Arabidopsis glutathione reductase 2 is indispensable in plastids, while mitochondrial glutathione is safeguarded by additional reduction and transport systems.

A highly negative glutathione redox potential (EGSH ) is maintained in the cytosol, plastids and mitochondria of plant cells to support fundamental processes, including antioxidant defence, redox regulation and iron-sulfur cluster biogenesis. Out of two glutathione reductase (GR) proteins in Arabidopsis, GR2 is predicted to be dual-targeted to plastids and mitochondria, but its differential roles in these organelles remain unclear. We dissected the role of GR2 in organelle glutathione redox homeostasis and plant development using a combination of genetic complementation and stacked mutants, biochemical activity studies, immunogold labelling and in vivo biosensing. Our data demonstrate that GR2 is dual-targeted to plastids and mitochondria, but embryo lethality of gr2 null mutants is caused specifically in plastids. Whereas lack of mitochondrial GR2 leads to a partially oxidised glutathione pool in the matrix, the ATP-binding cassette (ABC) transporter ATM3 and the mitochondrial thioredoxin system provide functional backup and maintain plant viability. We identify GR2 as essential in the plastid stroma, where it counters GSSG accumulation and developmental arrest. By contrast a functional triad of GR2, ATM3 and the thioredoxin system in the mitochondria provides resilience to excessive glutathione oxidation.

Regional and Racial Inequality in Infectious Disease Mortality in U.S. Cities, 1900-1948.

In the first half of the twentieth century, the rate of death from infectious disease in the United States fell precipitously. Although this decline is well-known and well-documented, there is surprisingly little evidence about whether it took place uniformly across the regions of the United States. We use data on infectious disease deaths from all reporting U.S. cities to describe regional patterns in the decline of urban infectious mortality from 1900 to 1948. We report three main results. First, urban infectious mortality was higher in the South in every year from 1900 to 1948. Second, infectious mortality declined later in southern cities than in cities in the other regions. Third, comparatively high infectious mortality in southern cities was driven primarily by extremely high infectious mortality among African Americans. From 1906 to 1920, African Americans in cities experienced a rate of death from infectious disease that was greater than what urban whites experienced during the 1918 flu pandemic.

Tenancy, Marriage, and the Boll Weevil Infestation, 1892-1930.

In the early twentieth century, the cotton-growing regions of the U.S. South were dominated by families of tenant farmers. Tenant farming created opportunities and incentives for prospective tenants to marry at young ages. These opportunities and incentives especially affected African Americans, who had few alternatives to working as tenants. Using complete-count Census of Population data from 1900-1930 and Census of Agriculture data from 1889-1929, we find that increases in tenancy over time increased the prevalence of marriage among young African Americans. We then study how marriage was affected by one of the most notorious disruptions to southern agriculture at the turn of the century: the boll weevil infestation of 1892-1922. Using historical Department of Agriculture maps, we show that the boll weevil's arrival reduced the share of farms worked by tenants as well as the share of African Americans who married at young ages. When the boll weevil infestation altered African Americans' opportunities and incentives to marry, the share of African Americans who married young fell accordingly. Our results provide new evidence about the effect of economic and political institutions on demographic transformations.

Geographic Variation in the Cumulative Risk of Imprisonment and Parental Imprisonment in the United States.

This article reports estimates of the cumulative risk of imprisonment and parental imprisonment for demographic groups in four regions and four states. Regional and state-level cumulative risks were markedly higher for African Americans and Latinos than for whites. African Americans faced the highest cumulative risks of imprisonment in the Midwest, Northeast, and two southern states. Latinos were most likely to serve time in state prison in the West, where their cumulative risk was comparable to that of African Americans. Latino children had a relatively high risk of having a parent imprisoned in the Northeast as well. Racial disparities in the cumulative risk of imprisonment and parental imprisonment did not increase linearly with increases in the cumulative risk for all groups.

Tenancy and African American Marriage in the Postbellum South.

The pervasiveness of tenancy in the postbellum South had countervailing effects on marriage between African Americans. Tenancy placed severe constraints on African American women's ability to find independent agricultural work. Freedwomen confronted not only planters' reluctance to contract directly with women but also whites' refusal to sell land to African Americans. Marriage consequently became one of African American women's few viable routes into the agricultural labor market. We find that the more counties relied on tenant farming, the more common was marriage among their youngest and oldest African American residents. However, many freedwomen resented their subordinate status within tenant marriages. Thus, we find that tenancy contributed to union dissolution as well as union formation among freedpeople. Microdata tracing individuals' marital transitions are consistent with these county-level results.

Autism gene variant causes hyperserotonemia, serotonin receptor hypersensitivity, social impairment and repetitive behavior.

Fifty years ago, increased whole-blood serotonin levels, or hyperserotonemia, first linked disrupted 5-HT homeostasis to Autism Spectrum Disorders (ASDs). The 5-HT transporter (SERT) gene (SLC6A4) has been associated with whole blood 5-HT levels and ASD susceptibility. Previously, we identified multiple gain-of-function SERT coding variants in children with ASD. Here we establish that transgenic mice expressing the most common of these variants, SERT Ala56, exhibit elevated, p38 MAPK-dependent transporter phosphorylation, enhanced 5-HT clearance rates and hyperserotonemia. These effects are accompanied by altered basal firing of raphe 5-HT neurons, as well as 5HT(1A) and 5HT(2A) receptor hypersensitivity. Strikingly, SERT Ala56 mice display alterations in social function, communication, and repetitive behavior. Our efforts provide strong support for the hypothesis that altered 5-HT homeostasis can impact risk for ASD traits and provide a model with construct and face validity that can support further analysis of ASD mechanisms and potentially novel treatments.

A perturbation in glutathione biosynthesis disrupts endoplasmic reticulum morphology and secretory membrane traffic in Arabidopsis thaliana.

To identify potentially novel and essential components of plant membrane trafficking mechanisms we performed a GFP-based forward genetic screen for seedling-lethal biosynthetic membrane trafficking mutants in Arabidopsis thaliana. Amongst these mutants, four recessive alleles of GSH2, which encodes glutathione synthase (GSH2), were recovered. Each allele was characterized by loss of the typical polygonal endoplasmic reticulum (ER) network and the accumulation of swollen ER-derived bodies which accumulated a soluble secretory marker. Since GSH2 is responsible for converting γ-glutamylcysteine (γ-EC) to glutathione (GSH) in the glutathione biosynthesis pathway, gsh2 mutants exhibited γ-EC hyperaccumulation and GSH deficiency. Redox-sensitive GFP revealed that gsh2 seedlings maintained redox poise in the cytoplasm but were more sensitive to oxidative challenge. Genetic and pharmacological evidence indicated that γ-EC accumulation rather than GSH deficiency was responsible for the perturbation of ER morphology. Use of soluble and membrane-bound ER markers suggested that the swollen ER bodies were derived from ER fusiform bodies. Despite the gross perturbation of ER morphology, gsh2 seedlings did not suffer from constitutive oxidative ER stress or lack of an unfolded protein response, and homozygotes for the weakest allele could be propagated. The link between glutathione biosynthesis and ER morphology and function is discussed.

Plant homologs of the Plasmodium falciparum chloroquine-resistance transporter, PfCRT, are required for glutathione homeostasis and stress responses.

In Arabidopsis thaliana, biosynthesis of the essential thiol antioxidant, glutathione (GSH), is plastid-regulated, but many GSH functions, including heavy metal detoxification and plant defense activation, depend on cytosolic GSH. This finding suggests that plastid and cytosol thiol pools are closely integrated and we show that in Arabidopsis this integration requires a family of three plastid thiol transporters homologous to the Plasmodium falciparum chloroquine-resistance transporter, PfCRT. Arabidopsis mutants lacking these transporters are heavy metal-sensitive, GSH-deficient, and hypersensitive to Phytophthora infection, confirming a direct requirement for correct GSH homeostasis in defense responses. Compartment-specific measurements of the glutathione redox potential using redox-sensitive GFP showed that knockout of the entire transporter family resulted in a more oxidized glutathione redox potential in the cytosol, but not in the plastids, indicating the GSH-deficient phenotype is restricted to the cytosolic compartment. Expression of the transporters in Xenopus oocytes confirmed that each can mediate GSH uptake. We conclude that these transporters play a significant role in regulating GSH levels and the redox potential of the cytosol.

Enhanced methane generation during theromophilic co-digestion of confectionary waste and grease-trap fats and oils with municipal wastewater sludge.

Recent interest in carbon-neutral biofuels has revived interest in co-digestion for methane generation. At wastewater treatment facilities, organic wastes may be co-digested with sludge using established anaerobic digesters. However, changes to organic loadings may induce digester instability, particularly for thermophilic digesters. To examine this problem, thermophilic (55 degrees C) co-digestion was studied for two food-industry wastes in semi-continuous laboratory digesters; in addition, the wastes' biochemical methane potentials were tested. Wastes with high chemical oxygen demand (COD) content were selected as feedstocks allowing increased input of potential energy to reactors without substantially altering volumetric loadings. Methane generation increased while reactor pH and volatile solids remained stable. Lag periods observed prior to methane stimulation suggested that acclimation of the microbial community may be critical to performance during co-digestion. Chemical oxygen demand mass balances in the experimental and control reactors indicated that all of the food industry waste COD was converted to methane.

A proof-of-concept experimental study for vacuum-driven anaerobic biosolids fermentation using the IntensiCarb technology.

This study demonstrates the potential of an innovative anaerobic treatment technology for municipal biosolids (IntensiCarb), which relies on vacuum evaporation to decouple solids and hydraulic retention times (SRT and HRT). We present proof-of-concept experiments using primary sludge and thickened waste activated sludge (50-50 v/v mixture) as feed for fermentation and carbon upgrading with the IntensiCarb unit. IntensiCarb fully decoupled the HRT and SRT in continuously stirred anaerobic reactors (CSAR) to achieve two intensification factors, that is, 1.3 and 2, while keeping the SRT constant at 3 days (including in the control fermenter). The intensified CSARs were compared to a conventional control system to determine the yields of particulate hydrolysis, VFA production, and nitrogen partitioning between fermentate and condensate. The intensified CSAR operating at an intensification factor 2 achieved a 65% improvement in particulate solubilization. Almost 50% of total ammonia was extracted without pH adjustment, while carbon was retained in the fermentate. Based on these results, the IntensiCarb technology allows water resource recovery facilities to achieve a high degree of plant-wide intensification while partitioning nutrients into different streams and thickening solids. PRACTITIONER POINTS: The IntensiCarb reactor can decouple hydraulic (HRT) and solids (SRT) retention times in anaerobic systems while also increasing particulate hydrolysis and overall plant capacity. Using vacuum as driving force of the IntensiCarb technology, the system could achieve thickening, digestion, and partial dewatering in the same unit-thus eliminating the complexity of multi-stage biosolids treatment lines. The ability to partition nutrients between particulate, fermentate, and condensate assigns to the IntensiCarb unit a key role in recovery strategies for value-added products such as nitrogen, phosphorus, and carbon, which can be recovered separately and independently.

Structure and function of the hetero-oligomeric cysteine synthase complex in plants.

Cysteine synthesis in bacteria and plants is catalyzed by serine acetyltransferase (SAT) and O-acetylserine (thiol)-lyase (OAS-TL), which form the hetero-oligomeric cysteine synthase complex (CSC). In plants, but not in bacteria, the CSC is assumed to control cellular sulfur homeostasis by reversible association of the subunits. Application of size exclusion chromatography, analytical ultracentrifugation, and isothermal titration calorimetry revealed a hexameric structure of mitochondrial SAT from Arabidopsis thaliana (AtSATm) and a 2:1 ratio of the OAS-TL dimer to the SAT hexamer in the CSC. Comparable results were obtained for the composition of the cytosolic SAT from A. thaliana (AtSATc) and the cytosolic SAT from Glycine max (Glyma16g03080, GmSATc) and their corresponding CSCs. The hexameric SAT structure is also supported by the calculated binding energies between SAT trimers. The interaction sites of dimers of AtSATm trimers are identified using peptide arrays. A negative Gibbs free energy (ΔG = -33 kcal mol(-1)) explains the spontaneous formation of the AtCSCs, whereas the measured SAT:OAS-TL affinity (K(D) = 30 nm) is 10 times weaker than that of bacterial CSCs. Free SAT from bacteria is >100-fold more sensitive to feedback inhibition by cysteine than AtSATm/c. The sensitivity of plant SATs to cysteine is further decreased by CSC formation, whereas the feedback inhibition of bacterial SAT by cysteine is not affected by CSC formation. The data demonstrate highly similar quaternary structures of the CSCs from bacteria and plants but emphasize differences with respect to the affinity of CSC formation (K(D)) and the regulation of cysteine sensitivity of SAT within the CSC.

Exclusion and exploitation: The incarceration of Black Americans from slavery to the present.

Understanding long-run patterns in the incarceration of Black Americans requires integrating the study of racial inequality with the study of political economy. I offer a parsimonious framework describing how the Black incarceration rate has been affected by the dynamics of exploitation and exclusion over time and across space. This framework helps to explain otherwise puzzling facts, like why the Black incarceration rate was lower in the South than in the North for much of the 20th century, why it was lowest in the South's cotton belt, and why it began to tick upward when it did. It also enables us to better understand recent changes in racial and class inequality in incarceration in the United States.

Non-invasive topology analysis of membrane proteins in the secretory pathway.

We present a novel method to experimentally visualize in vivo the topology of transmembrane proteins residing in the endoplasmic reticulum (ER) membrane or passing through the secretory pathway on their way to their final destination. This approach, so-called redox-based topology analysis (ReTA), is based on fusion of transmembrane proteins with redox-sensitive GFP (roGFP) and ratiometric imaging. The ratio images provide direct information on the orientation of roGFP relative to the membrane as the roGFP fluorescence alters with changes in the glutathione redox potential across the ER membrane. As proof of concept, we produced binary read-outs using oxidized roGFP inside the ER lumen and reduced roGFP on the cytosolic side of the membrane for both N- and C-terminal fusions of single and multi-spanning membrane proteins. Further, successive deletion of hydrophobic domains from the C-terminus of the K/HDEL receptor ERD2 resulted in alternating localization of roGFP and a topology model for AtERD2 with six transmembrane domains.

Microstructure and Compression Properties of VSS­V3B2 Eutectic Alloys in the V-Si-B System.

The present study reports on the microstructural evolution and room temperature plasticity of V(­Si)­B alloys with respect to the V solid solution (VSS)­V3B2 phase region. To investigate the occurring effects systematically, different binary V­B and ternary V­Si­B alloys were produced by conventional arc melting. Scanning electron microscope (SEM) analyses and X-ray diffraction (XRD) measurements were used to characterize the resulting as-cast microstructures. For the first time, the eutectic composition was systematically traced from the binary V­B domain to the ternary V­Si­B system. The observations discover that the binary eutectic trough (VSS­V3B2) seems to reach into the ternary system up to an alloy composition of V­5Si­9B. Room temperature compression tests were carried out in order to study the impact of single-phase and multi-phase microstructures on the strength and plasticity of binary and ternary alloys. The results indicate that the VSS phase controls the plastic deformability in the VSS­V3B2 eutectic microstructure whereas the intermetallic V3B2 acts as a strong hardening phase.

A comparative study of ultrasonic pretreatment and an internal recycle for the enhancement of mesophilic anaerobic digestion.

The objective of this study was to investigate the use of ultrasonic energy in an internal recycle and pretreatment mode of operation relative to a conventional mode of mesophilic anaerobic digestion. The primary focus was to determine if using ultrasonics in a pretreatment mode and in an internal recycle line produced changes in performance relative to each other and the control. Using a relatively low-energy sonication system, the data showed that the addition of ultrasonic energy, in either a recycle line or as a pretreatment technology, improved anaerobic digestion efficiency for waste-activated sludge. There was a 13 to 21% increase in biogas yield and an increase in total and volatile solids destruction of 3 to 10.3 additional percentage points, depending on the ultrasonic dose and location. Dewatering of the biosolids following ultrasonic treatment was poorer, as measured by an increase in the optimum polymer conditioning dose. The addition of ultrasonics to the digestion systems generated a more stable biosolids product, with a 2 to 58% reduction in organo-sulfur gas production from dewatered biosolids cakes.

5-HTTLPR and gender moderate changes in negative affect responses to tryptophan infusion.

Expression of the serotonin transporter is affected by the genotype of the 5-HTTLPR (short and long forms) as well as the genotype of the SNP rs25531 within this region. Based on the combined genotypes for these polymorphisms, we designated each allele as a high or low expressing allele according to established expression levels-resulting in HiHi, HiLo, & LoLo genotype groups for analysis. We evaluated effects of gender and the promoter genotype on induction of negative affect by intravenous infusion of L: -tryptophan (TRP). The protocol consisted of a day-1 sham saline infusion and a day-2 active TRP infusion. Models assessed 5-HTTLPR composite genotype and gender as predictors of change in ratings of negative emotion during TRP infusion. During sham infusion there were no significant changes from baseline in mood ratings. During TRP infusion all negative affect ratings increased significantly from baseline (P's < .02). The genotype x gender interaction was a significant predictor of depression-dejection (P = .013), and trended towards predicting anger-hostility (P = .084). Males in the HiHi group had greater increases in negative affect during infusion, compared to all groups except LoLo females, who also showed increased negative affect.

Analysis of neuroanatomical differences in mice with genetically modified serotonin transporters assessed by structural magnetic resonance imaging.

Background: The serotonin (5-HT) system has long been implicated in autism spectrum disorder (ASD) as indicated by elevated whole blood and platelet 5-HT, altered platelet and brain receptor and transporter binding, and genetic linkage and association findings. Based upon work in genetically modified mice, 5-HT is known to influence several aspects of brain development, but systematic neuroimaging studies have not previously been reported. In particular, the 5-HT transporter (serotonin transporter, SERT; 5-HTT) gene, Slc6a4, has been extensively studied. Methods: Using a 7-T MRI and deformation-based morphometry, we assessed neuroanatomical differences in an Slc6a4 knockout mouse on a C57BL/6 genetic background, along with an Slc6a4 Ala56 knockin mouse on two different genetic backgrounds (129S and C57BL/6). Results: Individually (same sex, same background, same genotype), the only differences found were in the female Slc6a4 knockout mouse; all the others had no significant differences. However, an analysis of variance across the whole study sample revealed a significant effect of Slc6a4 on the amygdala, thalamus, dorsal raphe nucleus, and lateral and frontal cortices. Conclusions: This work shows that an increase or decrease in SERT function has a significant effect on the neuroanatomy in 5-HT relevant regions, particularly the raphe nuclei. Notably, the Slc6a4 Ala56 knockin alone appears to have an insignificant, but suggestive, effect compared to the KO, which is consistent with Slc6a4 function. Despite the small number of 5-HT neurons and their localization to the brainstem, it is clear that 5-HT plays an important role in neuroanatomical organization.

Application of mechanical shear in an internal-recycle for the enhancement of mesophilic anaerobic digestion.

A combination of bench- and full-scale studies were conducted to determine the effectiveness of high-intensity mechanical shear in an internal recycle loop to enhance mesophilic anaerobic digestion and the implications of this process for routine operations of a digestion system. During short-term batch digestion (56 hours), a 46% increase in biogas production was observed. However, it was found that the degree of digestion enhancement was sludge-specific, with increases in volatile solids destruction ranging from 16.6 to 110%. A full-scale demonstration showed increased total and volatile solids destruction of 22 and 21% for the primary digester and 17.2 and 11% for the secondary digester, respectively. The data also suggest that increased protein degradation is one of the major mechanisms associated with the observed increases in volatile solids destruction. The full-scale demonstration also determined that shear enhanced digestion can be operated without process upset, based on volatile fatty acid profile and headspace biogas composition (methane and carbon dioxide). Dewatering properties, as measured by polymer demand, deteriorated in the primary digester, but there was improvement in the secondary digester. High-intensity shear does not appear to enhance pathogen reduction based on total and fecal coliform bacterial enumeration.

Impact of Maternal Serotonin Transporter Genotype on Placental Serotonin, Fetal Forebrain Serotonin, and Neurodevelopment.

Biomarker, neuroimaging, and genetic findings implicate the serotonin transporter (SERT) in autism spectrum disorder (ASD). Previously, we found that adult male mice expressing the autism-associated SERT Ala56 variant have altered central serotonin (5-HT) system function, as well as elevated peripheral blood 5-HT levels. Early in gestation, before midbrain 5-HT projections have reached the cortex, peripheral sources supply 5-HT to the forebrain, suggesting that altered maternal or placenta 5-HT system function could impact the developing embryo. We therefore used different combinations of maternal and embryo SERT Ala56 genotypes to examine effects on blood, placenta and embryo serotonin levels and neurodevelopment at embryonic day E14.5, when peripheral sources of 5-HT predominate, and E18.5, when midbrain 5-HT projections have reached the forebrain. Maternal SERT Ala56 genotype was associated with decreased placenta and embryonic forebrain 5-HT levels at E14.5. Low 5-HT in the placenta persisted, but forebrain levels normalized by E18.5. Maternal SERT Ala56 genotype effects on forebrain 5-HT levels were accompanied by a broadening of 5-HT-sensitive thalamocortical axon projections. In contrast, no effect of embryo genotype was seen in concepti from heterozygous dams. Blood 5-HT levels were dynamic across pregnancy and were increased in SERT Ala56 dams at E14.5. Placenta RNA sequencing data at E14.5 indicated substantial impact of maternal SERT Ala56 genotype, with alterations in immune and metabolic-related pathways. Collectively, these findings indicate that maternal SERT function impacts offspring placental 5-HT levels, forebrain 5-HT levels, and neurodevelopment.