Wild foods contribute to women's higher dietary diversity in India.

Wild foods, from forests and common lands, can contribute to food and nutrition security. Most previous studies have established correlations between wild food consumption and children's dietary diversity in Africa, but other groups and geographic contexts remain understudied. Here a rigorous quasi-experimental method was combined with monthly interval data to assess the contribution of wild foods to women's diets. We collected 24 h diet recall data monthly, from November 2016 to November 2017, from 570 households in East India. We found that wild foods contributed positively to diets, especially in June and July (when consumption of wild foods was highest). Women who consumed wild foods had higher average dietary diversity scores (13% and 9% higher in June and July, respectively) and were more likely to consume nutrient-dense, dark-green leafy vegetables than those who did not. Our results underscore the importance of policies that increase knowledge of wild foods and protect people's rights to access forests and other common lands for improved nutrition.

A guide to qualitative attribution methods for evaluation in conservation.

Knowledge of what conservation interventions improve biodiversity outcomes, and in which circumstances, is imperative. Experimental and quasi-experimental methods are increasingly used to establish causal inference and build the evidence base on the effectiveness of interventions, but their ability to provide insight into how and under what conditions an intervention should be implemented to improve biodiversity outcomes faces limitations. A suite of attribution methods that leverage qualitative methods for causal inference is available but underutilized in conversation impact evaluation. This article provides a guide to 5 such qualitative attribution methods: contribution analysis, process tracing, realist evaluation, qualitative comparative analysis, and most significant change. It defines and introduces each method and then illustrates how they could be applied through a case study of community conservancies in Namibia. This guide provides examples of how qualitative attribution methods can advance knowledge of what works, in which contexts, and why in biodiversity conservation.

Guía para los métodos de atribución cualitativa para la evaluación en conservación Resumen El conocimiento sobre cuáles intervenciones de conservación son las que aumentan los resultados en biodiversidad y en cuáles circunstancias sucede esto es imperativo. Los métodos experimentales y semiexperimentales se usan cada vez más para establecer la inferencia casual y construir la base de evidencias sobre la efectividad de las intervenciones, pero su capacidad para proporcionar conocimiento sobre cómo y bajo cuáles condiciones se debería implementar una intervención para aumentar los resultados en biodiversidad enfrenta limitaciones. Un conjunto de métodos de atribución que impulse los métodos cualitativos para la inferencia casual se encuentra disponible pero poco utilizado en la evaluación de los impactos de conservación. Este artículo proporciona una guía para cinco de los métodos de atribución cualitativa: análisis de contribución, rastreo de procesos, evaluación realista, análisis cualitativo comparativo y el cambio más significativo. La guía define e introduce cada método y después ilustra cómo podrían aplicarse mediante un estudio de caso de conservación en Namibia. Esta guía proporciona ejemplos de cómo los métodos de atribución cualitativa pueden incrementar el conocimiento de qué funciona, en cuáles contextos y por qué en la conservación d la biodiversidad.

Characterization of an active spore photoproduct lyase, a DNA repair enzyme in the radical S-adenosylmethionine superfamily.

The major photoproduct in UV-irradiated Bacillus spore DNA is a unique thymine dimer called spore photoproduct (SP, 5-thyminyl-5,6-dihydrothymine). The enzyme spore photoproduct lyase (SP lyase) has been found to catalyze the repair of SP dimers to thymine monomers in a reaction that requires S-adenosylmethionine. We present here the first detailed characterization of catalytically active SP lyase, which has been anaerobically purified from overexpressing Escherichia coli. Anaerobically purified SP lyase is monomeric and is red-brown in color. The purified enzyme contains approximately 3.1 iron and 3.0 acid-labile S(2-) per protein and has a UV-visible spectrum characteristic of iron-sulfur proteins (410 nm (11.9 mM(-1) cm(-1)) and 450 nm (10.5 mM(-1) cm(-1))). The X-band EPR spectrum of the purified enzyme shows a nearly isotropic signal (g = 2.02) characteristic of a [3Fe-4S]1+ cluster; reduction of SP lyase with dithionite results in the appearance of a new EPR signal (g = 2.03, 1.93, and 1.89) with temperature dependence and g values consistent with its assignment to a [4Fe-4S]1+ cluster. The reduced purified enzyme is active in SP repair, with a specific activity of 0.33 micromol/min/mg. Only a catalytic amount of S-adenosylmethionine is required for DNA repair, and no irreversible cleavage of S-adenosylmethionine into methionine and 5'-deoxyadenosine is observed during the reaction. Label transfer from [5'-3H]S-adenosylmethionine to repaired thymine is observed, providing evidence to support a mechanism in which a 5'-deoxyadenosyl radical intermediate directly abstracts a hydrogen from SP C-6 to generate a substrate radical, and subsequent to radical-mediated beta-scission, a product thymine radical abstracts a hydrogen from 5'-deoxyadenosine to regenerate the 5'-deoxyadenosyl radical. Together, our results support a mechanism in which S-adenosylmethionine acts as a catalytic cofactor, not a substrate, in the DNA repair reaction.

Direct H atom abstraction from spore photoproduct C-6 initiates DNA repair in the reaction catalyzed by spore photoproduct lyase: evidence for a reversibly generated adenosyl radical intermediate.

Spore photoproduct (SP) lyase, which catalyzes the direct reversal of SP (5-thyminyl-5,6-dihydrothymine) to thymine monomers, is the only identified nonphotoactivatable pyrimidine dimer lyase. Unlike DNA photolyase, SP lyase does not contain a flavin cofactor and does not require light for activation. Instead, preliminary studies point to the presence of an iron-sulfur cluster in SP lyase and the requirement for S-adenosylmethionine (AdoMet) for catalytic activity, suggesting that SP lyase belongs to the growing group of iron-sulfur cluster and AdoMet-dependent radical enzymes. Here we provide evidence for the role of AdoMet as a reversible deoxyadenosyl radical generator, which initiates repair by hydrogen atom abstraction from C-6 of SP. Reaction of 6-(3)H-SP, but not methyl-(3)H-SP, with SP lyase and AdoMet results in transfer of (3)H to AdoMet, while no tritiated 5'-deoxyadenosine is observed. When 5'-tritiated AdoMet is used in the reaction with unlabeled SP, transfer of (3)H into the repaired thymine monomers is observed. These results point to the reversible generation of a 5'-deoxyadenosyl radical intermediate, which reacts directly with the DNA lesion to initiate a radical-mediated beta-scission. We also demonstrate that AdoMet is a catalytic cofactor that is not consumed during turnover. Together, these results support a novel radical-based mechanism for the repair of UV-induced DNA damage.

Electron-nuclear double resonance spectroscopic evidence that S-adenosylmethionine binds in contact with the catalytically active [4Fe-4S](+) cluster of pyruvate formate-lyase activating enzyme.

Pyruvate formate-lyase activating enzyme (PFL-AE) is a representative member of an emerging family of enzymes that utilize iron-sulfur clusters and S-adenosylmethionine (AdoMet) to initiate radical catalysis. Although these enzymes have diverse functions, evidence is emerging that they operate by a common mechanism in which a [4Fe-4S](+) interacts with AdoMet to generate a 5'-deoxyadenosyl radical intermediate. To date, however, it has been unclear whether the iron-sulfur cluster is a simple electron-transfer center or whether it participates directly in the radical generation chemistry. Here we utilize electron paramagnetic resonance (EPR) and pulsed 35 GHz electron-nuclear double resonance (ENDOR) spectroscopy to address this question. EPR spectroscopy reveals a dramatic effect of AdoMet on the EPR spectrum of the [4Fe-4S](+) of PFL-AE, changing it from rhombic (g = 2.02, 1.94, 1.88) to nearly axial (g = 2.01, 1.88, 1.87). (2)H and (13)C ENDOR spectroscopy was performed on [4Fe-4S](+)-PFL-AE (S = (1)/(2)) in the presence of AdoMet labeled at the methyl position with either (2)H or (13)C (denoted [1+/AdoMet]). The observation of a substantial (2)H coupling of approximately 1 MHz ( approximately 6-7 MHz for (1)H), as well as hyperfine-split signals from the (13)C, manifestly require that AdoMet lie close to the cluster. (2)H and (13)C ENDOR data were also obtained for the interaction of AdoMet with the diamagnetic [4Fe-4S](2+) state of PFL-AE, which is visualized through cryoreduction of the frozen [4Fe-4S](2+)/AdoMet complex to form the reduced state (denoted [2+/AdoMet](red)) trapped in the structure of the oxidized state. (2)H and (13)C ENDOR spectra for [2+/AdoMet](red) are essentially identical to those obtained for the [1+/AdoMet] samples, showing that the cofactor binds in the same geometry to both the 1+ and 2+ states of PFL-AE. Analysis of 2D field-frequency (13)C ENDOR data reveals an isotropic hyperfine contribution, which requires that AdoMet lie in contact with the cluster, weakly interacting with it through an incipient bond/antibond. From the anisotropic hyperfine contributions for the (2)H and (13)C ENDOR, we have estimated the distance from the closest methyl proton of AdoMet to the closest iron of the cluster to be approximately 3.0-3.8 A, while the distance from the methyl carbon to the nearest iron is approximately 4-5 A. We have used this information to construct a model for the interaction of AdoMet with the [4Fe-4S](2+/+) cluster of PFL-AE and have proposed a mechanism for radical generation that is consistent with these results.