Trapping redox partnerships in oxidant-sensitive proteins with a small, thiol-reactive cross-linker

A broad range of redox-regulated proteins undergo reversible disulfide bond formation on oxidation-prone cysteine residues. Heightened reactivity of the thiol groups in these cysteines also increases susceptibility to modification by organic electrophiles, a property that can be exploited in the study of redox networks. Here, we explored whether divinyl sulfone (DVSF), a thiol-reactive bifunctional electrophile, cross-links oxidant-sensitive proteins to their putative redox partners in cells. To test this idea, previously identified oxidant targets involved in oxidant defense (namely, peroxiredoxins, methionine sulfoxide reductases, sulfiredoxin, and glutathione peroxidases), metabolism, and proteostasis were monitored for cross-link formation following treatment of Saccharomyces cerevisiae with DVSF. Several proteins screened, including multiple oxidant defense proteins, underwent intermolecular and/or intramolecular cross-linking in response to DVSF. Specific redox-active cysteines within a subset of DVSF targets were found to influence cross-linking; in addition, DVSF-mediated cross-linking of its targets was impaired in cells first exposed to oxidants. Since cross-linking appeared to involve redox-active cysteines in these proteins, we examined whether potential redox partners became cross-linked to them upon DVSF treatment. Specifically, we found that several substrates of thioredoxins were cross-linked to the cytosolic thioredoxin Trx2 in cells treated with DVSF. However, other DVSF targets, like the peroxiredoxin Ahp1, principally formed intra-protein cross-links upon DVSF treatment. Moreover, additional protein targets, including several known to undergo S-glutathionylation, were conjugated via DVSF to glutathione. Our results indicate that DVSF is of potential use as a chemical tool for irreversibly trapping and discovering thiol-based redox partnerships within cells

PNC-27, a Chimeric p53-Penetratin Peptide Binds to HDM-2 in a p53 Peptide-like Structure, Induces Selective Membrane-Pore Formation and Leads to Cancer Cell Lysis

PNC-27, a 32-residue peptide that contains an HDM-2 binding domain and a cell-penetrating peptide (CPP) leader sequence kills cancer, but not normal, cells by binding to HDM-2 associated with the plasma membrane and induces the formation of pores causing tumor cell lysis and necrosis. Conformational energy calculations on the structure of PNC-27 bound to HDM-2 suggest that 1:1 complexes form between PNC-27 and HDM-2 with the leader sequence pointing away from the complex. Immuno-scanning electron microscopy was carried out with cancer cells treated with PNC-27 and decorated with an anti-PNC-27 antibody coupled to 6 nm gold particles and an anti-HDM-2 antibody linked to 15 nm gold particles. We found multiple 6 nm- and 15 nm-labeled gold particles in approximately 1:1 ratios in layered ring-shaped structures in the pores near the cell surface suggesting that these complexes are important to the pore structure. No pores formed in the control, PNC-27-treated untransformed fibroblasts. Based on the theoretical and immuno-EM studies, we propose that the pores are lined by PNC-27 bound to HDM-2 at the membrane surface with the PNC-27 leader sequence lining the pores or by PNC-27 bound to HDM-2

Making Connections - Envisioning Springfield\u27s North End

This work explores a service learning strategy in the context of the senior Urban Design Studio taught in the Department of Landscape Architecture and Regional Planning at the University of Massachusetts Amherst. The primary goal of this project is to stimulate a conversation in the neighborhoods of the North End, to develop green design strategies, to improve services and businesses for residents and the employees of local businesses, and to foster cultural engagement and interaction in the North End that will enhance the vibrancy, resilience, and quality of life of this urban community. Making connections - Envisioning Springfield\u27s North End proposes improved connectivity in a physical, cultural, and social sense will be key to attaining these goals and to engaging and synergizing individuals and community groups in the North End - residents, businesses, schools, churches, employers, and employees. Six sustainable learning and planning principles have emerged from this studio: 1. Input and interaction – Visioning workshops connect campus and community 2. Community-building art - Expression of place and people 3. Healthy living - Urban agriculture and education 4. Urban greenways – Abandoned railways and urban rivers and streams 5. Green infrastructure - Green streets as networks and structural framework 6. Sustainable urban form – Mixed use and pedestrian friendly neighborhood

How students choose a major: The effect of a college major decision-making course on a student\u27s anxiety

The purpose of this study was to examine the effect of a college major decision-making course on a student\u27s level of anxiety about making the choice of major. This study investigated 18 undeclared, sophomore level college students (intervention group) at a four-year institution who enrolled in a course that was designed to educate them about their choices of major. This group was then compared to a control group of 22 undeclared, sophomore level college students (comparison group) at the same institution. The intervention group experienced an eight-week course for credit and a grade while the comparison group did not. The instrument used to determine the level of anxiety was the Career Thoughts Inventory (CTI). Results from the study were analyzed using a paired-samples t-test, analysis of co-variance (ANCOVA), analysis of variance (ANOVA), and Pearson correlation. The data revealed significant reduction in levels of anxiety for the intervention group while the data from the comparison group showed no significant changes from pretest to posttest. In addition, the intervention group reduced their anxiety at a greater rate than that of the comparison group. Although there is little literature regarding anxiety level as it pertains to declaring majors, the results of this study were consistent with the literature in that undeclared, sophomore level students reported feeling anxious about making a choice of major. ^ These findings suggest the need for more study about the anxiety level in students regarding their choice of college major in that anxiety level may influence a student\u27s level of satisfaction about their choice of major. These findings may also suggest the need for more college major decision-making courses being offered on a campus.

A Comprehensive Needs Assessment of Rural Syringe Services

The current opioid epidemic has resulted in a significant increase in opioid-related overdoses, and a corresponding rise in HIV and HCV transmission. Syringe Service Programs (SSPs) provide resources for people who inject drugs. There are fewer SSPs in rural U.S. and little research has been conducted to determine unique needs of SSP participants in rural areas.https://scholarworks.uvm.edu/comphp_gallery/1299/thumbnail.jp

The Epidemiology, Demographics, and Geographical Distribution of Human Non-Tuberculosis Mycobacteria (NTM) Disease in the Endemic Central Florida Region

Background: Of the \u3e100,000 people in the United States infected yearly with non-tuberculosis mycobacteria (NTM), Florida has the highest yearly incidence and prevalence of NTM disease. However, little has been documented on the epidemiology and distribution of NTM disease within Central Florida. Methods: A retrospective case review study was conducted from January, 2011 to December, 2017 at a large tertiary acute care medical center in Tampa, Florida to identify all NTM infection cases. Demographics (age, sex at birth, ethnicity), comorbidities, HIV testing status, residential zip code, NTM species, and specimen sources were collected. Results: Of the 507 isolates, Mycobacterium abscessus group was the most common (45.4%; n = 230), and contained M. abscessus spp. abscessus (34.5%; n = 175), M. abscessus spp. massilense (8.7%; n = 44), and M. abscessus spp. bolletii (1.18%; n = 6). Other rapid growers were M. fortuitum species (6.9%; n = 35) and M. chelonae (2.56%; n = 13). Of the slower growers, M. gordonae (19.9%; n = 101) and M. avium complex (8.28%; n = 42) were the most common. Of the M. avium complex, M. chimera was most common (4.9%; n = 25). Samples were mostly isolated from sputum (51.7%; n = 262), bronchial lavage (26%; n = 132), skin and soft tissue (11%; n = 58), and blood (7.1%; n = 36). Of the 361 unique patients, average age was 59.2 years (12 to 95 years), with 47.6% (n = 172) greater than 65 years of age, and mostly male 57.9% (n = 208). Caucasians represented 73.4% (n = 265) of our cohort, and African Americans and Hispanics represented 16.3% (n = 59) and 6.8% (n = 24), respectively. Most cases were in those residing outside the Tampa Bay metro area 81.2% (n = 293/361). Notable comorbidities included COPD (n = 83), cystic fibrosis (n = 41), lung transplant (n = 40), heart transplant (n = 12), pulmonary fibrosis (n = 12), and renal transplant (n = 7). A total of 145 individuals received HIV testing at the hospital facility, and of these 44 individuals were living with HIV. Conclusion: This study identified a diversity of NTM species across a wide geographical and demographic distribution in the endemic Central Florida region. M. abscessus group had the highest prevalence. This is valuable in understanding which populations are at risk for developing NTM infection in this area of Florida

The Epidemiology, Demographics, and Geographical Distribution of Human Non-Tuberculosis Mycobacteria (NTM) Disease in the Endemic Central Florida Region

Background: Of the \u3e100,000 people in the United States infected yearly with non-tuberculosis mycobacteria (NTM), Florida has the highest yearly incidence and prevalence of NTM disease. However, little has been documented on the epidemiology and distribution of NTM disease within Central Florida. Methods: A retrospective case review study was conducted from January, 2011 to December, 2017 at a large tertiary acute care medical center in Tampa, Florida to identify all NTM infection cases. Demographics (age, sex at birth, ethnicity), comorbidities, HIV testing status, residential zip code, NTM species, and specimen sources were collected. Results: Of the 507 isolates, Mycobacterium abscessus group was the most common (45.4%; n = 230), and contained M. abscessus spp. abscessus (34.5%; n = 175), M. abscessus spp. massilense (8.7%; n = 44), and M. abscessus spp. bolletii (1.18%; n = 6). Other rapid growers were M. fortuitum species (6.9%; n = 35) and M. chelonae (2.56%; n = 13). Of the slower growers, M. gordonae (19.9%; n = 101) and M. avium complex (8.28%; n = 42) were the most common. Of the M. avium complex, M. chimera was most common (4.9%; n = 25). Samples were mostly isolated from sputum (51.7%; n = 262), bronchial lavage (26%; n = 132), skin and soft tissue (11%; n = 58), and blood (7.1%; n = 36). Of the 361 unique patients, average age was 59.2 years (12 to 95 years), with 47.6% (n = 172) greater than 65 years of age, and mostly male 57.9% (n = 208). Caucasians represented 73.4% (n = 265) of our cohort, and African Americans and Hispanics represented 16.3% (n = 59) and 6.8% (n = 24), respectively. Most cases were in those residing outside the Tampa Bay metro area 81.2% (n = 293/361). Notable comorbidities included COPD (n = 83), cystic fibrosis (n = 41), lung transplant (n = 40), heart transplant (n = 12), pulmonary fibrosis (n = 12), and renal transplant (n = 7). A total of 145 individuals received HIV testing at the hospital facility, and of these 44 individuals were living with HIV. Conclusion: This study identified a diversity of NTM species across a wide geographical and demographic distribution in the endemic Central Florida region. M. abscessus group had the highest prevalence. This is valuable in understanding which populations are at risk for developing NTM infection in this area of Florida

The Impact of Recent Incarceration on Post-Release HIV Linkage to Care and Risk-Taking Behaviors in the Setting of Enhanced Linkage to Care Policies in Florida

Background: The United States has the largest incarcerated population in the world with 6.61 million adults in 2016.1 While incarceration is a known risk factor for difficulties in linkage to care2–3 and adverse health outcomes4–6, little is published on post-release incarcerated persons living with HIV (PLWH) in Florida. Methods: Data were acquired from the Florida Cohort, an ongoing, longitudinal, cross-sectional study of PLWH recruited across HIV clinics in the state of Florida, from 2014 to 2018. Chi-square and multiple regression analyses correlated recent incarceration (within last 12 months) with demographics, HIV care adherence, perceived barriers to care, and self-reported high-risk behaviors. Results: Of 936 participants, 6.4% (n = 60) reported recent incarceration within the last 12 months. Those recently incarcerated were more likely to report missing at least one appointment in the last 6 months (46.7% vs. 22.2%; P \u3c 0.0001), to have an excessively long travel time ( \u3e60 minutes) to a HIV provider (34.5% vs. 16.6%, P = 0.002; OR 2.66 [95% CI: 1.20–5.92]), and to lack reliable transportation (70% vs. 47.5%, P = 0.0007; OR 1.70 [95% CI: 0.82–3.52]) Those not recently incarcerated reported having completed a high school education (OR: 0.69 [95% CI: 0.5–0.97]) and stated they “never missed an appointment” (OR: 0.42 [95% CI: 0.22–0.81]). Recently incarcerated PLWH also had higher occurrence of high-risk behaviors such as receiving (40.4% vs. 8.7%; P = 0.001) or providing (30.4% vs. 10.4%; P = 0.000) money or drugs for sex, having used IV drugs (15% vs. 4%; P = 0.001), and not using condoms during exchange of drugs for sex (OR: 9.43 [95% CI: 3.78–23.52]). Conclusion: Recently incarcerated PLWH continue to have significant geographical and logistical barriers to care and self-report more high-risk behaviors than nonincarcerated peers. Enhanced case management and telehealth services may be useful in linkage to care when PLWH transition from correctional to community healthcare systems in the Florida setting

The Impact of Recent Incarceration on Post-Release HIV Linkage to Care and Risk-Taking Behaviors in the Setting of Enhanced Linkage to Care Policies in Florida

Background: The United States has the largest incarcerated population in the world with 6.61 million adults in 2016.1 While incarceration is a known risk factor for difficulties in linkage to care2–3 and adverse health outcomes4–6, little is published on post-release incarcerated persons living with HIV (PLWH) in Florida. Methods: Data were acquired from the Florida Cohort, an ongoing, longitudinal, cross-sectional study of PLWH recruited across HIV clinics in the state of Florida, from 2014 to 2018. Chi-square and multiple regression analyses correlated recent incarceration (within last 12 months) with demographics, HIV care adherence, perceived barriers to care, and self-reported high-risk behaviors. Results: Of 936 participants, 6.4% (n = 60) reported recent incarceration within the last 12 months. Those recently incarcerated were more likely to report missing at least one appointment in the last 6 months (46.7% vs. 22.2%; P \u3c 0.0001), to have an excessively long travel time ( \u3e60 minutes) to a HIV provider (34.5% vs. 16.6%, P = 0.002; OR 2.66 [95% CI: 1.20–5.92]), and to lack reliable transportation (70% vs. 47.5%, P = 0.0007; OR 1.70 [95% CI: 0.82–3.52]) Those not recently incarcerated reported having completed a high school education (OR: 0.69 [95% CI: 0.5–0.97]) and stated they “never missed an appointment” (OR: 0.42 [95% CI: 0.22–0.81]). Recently incarcerated PLWH also had higher occurrence of high-risk behaviors such as receiving (40.4% vs. 8.7%; P = 0.001) or providing (30.4% vs. 10.4%; P = 0.000) money or drugs for sex, having used IV drugs (15% vs. 4%; P = 0.001), and not using condoms during exchange of drugs for sex (OR: 9.43 [95% CI: 3.78–23.52]). Conclusion: Recently incarcerated PLWH continue to have significant geographical and logistical barriers to care and self-report more high-risk behaviors than nonincarcerated peers. Enhanced case management and telehealth services may be useful in linkage to care when PLWH transition from correctional to community healthcare systems in the Florida setting

PNC-27, a Chimeric p53-Penetratin Peptide Binds to HDM-2 in a p53 Peptide-like Structure, Induces Selective Membrane-Pore Formation and Leads to Cancer Cell Lysis

PNC-27, a 32-residue peptide that contains an HDM-2 binding domain and a cell-penetrating peptide (CPP) leader sequence kills cancer, but not normal, cells by binding to HDM-2 associated with the plasma membrane and induces the formation of pores causing tumor cell lysis and necrosis. Conformational energy calculations on the structure of PNC-27 bound to HDM-2 suggest that 1:1 complexes form between PNC-27 and HDM-2 with the leader sequence pointing away from the complex. Immuno-scanning electron microscopy was carried out with cancer cells treated with PNC-27 and decorated with an anti-PNC-27 antibody coupled to 6 nm gold particles and an anti-HDM-2 antibody linked to 15 nm gold particles. We found multiple 6 nm- and 15 nm-labeled gold particles in approximately 1:1 ratios in layered ring-shaped structures in the pores near the cell surface suggesting that these complexes are important to the pore structure. No pores formed in the control, PNC-27-treated untransformed fibroblasts. Based on the theoretical and immuno-EM studies, we propose that the pores are lined by PNC-27 bound to HDM-2 at the membrane surface with the PNC-27 leader sequence lining the pores or by PNC-27 bound to HDM-2