Aging and Intermittent Fasting Impact on Transcriptional Regulation and Physiological Responses of Adult Drosophila Neuronal and Muscle Tissues.

The progressive decline of the nervous system, including protein aggregate formation, reflects the subtle dysregulation of multiple functional pathways. Our previous work has shown intermittent fasting (IF) enhances longevity, maintains adult behaviors and reduces aggregates, in part, by promoting autophagic function in the aging Drosophila brain. To clarify the impact that IF-treatment has upon aging, we used high throughput RNA-sequencing technology to examine the changing transcriptome in adult Drosophila tissues. Principle component analysis (PCA) and other analyses showed ~1200 age-related transcriptional differences in head and muscle tissues, with few genes having matching expression patterns. Pathway components showing age-dependent expression differences were involved with stress response, metabolic, neural and chromatin remodeling functions. Middle-aged tissues also showed a significant increase in transcriptional drift-variance (TD), which in the CNS included multiple proteolytic pathway components. Overall, IF-treatment had a demonstrably positive impact on aged transcriptomes, partly ameliorating both fold and variance changes. Consistent with these findings, aged IF-treated flies displayed more youthful metabolic, behavioral and basal proteolytic profiles that closely correlated with transcriptional alterations to key components. These results indicate that even modest dietary changes can have therapeutic consequences, slowing the progressive decline of multiple cellular systems, including proteostasis in the aging nervous system.

The mitochondrial ATP synthase is a shared drug target for aging and dementia.

Aging is a major driving force underlying dementia, such as that caused by Alzheimer's disease (AD). While the idea of targeting aging as a therapeutic strategy is not new, it remains unclear how closely aging and age-associated diseases are coupled at the molecular level. Here, we discover a novel molecular link between aging and dementia through the identification of the molecular target for the AD drug candidate J147. J147 was developed using a series of phenotypic screening assays mimicking disease toxicities associated with the aging brain. We have previously demonstrated the therapeutic efficacy of J147 in several mouse models of AD. Here, we identify the mitochondrial α-F1 -ATP synthase (ATP5A) as a target for J147. By targeting ATP synthase, J147 causes an increase in intracellular calcium leading to sustained calcium/calmodulin-dependent protein kinase kinase ß (CAMKK2)-dependent activation of the AMPK/mTOR pathway, a canonical longevity mechanism. Accordingly, modulation of mitochondrial processes by J147 prevents age-associated drift of the hippocampal transcriptome and plasma metabolome in mice and extends lifespan in drosophila. Our results link aging and age-associated dementia through ATP synthase, a molecular drug target that can potentially be exploited for the suppression of both. These findings demonstrate that novel screens for new AD drug candidates identify compounds that act on established aging pathways, suggesting an unexpectedly close molecular relationship between the two.

Using Intervention Mapping for Program Design and Production of iCHAMPSS: An Online Decision Support System to Increase Adoption, Implementation, and Maintenance of Evidence-Based Sexual Health Programs.

In Texas and across the United States, unintended pregnancy, HIV, and sexually transmitted infections (STIs) among adolescents remain serious public health issues. Sexual risk-taking behaviors, including early sexual initiation, contribute to these public health problems. Over 35 sexual health evidence-based programs (EBPs) have been shown to reduce sexual risk behaviors and/or prevent teen pregnancies or STIs. Because more than half of these EBPs are designed for schools, they could reach and impact a considerable number of adolescents if implemented in these settings. Most schools across the U.S. and in Texas, however, do not implement these programs. U.S. school districts face many barriers to the successful dissemination (i.e., adoption, implementation, and maintenance) of sexual health EBPs, including lack of knowledge about EBPs and where to find them, perceived lack of support from school administrators and parents, lack of guidance regarding the adoption process, competing priorities, and lack of specialized training on sexual health. Therefore, this paper describes how we used intervention mapping (Steps 3 and 4, in particular), a systematic design framework that uses theory, empirical evidence, and input from the community to develop CHoosing And Maintaining Effective Programs for Sex Education in Schools (iCHAMPSS), an online decision support system to help school districts adopt, implement, and maintain sexual health EBPs. Guided by this systematic intervention design approach, iCHAMPSS has the potential to increase dissemination of sexual health EBPs in school settings.

iCHAMPSS: Usability and Psychosocial Impact for Increasing Implementation of Sexual Health Education.

INTRODUCTION: Diffusion of sexual health evidence-based programs (EBPs) in schools is a complex and challenging process. iCHAMPSS ( CHoosing And Maintaining effective Programs for Sex education in Schools) is an innovative theory- and Web-based decision support system that may help facilitate this process. The purpose of this study was to pilot-test iCHAMPSS for usability and short-term psychosocial impact. METHOD: School district stakeholders from across Texas were recruited ( N = 16) and given access to iCHAMPSS for 3 weeks in fall 2014. Pre- and posttests were administered to measure usability parameters and short-term psychosocial outcomes. Data were analyzed using descriptive statistics and the Wilcoxon signed-rank test. RESULTS: Most participants reported that iCHAMPSS was easy to use, credible, helpful, and of sufficient motivational appeal. iCHAMPSS significantly increased participants' self-efficacy to obtain approval from their board of trustees to implement a sexual health EBP. Positive, though nonsignificant, trends included increased knowledge to locate EBPs, skills to prioritize sexual health education at the district level, and ability to choose an EBP that best meets district needs. CONCLUSIONS: iCHAMPSS is an innovative decision support system that could accelerate uptake of EBPs by facilitating diffusion and advance the field of dissemination and implementation science for the promotion of sexual health EBPs.

Assessing Basal and Acute Autophagic Responses in the Adult Drosophila Nervous System: The Impact of Gender, Genetics and Diet on Endogenous Pathway Profiles.

The autophagy pathway is critical for the long-term homeostasis of cells and adult organisms and is often activated during periods of stress. Reduced pathway efficacy plays a central role in several progressive neurological disorders that are associated with the accumulation of cytotoxic peptides and protein aggregates. Previous studies have shown that genetic and transgenic alterations to the autophagy pathway impacts longevity and neural aggregate profiles of adult Drosophila. In this study, we have identified methods to measure the acute in vivo induction of the autophagy pathway in the adult fly CNS. Our findings indicate that the genotype, age, and gender of adult flies can influence pathway responses. Further, we demonstrate that middle-aged male flies exposed to intermittent fasting (IF) had improved neuronal autophagic profiles. IF-treated flies also had lower neural aggregate profiles, maintained more youthful behaviors and longer lifespans, when compared to ad libitum controls. In summary, we present methodology to detect dynamic in vivo changes that occur to the autophagic profiles in the adult Drosophila CNS and that a novel IF-treatment protocol improves pathway response in the aging nervous system.

Brain trauma and autophagy: What flies and mice can teach us about conserved responses.

Drosophila models have been successfully used to identify many genetic components that affect neurodegenerative disorders. Recently, there has been a growing interest in identifying innate and environmental factors that influence the individual outcomes following traumatic brain injury (TBI). This includes both severe TBI and more subtle, mild TBI (mTBI), which is common in people playing contact sports. Autophagy, as a clearance pathway, exerts protective effects in multiple neurological disease models. In a recent publication, we highlighted the development of a novel repetitive mTBI system using Drosophila, which recapitulates several phenotypes associated with trauma in mammalian models. In particular, flies subjected to mTBI exhibit an acute impairment of the macroautophagy/autophagy pathway that is restored 1 wk following traumatic injury exposure. These phenotypes closely resemble temporary autophagy defects observed in a mouse TBI model. Through these studies, we also identified methods to directly assess autophagic responses in the fly nervous system and laid the groundwork for future studies designed to identify genetic, epigenetic and environmental factors that have an impact on TBI outcomes.

Using Drosophila as an integrated model to study mild repetitive traumatic brain injury.

Traumatic brain injury (TBI) is a major cause of morbidity and mortality worldwide. In addition, there has been a growing appreciation that even repetitive, milder forms of TBI (mTBI) can have long-term deleterious consequences to neural tissues. Hampering our understanding of genetic and environmental factors that influence the cellular and molecular responses to injury has been the limited availability of effective genetic model systems that could be used to identify the key genes and pathways that modulate both the acute and long-term responses to TBI. Here we report the development of a severe and mild-repetitive TBI model using Drosophila. Using this system, key features that are typically found in mammalian TBI models were also identified in flies, including the activation of inflammatory and autophagy responses, increased Tau phosphorylation and neuronal defects that impair sleep-related behaviors. This novel injury paradigm demonstrates the utility of Drosophila as an effective tool to validate genetic and environmental factors that influence the whole animal response to trauma and to identify prospective therapies needed for the treatment of TBI.

Harm reduction as a complex adaptive system: A dynamic framework for analyzing Tanzanian policies concerning heroin use.

Contrary to popular belief, policies on drug use are not always based on scientific evidence or composed in a rational manner. Rather, decisions concerning drug policies reflect the negotiation of actors' ambitions, values, and facts as they organize in different ways around the perceived problems associated with illicit drug use. Drug policy is thus best represented as a complex adaptive system (CAS) that is dynamic, self-organizing, and coevolving. In this analysis, we use a CAS framework to examine how harm reduction emerged around heroin trafficking and use in Tanzania over the past thirty years (1985-present). This account is an organizational ethnography based on of the observant participation of the authors as actors within this system. We review the dynamic history and self-organizing nature of harm reduction, noting how interactions among system actors and components have coevolved with patterns of heroin us, policing, and treatment activities over time. Using a CAS framework, we describe harm reduction as a complex process where ambitions, values, facts, and technologies interact in the Tanzanian sociopolitical environment. We review the dynamic history and self-organizing nature of heroin policies, noting how the interactions within and between competing prohibitionist and harm reduction policies have changed with patterns of heroin use, policing, and treatment activities over time. Actors learn from their experiences to organize with other actors, align their values and facts, and implement new policies. Using a CAS approach provides researchers and policy actors a better understanding of patterns and intricacies in drug policy. This knowledge of how the system works can help improve the policy process through adaptive action to introduce new actors, different ideas, and avenues for communication into the system.

Aging and Autophagic Function Influences the Progressive Decline of Adult Drosophila Behaviors.

Multiple neurological disorders are characterized by the abnormal accumulation of protein aggregates and the progressive impairment of complex behaviors. Our Drosophila studies demonstrate that middle-aged wild-type flies (WT, ~4-weeks) exhibit a marked accumulation of neural aggregates that is commensurate with the decline of the autophagy pathway. However, enhancing autophagy via neuronal over-expression of Atg8a (Atg8a-OE) reduces the age-dependent accumulation of aggregates. Here we assess basal locomotor activity profiles for single- and group-housed male and female WT flies and observed that only modest behavioral changes occurred by 4-weeks of age, with the noted exception of group-housed male flies. Male flies in same-sex social groups exhibit a progressive increase in nighttime activity. Infrared videos show aged group-housed males (4-weeks) are engaged in extensive bouts of courtship during periods of darkness, which is partly repressed during lighted conditions. Together, these nighttime courtship behaviors were nearly absent in young WT flies and aged Atg8a-OE flies. Previous studies have indicated a regulatory role for olfaction in male courtship partner choice. Coincidently, the mRNA expression profiles of several olfactory genes decline with age in WT flies; however, they are maintained in age-matched Atg8a-OE flies. Together, these results suggest that middle-aged male flies develop impairments in olfaction, which could contribute to the dysregulation of courtship behaviors during dark time periods. Combined, our results demonstrate that as Drosophila age, they develop early behavior defects that are coordinate with protein aggregate accumulation in the nervous system. In addition, the nighttime activity behavior is preserved when neuronal autophagy is maintained (Atg8a-OE flies). Thus, environmental or genetic factors that modify autophagic capacity could have a positive impact on neuronal aging and complex behaviors.

Mitophagy is required for acute cardioprotection by simvastatin.

AIMS: We have shown that autophagy and mitophagy are required for preconditioning. While statin's cardioprotective effects are well known, the role of autophagy/mitophagy in statin-mediated cardioprotection is not. In this study, we used HL-1 cardiomyocytes and mice subjected to ischemia/reperfusion to elucidate the mechanism of statin-mediated cardioprotection. RESULTS: HL-1 cardiomyocytes exposed to simvastatin for 24 h exhibited diminished protein kinase B (Akt)/mammalian target of rapamycin (mTOR) signaling, increased activation of unc-51-like kinase 1, and upregulation of autophagy and mitophagy. Similar findings were obtained in hearts of mice given simvastatin. Mevalonate abolished simvastatin's effects on Akt/mTOR signaling and autophagy induction in HL-1 cells, indicating that the effects are mediated through inhibition of 3-hydroxy-3-methylglutaryl coenzyme A (HMG-CoA) reductase. Simvastatin-treated HL-1 cells exhibited mitochondrial translocation of Parkin and p62/SQSTM1, fission, and mitophagy. Because Parkin is required for mitophagy and is expressed in heart, we investigated the effect of simvastatin on infarct size in Parkin knockout mice. Simvastatin reduced infarct size in wild-type mice but showed no benefit in Parkin knockout mice. Inhibition of HMG-CoA reductase limits mevalonate availability for both cholesterol and coenzyme Q10 (CoQ) biosynthesis. CoQ supplementation had no effect on statin-induced Akt/mTOR dephosphorylation or macroautophagy in HL-1 cells, but it potently blocked mitophagy. Importantly, CoQ supplementation abolished statin-mediated cardioprotection in vivo. INNOVATION AND CONCLUSION: Acute simvastatin treatment suppresses mTOR signaling and triggers Parkin-dependent mitophagy, the latter which is required for cardioprotection. Coadministration of CoQ with simvastatin impairs mitophagy and cardioprotection. These results raise the concern that CoQ may interfere with anti-ischemic benefits of statins mediated through stimulation of mitophagy.

An Overview of HIV Prevention Interventions for People Who Inject Drugs in Tanzania.

In the past decade, Tanzania has seen a rapid rise in the number of people who inject drugs (PWID), specifically heroin. While the overall HIV prevalence in Tanzania has declined recently to 5.6%, in 2009, the HIV prevalence among PWID remains alarmingly high at 35%. In this paper, we describe how the Tanzania AIDS Prevention Program (TAPP), Médecins du Monde France (MdM-F), and other organisations have been at the forefront of addressing this public health issue in Africa, implementing a wide array of harm reduction interventions including medication-assisted treatment (MAT), needle and syringe programs (NSP), and "sober houses" for residential treatment in the capital, Dar es Salaam, and in Zanzibar. Looking toward the future, we discuss the need to (1) extend existing services and programs to reach more PWID and others at risk for HIV, (2) develop additional programs to strengthen existing programs, and (3) expand activities to include structural interventions to address vulnerabilities that increase HIV risk for all Tanzanians.

Preconditioning involves selective mitophagy mediated by Parkin and p62/SQSTM1.

Autophagy-dependent mitochondrial turnover in response to cellular stress is necessary for maintaining cellular homeostasis. However, the mechanisms that govern the selective targeting of damaged mitochondria are poorly understood. Parkin, an E3 ubiquitin ligase, has been shown to be essential for the selective clearance of damaged mitochondria. Parkin is expressed in the heart, yet its function has not been investigated in the context of cardioprotection. We previously reported that autophagy is required for cardioprotection by ischemic preconditioning (IPC). In the present study, we used simulated ischemia (sI) in vitro and IPC of hearts to investigate the role of Parkin in mediating cardioprotection ex vivo and in vivo. In HL-1 cells, sI induced Parkin translocation to mitochondria and mitochondrial elimination. IPC induced Parkin translocation to mitochondria in Langendorff-perfused rat hearts and in vivo in mice subjected to regional IPC. Mitochondrial depolarization with an uncoupling agent similarly induced Parkin translocation to mitochondria in cells and Langendorff-perfused rat hearts. Mitochondrial loss was blunted in Atg5-deficient cells, revealing the requirement for autophagy in mitochondrial elimination. Consistent with previous reports indicating a role for p62/SQSTM1 in mitophagy, we found that depletion of p62 attenuated mitophagy and exacerbated cell death in HL-1 cardiomyocytes subjected to sI. While wild type mice showed p62 translocation to mitochondria and an increase in ubiquitination, Parkin knockout mice exhibited attenuated IPC-induced p62 translocation to the mitochondria. Importantly, ablation of Parkin in mice abolished the cardioprotective effects of IPC. These results reveal for the first time the crucial role of Parkin and mitophagy in cardioprotection.

Diminished AMPK signaling response to fasting in thioredoxin-interacting protein knockout mice.

Thioredoxin-interacting protein (Txnip) knockout (TKO) mice exhibit impaired response to fasting. Herein, we showed that activation of adenine monophosphate-activated protein kinase and cellular AMP levels were diminished in the heart and soleus muscle but not in gastrocnemius muscle of fasting TKO mice. Similarly, glycogen content in fasted TKO mice was increased in oxidative muscles but was not different in glycolytic muscles. These data suggest Txnip deficiency has a higher impact on oxidative muscle than glycolytic muscles and provide new insights into the metabolic role of Txnip.

A psychosocial comparison of New Orleans and Houston crack smokers in the wake of Hurricane Katrina.

The purpose of this study was to compare psychological distress in a sample of African American crack cocaine users who relocated to Houston from New Orleans after Hurricane Katrina to African American drug users resident in Houston. Fifty-four African Americans from New Orleans were compared to a sample of 162 people in Houston. Data were collected between June 2002 and December 2005. There were no significant differences between the two groups on either depression or anxiety, but the New Orleans sample scored higher on the self-esteem scale and scored slightly lower on the risk-taking scale.

Male homosexual identities, relationships, and practices among young men who have sex with men in Vietnam: implications for HIV prevention.

Rapid socioeconomic transformation in Vietnam in last 15 years has been followed by more liberation of sexual expression and representation of sexual identity among young people. There has been an increase in the visibility of homosexual men in major cities of Vietnam who were largely an unknown population until the emergence of the HIV epidemic. Men who have sex with men (MSM) are now considered as one of the target groups in many HIV prevention programs. This qualitative study examines local identities, relationships, and sexual practices among young MSM aged 15-24 in the cities of Hanoi and Ho Chi Minh City. Our analyses were based on 26 in-depth interviews and 10 focus group discussions with young MSM recruited through public place intercepts and cruising areas. Data document the linguistic classification, sexual relationships and behaviors, identity and process of homosexual identification, and the potential linkage between sexual identity and sexual behaviors of MSM in Vietnam. Data also highlight the stages of homosexual community development in urban Vietnam and important differences between Vietnam and the West in the representation of homosexual identity, relationships, and practices. In light of the findings, we suggest that the continuing development and elaboration of a homosexual community in Hanoi and Ho Chi Minh City offers significant opportunities for targeted HIV/AIDS prevention activities in the Vietnamese MSM population.

Improving Condom Self-Efficacy and Use among Individuals Living with HIV: The Positive Choices Mapping Intervention.

Few interventions have been designed to improve behavioral outcomes and reduce risk of HIV transmission of individuals living with HIV, most focusing on preventative efforts directed at individuals who are HIV-negative. However, people living with HIV present individual and public health risks (infection with a different strain of HIV, health complications from contracting STD's, continued sexual activity with individuals with unknown HIV status) that have become the focus of intervention efforts. The current paper explores a promising new intervention, The Positive Choices Mapping intervention (PCM), designed to increase condom self-efficacy and use among African American crack cocaine smokers who are living with HIV. The intervention was grounded in Social Cognitive Theory and incorporated an empirically backed visual representation strategy (node-link mapping). The focus of the current paper is on the main components of the intervention.

Perceptions of risk in research participation among underserved minority drug users.

Research with underserved minority drug users is essential to quality health care and prevention. Understanding how participants perceive risk in research is necessary to inform research regulators so that research protections are neither lax, exposing participants to harm, nor overly stringent, thereby denying access to beneficial research. Data from 37 semistructured interviews of underserved, African-American crack cocaine users, collected from February to May 2006 in a large, urban setting, were analyzed using content analysis. In three hypothetical studies, participants recognized risks as relative and articulated and evaluated specific risks. Research regulators may enhance the accuracy of risk assessment in research by incorporating the views of participants. Study implications and limitations are noted. Future research on risk perception in research participation is suggested.

Decisions to participate in research: views of underserved minority drug users with or at risk for HIV.

Under-representation of minority populations, particularly African Americans, in HIV/AIDS research is problematic because African Americans bear a greater disease burden from HIV/AIDS. Studies of motivations for participating in research have emphasized factors affecting individuals' willingness to participate and barriers to participation, especially in regard to HIV vaccine research. Little is known about how underserved minority drug users perceive research and their decisions to participate. This study describes African American drug users' perceptions of research participation and their decisions to participate based on three kinds of hypothetical HIV/AIDS-related clinical studies. In-depth, qualitative interviews were conducted with 37 underserved, African American crack cocaine users, recruited from participants already enrolled in three different behavioral HIV prevention studies. Interviews were recorded, transcribed, coded for themes and sub-themes and analyzed using directed and conventional content analysis. Participants' decisions to take part in research often involved multiple motivations for participating. In addition, decisions to participate were characterized by four themes: a desire for information; skepticism and mistrust of research and researchers; perceptions of medical care and monitoring within a study; and participant control in decisions to participate or decline participation. Lack of adequate information and/or medical care and monitoring within a study were related to mistrust, while the provision of information was viewed by some individuals as a right and acknowledgement of the participant's contribution to the study. Participants perceived, rightly or wrongly, that medical monitoring would control some of the risks of a study. Participants also described situations of exerting control over decisions to enter or withdraw from a research study. Preliminary findings suggest that continuous communication and provision of information may enhance enrollment and adherence. Further exploration of decisions to participate in research will add to the understanding of this complex phenomenon and enhance the ability of individuals with HIV/AIDS to benefit from research.

Internet influences on sexual practices among young people in Hanoi, Vietnam.

While the influences of the Internet on adult sexuality are well recognized, research on the potential connection between the Internet and young people's sexuality is still limited. We conducted a qualitative study to examine how young people (aged 15-19 years) in Hanoi, Vietnam used the Internet to develop sexual practices and identities. Our analysis of texts from focus groups, in-depth interviews, chat scripts and field notes reveals how the Internet is used to assemble sexual information that was not available from other sources such as the family and school. Young people's narratives also show how they use the Internet as a medium for expressing sexual identities and desires. In the light of these findings, we suggest expanding sex education to include issues that are important to young people such as emotions and relationships, rather than simply focusing more narrowly on reproduction, public health and other interests of the state.

The lives of female sex workers in Vietnam: Findings from a qualitative study.

To facilitate better understanding of the environment and power structures in which sex work in Vietnam takes place, this study examined the sex workers' social and economic lives, their working environment, social relationships and presentation of self in everyday social contacts and interactions. Thirty in-depth interviews and 14 focus groups were conducted with street-based and venue-based sex workers in the cities of Da Nang and Hanoi. Results show that sex workers live and work within a complex system involving multiple relationships. In any of these relations, women have limited power to protect their personal security and secure payment for services rendered. Economic hardship is a major problem facing street-level sex workers and contributes to unsafe sexual practices. Venue-based sex workers worry less about economic hardships as such, but frequently incur gambling debts. Women also reported incidents of abuse and experiences of social stigma. Although many women exhibited a strong desire to leave sex work, they found themselves trapped in the sex industry by the lack of alternative employment options. This study provides evidence that socio-psychological factors must be addressed along with risky behaviours to promote women's well-being and social integration.