RGC1/RGC2 deletions cause increased sensitivity to oxidative stress in Saccharomyces cerevisiae, which can be overcome by constitutive nuclear Yap1 expression

Oxidative stress mechanism in yeast presents an innovative pathway to understand in creating the next generation of antifungal drugs. Rgc1 and Rgc2 are paralogous proteins that regulate the Fps1 glycerol channel in hyperosmotic stress. Hyperosmotic conditions lead Hog1 MAP kinase to phosphorylate Rgc2 and cause its dissociation from Fps1, allowing the channel to close and protect the cell from damage. Rgc2 contains pleckstrin homology (PH) domains broken up by long insertions and more phosphorylation sites than targeted by Hog1 in response to hyperosmotic stress. Since none of the other MAP kinases in yeast were seen to phosphorylate Rgc2 during oxidative stress, it is thought that Rgc2 may bind to other proteins. In this study, the sensitivity of a strain deleted for both RGC1 and RGC2 was compared to strains with single deletions in either gene in response to oxidative stress. Having deletions in both RGC1 and RGC2 caused increased sensitivity to hydrogen peroxide whereas strains with deletions in either gene seemed unaffected, correlating with the fact that Rgc1 and Rgc2 are paralogous proteins, able to recover each other's functions. A second analysis compared mutated Fps1 (fps1∆-FKSV) and a strain with deletions for both RGC1 and RGC2 (rgc1/2∆). The fps1∆-FKSV strain has four amino acid substitutions in the C-terminal region where Rgc2 binds to Fps1. While both strains grew less than wild-type in hydrogen peroxide, the rgc1/2∆ strain was more sensitive suggesting that Rgc1/2 has an additional role in oxidative stress. To identify the oxidative stress function of Rgc1/2, a genomic overexpression library was transformed into the rgc1/2∆ strain and used for a suppressor screen in the presence of hydrogen peroxide. Although the screen revealed a manageable amount of 49 candidates, only four produced sequences that spanned a protein-encoding region. The candidate plasmids were transformed back into the rgc1/2∆ strain for preparation of a sensitivity assay which showed that the colonies did not survive any better than the starting rgc1/2∆ strain. Without a plausible plasmid candidate, we decided to look into the effect of YAP1 on the rgc1/2∆ strain. Yap1 is a transcription factor known to activate many genes in oxidative stress. Two forms of YAP1 were transformed into rgc1/2∆: wild-type YAP1 and YAP1-A627E which contains a mutation in the nuclear export signal. Compared to the controls, YAP1-A627E allowed the rgc1/2∆ strain to grow at 1.5mM H2O2 while wild-type YAP1 did not. This result showed that a constitutively nuclear Yap1 can overcome deletions in RGC1 and RGC2. It also suggested that an increased activity in the nucleus was important in hydrogen peroxide resistance and another suppressor screen of rgc1/2∆ was performed looking for spontaneous mutations in the genomic DNA. The screened colonies were tested for their survival on hydrogen peroxide but their resistance appeared to be transient. We have shown Rgc1 and Rgc2 to be important cellular components in oxidative stress in addition to hyperosmotic stress. Further research on Rgc1/2 would provide invaluable knowledge on oxidative stress protection in yeast and a better foundation on which to build antifungal drugs

Unanticipated differences between α- and γ-diaminobutyric acid-linked hairpin polyamide-alkylator conjugates

Hairpin polyamide–chlorambucil conjugates containing an {alpha}-diaminobutyric acid ({alpha}-DABA) turn moiety are compared to their constitutional isomers containing the well-characterized {gamma}-DABA turn. Although the DNA-binding properties of unconjugated polyamides are similar, the {alpha}-DABA conjugates display increased alkylation specificity and decreased rate of reaction. Treatment of a human colon carcinoma cell line with {alpha}-DABA versus {gamma}-DABA hairpin conjugates shows only slight differences in toxicities while producing similar effects on cell morphology and G2/M stage cell cycle arrest. However, striking differences in animal toxicity between the two classes are observed. Although mice treated with an {alpha}-DABA hairpin polyamide do not differ significantly from control mice, the analogous {gamma}-DABA hairpin is lethal. This dramatic difference from a subtle structural change would not have been predicted

Recovery-Oriented Training and Staff Attitudes Over Time in Two State Hospitals

Recovery attitudes and concepts are often promoted to community mental health staff through educational and in-service trainings, but no study found has examined this in state hospitals. The current observational study aimed to examine the types of recovery-oriented trainings that occurred at two state hospitals over 1 year and subsequent changes in staff recovery attitudes. A total of 184 state hospital staff completed questionnaires assessing their personal optimism, consumer optimism, and agency recovery orientation at baseline and 1 year later. The types of recovery-oriented trainings staff received were categorized as general/inspirational or specific/practical training. Results found that the majority of staff at the two state hospitals received some recovery-oriented training, mostly general/inspirational training. Staff who received specific/practical training had a greater increase in agency recovery attitudes than staff who received only general/inspirational training or no training. However, the more trainings staff had, the higher their consumer optimism. These results suggest state hospitals are incorporating recovery-oriented staff trainings, but more specific trainings may be needed and all staff involved in different levels of care need to be included

Master\u27s Recital

Program listing performers and works performe

α-Diaminobutyric acid-linked hairpin polyamides

A hairpin polyamide-chlorambucil conjugate linked by α-diaminobutyric acid (α-DABA) has been shown to have interesting biological properties in cellular and small animal models. Remarkably, this new class of hairpin polyamides has not been previously characterized with regard to energetics and sequence specificity. Herein we present a series of pyrrole-imidazole hairpin polyamides linked by α-DABA and compare them to polyamides containing the standard γ-DABA turn unit. The α-DABA hairpins have overall decreased binding affinities. However, α-DABA polyamide-chlorambucil conjugates are sequence-specific DNA alkylators with increased specificities. Affinity cleavage studies of α-DABA polyamide-EDTA conjugates confirmed their preference for binding DNA in a forward hairpin conformation. In contrast, an unsubstituted glycine-linked polyamide prefers to bind in an extended binding mode. Thus, substitution on the turn unit locks the α-DABA polyamide into the forward hairpin binding motif

Staff Turnover in Statewide Implementation of ACT: Relationship with ACT Fidelity and Other Team Characteristics

Staff turnover on assertive community treatment (ACT) teams is a poorly understood phenomenon. This study examined annual turnover and fidelity data collected in a statewide implementation of ACT over a 5-year period. Mean annual staff turnover across all observations was 30.0%. Turnover was negatively correlated with overall fidelity at Year 1 and 3. The team approach fidelity item was negatively correlated with staff turnover at Year 3. For 13 teams with 3 years of follow-up data, turnover rates did not change over time. Most ACT staff turnover rates were comparable or better than other turnover rates reported in the mental health and substance abuse literature

Clinical and Psychological Correlates of Two Domains of Hopelessness in Schizophrenia

Hopelessness is a widely observed barrier to recovery from schizophrenia spectrum disorders. Yet little is known about how clinical, social, and psychological factors independently affect hope. Additionally, the relationships that exist between these factors and different kinds of hope are unclear. To explore both issues, we correlated two aspects of hope, expectations of the future and agency, with stigma, clinical symptoms, anxiety, and coping preferences in 143 persons with a schizophrenia spectrum disorder. Multiple regressions revealed that hope for the future was predicted by lesser alienation, lesser preference for ignoring stressors, and lesser emotional discomfort and negative symptoms, accounting for 43% of the variance. A greater sense of agency was linked to lesser endorsement of mental illness stereotypes, fewer negative symptoms, lesser social phobia, and lesser preference for ignoring stressors, accounting for 44% of the variance. Implications for research and interventions are discussed

Housing Preferences and Choices Among Adults with Mental Illness and Substance Use Disorders: A Qualitative Study

Housing is a crucial issue for adults with severe mental illness and co-occurring substance use disorders, as this population is particularly susceptible to housing instability and homelessness. We interviewed 40 adults with dual disorders, living in either supervised or independent housing arrangements, to examine housing preferences, decision making processes surrounding housing choices, and perceived barriers to housing. We found that many clients indicated their housing preferences had changed over time, and some clients related housing preferences to recovery. Although the majority of clients preferred independent housing, many also described benefits of supervised housing. Clients' current living situations appeared to be driven primarily by treatment provider recommendations and availability of housing. Common barriers to obtaining desired housing were lack of income and information. These findings have implications for supported housing models and approaches to providing housing for clients

Promoting tau secretion and propagation by hyperactive p300/CBP via autophagy-lysosomal pathway in tauopathy.

BackgroundThe trans-neuronal propagation of tau has been implicated in the progression of tau-mediated neurodegeneration. There is critical knowledge gap in understanding how tau is released and transmitted, and how that is dysregulated in diseases. Previously, we reported that lysine acetyltransferase p300/CBP acetylates tau and regulates its degradation and toxicity. However, whether p300/CBP is involved in regulation of tau secretion and propagation is unknown.MethodWe investigated the relationship between p300/CBP activity, the autophagy-lysosomal pathway (ALP) and tau secretion in mouse models of tauopathy and in cultured rodent and human neurons. Through a high-through-put compound screen, we identified a new p300 inhibitor that promotes autophagic flux and reduces tau secretion. Using fibril-induced tau spreading models in vitro and in vivo, we examined how p300/CBP regulates tau propagation.ResultsIncreased p300/CBP activity was associated with aberrant accumulation of ALP markers in a tau transgenic mouse model. p300/CBP hyperactivation blocked autophagic flux and increased tau secretion in neurons. Conversely, inhibiting p300/CBP promoted autophagic flux, reduced tau secretion, and reduced tau propagation in fibril-induced tau spreading models in vitro and in vivo.ConclusionsWe report that p300/CBP, a lysine acetyltransferase aberrantly activated in tauopathies, causes impairment in ALP, leading to excess tau secretion. This effect, together with increased intracellular tau accumulation, contributes to enhanced spreading of tau. Our findings suggest that inhibition of p300/CBP as a novel approach to correct ALP dysfunction and block disease progression in tauopathy