Genetic and spectrally distinct in vivo imaging: embryonic stem cells and mice with widespread expression of a monomeric red fluorescent protein

BACKGROUND: DsRed the red fluorescent protein (RFP) isolated from Discosoma sp. coral holds much promise as a genetically and spectrally distinct alternative to green fluorescent protein (GFP) for application in mice. Widespread use of DsRed has been hampered by several issues resulting in the inability to establish and maintain lines of red fluorescent protein expressing embryonic stem cells and mice. This has been attributed to the non-viability, or toxicity, of the protein, probably as a result of its obligate tetramerization. A mutagenesis approach directing the stepwise evolution of DsRed has produced mRFP1, the first true monomer. mRFP1 currently represents an attractive autofluorescent reporter for use in heterologous systems. RESULTS: We have used embryonic stem cell-mediated transgenesis to evaluate mRFP1 in embryonic stem cells and mice. We find that mRFP1 exhibits the most spatially homogenous expression when compared to the native (tetrameric) and variant dimeric forms of DsRed. High levels of mRFP1 expression do not affect cell morphology, developmental potential or viability and fertility of animals. High levels of widespread mRFP1 expression are maintained in a constitutive manner in embryonic stem cells in culture and in transgenic animals. We have used various optical imaging modalities to visualize mRFP1 expressing cells in culture, in embryos and adult mice. Moreover co-visualization of red, green and cyan fluorescent cells within a sample is easily achieved without the need for specialized methodologies, such as spectral deconvolution or linear unmixing. CONCLUSION: Fluorescent proteins with excitation and/or emission profiles in the red part of the visible spectrum represent distinct partners, or longer wavelength substitutes for GFP. Not only do DsRed-based RFPs provide a genetically and spectrally distinct addition to the available repertoire of autoflorescent proteins, but by virtue of their spectral properties they permit deeper tissue imaging. Our work in generating CAG::mRFP1 transgenic ES cells and mice demonstrates the developmental neutrality of mRFP1 in an organismal context. It paves the way for the use of DsRed-based monomeric RFPs in transgenic and gene targeted approaches which often necessitate spatially and/or temporally restricted reporter expression. Moreover animals of the CAG::mRFP1 transgenic strain serve as a source of RFP tagged tissue for the derivation of cell lines and explant, transplant and embryo chimera experiments

Playing to our Strengths: An Academic/Health Department Collaboration to Facilitate Public Protections Against Tuberculosis

Background: Economic modeling and analysis can facilitate evidence-based policy and practice. Such analyses may exceed the technical capacity and mission of public health agencies, yet may be critical to sustain health protections such as tuberculosis (TB) control. The net effect of TB prevention is incompletely understood, hampering objective value judgments of national TB elimination policies. This may promote inefficiencies and threaten individual and public health protections. We describe how a HRSA-funded Texas Public Health Training Center (TPHTC) coordinated specialized analytical skills with the needs of a state public health department to create a user-friendly tool to inform planning and resource allocation; and to identify risks, benefits, opportunities, and value in the context of Texas’ Department of State Health Services (TDSHS) TB control programs. Methods: Key informants within TDSHS and faculty from the University of North Texas Health Science Center (UNTHSC) and its associated TPHTC collaborated to develop a combined health, cost, and outcome model to answer questions of interest to TDSHS. The model incorporates standard techniques, TB specific logic and data, and Texas specific population, epidemiologic, and programmatic data into a Microsoft Excel platform. Outcome measures include 2013 USD costs from a public agency perspective and health losses denominated in Quality Adjusted Life Years (QALYs) (but not assigned a dollar value). Outcomes represent the 10-year accrued net present value (NPV) of variable costs associated with each incident TB case. The model used only publicly available, non-confidential data. Results: Under current public TB prevention and control practices we estimate public spending for acute medical care and health losses for Texas’ 1325 TB cases in 2011 will exceed a NPV $100,000,000 and 4800 QALYs by 2022; societal costs would likely be much higher. Public TB control is relatively cost effective and preserves 3.7 QALYs/case averted at$54,000/QALY. Implications: Partnerships between academic institutions and public health departments offer the potential to produce the economic analyses needed for judicious use of public resources. We used accessible methods, data, and technology to create a model to analyze the relative value of public TB control in Texas as well as to identify potential efficiencies within activities. Our analysis suggests public efforts against latent TB infection effectively and efficiently mitigate some of the substantial health and cost impact of TB to Texas. Most of TB’s burden remains, however, and more careful targeting of prevention by risk promises rich rewards

Oxytocin Attenuates Yohimbine-Induced Reinstatement of Alcohol-Seeking in Female Rats via the Central Amygdala

Alcohol use disorder is a significant public health concern, further exacerbated by an increased risk of relapse due to stress. In addition, factors such as biological sex may contribute to the progression of addiction, as females are especially susceptible to stress-induced relapse. While there have been many studies surrounding potential pharmacological interventions for male stress-induced ethanol reinstatement, research regarding females is scarce. Recently, the neuropeptide oxytocin has gained interest as a possible pharmacological intervention for relapse. The present study examines how oxytocin affects yohimbine-induced reinstatement of ethanol-seeking in female rats using a self-administration paradigm. Adult female rats were trained to press a lever to access ethanol in daily self-administration sessions. Rats then underwent extinction training before a yohimbine-induced reinstatement test. Rats administered with yohimbine demonstrated significantly higher lever response indicating a reinstatement of ethanol-seeking behavior. Oxytocin administration, both systemically and directly into the central amygdala, attenuated the effect of yohimbine-induced reinstatement of ethanol-seeking behavior. The findings from this study establish that oxytocin is effective at attenuating alcohol-relapse behavior mediated by the pharmacological stressor yohimbine and that this effect is modulated by the central amygdala in females. This provides valuable insight regarding oxytocin’s potential therapeutic effect in female stress-induced alcohol relapse

Trends in healthcare utilization among older Americans with colorectal cancer: A retrospective database analysis

<p>Abstract</p> <p>Background</p> <p>Analyses of utilization trends (cost drivers) allow us to understand changes in colorectal cancer (CRC) costs over time, better predict future costs, identify changes in the use of specific types of care (eg, hospice), and provide inputs for cost-effectiveness models. This retrospective cohort study evaluated healthcare resource use among US Medicare beneficiaries diagnosed with CRC between 1992 and 2002.</p> <p>Methods</p> <p>Cohorts included patients aged 66+ newly diagnosed with adenocarcinoma of the colon (n = 52,371) or rectum (n = 18,619) between 1992 and 2002 and matched patients from the general Medicare population, followed until death or December 31, 2005. Demographic and clinical characteristics were evaluated by cancer subsite. Resource use, including the percentage that used each type of resource, number of hospitalizations, and number of hospital and skilled nursing facility days, was evaluated by stage and subsite. The number of office, outpatient, and inpatient visits per person-year was calculated for each cohort, and was described by year of service, subsite, and treatment phase. Hospice use rates in the last year of life were calculated by year of service, stage, and subsite for CRC patients who died of CRC.</p> <p>Results</p> <p>CRC patients (mean age: 77.3 years; 44.9% male) used more resources than controls in every category (<it>P </it>< .001), with the largest differences seen in hospital days and home health use. Most resource use (except hospice) remained relatively steady over time. The initial phase was the most resource intense in terms of office and outpatient visits. Hospice use among patients who died of CRC increased from 20.0% in 1992 to 70.5% in 2004, and age-related differences appear to have evened out in later years.</p> <p>Conclusion</p> <p>Use of hospice care among CRC decedents increased substantially over the study period, while other resource use remained generally steady. Our findings may be useful for understanding CRC cost drivers, tracking trends, and forecasting resource needs for CRC patients in the future.</p

The elusive MAESTRO gene: Its human reproductive tissue-specific expression pattern

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United States Acculturation and Cancer Patients' End-of-Life Care

Background: Culture shapes how people understand illness and death, but few studies examine whether acculturation influences patients' end-of-life treatment preferences and medical care. Methods and Findings: In this multi-site, prospective, longitudinal cohort study of terminally-ill cancer patients and their caregivers (n = 171 dyads), trained interviewers administered the United States Acculturation Scale (USAS). The USAS is a 19-item scale developed to assess the degree of "Americanization" in first generation or non-US born caregivers of terminally-ill cancer patients. We evaluated the internal consistency, concurrent, criterion, and content validity of the USAS. We also examined whether caregivers' USAS scores predicted patients' communication, treatment preferences, and end-of-life medical care in multivariable models that corrected for significant confounding influences (e.g. education, country of origin, English proficiency). The USAS measure was internally consistent (Cronbach α = 0.98); and significantly associated with US birthplace (r = 0.66, P<0.0001). USAS scores were predictive of patients' preferences for prognostic information (AOR = 1.31, 95% CI:1.00-1.72), but not comfort asking physicians' questions about care (AOR 1.23, 95% CI:0.87-1.73). They predicted patients' preferences for feeding tubes (AOR = 0.68, 95% CI:0.49-0.99) and wish to avoid dying in an intensive care unit (AOR = 1.36, 95% CI:1.05-1.76). Scores indicating greater acculturation were also associated with increased odds of patient participation in clinical trials (AOR = 2.20, 95% CI:1.28-3.78), compared with lower USAS scores, and greater odds of patients receiving chemotherapy (AOR = 1.59, 95% CI:1.20-2.12). Conclusion: The USAS is a reliable and valid measure of "Americanization" associated with advanced cancer patients' end-of-life preferences and care. USAS scores indicating greater caregiver acculturation were associated with increased odds of patient participation in cancer treatment (chemotherapy, clinical trials) compared with lower scores. Future studies should examine the effects of acculturation on end-of-life care to identify patient and provider factors that explain these effects and targets for future interventions to improve care (e.g., by designing more culturally-competent health education materials). © 2013 Wright et al

A Woman's Territory: Female Protagonists in 21st Century Road Movie–Based Fairy Tale Films

This paper closely examines fairy tale films with road movie components, in particular those films featuring female protagonists. The study’s objective is twofold: first, to further develop existing research on the road movie by exploring one of the lesser known constituents of this broad genre and, second, to address gaps in scholarly literature on road movies when it comes to themes in female-led trips and the characterization of travelling females. \ud Through a detailed analysis of the journeys of female characters in recent voyage-oriented fairy tale films—Peter Pan (P.J. Hogan, 2003), Alice in Wonderland (Tim Burton, 2010) and Tangled (Byron Howard and Nathan Greno, 2010)—the investigation shows that these new heroines significantly differ from those of old. Their travels are portrayed as being less difficult, and the traits that they exhibit while on the road, namely fearlessness, rationality and an undying optimism, are rather favourable when compared to those exhibited by former road heroines. \ud Although these protagonists still face characters who wish to impede their movement away from a domestic setting, it is demonstrated that the protagonists are ultimately successful in not only acquiring power in the surroundings that they find themselves in, but also in carrying over their goals and dreams to their own worlds upon their return, privileges that most former road heroines did not have. Ultimately, the study shows that females can be as efficient travellers as males, and in some cases are portrayed as even more competent than their male counterparts