Conditionally-live attenuated SIV upregulates global T effector memory cell frequency under replication permissive conditions.

Background: Live attenuated SIV induces potent protection against superinfection with virulent virus; however the mechanism of this vaccine effect is poorly understood. Such knowledge is important for the development of clinically acceptable vaccine modalities against HIV. Results: Using a novel, doxycycline dependent, replication-competent live-attenuated SIVmac239Δnef (SIVrtTAΔnef), we show that under replication-permissive conditions SIV-rtTAΔnef is fully viable. Twelve rhesus macaques were infected with a peak plasma vRNA on average two log10 lower than in 6 macaques infected with unconditionally replication-competent SIVΔnef. Consistent with the attenuated phenotype of the viruses the majority of animals displayed low or undetectable levels of viraemia by 42-84 days after infection. Next, comparison of circulating T cells before and after chronic infection with parental SIVΔnef revealed a profound global polarisation toward CD28-CCR7- T-effector memory 2 (TEM2) cells within CD95+CD4+ and CD95+CD8+ populations. Critically, a similar effect was seen in the CD95+ CD4+ population and to somewhat lesser extent in the CD95+ CD8+ population of SIV-rtTAΔnef chronically infected macaques that were maintained on doxycycline, but was not seen in animals from which doxycycline had been withdrawn. The proportions of gut-homing T-central memory (TCM) and TEM defined by the expression of α4β7 and CD95 and differential expression of CD28 were increased in CD4 and CD8 cells under replication competent conditions and gut-homing CD4 TCM were also significantly increased under non-permissive conditions. TEM2 polarisation was seen in the small intestines of animals under replication permissive conditions but the effect was less pronounced than in the circulation. Intracellular cytokine staining of circulating SIV-specific T cells for IL-2, IFN-γ, TNF-α and IL-17 showed that the extent of polyfunctionality in CD4 and CD8 T cells was associated with replication permissivity; however, signature patterns of cytokine combinations were not distinguishable between groups of macaques. Conclusion: Taken together our results show that the global T memory cell compartment is profoundly skewed towards a mature effector phenotype by attenuated SIV. Results with the replication-conditional mutant suggest that maintenance of this effect, that may be important in vaccine design, might require persistence of replicating virus

Synthetic Lethality of Chk1 Inhibition Combined with p53 and/or p21 Loss During a DNA Damage Response in Normal and Tumor Cells

Cell cycle checkpoints ensure genome integrity and are frequently compromised in human cancers. A therapeutic strategy being explored takes advantage of checkpoint defects in p53-deficient tumors in order to sensitize them to DNA-damaging agents by eliminating Chk1-mediated checkpoint responses. Using mouse models, we demonstrated that p21 is a key determinant of how cells respond to the combination of DNA damage and Chk1 inhibition (combination therapy) in normal cells as well as in tumors. Loss of p21 sensitized normal cells to the combination therapy much more than did p53 loss and the enhanced lethality was partially blocked by CDK inhibition. In addition, basal pools of p21 (p53 independent) provided p53 null cells with protection from the combination therapy. Our results uncover a novel p53-independent function for p21 in protecting cells from the lethal effects of DNA damage followed by Chk1 inhibition. As p21 levels are low in a significant fraction of colorectal tumors, they are predicted to be particularly sensitive to the combination therapy. Results reported in this study support this prediction

The Effect of Race and Chronic Obstructive Pulmonary Disease on Long-Term Survival after Coronary Artery Bypass Grafting

Background: Chronic obstructive pulmonary disease (COPD) is a known predictor of decreased long-term survival after coronary artery bypass grafting (CABG). Differences in survival by race have not been examined. Methods: A retrospective cohort study was conducted of CABG patients between 2002 and 2011. Long-term survival was compared in patients with and without COPD and stratified by race. Hazard ratios (HR) and 95% confidence intervals (CI) were computed using a Cox regression model. Results: A total of 984 (20%) patients had COPD (black n = 182; white n = 802) at the time of CABG (N = 4,801). The median follow-up for study participants was 4.4 years. COPD was observed to be a statistically significant predictor of decreased survival independent of race following CABG (no COPD: HR = 1.0; white COPD: adjusted HR = 1.9, 95% CI = 1.7–2.3; black COPD: adjusted HR = 1.6, 95% CI = 1.1–2.2). Conclusion: Contrary to the expected increased risk of mortality among black COPD patients in the general population, a similar survival disadvantage was not observed in our CABG population

Impact of Community-Based Larviciding on the Prevalence of Malaria Infection in Dar es Salaam, Tanzania.

The use of larval source management is not prioritized by contemporary malaria control programs in sub-Saharan Africa despite historical success. Larviciding, in particular, could be effective in urban areas where transmission is focal and accessibility to Anopheles breeding habitats is generally easier than in rural settings. The objective of this study is to assess the effectiveness of a community-based microbial larviciding intervention to reduce the prevalence of malaria infection in Dar es Salaam, United Republic of Tanzania. Larviciding was implemented in 3 out of 15 targeted wards of Dar es Salaam in 2006 after two years of baseline data collection. This intervention was subsequently scaled up to 9 wards a year later, and to all 15 targeted wards in 2008. Continuous randomized cluster sampling of malaria prevalence and socio-demographic characteristics was carried out during 6 survey rounds (2004-2008), which included both cross-sectional and longitudinal data (N = 64,537). Bayesian random effects logistic regression models were used to quantify the effect of the intervention on malaria prevalence at the individual level. Effect size estimates suggest a significant protective effect of the larviciding intervention. After adjustment for confounders, the odds of individuals living in areas treated with larviciding being infected with malaria were 21% lower (Odds Ratio = 0.79; 95% Credible Intervals: 0.66-0.93) than those who lived in areas not treated. The larviciding intervention was most effective during dry seasons and had synergistic effects with other protective measures such as use of insecticide-treated bed nets and house proofing (i.e., complete ceiling or window screens). A large-scale community-based larviciding intervention significantly reduced the prevalence of malaria infection in urban Dar es Salaam

Role of Occult and Post-acute Phase Replication in Protective Immunity Induced with a Novel Live Attenuated SIV Vaccine

In order to evaluate the role of persisting virus replication during occult phase immunisation in the live attenuated SIV vaccine model, a novel SIVmac239Δnef variant (SIVrtTA) genetically engineered to replicate in the presence of doxycycline was evaluated for its ability to protect against wild-type SIVmac239. Indian rhesus macaques were vaccinated either with SIVrtTA or with SIVmac239Δnef. Doxycycline was withdrawn from 4 of 8 SIVrtTA vaccinates before challenge with wild-type virus. Unvaccinated challenge controls exhibited ~107 peak plasma viral RNA copies/ml persisting beyond the acute phase. Six vaccinates, four SIVmac239Δnef and two SIVrtTA vaccinates exhibited complete protection, defined by lack of wild-type viraemia post-challenge and virus-specific PCR analysis of tissues recovered post-mortem, whereas six SIVrtTA vaccinates were protected from high levels of viraemia. Critically, the complete protection in two SIVrtTA vaccinates was associated with enhanced SIVrtTA replication in the immediate post-acute vaccination period but was independent of doxycycline status at the time of challenge. Mutations were identified in the LTR promoter region and rtTA gene that do not affect doxycycline-control but were associated with enhanced post-acute phase replication in protected vaccinates. High frequencies of total circulating CD8+T effector memory cells and a higher total frequency of SIV-specific CD8+ mono and polyfunctional T cells on the day of wild-type challenge were associated with complete protection but these parameters were not predictive of outcome when assessed 130 days after challenge. Moreover, challenge virus-specific Nef CD8+ polyfunctional T cell responses and antigen were detected in tissues post mortem in completely-protected macaques indicating post-challenge control of infection. Within the parameters of the study design, on-going occult-phase replication may not be absolutely required for protective immunity

The development and effect of a multimedia presentation upon knowledges about a university general physical education program

This investigation sought to develop a physical education multimedia presentation designed to inform students about the UNC-G General Physical Education Program, and to determine the effect of the presentation on the program knowledges of incoming freshmen students. The preliminary preparation included the following steps: (a) informal interviews with the UNC-G physical education faculty, (b) the development of a knowledge test, and (c) the development of a physical education multimedia presentation. Twelve members of the UNC-G physical education faculty were interviewed during the spring semester, 1977. The results of the interviews were utilized in determining the content of the knowledge test and the multimedia presentation. A criterion-referenced mastery test consisting of 34 true/false items was constructed to measure student knowledges about the UNC-G General Physical Education Program. Content validity was established based upon the judgment of three selected test judges. The physical education multimedia presentation consisted of: (a) a slide/tape series, (b) a set of five activity booklets, and (c) an activity brochure

Rat killing, Two prisoners and Molting : three short stories

These three short stories are united in an exploration of the possibilities and limitations of behavioral observation in revealing emotions. As much as possible, observable details reveal the characters' emotional states in order to create, through images, the tangible stimuli from which conclusions about a person's feelings naturally derive. The problem was to select what a character sees and does ostensibly that reveal his inner feelings. By revealing his emotions in this indirect manner, through observable reality, his inner self becomes tangible and, hopefully, more capable of evoking emotive responses in readers. Through this technique, though structured in a straightforward, traditional manner, the stories are intended to become more concrete, vivid and dramatic

The effects of maternal education on pregnancy knowledge and behaviors using the Pregnancy Risk Assessment Monitoring System data

Background: It is estimated that 90 million Americans have trouble understanding and using health information. Specific Aims: Examine the: (1) association between education and the health knowledge and behaviors of maternal women in two different time periods (1997-1999 and 2006, 2008, & 2009) and (2) trends in maternal health knowledge and behaviors, separately, stratified by education. Study Design: A retrospective design was conducted, using the North Carolina PRAMS data. Analyses: Multiple logistic regression models estimated the likelihood that (1) education would affect behavior and knowledge in the areas of the folic acid consumption, breastfeeding, and infant sleep position and (2) there would be a positive trend in these three areas. Results: Women with <12th grade education were less likely to (1) have knowledge of folic acid (OR=0.64, 95% CI=0.48- 0.84) and (2) use the correct infant sleep position (OR=0.54, 95% CI=0.34-0.85) when compared with high school graduates. The trend found that women with <12 th grade education in the latter time period were more likely to have knowledge about folic acid. Women of all educational backgrounds were more likely to breastfeed in the latter phase compared to earlier phase and women with between 9th and 15th grades education were more likely to place their infant to sleep on his/her back. Significance: The results suggest that improvements over time are occurring in some key maternal health areas, such as folic acid knowledge, breastfeeding, and correct infant sleep position

Role for dual-specificity phosphatases on mitogen-activated protein kinase signaling in adipocytes

Obesity and diabetes are major public health concerns that contribute to cardiovascular disease, hypertension, and stroke. It is now widely accepted that chronic inflammation is an important element of pathogenic mechanisms linking obesity to diabetes. Moreover, clinical and experimental evidence has established the mitogen-activated protein kinase (MAPK) signaling pathway as a pivotal mediator during inflammatory stress in coupling obesity to insulin resistance (IR). While numerous studies have examined the upstream kinase activation of MAPKs, few have examined mechanisms that dephosphorylate, and thus, deactivate these pathways, potentially affording protection against adipose tissue (AT) inflammation and obesity-induced IR. Data presented in this dissertation demonstrate that several MAPK-specific dual-specificity phosphatases (DUSPs) are induced in AT under conditions of genetic and diet-induced obesity that is associated with increased inflammation and IR. While AT is composed of multiple cell types, evidence suggests an essential role for preadipocytes (PAs) and adipocytes (ADs) in the development of AT inflammation and IR. Therefore, we further report phenotypic differences in DUSP expression where four of the ten MAPK-specific DUSPs are more abundant in PAs compared to ADs while two of the ten DUSPs are more abundant in ADs compared to PAs, suggesting a regulatory role for these phosphatases that is cell type specific. Moreover, phenotypic differences were observed regarding MAPK signaling and DUSP expression in PAs and ADs exposed to TNFalpha-mediated inflammatory stress, where ERK, JNK, and p38 phosphorylation was markedly elevated and transient in PAs while ERK and JNK phosphorylation was prolonged in ADs concomitant with the phenotypic differences in inducible DUSPs. As induction of DUSPs in PAs kinetically mirrored MAPK dephosphorylation, we further show that de novo mRNA synthesis was essential for MAPK dephosphorylation, suggesting a role for inducible DUSPs in the modulation of MAPK signaling. Based on these data, we present empirical evidence that DUSP knockdown markedly increased the magnitude and duration of ERK, JNK, and p38 phosphorylation in response to inflammatory stress, subsequently elevating MAPK-dependent pro-inflammatory cytokine and chemokine gene expression. Collectively, these findings demonstrate an essential role for DUSPs in the timely modulation of MAPK signaling, highlighting prospective therapeutic targets linking obesity with metabolic inflammatory diseases

Impaired decisional impulsivity in pathological videogamers

Abstract Background Pathological gaming is an emerging and poorly understood problem. Impulsivity is commonly impaired in disorders of behavioural and substance addiction, hence we sought to systematically investigate the different subtypes of decisional and motor impulsivity in a well-defined pathological gaming cohort. Methods Fifty-two pathological gaming subjects and age-, gender- and IQ-matched healthy volunteers were tested on decisional impulsivity (Information Sampling Task testing reflection impulsivity and delay discounting questionnaire testing impulsive choice), and motor impulsivity (Stop Signal Task testing motor response inhibition, and the premature responding task). We used stringent diagnostic criteria highlighting functional impairment. Results In the Information Sampling Task, pathological gaming participants sampled less evidence prior to making a decision and scored fewer points compared with healthy volunteers. Gaming severity was also negatively correlated with evidence gathered and positively correlated with sampling error and points acquired. In the delay discounting task, pathological gamers made more impulsive choices, preferring smaller immediate over larger delayed rewards. Pathological gamers made more premature responses related to comorbid nicotine use. Greater number of hours played also correlated with a Motivational Index. Greater frequency of role playing games was associated with impaired motor response inhibition and strategy games with faster Go reaction time. Conclusions We show that pathological gaming is associated with impaired decisional impulsivity with negative consequences in task performance. Decisional impulsivity may be a potential target in therapeutic management