Changes In Relative Activity Research Articles

Correlating structure and activity of pepsin enzyme in H2O and D2O for the study of gastric digestion

D2O, an isotope of H2O, is commonly used as a solvent in neutron scattering; the large difference in scattering length density between H and D can provide better contrast between the sample and the solvent. However, this is of concern for studies using enzymes as the use of D2O can influence protein interactions (due to differences in hydrogen bonding) and is therefore expected to affect the function, activity and solubility of enzymes. Neutron-based in vitro digestion assays on proteins, including those found in food or as oral protein and peptide drugs, often involve different solvents or pH conditions where the activity of the digestive enzyme may not be optimal. Herein, we investigate the structure and activity of the main gastric protease, porcine pepsin, in both H2O and D2O at pH values in the range 1 – 8. We showed that the activity of pepsin was lower in D2O, although the relative change in activity with pH was similar for both solvents. We demonstrated using a combination of SAXS and CD that this relative change in activity was not related to any structural change within the protein but was, rather, linked to relative changes in solubility of the protein.

Fox and cat responses to fox baiting intensity, rainfall and prey abundance in the Upper Warren, Western Australia

Context Invasive predators are major drivers of global biodiversity loss. Red foxes (Vulpes vulpes) and feral cats (Felis catus) have contributed to the decline and extinction of many native species in Australia. The deployment of poison baits to control fox populations is a widespread conservation tool, but the effects of baiting intensity, rainfall and prey abundance on baiting effectiveness remain poorly understood. Aims We aimed to understand what influences the association between fox baiting intensity, red fox activity and feral cat activity, to provide inferences about what might affect the effectiveness of fox baiting in reducing fox activity. Methods We used generalised linear models to assess how fox and cat activity changes in relation to fox baiting intensity, rainfall, native prey availability and distance to agricultural land over a 6-year period (2006–13) in the forest ecosystems of the Upper Warren region of south-western Australia. Key results We found that fox activity was negatively associated with rainfall in the previous 12 months and positively associated with prey abundance and fox baiting intensity. We also found an interaction between fox baiting and prey abundance, with fox activity increasing with prey activity in areas of low and moderate baiting intensity, but remaining constant in areas of high baiting intensity. Feral cat activity was positively associated with prey abundance and fox baiting intensity. We found no clear relationship between fox and cat activity. Conclusions The drivers of the association between fox baiting and fox activity are unclear because intense fox baiting was targeted at areas of known high fox abundance. However, our results indicate that intense fox baiting may be effective at decoupling the positive association between fox activity and prey abundance. Our results also suggest a positive association between fox baiting intensity and feral cat activity, thus supporting the case for integrated fox and cat management. Implications We caution interpretation of our results, but note that management of invasive predators could be improved by adjusting the intensity of management in response to changes in environmental conditions and local context (e.g. strategically conducting intense predator management where prey abundance is highest). Improved understanding of these associations requires a monitoring program with sufficient replication and statistical power to detect any treatment effects.

Impact of lockdown during COVID‐19 pandemic on physical activity and arrhythmia burden in heart failure patients

Restricted outdoor activity during COVID-19 related lockdown may accelerate heart failure (HF) progression and thereby increase cardiac arrhythmias. We analyzed the impact of March/April 2020 lockdown on physical activity and arrhythmia burden in HF patients treated with cardiac resynchronization therapy (CRT) devices with daily, automatic remote monitoring (RM) function. The study cohort included 405 HF patients enrolled in Observation of Clinical Routine Care for Heart Failure Patients Implanted with BIOTRONIK CRT Devices (BIO|STREAM.HF) registry in 16 countries, who had left ventricular ejection fraction (LVEF) ≤40% (mean 28.2±6.6%) and NYHA class II/III/IV (47.9%/49.6%/2.5%) before CRT pacemaker/defibrillator implantation. The analyzed RM data comprised physical activity detected by accelerometer, mean heart rate and nocturnal rate, PP variability, percentage of biventricular pacing, atrial high rate episode (AHRE) burden, ventricular extrasystoles and tachyarrhythmias, defibrillator shocks, and number of implant interrogations (i.e., follow-ups). Intraindividual differences in RM parameters before (4-week period) versus during (4-week period) lockdown were tested for statistical significance and independent predictors were identified. There was a significant relative change in activity (mean -6.5%, p<.001), AHRE burden (+17%, p=.013), and follow-up rate (-75%, p<.001) during lockdown, with no significant changes in other RM parameters. Activity decreased by ≥8min/day in 46.5% of patients; predictors were higher LVEF, lower NYHA class, no defibrillator indication, and more activity before lockdown. AHRE burden increased by ≥17min/day in 4.7% of patients; predictors were history of atrial fibrillation, higher LVEF, higher body mass index, and activity decrease during lockdown. Unfavorable changes in physical activity, AHRE burden, and follow-up rate were observed during lockdown, but not in ventricular arrhythmia.

Predicting Subjective Recovery from Lower Limb Surgery Using Consumer Wearables

Introduction: A major challenge in the monitoring of rehabilitation is the lack of long-term individual baseline data which would enable accurate and objective assessment of functional recovery. Consumer-grade wearable devices enable the tracking of individual everyday functioning prior to illness or other medical events which necessitate the monitoring of recovery trajectories. Methods: For 1,324 individuals who underwent surgery on a lower limb, we collected their Fitbit device data of steps, heart rate, and sleep from 26 weeks before to 26 weeks after the self-reported surgery date. We identified subgroups of individuals who self-reported surgeries for bone fracture repair (n = 355), tendon or ligament repair/reconstruction (n = 773), and knee or hip joint replacement (n = 196). We used linear mixed models to estimate the average effect of time relative to surgery on daily activity measurements while adjusting for gender, age, and the participant-specific activity baseline. We used a sub-cohort of 127 individuals with dense wearable data who underwent tendon/ligament surgery and employed XGBoost to predict the self-reported recovery time. Results: The 1,324 study individuals were all US residents, predominantly female (84%), white or Caucasian (85%), and young to middle-aged (mean age 36.2 years). We showed that 12 weeks pre- and 26 weeks post-surgery trajectories of daily behavioral measurements (steps sum, heart rate, sleep efficiency score) can capture activity changes relative to an individual’s baseline. We demonstrated that the trajectories differ across surgery types, recapitulate the documented effect of age on functional recovery, and highlight differences in relative activity change across self-reported recovery time groups. Finally, using a sub-cohort of 127 individuals, we showed that long-term recovery can be accurately predicted, on an individual level, only 1 month after surgery (AUROC 0.734, AUPRC 0.8). Furthermore, we showed that predictions are most accurate when long-term, individual baseline data are available. Discussion: Leveraging long-term, passively collected wearable data promises to enable relative assessment of individual recovery and is a first step towards data-driven intervention for individuals.

Separate Sets of Mutations Enhance Activity and Substrate Scope of Amine Dehydrogenase

AbstractMutations were introduced into the leucine amine dehydrogenase (L‐AmDH) derived from G. stearothermophilus leucine dehydrogenase (LeuDH) with the goals of increased activity and expanded substrate acceptance. A triple variant (L‐AmDH‐TV) including D32A, F101S, and C290V showed an average of 2.5‐fold higher activity toward aliphatic ketones and an 8.0 °C increase in melting temperature. L‐AmDH‐TV did not show significant changes in relative activity for different substrates. In contrast, L39A, L39G, A112G, and T133G in varied combinations added to L‐AmDH‐TV changed the shape of the substrate binding pocket. L‐AmDH‐TV was not active on ketones larger than 2‐hexanone. L39A and L39G enabled activity for straight‐chain ketones as large as 2‐decanone and in combination with A112G enabled activity toward longer branched ketones including 5‐methyl‐2‐octanone.

Exploring the Utility of IASI for Monitoring Volcanic SO2 Emissions

AbstractSatellite remote sensing is a valuable method for detecting and quantifying sulfur dioxide (SO2) emissions at volcanoes. The use of ultraviolet satellite instruments for monitoring purposes has been assessed in numerous studies, but there are advantages to using infrared measurements, including that they can operate at night and during high‐latitude winters. This study focuses on the Infrared Atmospheric Sounding Interferometer (IASI). Retrievals developed for this instrument have been shown to be successful when applied to large eruptions, but little has been done to explore their potential for detecting and quantifying emissions from smaller and lower altitude emissions or for the assessment of ongoing activity. Here a “fast” linear retrieval has been applied across the globe to detect volcanic sources of SO2. The results are dominated by emissions from explosive eruptions, but signals are also evident from weak eruptions, passive degassing, and anthropogenic activity. Ecuador and Kamchatka were selected for further study with a more processing intensive iterative retrieval which can quantify the SO2 amount. At Tungurahua in Ecuador, good agreement was seen between IASI, the Ozone Monitoring Instrument (OMI) and ground‐based flux data, demonstrating that the retrieval is capable of capturing relative changes in activity. Similarly, good agreement was found between IASI and OMI in Kamchatka. In this high‐latitude region, OMI is unable to operate for 3 or 4 months in each year. It is therefore suggested that IASI could be used alongside other instruments for evaluating changes in volcanic activity.

Ocular stability and set-point adaptation.

A fundamental challenge to the brain is how to prevent intrusive movements when quiet is needed. Unwanted limb movements such as tremor impair fine motor control and unwanted eye drifts such as nystagmus impair vision. A stable platform is also necessary to launch accurate movements. Accordingly, nature has designed control systems with agonist (excitation) and antagonist (inhibition) muscle pairs functioning in push-pull, around a steady level of balanced tonic activity, the set-point Sensory information can be organized similarly, as in the vestibulo-ocular reflex, which generates eye movements that compensate for head movements. The semicircular canals, working in coplanar pairs, one in each labyrinth, are reciprocally excited and inhibited as they transduce head rotations. The relative change in activity is relayed to the vestibular nuclei, which operate around a set-point of stable balanced activity. When a pathological imbalance occurs, producing unwanted nystagmus without head movement, an adaptive mechanism restores the proper set-point and eliminates the nystagmus. Here we used 90 min of continuous 7 T magnetic field labyrinthine stimulation (MVS) in normal humans to produce sustained nystagmus simulating vestibular imbalance. We identified multiple time-scale processes towards a new zero set-point showing that MVS is an excellent paradigm to investigate the neurobiology of set-point adaptation.This article is part of the themed issue 'Movement suppression: brain mechanisms for stopping and stillness'.

Patterns of US air transport across the economic unevenness of 2003–2013

ABSTRACTThis map summarizes the relative change in activity at 379 airports during the tumultuous economic period that lasted from 2003 to 2013 in the conterminous USA. Rather than treating airports only as individual nodes, the work identifies relative regional spatial change in airport activity based upon the combination of the percentage changes in three factors: departures, passenger levels, and available seats. The geographic results, calculated by kriging, show that the outcome over the period is not spatially uniform. In particular, the map shows that parts of the Rust Belt, Appalachia, and the Intermountain West fared relatively worse while the plains and coasts did somewhat better. The analysis expresses the fact that while footloose in the short-run, long-term adjustments in the airline industry, like those experienced across 2003–2013, did so in a spatially coherent way.

Variable interaction specificity and symbiont performance in Panamanian Trachymyrmex and Sericomyrmex fungus-growing ants.

BackgroundCooperative benefits of mutualistic interactions are affected by genetic variation among the interacting partners, which may have consequences for interaction-specificities across guilds of sympatric species with similar mutualistic life histories. The gardens of fungus-growing (attine) ants produce carbohydrate active enzymes that degrade plant material collected by the ants and offer them food in exchange. The spectrum of these enzyme activities is an important symbiont service to the host but may vary among cultivar genotypes. The sympatric occurrence of several Trachymyrmex and Sericomyrmex higher attine ants in Gamboa, Panama provided the opportunity to do a quantitative study of species-level interaction-specificity.ResultsWe genotyped the ants for Cytochrome Oxidase and their Leucoagaricus fungal cultivars for ITS rDNA. Combined with activity measurements for 12 carbohydrate active enzymes, these data allowed us to test whether garden enzyme activity was affected by fungal strain, farming ants or combinations of the two. We detected two cryptic ant species, raising ant species number from four to six, and we show that the 38 sampled colonies reared a total of seven fungal haplotypes that were different enough to represent separate Leucoagaricus species. The Sericomyrmex species and one of the Trachymyrmex species reared the same fungal cultivar in all sampled colonies, but the remaining four Trachymyrmex species largely shared the other cultivars. Fungal enzyme activity spectra were significantly affected by both cultivar species and farming ant species, and more so for certain ant-cultivar combinations than others. However, relative changes in activity of single enzymes only depended on cultivar genotype and not on the ant species farming a cultivar.ConclusionsAnt cultivar symbiont-specificity varied from almost full symbiont sharing to one-to-one specialization, suggesting that trade-offs between enzyme activity spectra and life-history traits such as desiccation tolerance, disease susceptibility and temperature sensitivity may apply in some combinations but not in others. We hypothesize that this may be related to ecological specialization in general, but this awaits further testing. Our finding of both cryptic ant species and extensive cultivar diversity underlines the importance of identifying all species-level variation before embarking on estimates of interaction specificity.Electronic supplementary materialThe online version of this article (doi:10.1186/s12862-014-0244-6) contains supplementary material, which is available to authorized users.

Enhanced activity of sol–gel prepared SnOx–TiO2 in photocatalytic methanol reforming

ObjectiveIn this contribution a simple method is elaborated for the preparation of Pt/SnOx–TiO2 catalysts which are active in the photocatalytic reforming of methanol. The region of low tin content (Sn/Ti atomic ratio: 0–0.015) is explored. Tin content related activity changes of the catalysts are discussed in terms of the results of structural characterization. MethodsTin-modified anatase TiO2 samples were obtained from sol–gel method by use of Ti-isopropoxide, SnCl4 or Sn(OBu)4 and citric acid in absolute ethanol solution followed by calcinations at 400°C in air. After loading Pt by impregnation the catalytic activity has been tested irradiating the slurry of the catalysts suspended in mixture of water and methanol by means of two different light sources, one working exclusively in the visible region and one in a broader spectral range including a UV-component, respectively. Hydrogen production activity has been correlated with structural data obtained from different characterization techniques such as ESR spectroscopy, XRD, XPS, SEM and Raman spectroscopy as well as diffuse reflectance UV–Vis spectrophotometry. ResultsAccording to ESR spectroscopic measurements the presence of oxygen vacancies was responsible for the slight visible light absorption. XRD, XPS, SEM and Raman spectroscopic results indicated that highly dispersed SnOx accumulated in the surface region of TiO2, no tin incorporation into the lattice was observed. The fundamental band gap was not influenced by the presence of tin. Loading of the tin-modified samples by 0.5wt% Pt increased the hydrogen production in comparison to 0.5wt% Pt/TiO2 samples. ConclusionsThe improvement in the catalytic activity was explained by formation of new tin related charge carrier traps leading to (i) prevention of electron–hole recombination and (ii) additional modification of the adsorption of reactants. Details of the two different working mechanisms are described. Practice implicationThe increased hydrogen production over the tin modified samples provides a promising step toward practical usage of light for hydrogen production.

Stabilization of Ferrochelatase Via Lysine Residues on the Carboxyl Terminal Extension

There are accumulating evidences suggesting that ferrochelatase is involved in different cellular functions other than heme biosynthesis. The carboxyl-terminal extension of the enzyme may play a role associated with its stability and signaling process. Two transcript variants encoding different isoforms have been found for this ferrochelatase-coding gene. Variants encoding different isoforms have been found for this gene and were implicated in novel cellular activities including gene expression. Deficiency in ferrochelatase variants is believed to be associated with its stability. In this study, mutagenesis study of the lysine residues at 357, 360 and 365 was conducted. The relative change in activity by measurement of fluorescence intensity of the mutant enzyme revealed that K365L retained > 40% activity, while the mutant K365I retained 24% activity. Other mutants with K357S and K360Q showed little enzyme activity (< 4%). The change in the tryptophan fluorescence intensity after Guanidinium chloride-induced denaturation of the mutants indicated that the enzyme become unfolded under the assay condition. The K365L mutation suggests a structural role of tryptophan in the enzyme. The stability of the enzyme attributed to conservation of lysine residues in the C-terminal extension of the enzyme. The lysine and tryptophan residues serve as a structure determinant of ferrochelatase.

Evidence for a Role of a Cortico-Subcortical Network for Automatic and Unconscious Motor Inhibition of Manual Responses

It is now clear that non-consciously perceived stimuli can bias our decisions. Although previous researches highlighted the importance of automatic and unconscious processes involved in voluntary action, the neural correlates of such processes remain unclear. Basal ganglia dysfunctions have long been associated with impairment in automatic motor control. In addition, a key role of the medial frontal cortex has been suggested by administrating a subliminal masked prime task to a patient with a small lesion restricted to the supplementary motor area (SMA). In this task, invisible masked arrows stimuli were followed by visible arrow targets for a left or right hand response at different interstimuli intervals (ISI), producing a traditional facilitation effect for compatible trials at short ISI and a reversal inhibitory effect at longer ISI. Here, by using fast event-related fMRI and a weighted parametric analysis, we showed BOLD related activity changes in a cortico-subcortical network, especially in the SMA and the striatum, directly linked to the individual behavioral pattern. This new imaging result corroborates previous works on subliminal priming using lesional approaches. This finding implies that one of the roles of these regions was to suppress a partially activated movement below the threshold of awareness.

Prefrontal electroencephalographic activity during the working memory processes involved in a sexually motivated task in male rats

The prefrontal cortex is involved in working memory functions, and several studies using food or drink as rewards have demonstrated that the rat is capable of performing tasks that involve working memory. Sexual activity is another highly-rewarding, motivated behaviour that has proven to be an efficient incentive in classical operant tasks. The objective of this study was to determine whether the functional activity of the medial prefrontal cortex (mPFC) changes in relation to the working memory processes involved in a sexually motivated task performed in male rats. Thus, male Wistar rats implanted in the mPFC were subjected to a nonmatching-to-sample task in a T-maze using sexual interaction as a reinforcer during a 4-day training period. On the basis of their performance during training, the rats were classified as 'good-learners' or 'bad-learners'. Only the good-learner rats showed an increase in the absolute power of the 8-13Hz band during both the sample and test runs; a finding that could be related to learning of the working memory elements entailed in the task. During the maintenance phase only (i.e., once the rule had been learned well), the good-learner rats also showed an increased correlation of the 8-13Hz band during the sample run, indicating that a high degree of coupling between the prefrontal cortices is necessary for the processing required to allow the rats to make correct decisions in the maintenance phase. Taken together, these data show that mPFC activity changes in relation to the working memory processes involved in a sexually motivated task in male rats.

Aging effects on functional auditory and visual processing using fMRI with variable sensory loading

IntroductionTraditionally, studies investigating the functional implications of age-related structural brain alterations have focused on higher cognitive processes; by increasing stimulus load, these studies assess behavioral and neurophysiological performance. In order to understand age-related changes in these higher cognitive processes, it is crucial to examine changes in visual and auditory processes that are the gateways to higher cognitive functions. This study provides evidence for age-related functional decline in visual and auditory processing, and regional alterations in functional brain processing, using non-invasive neuroimaging. MethodsUsing functional magnetic resonance imaging (fMRI), younger (n = 11; mean age = 31) and older (n = 10; mean age = 68) adults were imaged while observing flashing checkerboard images (passive visual stimuli) and hearing word lists (passive auditory stimuli) across varying stimuli presentation rates. ResultsYounger adults showed greater overall levels of temporal and occipital cortical activation than older adults for both auditory and visual stimuli. The relative change in activity as a function of stimulus presentation rate showed differences between young and older participants. In visual cortex, the older group showed a decrease in fMRI blood oxygen level dependent (BOLD) signal magnitude as stimulus frequency increased, whereas the younger group showed a linear increase. In auditory cortex, the younger group showed a relative increase as a function of word presentation rate, while older participants showed a relatively stable magnitude of fMRI BOLD response across all rates. When analyzing participants across all ages, only the auditory cortical activation showed a continuous, monotonically decreasing BOLD signal magnitude as a function of age. ConclusionsOur preliminary findings show an age-related decline in demand-related, passive early sensory processing. As stimulus demand increases, visual and auditory cortex do not show increases in activity in older compared to younger people. This may negatively impact on the fidelity of information available to higher cognitive processing. Such evidence may inform future studies focused on cognitive decline in aging.

Influence of late-life cognitive activity on cognitive health

To test the hypothesis that late-life participation in mentally stimulating activities affects subsequent cognitive health. Analyses are based on 1,076 older persons without dementia at study onset participating in a longitudinal cohort study. They completed annual clinical evaluations for a mean of 4.9 years. Each evaluation included administration of a self-report scale about participation in mentally stimulating activities and a battery of cognitive performance tests. Previously established measures of cognitively stimulating activity and cognitive function were derived. We assessed the temporal sequence of activity changes in relation to functional changes in a series of cross-lagged panel models adjusted for age, sex, and education. During the observation period, cognitive activity participation (estimate of mean annual change = -0.066, SE = 0.005, p < 0.001) and cognitive functioning (estimate = -0.077, SE = 0.005, p < 0.001) declined at rates that were moderately correlated (r = 0.44, p < 0.001). The level of cognitive activity in a given year predicted the level of global cognitive function in the following year, but the level of global cognition did not predict the subsequent level of cognitive activity participation. Cognitive activity showed the same pattern of unidirectional associations with measures of episodic and semantic memory, but its associations with working memory were bidirectional. The results suggest that more frequent mental stimulation in old age leads to better cognitive functioning.

The Activity and Cofactor Preferences of N-Acetyl-1-d-myo-inosityl-2-amino-2-deoxy-α-d-glucopyranoside Deacetylase (MshB) Change Depending on Environmental Conditions

Actinomycetes, such as Mycobacterium species, are Gram-positive bacteria that utilize the small molecule mycothiol (MSH) as their primary reducing agent. Consequently, the enzymes involved in MSH biosynthesis are targets for drug development. The metal-dependent enzyme N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-α-D-glucopyranoside deacetylase (MshB) catalyzes the hydrolysis of N-acetyl-1-D-myo-inosityl-2-amino-2-deoxy-α-D-glucopyranoside to form 1-D-myo-inosityl-2-amino-2-deoxy-α-D-glucopyranoside and acetate, the fourth overall step in MSH biosynthesis. Inhibitors of metalloenzymes typically contain a group that binds to the active site metal ion; thus, a comprehensive understanding of the native cofactor(s) of metalloenzymes is critical for the development of biologically effective inhibitors. Herein, we examined the effect of metal ions on the overall activity of MshB and probed the identity of the native cofactor. We found that the activity of MshB follows the trend Fe(2+) > Co(2+) > Zn(2+) > Mn(2+) and Ni(2+). Additionally, our results show that the identity of the cofactor bound to purified MshB is highly dependent on the purification conditions used (aerobic versus anaerobic), as well as the metal ion content of the medium during protein expression. MshB prefers Fe(2+) under anaerobic conditions regardless of the metal ion content of the medium and switches between Fe(2+) and Zn(2+) under aerobic conditions as the metal content of the medium is altered. These results indicate that the cofactor bound to MshB under biological conditions is dependent on environmental conditions, suggesting that MshB may be a cambialistic metallohydrolase that contains a dynamic cofactor. Consequently, biologically effective inhibitors will likely need to dually target Fe(2+)-MshB and Zn(2+)-MshB.

Kinetic studies of unmodified individual Escherichia coli β-galactosidase molecules in free solution

Assays were performed on individual Escherichia coli beta-galactosidase molecules at 2 different concentrations of the substrate DDAO-beta-D-galactoside using a free zone capillary electrophoresis-based protocol with an in-laboratory-constructed instrument utilizing laser-induced fluorescence detection. In a typical run, 2 enzyme molecules were injected into the capillary. They were separated from each other by a brief period of electrophoresis and incubated on the capillary in the presence of the substrate. They were then mobilized on the capillary into a zone of substrate at a different concentration, re-incubated, and the product peaks mobilized past the detector . The relative change in activity as the concentration was increased differed between molecules, suggesting differences in Km. In a different experiment, the capillary was filled with on average 13 enzyme molecules per run, incubated, and the activities of the individual molecules determined. The shapes of the distribution curves of single molecule activities obtained at different concentrations of the substrate resorufin-beta-D-galactoside were indistinguishable, suggesting a homogeneous Km. To explain why individual enzyme molecules behaved as if they were heterogeneous with respect to Km but the population behaved as if it were homogeneous, theoretical Michaelis-Menten curves were constructed. The curves for populations with heterogeneous Km values were found to be indistinguishable from that of a homogeneous population.

Escherichia coli Arabinose Isomerase and Staphylococcus aureus Tagatose-6-Phosphate Isomerase: Which is a Better Template for Directed Evolution of Non-Natural Substrate Isomerization?

Metallic and non-metallic isomerases can be used to produce commercially important monosaccharides. To determine which category of isomerase is more suitable as a template for directed evolution to improve enzymes for galactose isomerization, L-arabinose isomerase from Escherichia coli (ECAI; E.C. 5.3.1.4) and tagatose-6-phosphate isomerase from Staphylococcus aureus (SATI; E.C. 5.3.1.26) were chosen as models of a metallic and non-metallic isomerase, respectively. Random mutations were introduced into the genes encoding ECAI and SATI at the same rate, resulting in the generation of 515 mutants of each isomerase. The isomerization activity of each of the mutants toward a non-natural substrate (galactose) was then measured. With an average mutation rate of 0.2 mutations/kb, 47.5% of the mutated ECAIs showed an increase in activity compared with wild-type ECAI, and the remaining 52.5% showed a decrease in activity. Among the mutated SATIs, 58.6% showed an increase in activity, whereas 41.4% showed a decrease in activity. Mutant clones showing a significant change in relative activity were sequenced and specific increases in activity were measured. The maximum increase in activity achieved by mutation of ECAI was 130%, and that for SATI was 190%. Based on these results, the characteristics of the different isomerases are discussed in terms of their usefulness for directed evolution of non-natural substrate isomerization.

Regional cerebral glucose metabolism and anxiety symptoms in bipolar depression: Effects of levothyroxine

We examined the relationships between regional brain activity and anxiety in bipolar depressed patients receiving adjunctive treatment with levothyroxine. Regional brain activity was assessed with positron emission tomography and [ 18F]fluorodeoxyglucose in 10 euthyroid, depressed bipolar women before and after 7 weeks of adjunctive therapy with levothyroxine. The primary biological measures were relative (to global) regional radioactivity as a surrogate index of glucose metabolism in pre-selected brain regions. Relationships were assessed between regional brain activity and anxiety symptoms while controlling for depression severity. At baseline, Trait Anxiety Inventory measures covaried positively with relative brain activity bilaterally in the dorsal anterior cingulate, superior temporal gyri, parahippocampal gyri, amygdala, hippocampus, ventral striatum, and right insula; state anxiety showed a similar pattern. After treatment anxiety was improved significantly. Change in trait anxiety covaried positively with changes in relative activity in right amygdala and hippocampus. Change in state anxiety covaried positively with changes in relative activity in the hippocampus bilaterally and left thalamus, and negatively with changes in left middle frontal gyrus and right dorsal anterior cingulate. Results indicate that comorbid anxiety symptoms have specific regional cerebral metabolic correlates in bipolar depression and cannot only be explained exclusively by the depressive state of the patients.

Cell Surface Biliverdin Reductase Mediates Biliverdin-induced Anti-inflammatory Effects via Phosphatidylinositol 3-Kinase and Akt

Biliverdin reductase A (BVR) catalyzes the reduction of biliverdin (BV) to bilirubin (BR) in all cells. Others and we have shown that biliverdin is a potent anti-inflammatory molecule, however, the mechanism by which BV exerts its protective effects is unclear. We describe and elucidate a novel finding demonstrating that BVR is expressed on the external plasma membrane of macrophages (and other cells) where it quickly converts BV to BR. The enzymatic conversion of BV to BR on the surface by BVR initiates a signaling cascade through tyrosine phosphorylation of BVR on the cytoplasmic tail. Phosphorylated BVR in turn binds to the p85alpha subunit of phosphatidylinositol 3-kinase and activates downstream signaling to Akt. Using bacterial endotoxin (lipopolysaccharide) to initiate an inflammatory response in macrophages, we find a rapid increase in BVR surface expression. One of the mechanisms by which BV mediates its protective effects in response to lipopolysaccharide is through enhanced production of interleukin-10 (IL-10) the prototypical anti-inflammatory cytokine. IL-10 regulation is dependent in part on the activation of Akt. The effects of BV on IL-10 expression are lost with blockade of Akt. Inhibition of surface BVR with RNA interference attenuates BV-induced Akt signaling and IL-10 expression and in vivo negates the cytoprotective effects of BV in models of shock and acute hepatitis. Collectively, our findings elucidate a potentially important new molecular mechanism by which BV, through the enzymatic activity and phosphorylation of surface BVR (BVR)(surf) modulates the inflammatory response.