Disturbance theory for ecosystem ecologists: A primer.

Understanding what regulates ecosystem functional responses to disturbance is essential in this era of global change. However, many pioneering and still influential disturbance-related theorie proposed by ecosystem ecologists were developed prior to rapid global change, and before tools and metrics were available to test them. In light of new knowledge and conceptual advances across biological disciplines, we present four disturbance ecology concepts that are particularly relevant to ecosystem ecologists new to the field: (a) the directionality of ecosystem functional response to disturbance; (b) functional thresholds; (c) disturbance-succession interactions; and (d) diversity-functional stability relationships. We discuss how knowledge, theory, and terminology developed by several biological disciplines, when integrated, can enhance how ecosystem ecologists analyze and interpret functional responses to disturbance. For example, when interpreting thresholds and disturbance-succession interactions, ecosystem ecologists should consider concurrent biotic regime change, non-linearity, and multiple response pathways, typically the theoretical and analytical domain of population and community ecologists. Similarly, the interpretation of ecosystem functional responses to disturbance requires analytical approaches that recognize disturbance can promote, inhibit, or fundamentally change ecosystem functions. We suggest that truly integrative approaches and knowledge are essential to advancing ecosystem functional responses to disturbance.

How populist attitudes scales fail to capture support for populists in power.

Populist attitudes are generally measured in surveys through three necessary and non-compensatory elements of populism, namely anti-elitism, people-centrism, and Manicheanism. Using Comparative Study of Electoral Systems Module 5 (2016-2020) data for 30 countries, we evaluate whether this approach explains voting for populist parties across countries in Asia, Europe and the Americas. We show that the existing scales of populist attitudes effectively explain voting for populists in countries where populist leaders and parties are in opposition but fail to explain voting for populist parties in countries where they are in power. We argue that current approaches assume "the elite" to mean "politicians", thus failing to capture attitudes towards "non-political elites" often targeted by populists in office-in particular, journalists, academics/experts, bureaucrats, and corporate business leaders. The results reveal limits to the usefulness of existing survey batteries in cross-national studies of populism and emphasize the need to develop approaches that are more generalizable across political and national contexts.

Disturbance-accelerated succession increases the production of a temperate forest.

Many secondary deciduous forests of eastern North America are approaching a transition in which mature early-successional trees are declining, resulting in an uncertain future for this century-long carbon (C) sink. We initiated the Forest Accelerated Succession Experiment (FASET) at the University of Michigan Biological Station to examine the patterns and mechanisms underlying forest C cycling following the stem girdling-induced mortality of >6,700 early-successional Populus spp. (aspen) and Betula papyrifera (paper birch). Meteorological flux tower-based C cycling observations from the 33-ha treatment forest have been paired with those from a nearby unmanipulated forest since 2008. Following over a decade of observations, we revisit our core hypothesis: that net ecosystem production (NEP) would increase following the transition to mid-late-successional species dominance due to increased canopy structural complexity. Supporting our hypothesis, NEP was stable, briefly declined, and then increased relative to the control in the decade following disturbance; however, increasing NEP was not associated with rising structural complexity but rather with a rapid 1-yr recovery of total leaf area index as mid-late-successional Acer, Quercus, and Pinus assumed canopy dominance. The transition to mid-late-successional species dominance improved carbon-use efficiency (CUE = NEP/gross primary production) as ecosystem respiration declined. Similar soil respiration rates in control and treatment forests, along with species differences in leaf physiology and the rising relative growth rates of mid-late-successional species in the treatment forest, suggest changes in aboveground plant respiration and growth were primarily responsible for increases in NEP. We conclude that deciduous forests transitioning from early to middle succession are capable of sustained or increased NEP, even when experiencing extensive tree mortality. This adds to mounting evidence that aging deciduous forests in the region will function as C sinks for decades to come.

COVID-19, digital privacy, and the social limits on data-focused public health responses.

The implementation of digital contact tracing applications around the world to help reduce the spread of the COVID-19 pandemic represents one of the most ambitious uses of massive-scale citizen data ever attempted. There is major divergence among nations, however, between a "privacy-first" approach which protects citizens' data at the cost of extremely limited access for public health authorities and researchers, and a "data-first" approach which stores large amounts of data which, while of immeasurable value to epidemiologists and other researchers, may significantly intrude upon citizens' privacy. The lack of a consensus on privacy protection in the contact tracing process creates risks of non-compliance or deliberate obfuscation from citizens who fear revealing private aspects of their lives - a factor greatly exacerbated by recent major scandals over online privacy and the illicit use of citizens' digital information, which have heightened public consciousness of these issues and created significant new challenges for any collection of large-scale public data. While digital contact tracing for COVID-19 remains in its infancy, the lack of consensus around best practices for its implementation and for reassuring citizens of the protection of their privacy may already have impeded its capacity to contribute to the pandemic response.

Forest structure, diversity, and primary production in relation to disturbance severity.

Differential disturbance severity effects on forest vegetation structure, species diversity, and net primary production (NPP) have been long theorized and observed. Here, we examined these factors concurrently to explore the potential for a mechanistic pathway linking disturbance severity, changes in light environment, leaf functional response, and wood NPP in a temperate hardwood forest.Using a suite of measurements spanning an experimental gradient of tree mortality, we evaluated the direction and magnitude of change in vegetation structural and diversity indexes in relation to wood NPP. Informed by prior observations, we hypothesized that forest structural and species diversity changes and wood NPP would exhibit either a linear, unimodal, or threshold response in relation to disturbance severity. We expected increasing disturbance severity would progressively shift subcanopy light availability and leaf traits, thereby coupling structural and species diversity changes with primary production.Linear or unimodal changes in three of four vegetation structural indexes were observed across the gradient in disturbance severity. However, disturbance-related changes in vegetation structure were not consistently correlated with shifts in light environment, leaf traits, and wood NPP. Species diversity indexes did not change in response to rising disturbance severity.We conclude that, in our study system, the sensitivity of wood NPP to rising disturbance severity is generally tied to changing vegetation structure but not species diversity. Changes in vegetation structure are inconsistently coupled with light environment and leaf traits, resulting in mixed support for our hypothesized cascade linking disturbance severity to wood NPP.

Covariance in diurnal patterns of suicide-related expressions on Twitter and recorded suicide deaths.

Social media data is increasingly used to gain insights into trends in mental health, but prior studies aimed at confirming a link between online expression of suicidal ideation on social media and actual suicide deaths have been inconclusive. Using comprehensive six-year data sets of Twitter posts and suicide deaths in Japan, we examine the diurnal relationship between the proportional incidence of a suicide-related keyword, "kietai" ("I want to disappear"), and suicide deaths with an OLS regression model. We also use co-occurrence analysis to reveal changes in the linguistic context of the suicide-related keyword at different hours of the day. We find a clear diurnal pattern in the use of this suicide-related keyword, peaking between 1am and 5am. This diurnal trend is positively correlated with suicide deaths among younger cohorts (ages 15 to 44), but the correlation is negative among older adults (45 and over). The correlation among young adults strengthens when a delay between tweet incidence and suicide deaths is included. Compared to daytime tweets, nighttime tweets exhibited a stronger relationship between words related to self-disgust and words directly indicating suicidal intent. This study confirms the hypothesised link between online suicidal ideation and suicide death. Despite frequent flippant use of the keyword, the consistent correlation and the diurnal changes in the context of the keyword's usage demonstrate the value of social media data to the study of mental health trends in groups at risk of suicide.

Defining a spectrum of integrative trait-based vegetation canopy structural types.

Vegetation canopy structure is a fundamental characteristic of terrestrial ecosystems that defines vegetation types and drives ecosystem functioning. We use the multivariate structural trait composition of vegetation canopies to classify ecosystems within a global canopy structure spectrum. Across the temperate forest sub-set of this spectrum, we assess gradients in canopy structural traits, characterise canopy structural types (CST) and evaluate drivers and functional consequences of canopy structural variation. We derive CSTs from multivariate canopy structure data, illustrating variation along three primary structural axes and resolution into six largely distinct and functionally relevant CSTs. Our results illustrate that within-ecosystem successional processes and disturbance legacies can produce variation in canopy structure similar to that associated with sub-continental variation in forest types and eco-climatic zones. The potential to classify ecosystems into CSTs based on suites of structural traits represents an important advance in understanding and modelling structure-function relationships in vegetated ecosystems.

High rates of primary production in structurally complex forests.

Structure-function relationships are central to many ecological paradigms. Chief among these is the linkage of net primary production (NPP) with species diversity and canopy structure. Using the National Ecological Observatory Network (NEON) as a subcontinental-scale research platform, we examined how temperate-forest NPP relates to several measures of site-level canopy structure and tree species diversity. Novel multidimensional canopy traits describing structural complexity, most notably canopy rugosity, were more strongly related to site NPP than were species diversity measures and other commonly characterized canopy structural features. The amount of variation in site-level NPP explained by canopy rugosity alone was 83%, which was substantially greater than that explained individually by vegetation area index (31%) or Shannon's index of species diversity (30%). Forests that were more structurally complex, had higher vegetation-area indices, or were more diverse absorbed more light and used light more efficiently to power biomass production, but these relationships were most strongly tied to structural complexity. Implications for ecosystem modeling and management are wide ranging, suggesting structural complexity traits are broad, mechanistically robust indicators of NPP that, in application, could improve the prediction and management of temperate forest carbon sequestration.

An analysis of enhanced tree trimming effectiveness on reducing power outages.

We evaluated the effectiveness of an enhanced tree trimming (ETT) program for its ability to reduce tree-related power outages, and thereby improve resilience, on an electric utility distribution system during storm events. Evaluations encompassed thirteen years of trimming (i.e. 2005-2017) data and were performed for both backbone and lateral utility lines. Backbones included all three phase lines between a substation and a faultable device whereas all other lines were considered laterals. The study site spanned the entire state of Connecticut, where the dominant vegetation is temperate deciduous forest. We controlled for variations in weather, tree cover, and wire type, by pairing ETT-treated zones with nearby untreated zones. ETT-treated conductors had storm outage rates that were 0.07-0.36 outages/km/year lower than untreated conductors or 35-180% lower than the service-area's average annual outage rate for untreated conductors. ETT-treatment was associated with lower outage rates for "minor" outage types (i.e., blown fuse, tripped recloser, etc.) but the treatment effect was not statistically significant for "major" outage types (damaged poles or wires). System-wide ETT application, for the approximately 27,000 km of conductors in the study area, was predicted to reduce annual storm-related outages by an average of 81-104 and 318-759 outages/year for backbone and lateral lines, respectively. Our study provided a robust empirical evaluation of ETT and also proposes a geospatial methodology that controls for variations in weather and environment.

Patterns and predictors of fleshy fruit phenology at five international botanical gardens.

PREMISE OF THE STUDY: To improve our understanding of the patterns and drivers of fleshy fruit phenology, we examined the sequence, patterns across years and locations, and drivers of fruiting times at five botanical gardens on three continents. METHODS: We monitored four stages of fruit phenology for 406 temperate, fleshy-fruited, woody plant species in 2014 and 2015. KEY RESULTS: Across all gardens, ripe fruits were present from May to March of the following year, with peak fruiting durations ranging from under 1 week to over 150 days. Species-level first fruiting and onset of peak fruiting dates were strongly associated with one another within sites and were more consistent between years and sites than the end of peak fruiting and last fruiting date. The order of fruiting among species between years and gardens was moderately consistent, and both peak fruiting times and fruiting durations were found to be phylogenetically conserved. CONCLUSIONS: The consistent order of fruiting among species between years and locations indicates species-specific phenological responses to environmental conditions. Wide variation in fruiting times across species and in the duration of peak fruiting reinforces the importance of understanding how plant phenology impacts dispersers and monitoring the health and consistency of these interactions.

Tracking the Werther Effect on social media: Emotional responses to prominent suicide deaths on twitter and subsequent increases in suicide.

Rises in suicide rates following media reports of the deaths by suicide of public figures are a well-documented phenomenon. However, it remains unclear why, or by what exact mechanism, celebrity suicides act to increase suicidal risk in the wider public due to the lack of data showing how the public understands and reacts to the suicide of well-known figures. This study used a supervised machine learning approach to investigate the emotional content of almost 1 million messages sent on Twitter related to the suicides of 18 prominent individuals in Japan between 2010 and 2014. The results revealed that different demographic characteristics of the deceased person (age, gender, and occupation) resulted in significant differences in emotional response; notably that the suicides of younger people, of women and of people in entertainment careers created more emotional responses (measured as a ratio of emotionally-coded tweets within the overall volume of tweets for each case) than for older people, men, and those in other careers. Moreover, certain types of emotional response were shown to correlate to subsequent rises in the national suicide counts, with "surprised" reactions being positively correlated with the suicide counts, while a high proportion of polite messages of condolence were negatively correlated. The study demonstrates the importance of, and describes a methodology for, considering the content of social media messages, not just their volume, in research into the mechanism by which these widely-reported deaths increase suicide risk in the broader public.

Effects of canopy structure and species diversity on primary production in upper Great Lakes forests.

Canopy structure and tree species diversity, shaped by succession, disturbance, and community composition, are linked to numerous ecosystem functions, including net primary production (NPP). Understanding of how ecosystem structural metrics are interrelated and mechanistically link to NPP, however, is incomplete. We characterized leaf area index (LAI), Simpson's index of Diversity (D', a measure of species diversity), and canopy rugosity (Rc, a measure of canopy physical complexity) in 11 forest stands comprising two chronosequences varying in establishing disturbance, and in three late successional communities. We related LAI, D', and Rc to wood NPP (NPPw), and examined whether absorption of photosynthetically active radiation and light use-efficiency (LUE) link NPPw with ecosystem structure. We found that recovery of LAI and D' was delayed following more severe establishing disturbances, but that the development of Rc was strikingly conserved regardless of disturbance, converging on a common mean value in late-successional stands irrespective of differences in leaf area index and species diversity. LAI was significantly correlated with NPPw in each stage of ecosystem development, but NPPw was only correlated with Rc in early successional stages and with D' in late successional stages. Across all stands, NPPw was coupled with LAI and Rc, (but not D') through positive relationships with light absorption and LUE. We conclude by advocating for better integration of ecological disciplines investigating structure-function interactions, suggesting that improved understanding of such relationships will require ecologists to traverse disciplinary boundaries.

Net primary production of a temperate deciduous forest exhibits a threshold response to increasing disturbance severity.

The global carbon (C) balance is vulnerable to disturbances that alter terrestrial C storage. Disturbances to forests occur along a continuum of severity, from low-intensity disturbance causing the mortality or defoliation of only a subset of trees to severe stand- replacing disturbance that kills all trees; yet considerable uncertainty remains in how forest production changes across gradients of disturbance intensity. We used a gradient of tree mortality in an upper Great Lakes forest ecosystem to: (1) quantify how aboveground wood net primary production (ANPP,) responds to a range of disturbance severities; and (2) identify mechanisms supporting ANPPw resistance or resilience following moderate disturbance. We found that ANPPw declined nonlinearly with rising disturbance severity, remaining stable until >60% of the total tree basal area senesced. As upper canopy openness increased from disturbance, greater light availability to the subcanopy enhanced the leaf-level photosynthesis and growth of this formerly light-limited canopy stratum, compensating for upper canopy production losses and a reduction in total leaf area index (LAI). As a result, whole-ecosystem production efficiency (ANPPw/LAI) increased with rising disturbance severity, except in plots beyond the disturbance threshold. These findings provide a mechanistic explanation for a nonlinear relationship between ANPPw, and disturbance severity, in which the physiological and growth enhancement of undisturbed vegetation is proportional to the level of disturbance until a threshold is exceeded. Our results have important ecological and management implications, demonstrating that in some ecosystems moderate levels of disturbance minimally alter forest production.

Quantifying canopy complexity and effects on productivity and resilience in late-successional hemlock-hardwood forests.

The regrowing forests of eastern North America have been an important global C sink over the past 100+ years, but many are now transitioning into late succession. The consequences of this transition are unclear due to uncertainty around the C dynamics of old- growth forests. Canopy structural complexity (CSC) has been shown to be an important source of variability in C dynamics in younger forests (e.g., in productivity and resilience to disturbance), but its role in late-successional forests has not been widely addressed. We investigated patterns of CSC in two old-growth forest landscapes in the Upper Peninsula of Michigan, USA, to assess factors associated with CSC and its influence on productivity and disturbance resilience (to moderate-severity windstorm). CSC was quantified using a portable below-canopy LiDAR (PCL) system in 65 plots that also had long-term (50-70+ years). inventory data, which were used to quantify aboveground net primary productivity (ANPP), disturbance history, and stand characteristics. We found high and variable CSC relative to younger forests across a suite of PCL-derived metrics. Variation in CSC was driven by species composition and size structure, rather than disturbance history or site characteristics. Recent moderate severity wind disturbance decreased plot-scale CSC, but increased stand-scale variation in CSC. The strong positive correlation between CSC and productivity illustrated in younger forests was not present in undisturbed portions of these late-successional ecosystems. Moderate severity disturbance appeared to reestablish the positive link between CSC and productivity, but this relationship was scale and severity dependent. A positive CSC-productivity relationship was evident at the plot scale with low-severity, dispersed disturbance, but only at a patch scale in more severely disturbed areas. CSC does not appear to strongly correlate With variation in productivity in undisturbed old-growth forests, but may play a very important (and scale/severity-dependent) role in their response to disturbance. Understanding potential, drivers and consequences of CSC in late-successional forests will inform management focused on promoting complexity and old-growth conditions, and illustrate potential inipacts of such treatments on regional C dynamics.

Substantial variation in leaf senescence times among 1360 temperate woody plant species: implications for phenology and ecosystem processes.

BACKGROUND AND AIMS: Autumn leaf senescence marks the end of the growing season in temperate ecosystems. Its timing influences a number of ecosystem processes, including carbon, water and nutrient cycling. Climate change is altering leaf senescence phenology and, as those changes continue, it will affect individual woody plants, species and ecosystems. In contrast to spring leaf out times, however, leaf senescence times remain relatively understudied. Variation in the phenology of leaf senescence among species and locations is still poorly understood. METHODS: Leaf senescence phenology of 1360 deciduous plant species at six temperate botanical gardens in Asia, North America and Europe was recorded in 2012 and 2013. This large data set was used to explore ecological and phylogenetic factors associated with variation in leaf senescence. KEY RESULTS: Leaf senescence dates among species varied by 3 months on average across the six locations. Plant species tended to undergo leaf senescence in the same order in the autumns of both years at each location, but the order of senescence was only weakly correlated across sites. Leaf senescence times were not related to spring leaf out times, were not evolutionarily conserved and were only minimally influenced by growth habit, wood anatomy and percentage colour change or leaf drop. These weak patterns of leaf senescence timing contrast with much stronger leaf out patterns from a previous study. CONCLUSIONS: The results suggest that, in contrast to the broader temperature effects that determine leaf out times, leaf senescence times are probably determined by a larger or different suite of local environmental effects, including temperature, soil moisture, frost and wind. Determining the importance of these factors for a wide range of species represents the next challenge for understanding how climate change is affecting the end of the growing season and associated ecosystem processes.

Leaf out times of temperate woody plants are related to phylogeny, deciduousness, growth habit and wood anatomy.

Leaf out phenology affects a wide variety of ecosystem processes and ecological interactions and will take on added significance as leaf out times increasingly shift in response to warming temperatures associated with climate change. There is, however, relatively little information available on the factors affecting species differences in leaf out phenology. An international team of researchers from eight Northern Hemisphere temperate botanical gardens recorded leaf out dates of c. 1600 woody species in 2011 and 2012. Leaf out dates in woody species differed by as much as 3 months at a single site and exhibited strong phylogenetic and anatomical relationships. On average, angiosperms leafed out earlier than gymnosperms, deciduous species earlier than evergreen species, shrubs earlier than trees, diffuse and semi-ring porous species earlier than ring porous species, and species with smaller diameter xylem vessels earlier than species with larger diameter vessels. The order of species leaf out was generally consistent between years and among sites. As species distribution and abundance shift due to climate change, interspecific differences in leaf out phenology may affect ecosystem processes such as carbon, water, and nutrient cycling. Our open access leaf out data provide a critical framework for monitoring and modelling such changes going forward.

Glutathione analogs in prokaryotes.

BACKGROUND: Oxygen is both essential and toxic to all forms of aerobic life and the chemical versatility and reactivity of thiols play a key role in both aspects. Cysteine thiol groups have key catalytic functions in enzymes but are readily damaged by reactive oxygen species (ROS). Low-molecular-weight thiols provide protective buffers against the hazards of ROS toxicity. Glutathione is the small protective thiol in nearly all eukaryotes but in prokaryotes the situation is far more complex. SCOPE OF REVIEW: This review provides an introduction to the diversity of low-molecular-weight thiol protective systems in bacteria. The topics covered include the limitations of cysteine as a protector, the multiple origins and distribution of glutathione biosynthesis, mycothiol biosynthesis and function in Actinobacteria, recent discoveries involving bacillithiol found in Firmicutes, new insights on the biosynthesis and distribution of ergothioneine, and the potential protective roles played by coenzyme A and other thiols. MAJOR CONCLUSIONS: Bacteria have evolved a diverse collection of low-molecular-weight protective thiols to deal with oxygen toxicity and environmental challenges. Our understanding of how many of these thiols are produced and utilized is still at an early stage. GENERAL SIGNIFICANCE: Extensive diversity existed among prokaryotes prior to evolution of the cyanobacteria and the development of an oxidizing atmosphere. Bacteria that managed to adapt to life under oxygen evolved, or acquired, the ability to produce a variety of small thiols for protection against the hazards of aerobic metabolism. Many pathogenic prokaryotes depend upon novel thiol protection systems that may provide targets for new antibacterial agents. This article is part of a Special Issue entitled Cellular functions of glutathione.

Characterization of BshA, bacillithiol glycosyltransferase from Staphylococcus aureus and Bacillus subtilis.

The first step during bacillithiol (BSH) biosynthesis involves the formation of N-acetylglucosaminylmalate from UDP-N-acetylglucosamine and l-malate and is catalyzed by a GT4 class glycosyltransferase enzyme (BshA). Recombinant Staphylococcus aureus and Bacillus subtilis BshA were highly specific and active with l-malate but the former showed low activity with d-glyceric acid and the latter with d-malate. We show that BshA is inhibited by BSH and similarly that MshA (first enzyme of mycothiol biosynthesis) is inhibited by the final product MSH.

Detoxification of toxins by bacillithiol in Staphylococcus aureus.

Bacillithiol (BSH), an α-anomeric glycoside of l-cysteinyl-d-glucosaminyl-l-malate, is a major low-molecular-mass thiol found in bacteria such as Bacillus sp., Staphylococcus aureus and Deinococcus radiodurans. Like other low-molecular-mass thiols such as glutathione and mycothiol, BSH is likely to be involved in protection against environmental toxins including thiol-reactive antibiotics. We report here a BSH-dependent detoxification mechanism in S. aureus. When S. aureus Newman strain was treated with monobromobimane and monochlorobimane, the cellular BSH was converted to the fluorescent S-conjugate BS-bimane. A bacillithiol conjugate amidase activity acted upon the BS-bimane to produce Cys-bimane, which was then acetylated by an N-acetyltransferase to generate N-acetyl-Cys-bimane, a mercapturic acid. An S. aureus mutant lacking BSH did not produce mercapturic acid when treated with monobromobimane and monochlorobimane, confirming the involvement of bacillithiol. Furthermore, treatment of S. aureus Newman with rifamycin, the parent compound of the first-line anti-tuberculosis drug, rifampicin, indicated that this thiol-reactive antibiotic is also detoxified in a BSH-dependent manner, since mercapturic acids of rifamycin were observed in the culture medium. These data indicate that toxins and thiol-reactive antibiotics are detoxified to less potent mercapturic acids in a BSH-dependent manner and then exported out of the cell in S. aureus.

The DinB superfamily includes novel mycothiol, bacillithiol, and glutathione S-transferases.

The superfamily of glutathione S-transferases has been the subject of extensive study; however, Actinobacteria produce mycothiol (MSH) in place of glutathione, and no mycothiol S-transferase (MST) has been identified. Using mycothiol and monochlorobimane as substrates, an MST activity was detected in extracts of Mycobacterium smegmatis and purified sufficiently to allow identification of MSMEG_0887, a member the DUF664 family of the DinB superfamily, as the MST. The identity of the M. smegmatis and homologous Mycobacterium tuberculosis (Rv0443) enzymes was confirmed by cloning, and the expressed proteins were found to be active with MSH but not bacillithiol (BSH) or glutathione (GSH). Bacillus subtilis YfiT is another member of the DinB superfamily, but this bacterium produces BSH. The YfiT protein was shown to have S-transferase activity with monochlorobimane when assayed with BSH but not with MSH or GSH. Enterococcus faecalis EF_3021 shares some homology with MSMEG_0887, but En. faecalis produces GSH but not MSH or BSH. Cloned and expressed EF_0321 was active with monochlorobimane and GSH but not with MSH or BSH. MDMPI_2 is another member of the DinB superfamily and has been previously shown to have mycothiol-dependent maleylpyruvate isomerase activity. Three of the eight families of the DinB superfamily include proteins shown to catalyze thiol-dependent metabolic or detoxification activities. Because more than two-thirds of the sequences assigned to the DinB superfamily are members of these families, it seems likely that such activity is dominant in the DinB superfamily.