environment issues are key drivers for process improvements in the chemical industry, the development of reactions or pathways replacing hazardous reagents is another major factor determining the sequence of molecular transformations from raw material to product. Biocatalyst production technologies and integrated process engineering have been instrumental in the establishment of biocatalytic reaction steps in chemical synthesis. The inherent properties of biocatalysts make them the privileged catalysts for highly selective asymmetric molecular transformations like e.g. hydrolysis reactions, oxidation reactions, carbon-carbon bond formation reactions as well as molecular unit transfer reactions. The universe of six enzyme classes provides a tremendous goldmine for discovering improved versions of enzymes with known functions as well as for finding completely novel enzymes. With the growing collection of biocatalytic reactions, the retrosynthetic thinking from chemical synthesis can be applied to biocatalysis as well. Once the feasibility of a biocatalytic reaction has been proven, up- and downscaling experiments have been useful for engineering the most adequate process design. In the case of the first large-scale biocatalytic Baeyer-Villiger oxidation, the debottlenecking of the substrate feed and product recovery, final purification and overcoming thermodynamic limitations have been essential in establishing bioprocesses with high yields of enantiopure products. These downscaling experiments in conjunction with new analytical techniques have proven useful also in the case of asymmetric synthesis of natural compounds. Spatial and temporal organisation of biocatalysts, reactants or products is another interesting engineering option for biocatalytic process design. The interdisciplinary character of the dead ends and locks between chemistry, biology and engineering requires investigations of the interfaces. Communication across scientific and technological disciplines including the value creation perspective is important for the development of a better synthesis for the final product-in-the-bottle. Whether the successful problem solution will come from the engineering of substrates, reaction media, process conditions or from the search for better and new enzymes, progress in the understanding of the molecular mechanisms of enzyme action will be key for the further development of the science of synthesis with its challenges towards the more difficult and more complex target molecules

Enzymatic synthesis of chiral amino-alcohols by coupling transketolase and transaminase-catalyzed reactions in a cascading continuous-flow microreactor system

Rapid biocatalytic process development and intensification continues to be challenging with currently available methods. Chiral amino-alcohols are of particular interest as they represent key industrial synthons for the production of complex molecules and optically pure pharmaceuticals. (2S,3R)-2-amino-1,3,4-butanetriol (ABT), a building block for the synthesis of protease inhibitors and detoxifying agents, can be synthesized from simple, non-chiral starting materials, by coupling a transketolase- and a transaminase-catalyzed reaction. However, until today, full conversion has not been shown and, typically, long reaction times are reported, making process modifications and improvement challenging. In this contribution, we present a novel microreactor-based approach based on free enzymes, and we report for the first time full conversion of ABT in a coupled enzyme cascade for both batch and continuous-flow systems. Using the compartmentalization of the reactions afforded by the microreactor cascade, we overcame inhibitory effects, increased the activity per unit volume, and optimized individual reaction conditions. The transketolase-catalyzed reaction was completed in under 10 min with a volumetric activity of 3.25 U ml-1 . Following optimization of the transaminase-catalyzed reaction, a volumetric activity of 10.8 U ml-1 was attained which led to full conversion of the coupled reaction in 2 hr. The presented approach illustrates how continuous-flow microreactors can be applied for the design and optimization of biocatalytic processes

A compact statistical model of the song syntax in Bengalese finch

Songs of many songbird species consist of variable sequences of a finite number of syllables. A common approach for characterizing the syntax of these complex syllable sequences is to use transition probabilities between the syllables. This is equivalent to the Markov model, in which each syllable is associated with one state, and the transition probabilities between the states do not depend on the state transition history. Here we analyze the song syntax in a Bengalese finch. We show that the Markov model fails to capture the statistical properties of the syllable sequences. Instead, a state transition model that accurately describes the statistics of the syllable sequences includes adaptation of the self-transition probabilities when states are repeatedly revisited, and allows associations of more than one state to the same syllable. Such a model does not increase the model complexity significantly. Mathematically, the model is a partially observable Markov model with adaptation (POMMA). The success of the POMMA supports the branching chain network hypothesis of how syntax is controlled within the premotor song nucleus HVC, and suggests that adaptation and many-to-one mapping from neural substrates to syllables are important features of the neural control of complex song syntax