Fanconi anemia (FA) is a genetically inherited disease that frequently results in bone marrow failure, a high prevalence of leukemia, and solid tumors of the head and neck that are associated with genomic instability. Abnormal chromosome configurations produced in FA cells, called radials, are diagnostic of the disease. The mechanism of radial formation is not fully understood. However, they can be induced to form by exposure of cells to agents that promote a type of DNA damage called interstrand crosslinks (ICLs), covalently bonded DNA strands caused by exposure to ICL inducing agents such as those used in chemotherapy or endogenously produced metabolic byproducts. Radials can occur spontaneously in the cells of FA patients, therefore, understanding the formation of radials, as well as their fate during the cell cycle, may help in designing effective therapies to counteract the genomic instability. The current aim is to investigate radial formation and cell recovery afterwards through careful examination and identification of structural abnormalities in chromosomes that may be a result of prior radial formation. To carry out this goal, a population study will be performed using the primary PD20 (FANCD2) cell line. Mitomycin C (MMC) will be used to induce ICLs and cells will be harvested with paired controls at multiple time points following damage. Cells will be analyzed using classical G-banding for the presence of structural and numerical chromosome abnormalities. A significant increase in chromosome abnormalities structurally consistent with radials that have gone through mitosis is expected to be observed during analysis.

Cocaine and other drugs of abuse have destroyed the lives of many individuals as those who take such drugs struggle with the powerful vice of addiction. Such drugs have been shown to cause changes in the brain, specifically in the cortex and the striatum, which has been linked to the manifestation and maintenance of addiction. Within the striatum there exists cell populations of neurons, which vary in their outputs to the brain and are regulated by glutamate inputs from the cortex. The involvement of these individual cell populations in addiction, including how they are regulated by cortical glutamate, is not yet fully understood. In order to investigate this issue, I collect data in and run rat behavior experiments with rats that have been treated with viral vectors, which temporarily decrease neuron activity in specific cell populations when activated by specific ligands. After the behavioral data has been collected I process the tissues by slicing the brains and stain them with techniques such as immunohistochemistry. Then I determine if the vectors were actually present in the expected cells due to their fluorescence after immunohistochemistry procedures. In some of the work that I have done with Kerry Kerstetter we have found that decreasing activity in cortical glutamate projections to the striatum resulted in an increase in motivation for cocaine. A better understanding of the intricate pathways involved in drug motivation will allow for better treatment and care for those suffering from addiction. Our hope is to provide more information on such pathways to help progress the development of future treatments for victims of addiction.

Autism spectrum disorders (ASD) are neurodevelopmental disorders with symptoms related to social communication and repetitive behavior. One aspect of social communication that is particularly challenging for children with ASD is pragmatic language including the ability to clearly convey narrative information to others. Narrative language, such as story telling, is a type of pragmatic ability that utilizes cognitive-linguistic planning and coordination of information. The first objective of our study is to compare the narrative language abilities of typically developing children to children with ASD. Our second objective is to test the relation between narrative abilities and severity of symptoms within the group with ASD. Seventy-five 6 to 11 year old children with idiopathic ASD or typical development participated. Groups matched in age, IQ, and sex ratio. Narrative language samples were extracted from a longer diagnostic interview, the Autism Diagnostic Observation Schedule (ADOS). Scoring of narrative language skills was based on the coding system developed by Reilly and colleagues. Symptoms were assessed via the ADOS, the Autism Diagnostic Interview-Revised, Social Responsiveness Scale-2, and Repetitive Behavior Scale-Revised. We expect to find a lower level of narrative ability in the group with ASD, given previous reports of pragmatic errors , misinterpretation of key events, and dysfluencies among children with ASD. For the second objective, we predict that there will be a correlation between the narrative level and social symptoms, such that better narrative ability relates to fewer symptoms. In the repetitive domain, we expect to see more repetitive behaviors with relation to lower level of narrative ability by looking at dysfluent speech, which entails repetitions and self-corrections. Our results may lead to future studies regarding narrative ability in speech interventions towards children with ASD. 

While language delay and speech impairment in individuals with Autism Spectrum Disorders (ASD) have been widely investigated, few studies have analyzed the role that non-verbal forms of communication, such as gestures, may play in augmenting impaired speech in individuals with ASD. One system that has been heavily studied, which has the potential to enhance or inhibit communication, is the Mirror Neuron System (MNS). While the MNS could serve to augment speech, the dysfunction of this system could prove detrimental to those who already possess impaired language skills. If the MNS were to be equally affected by ASD, individuals with this disorder would be unable to compensate for impairments in verbal communication with non-verbal forms of interaction. Although reduced activity in MNS areas in ASD individuals has been identified, the implications of this dysfunction remain unclear. In order to identify if the dysfunction of the MNS impairs non-verbal forms of communication, we recorded 16 typically developing (TD) adolescents and 13 ASD adolescents as they played a game of charades. We utilized charades that were not associated with speech centers, but instead have been shown to rely heavily on an intact MNS and motor cortex to mimic the action. This allowed us to identify whether reduced activation of mirror neurons impacts neural centers that serve to augment language through nonverbal means. Using video footage, two blind coders scored the quality of gestures based upon a 0-8 point scale that was unique to each gesture. Our preliminary results suggest that adolescents with ASD do not have significantly lower scores of functional gesture quality compared to TD individuals. One possible implication of these data is that other regions of the brain may compensate for MNS dysfunction and, importantly, may serve as potential therapeutic targets for strategic interventions to minimize the language impairments associated ASD.

Glutamate within the nucleus accumbens (NAc) regulates the development of psychomotor sensitization, a model of addiction. Two prominent sources of glutamate are the prefrontal cortex (PFC) and thalamus neurons projecting to the NAc, and previous studies have shown these two regions contribute to amphetamine sensitization. We developed a combinatorial viral-mediated gene transfer technique to alter intracellular signaling cascades and neuron activity in either PFC or thalamic neurons projecting to NAc. We hypothesized that both PFC and thalamic glutamatergic projection neurons would regulate sensitization in a similar manner. Rats received bilateral injections of a retrogradely transported canine adenovirus expressing cre-recombinase into the NAc and an adenoassociated virus expressing Gi/o-Designer Receptors Exclusively Activated by Designer Drugs (hM4Di-DREADD) in either PFC or thalamus. This allowed for transient reduction in activity of specifically PFC or thalamic neurons projecting to NAc. I gave rats 4 injections of amphetamine or saline, and I then recorded locomotor activity and stereotypies. My results indicate that decreasing the activity of PFC projections to NAc enhanced amphetamine sensitization, whereas decreasing activity of the thalamic projections to the NAc blocked sensitization. These results suggest that PFC and thalamic glutamatergic input to the NAc differentially regulate amphetamine sensitization. It is possible that the projections from the PFC to NAc may preferentially affect the indirect pathway medium spiny neurons, whereas the thalamus projection to NAc would affect the direct pathway medium spiny neurons. Future studies will begin to address this possibility.

Background: Chronically homeless individuals with alcohol dependence experience serious alcohol-related consequences (Collins et al., 2012; Hwang, Wilkins, Tjepkema, O’Campo, & Dunn, 2009). It is therefore important to identify ways to reduce alcohol-related harm, such as the use of safer drinking strategies. No studies to date have explored this topic among chronically homeless individuals with alcohol dependence. Aims: This study aimed to document participants’ endorsement of safer drinking strategies over the course of a longitudinal pilot study of a combined pharmacobehavioral intervention to reduce alcohol-related harm (Collins et al., 2013). Methods: Participants (N=31; 12.9% women) were currently or formerly chronically homeless individuals with alcohol dependence. Participants’ use of safer drinking strategies was elicited using a list of safer drinking strategies and open-ended questions (e.g., “What strategies on this list might you be willing to try over the next week?”). A qualitative methodology called conventional content analysis was used to explore, categorize and document safer drinking strategies used by this population. The safer drinking strategies that participants chose to try were recorded for weeks 0 and 8. Results: At week 0, strategies that buffered the physical effects of alcohol represented the most highly endorsed category (51.1%), such as eating more often. The next most endorsed category was altering the manner of drinking (36.17%), for example spacing drinks. The final category represented reducing alcohol use (12.76%), such as drinking to avoid withdrawal. Discussion: Preliminary findings indicate that chronically homeless individuals are interested in using safer drinking strategies to reduce harm associated with their drinking. They also indicate that the most relevant ways to reduce harm for this population might be to help individuals buffer the effects of alcohol and alter their manner of drinking.

Most treatment programs for alcohol dependence have prioritized abstinence from substances as the primary treatment goal. However, abstinence-based goals are not always considered desirable or effective by more severely affected populations, such as chronically homeless people with alcohol dependence. Because these individuals comprise a multimorbid and high-utilizing population, they are in need of more focused research attention to elucidate their preferred treatment goals. The aim of this secondary study was therefore to qualitatively and quantitatively document participant-generated treatment goals. Participants were currently or formerly chronically homeless individuals (N=31) with alcohol dependence who were participating in a pharmacobehavioral pilot of extended-release naltrexone and harm-reduction counseling. During the week 0 assessment, study interventionists elicited participants’ treatment goals and recorded them on an open-ended grid. Progress and achievement of goals was confirmed via self-report and recorded by study interventionists at the week 8 follow-up. Conventional content analysis was performed to classify participant-generated treatment goals at weeks 0 and 8. Representation of the three top categories remained stable over the course of the treatment. In the order of their frequency they included drinking-specific goals, quality-of-life goals and health-related goals. Quantitative analyses indicated participants generated an increasing number of goals over the course of treatment; however, the proportions of goals achieved and made progress towards did not increase significantly. Findings confirm hypotheses that participants can independently generate and achieve treatment goals toward alcohol harm reduction and quality-of-life improvement. Larger RCTs are necessary to confirm the role of harm-reduction treatment in helping individuals achieve these goals.

Maltreatment occurring early in life has been shown to cause life-long emotional and physical problems. The topic of Infant and Toddler Mental Health (ITMH) is therefore important in ensuring that infants and toddlers develop into healthy, well-adjusted children and adults. Properly trained nurses can make an immense impact in recognizing and encouraging relational health for this age group. The problem is that there is a gap in the baccalaureate nursing curriculum in regards to training nurses to recognize concepts of ITMH, subsequently affecting the way nurses are able to work with children ages 0-3 and their caregivers. This BSN honors project addressed the question: what are the key ITMH principles and concepts that should be included in baccalaureate nursing curriculum? Semi-structured interviews were conducted with 2-5 Infant Mental Health Specialists from a variety of nursing backgrounds with open-ended questions about ITMH in the context of BSN curriculum. Data from the interviews was analyzed to identify themes, which comprised the essential concepts of ITMH to be included in BSN curriculum. Interviewees were notified of the identified concepts so they may endorse the results and a final literature review was conducted for validation. In addition, a survey of UW BSN nursing students, and new pediatric nurses with less than one year of experience at Seattle Children’s Hospital, was conducted to gauge their knowledge of ITMH concepts. The survey featured a short video depicting a relational health challenge followed by an opportunity for respondents to provide open-ended written reflections. Survey results were analyzed to determine which ITMH concepts the surveyees recognize and where BSN curriculum could be strengthened. The results of this two-part research project is being used to suggest potential improvements to nursing curricula, which in turn will improve the quality of life of infants, toddlers and their families.

Asians have been one of the fastest-growing populations to immigrate to America for over a century. As a marginalized minority, they still face a variety of problems, including, but not limited to, dehumanizing stereotypes, questioning of citizenship, and alienation from social expectations. Studies on ethnic populations have divulged Asian American and other minority experiences, focusing on what it means to be discriminated against. In discrimination against Asian Americans, certain stereotypes can easily be misinterpreted as positive, painting Asians as ideal students, laborers, or spouses. However, such stereotypes are too incorrect to be useful, in that they put unreasonable ideals and expectations on Asians, meanwhile glossing over the realities of anti-Asian racism. American colleges and universities have taken measures of varying degree to address and better accommodate minority groups. Even with universities implementing diversity pedagogies and equal opportunity policies, problems with racism and alienation for Asian Americans may remain salient. If Asian American college students experience racism, are there common themes of discrimination that come up when they talk about their racial experiences? Additionally, are these stereotypes and themes shared amongst ethnic groups? This qualitative research takes an ethnographic approach by interviewing Asian American college students about their experience as self-identified Asian Americans. These interviews are transcribed and coded, by highlighting themes consistent with those found in past literature, including common stereotypes and degrees of racism severity. Ascription to the popular stereotypes, including the “model minority” and “perpetual foreigner” stereotypes, are predictably a commonplace occurrence throughout the lives of Asian American students, while degrees to which the racism occurs – from aversive to more overt actions – may vary amongst individuals. This ethnography is inspired by the need to further address experiences of discrimination within Asian American populations, in order to better define their place in the racism literature.

The toll-like receptor (TLR) proteins are expressed on cells of the innate immune system and recognize structural motifs present on microbial pathogens. Recognition of pathogens by TLRs activates an intracellular signaling cascade and activation of NF-kB, a transcription factor that is critical for expression of inflammatory mediators. Previous studies from our laboratory have shown that single nucleotide variations present in human TLR1 lead to altered expression of TLR1 and are associated with increased risk of death in patients with sepsis. We tested the hypothesis that variations of mouse Tlr1 which correspond to common human variants would result in comparable changes in TLR1 activity. We used site-directed mutagenesis to introduce coding variations in the Tlr1 cDNA. After transfecting the variants into cultured human cells, we stimulated them with Pam3CSK4, a triacylated lipoprotein known to be recognized by the TLR1/TLR2 complex. We then used a bioluminescent luciferase reporter system to measure the levels of NF-kB transcriptional activity. Tlr1 with an Ile allele at position 605 showed increased activity relative to a Leu allele, and the Tlr1 with a Ser allele at position 605 showed increased activity compared to Leu, but to a lesser degree than the Ile allele. Tlr1 with a Ser allele at position 251 showed increased activity compared to Asn, but elucidating its specific effect will require further investigation. These results indicate that variations in the transmembrane domain of mouse TLR1 result in alterations in function comparable to that of human variants in the same region. These findings suggest that mouse models may be used to study TLR1 variants’ effect on the immune response in humans. This could lead to a better understanding of the mechanisms leading to poor clinical outcomes for patients with sepsis and other inflammatory diseases.

It’s important that the public heeds warnings issued by the local government during a natural disaster, despite costs associated with taking preventative action which are not guaranteed to have been necessary if the threat fails to occur. This can lead to a low compliance rate for warnings issued for natural disasters. To begin to understand the characteristics of warning-compliant citizens, we aim to see if previously experiencing a catastrophe influences future behavior. We believe the disaster would be more salient and easily recalled the next time a warning is issued, potentially making people more cautious. This is called the availability heuristic (Tversky & Kahneman, 1974). In a controlled computer simulation, we ask participants to act as a road maintenance company president and judge whether to take precautionary action in preparation for freezing roads. Each of 90 trials will represent a single night, and one or three of these trials will be manipulated in the middle of the experimental sequence so participants experience a large and sudden temperature drop from what was forecasted. For those trials, we expect participants not to take precautionary action, leading to a large penalty. We then anticipate that after experiencing these “Big Miss” trials that the participants will take precautionary measures more frequently, because of the activation of the availability heuristic. If this happens, we may propose that victims of previous disasters are more cautious with similar scenarios in the future, and we may then move forward in finding ways to create the same mentality in residents of disaster warning zones.

BTX/A is often used in human masticatory muscles to treat TMJ disorders or reduce muscular mass. Although its multiple applications are common practice, the consequences of its neuromuscular blockage in masticatory muscles are unknown. The goal of this study is to investigate these consequences by analyzing the compound muscle action potential or evoked electromyography (eEMG) of bilateral masseter muscles (MAs) in rabbits. Eighteen female rabbits were injected with either 10 units of BTX/A (n=13) or the equivalent 0.9% saline (n=5) into three locations on one masseter at weeks 0, 12 and 24. At week 36, bilateral MA nerve trunks were exposed through surgery, and eEMGs from 11 sites of injected and non-injected MAs were examined through inserted fine-wire electrodes when the nerve was electrically stimulated at the threshold intensity (5-6V) with 2Hz frequency. Ten consecutive evoked pulses from each recording site were identified and filtered. Time domains were analyzed to calculate onset, duration, peak and integral values of these pulses. Paired t tests and non-parametric statistics were used to test differences between injected and non-injected MAs, and between BTX/A and saline injected MAs, respectively. The preliminary results show that although the overall values of stimulated pulses were smaller in the BTX/A injected than non-injected MAs, significant differences were only found in a few sites, and these sites were mainly located at the middle portion of MAs. However, there was no difference between saline injected and non-injected MAs. The comparison between BTX/A and saline injected MAs are ongoing. These results suggest that multiple applications of BTX/A in MAs may not lead to severe neuromuscular blockages over a long term as seen in the single application of BTX/A over a short term. The new information provided by these findings can lead to a better understanding and use of BTX/A in clinical dentistry.