A bacteriophage integrase regulates virulence factor production

in Pseudomonas aeruginosa

Pseudomonas aeruginosa is a bacterial pathogen that causes hospital-acquired infections and is very difficult to kill with antibiotics. This is especially true when P. aeruginosa grows as a biofilm—a complex community of bacteria encased in a protective extracellular matrix. P. aeruginosa biofilms produce large amounts of filamentous Pf bacteriophage, which are viruses that infect P. aeruginosa. Pf bacteriophage are known to increase the virulence of P. aeruginosa. However, the underlying mechanisms that cause this are unknown. Our preliminary results suggest that when the Pf bacteriophage integrase gene intP is overexpressed, production of the virulence factor pyocyanin is enhanced. When intP is deleted, pyocyanin production is repressed in P. aeruginosa. Like other bacteriophage integrases, intP inserts bacteriophage DNA into the bacterial chromosome. We hypothesize that intP integrates bacteriophage DNA into bacterial genes that regulate pyocyanin production. To test this hypothesis, we will use genetic approaches to disable the integrase activity of intP followed by biochemical assays to measure pyocyanin production. Understanding how Pf bacteriophage manipulate the virulence of P. aeruginosa may result in new ways to treat or prevent infections caused by this ubiquitous bacterial pathogen.

Ski towns across the Rocky Mountain West are facing the inter-connected problems of rising housing prices and shortages of labor. These local issues have led to severe challenges associated with the lack of affordable housing. My study focuses on three key elements that contribute to the problem of affordable housing in the booming ski town of Whitefish, Montana. These elements include: (1) the geographical and historical context that makes this community similar to other ski towns in this region; (2) the economic and land use planning factors that constrain the affordable housing market; and (3) the limited sustainable solutions to this housing crisis. As part of this study, I propose a site plan to convert a city-owned parcel (known locally as “The Snow Lot”) into a multi-family use community. My site design is guided by the objectives of affordability, sustainability, practicality (e.g., meeting parking needs), and energy efficiency. I employ collaboration with the town of Whitefish and data collection through site visits and planning documents. The intention is that my site design will help the City of Whitefish expand its low-cost sustainable affordable housing solutions in the future.

The paper Intervention for a Child with Persisting Speech and Literacy Difficulties: A Psycholinguistic Approach Joy Stackhouse, Michelle Pascoe, and Hilary Gardner illustrates a psycholinguistic approach to investigating childhood speech and literacy difficulties. A psycholinguistic approach claims that that speech and literacy development is the product of an intact speech processing system composed of speech input processing, stored word representations, and speech output processing.

The study examined the relationship among measures (PCC(percentage consonants correct), sound inventory, and word shapes) used in assessing the connected speech samples of children ages 3-7 years old who have difficulty learning speech sounds. Speech-language therapists routinely utilize connected speech samples as part of their assessment and diagnostic process, because they provide an accurate representation of a child’s natural speech patterns. Furthermore, connected speech samples Relationships among the results of PCC, sound inventory, and word shapes and client variables will be explored.

When clinicians can pin-point exactly where a speech issue is originating in a child’s speech-language system, they can help the child learn more efficiently; this results in a quicker therapy process. Stackhouse, Pascoe, and Gardner (2006), presented an approach to speech therapy intervention which incorporated a three-way method. In their psycholinguistic approach, they gathered information about incoming speech, how the information was stored and processed, and the resulting production of speech by one child with a speech delay. This approach offered a more effective way to plan treatment. The current study’s goal was to explore the relationships among measures that could support the verbal aspect of the psycholinguistic model. Elements of both static and dynamic assessment methods were studied to foster better understanding of speech skills. This method was executed by comparing data from previous testing sessions of children, ages 3-7, specifically those sessions which used a standardized (static) test: the Hodson Assessment of Phonological Processes (HAPP), along with a test that measured the amount of help needed to say a sound (dynamic test): Glaspey Dynamic Assessment of Phonology (GDAP). By comparing the scores of these two tests with each other along with other client variables, the data were used to inform treatment plans for future use. Having enhanced measures that address specific areas in the psycholinguistic approach (input, storing, and output) could allow clinicians to create more appropriate treatment plans. The intention of this study was to discover how much more effectively these two kinds of measures could improve understanding of the verbal component of the approach. In using the HAPP assessment and by guiding treatment with the GDAP, the enhanced results of this “hybrid” psycholinguistic approach will be seen and described in this poster presentation.

There has been a fair amount of research over the past several decades on teachers’ understanding of the multiplicative structure of integers. What can easily be discerned from the literature is a lack of understanding on the part of these educational professionals. It would be easy to assume that this lack of understanding is thereby held by the students in these classrooms. Yet, very little research has examined children's understanding of this mathematical idea. In this quasi-experimental study, we focus the effects of the use of a manipulative, the prime towers, in a three-day teaching experiment carried out in a fourth grade classroom. Students “build” towers of blocks that represent each number 2-100 as a product of prime factors. Towers are studied, compared, and contrasted to build understanding of the significance of prime factorization in predicting a number’s multiplicative structure. The experiment measured students’ ability to identify use prime factorization as a tool to find all the factor pairs, multiples, prime, and composite numbers for natural numbers 1-100 (Common Core Standard 4.OA.4). The results demonstrated represent four classes of fourth grade students spanning two schools. The conclusions drawn will help to identify and refine instructional practices that promote the understanding of the Fundamental Theorem of Arithmetic at a fourth grade level. In connection, both qualitative and quantitative data are presented to insure the practices promoted are both instructional and engaging.

Purpose: Caregivers of stroke survivors who have language impairments (i.e., aphasia) are known to have decreased psychosocial well-being, including increased depression and anxiety. Communication impairments from aphasia increase caregiver burden (i.e., third-party disability) and place strain on the relationship between stroke survivors and their caregiver(s). Intensive Comprehensive Aphasia Programs (ICAPs) are an emerging health care model providing holistic, intensive treatment to improve the stroke survivor's communication skills and the psychosocial well-being of both the stroke survivor and their caregiver(s). This cohort-based model targets the impairment, activity, and participation domains of the WHO-IFC model. This study will examine psychosocial outcomes of caregivers who participated in an ICAP at the University of Montana during the summer of 2017.

Methods: Eight non-paid, family caregivers of stroke survivors with aphasia participated in an ICAP lasting four weeks. Caregiver intervention included: (1) twice weekly (1.5 hours/session) group counseling provided by a licensed family counselor, and (2) weekly (1.5 hours/session) caregiver education group provided by graduate student clinicians in speech-language pathology and their licensed supervisors. Prior to and immediately following the ICAP, caregivers were administered the following self-report measures: (1) Beck Depression Inventory-II, (2) Beck Hopelessness Scale, (3) Multidimensional Scale of Perceived Social Support, (4) Perceived Stress Scale, (5) General Self-Efficacy Scale, (6) Caregiver Reaction Scale, (7) Bakas Caregiving Outcomes Scale, and (8) Family Crisis Oriented Personal Evaluation Scales. Data from these outcome measures is currently being organized for analysis. Preliminary analyses will be presented.

Significance: ICAP interventions offer unique opportunities for caregivers to participate in education, training, and social support with other caregivers. ICAP's have potential to increase communication, improve psychosocial well-being, and overall quality of life for both the stroke survivors and their caregivers.

The purpose of this study was to design a test of soft sounds that would be beneficial to patients with decreased hearing. Hearing aid fitting strategies concentrate more on hearing conversational speech levels rather than soft sounds. These soft sounds often allow a full auditory experience for the patient. Soundscape stimuli are sound exemplars that are constructed with high quality sound recordings in a specific fashion to create a real-life sounding experience. Soundscapes are a relatively new technology that are used in the field of acoustics. Little use of these stimuli have been utilized in audiology. For this study we made a series of ten one minute long soundscapes to present to participants. They were made with very strict criteria. We established ten themes to construct them and chose a background sound that may be present in the environment we were simulating. For example, one of our topics was camping, and the background sound was a fire crackling. We then proceeded to choose six micro-sounds per theme to present intermittently throughout the background clips. These were sounds that would play within the background for approximately one to five seconds at fifteen second intervals. Each background (ten themes, were repeated twice for a total of twenty), had three of the six designated sounds embedded in them while the other corresponding background sound had the remaining three. The micro-sounds were embedded at specific intensity levels that would be audible only near the sound awareness level for the micro-sound (20 dB). The test procedure should allow those with hearing loss to hear soft sounds with a hearing aid.

The Bitterroot Valley, located south of Missoula, Montana along US Hwy 93, has a diverse geological history including significant glacial carved canyons and evidence of Glacial Lake Missoula. Just a couple miles south of Hamilton (46-9-36 N, 114-11-29 W) is a peatland field about three hundred acres in size. In the Bitterroot we are interested in dating the oldest lower portion of this deposit out of curiosity to better understanding the geology of the valley. Core samples were obtained in early February 2018. This particular area has a history of peat mining, and we know the top two to three feet of material was removed in the 1970s. Using a hand soil auger, we removed and saved three samples at: 1.8 ft., 2.5 ft., and 3.5 ft. depth. The total thickness of the peat deposit may have been originally five feet or more where the sample was taken. From the surface we drilled through continuous peat to the lowest depth, where we recovered one-inch diameter smooth stones, mixed with smaller pebbles and sand, indicating the bottom of the peat layer. The samples were dried and the sample from the 2.5 ft. depth was sent to Beta Analytic, Inc. in Miami, FL, for radiocarbon dating. Our goal was to determine whether the peat deposit is old enough to be considered ice age or whether it dates to the post ice age Holocene. Either way this information will improve our knowledge of the geological history of the Bitterroot Valley.

Projections of climate change show that western Montana will experience hotter and drier summers that may extend already drastic fire seasons. Extended fire seasons can lead to extreme smoke events, which are known to have harmful impacts on human health. However, there is not extensive research on these human health effects or on adaptation strategies pertaining to western Montana. Research in this project was conducted through literature reviews as well as personal interviews. A final report draws on available research to provide readers with a clear understanding of the relationship between climate change, extreme smoke, and human health impacts, as well as exploring possible adaptation strategies. The interviews in this study supplement the literary research with personal experience, and ensure that research is grounded with community insight. This report can act as a resource for individuals living in areas affected by fire. It is incredibly important for community and public health workers, citizens, organizations, and policymakers to understand how climate change can influence smoke emergencies, what the actual and perceived health impacts are of these events, and the options for adapting to smoke and preventing negative health outcomes.

Astrophotography is the photographing of celestial objects and phenomena commonly used by amerature and professional astronomers. Due to Earth’s rotation, the stars and constellations appear to move overhead and therefore become blurred in long exposure photographs. To counteract these ‘star trails’ a barn door tracker can be used to move the camera synchronously with the sky overhead. These trackers can be as simple as two pieces of wood with a hinge or as complex as an expensive commercial product. The purpose of this project is to manufacture a barn door tracker inspired by existing methods. Each prototype progressed toward efficiency, stability, and sophistication, moving from cardboard and motor to 3D printed material and circuitry to metal and microcontroller. The design and manufacturing techniques are subject to change to enhance reliability, performance, and portability as needed in response to field experiments. This ongoing project explores design and manufacturing practices while providing means of capturing long exposure astrophotographs without star trails.

In most Western U.S. states, including Montana, water rights determine the distribution, use, and management of water across the landscape by creating a system of private and public flow allocations based on priority dates for water use. For many streams in Montana, the state has issued more water rights than there is actual water available in streams during some parts of the year. Under potential climate change scenarios of less available water or changing distribution and timing patterns, it is important to understand if ‘paper’ water rights (priority dates and flow volumes) can accurately predict when irrigators will need to stop using water, as this information can influence how water rights will be managed by the state. In this research, I will use current, publically-available geospatial and hydrologic information on water rights in Montana to construct a model for water use and availability in an agriculturally-based watershed dependent on streamflow for irrigation. Using data from Montana.gov and the MT Department of Natural Resources and Conservation (DNRC) Water Right Query System, I will construct an account of water allocation via water rights on Bass Creek in the Bitterroot watershed. I will then compare this account with actual flow volumes (from USGS flow data) to determine if Bass Creek is over allocated in some years. I hope to also compare the allocation scheme and flow volumes with remotely sensed data indicating water use on the landscape, and to determine if and when water users stopped using water due to over allocation. I expect that the results will show that current information on water rights and water use does not accurately reflect the flow rates and cannot be used to predict over allocation of streamflow in agriculturally-based watersheds in Montana.

Purpose: Intensive comprehensive aphasia programs (ICAPs) are community-based rehabilitation programs designed to improve the speech, language, cognition, and psychosocial well-being of stroke survivors. ICAPs integrate individual and group therapy, current technologies, and client/family wellness and education. Although many aphasia treatment programs exist, traditional therapy models lack the therapeutic intensity and holistic intervention approach that ICAPs provide. The intensity of ICAPs allows the patients with aphasia (PWA) to engage in 72 hours of therapy in four weeks compared to traditional aphasia therapy which offers 30 hours in 10 weeks. The purpose of this retrospective study is to examine language impairment and psychosocial outcomes of PWA following participation in an ICAP at the University of Montana.

Methods: Approximately 40 PWA participated in at least one of six ICAP sessions that occurred between 2014 and 2017. Treatment delivery included: individual evidence-based language impairment treatment, conversation groups, a weekly large group with focus on psychosocial well-being, and weekly community outings to facilitate social communication. The treatment intensity was 4-4.5 hours of therapy per day, 4 days per week, for 4 weeks. Both pre- and post-treatment assessments of language and psychosocial well-being were administered including: the Western Aphasia Battery- Revised, the Boston Naming Test-second edition, the Assessment for Living with Aphasia, and the Geriatric Depression Scale. Outcome data from the six ICAPs is currently being organized for analysis. Preliminary analyses will be presented.

Significance: Evidence suggests that the therapeutic intensity and the holistic intervention approach offered by ICAPs are beneficial to improving quality of life and communicative rehabilitation for stroke survivors with aphasia. Future service delivery models should consider this multifaceted approach as well as ways to better support autonomy and sense of respect and dignity throughout therapy.

In 2016, over 10.4 million people contracted Tuberculosis (TB) infections for the first time, resulting in the deaths of 1.7 million people worldwide. Despite the existence of a vaccine, TB remains the number one cause of death from a single infectious disease according to the 2017 report from the World Health Organization. As a result, research and development of a new TB vaccine is ongoing, as it is suspected the currently available formulation does not contain an adjuvant which produces the necessary TH-17 response. In the search for new drugs to combat this epidemic, knowledge of how a compound triggers an immune response is paramount. One method for identifying an activated immune protein complex is via immunoprecipitation, colloquially referred to as a “pulldown.”

In this study, a pelletable, beaded support of polystyrene (PS) with surface-absorbed trehalose diester (TDE) has been developed for use as a tool for identification of C-type lectin receptors. TDE was used as a stand-in for trehalose dimycolate, a glycolipid found in the cell wall of Mycobacterium tuberculosis, which is recognized by C-type lectin. Confirmation of surface-absorption and characterization of the modified beads was performed via dynamic light scattering and high-performance liquid chromatography.

Purpose: The purpose of this study is to examine school-based pre-service professionals’ knowledge of speech-language pathologists’ (SLPs) literacy assessment and intervention practices in K-12 students before and following participation in an interprofessional education (IPE) workshop.

Methods: A pre-/post-workshop survey of school-based SLP’s literacy practices will be distributed to the attendees of the IPE workshop. Participation is voluntary and anonymous. Descriptive statistics will be analyzed and reported.

Originality: A growing body of literature suggests that collaborative interprofessional practice (IPP) is more likely to be successfully conducted when professionals have participated in IPE experiences when they were enrolled in their pre-service professional training programs. In particular, knowledge of the roles, responsibilities, and scope of practice of the other professionals with whom they will interact has been identified as a significant predictor of successful IPP.

Significance: Results of this study will provide preliminary data of the effectiveness of an interprofessional education (IPE) workshop with respect to informing school-based pre-service professionals on the scope of the school-based SLP’s practice in literacy assessment and intervention. This is significant in that while there are numerous studies of IPE practices in medical-based fields, such as nursing and pharmacy, few such studies exist that examine the IPE experiences of school-based pre-service professionals.

Imperfect detection is a known issue with conducting wildlife surveys. False positive detections, where an individual is counted as present when it truly is not, are often assumed to not occur. This assumption can skew detection rates and create misleading results when calculating population estimates. Survey methods such as the dependent double-observer method developed by Nichols et al. (2000) are suggested to reduce the occurrence of false positives by using two collaborating observers. This study quantified and compared rates of false positives between a single-observer method and the dependent double-observer method. This was accomplished with auditory surveys of ten grassland songbird species native to central Montana. Both inexperienced and experienced volunteer observers were asked to listen to randomly-generated surveys containing the vocalizations of these ten songbirds and identify the species. The decrease in false positive rates using the dependent double-observer method is substantial. Further evaluation will provide information on the effectiveness of the dependent double-observer method in providing more precise and less biased population estimates.

This research project seeks to explain the use of lithics found at the Bridge River site in British Columbia through the extraction and analysis of ancient DNA (aDNA) found on the surface of stone tools. The methods used for extraction were nondestructive. Using sonication to release the trapped aDNA from microcracks on the tool’s surface, and the amplification of mitochondrial DNA regions Cytochrome B and 16S in order to determine what species the tools were used to process. The findings of this project have the potential to further refine the extraction process for ancient DNA present on lithic material, as well as end archaeologists’ longtime debate over whether or not certain tools were used explicitly for one particular organic material, such as with the making of bone tools, and whether or not scrapers were specifically used for one species at the Bridge River site. To date, we have worked with over 65 tools, and extracted both Puma (Puma concolor) and Dog (Canis lupis familiaris) DNA from our samples. This project provides us with a unique opportunity to both enhance our knowledge of lithic use at archaeological sites, and successfully extract more genetic material moving forward.

The goal of this study was to study social learning in an animal model. Social learning is essential for survival across most animal species, and an animal model of social learning would provide a tool for understanding the brain functions that support these behaviors. Social learning was evaluated using a paradigm known as the “tube test” (Lindzey et al. 1961) which measures dominance relationships based on whether a degu advances or retreats when they encounter one-another in a tube, as well as the amount of time this interaction takes to proceed. While previous studies have examined dominance relationships using this test, these studies do not normally examine changes over time. Dyads of either cagemate (familiar) or stranger (unfamiliar) male degus were tested every day for 5 days in the tube test. One month after the first round of trials, a second round of trials will explore how the social relationships are retained over time. Preliminary observations suggest that the latencies between when the degus met in the center of the tube and one retreated did not differ between cagemate and stranger groups and did not appear to change over days. These results suggest that the tube test method may need to be further refined before it can be used as a tool to study social learning.

On the cell surface are G protein coupled receptors that bind to agonists, causing activation of intracellular G proteins, by catalyzing exchange of GTP for GDP at the G protein alpha subunit (Ga). G proteins are also activated by guanine nucleotide exchange factors (GEF) inside of the cell; these include Ric-8A and Ric-8B. GTP-bound Ga can stimulate the activity of intracellular enzymes. For example, Gas activates adenylyl cyclase, while Gai1 inhibits the activity of this enzyme. Biochemical studies have shown that Ric-8A is a GEF towards Gai1 whereas its isoform Ric-8B acts on Gas. Previous studies in our laboratory and others have shown that a region in Gαi1 called switch II binds to Ric-8A. In this study, we test the hypothesis that differences in amino acid sequence between Gai1 and Gas in switch II are responsible for the ability of these G proteins to discriminate between Ric-8A and Ric-8B. The switch regions of Gai1 and Gas differ in only three amino acids. We predict that, by mutating these amino acids in Gai1 to their corresponding residues in Gas, affinity for Ric-8A will be impaired. Single mutation primers (S206D, K209R and H213Q) were made, transformed and amplified through a polymerase chain reaction (PCR). Once mutant plasmid was expressed in E. coli cells, it is purified, and a tryptophan fluorescence assay is performed. This assay technique detects changes in the fluorescence of tryptophan 211, a side chain in switch II that is sensitive to the exchange of GTP for GDP. Our research sheds light on how mutants in Gai1 in the switch II region plays important role in specificity of binding for Ric-8A.

Microaggressions are a relatively understudied phenomena in psychology and are often not immediately recognized by even keen observers (Sue, 2010). Microaggressions are defined as any verbal, behavioral, or environmental statements or actions that are derogatory or hostile and insulting in nature, specifically targeted at a person or people (Sue, Capodilupo, Torino, Bucceri, Holder, Nadal, and Esquilin, 2007). Microaggressions are a form of racism that gradually chip away at a person’s well-being in a nearly unseen manner (Sue, 2010). Furthermore, Sue (2010) separates microaggressions into three types: microinvalidations, microinsults, and microassaults. However, it should be noted that an individual does not need to be a “racist” to commit a microaggression (Sue et al., 2007).

The current study examines how the number of microaggressions in eighth grade Montana history books have evolved over time. Studying existing data from within the lab revealed the occurrence of many microaggressions against Native Americans in eighth grade Montana specific history books (some published as early as 1951 and the most recent being published in 2008). However, the reviewed study did not examine the specific changes in microaggressions over time. Consequently, this study seeks to enhance the growing body of research by examining the shift in microaggressions in textbooks over time and further analyzing how these microaggressive statements have evolved. Previous research has identified that having a history curriculum that includes biased information about a person’s ethnic origins can be damaging to the way they perceive their school climate (Banks, 2016). This study will be beneficial to educators to help them recognize institutional racism within textbooks across time and combat this racism in their classrooms.

Method

This qualitative research project will use previously coded data and nVivo software to categorize the types of microaggressions presented across time in eighth grade Montana history textbooks.

FBF-1 and FBF-2 are proteins within the PUF protein family that are required for stem cell maintenance in Caenorhabditis elegans. FBF-1 and FBF-2 exhibit mRNA binding activity and are involved in localization, activation and repression of their target mRNA’s. The two are similar in sequence with the exception of four Variable Regions (VR’s). FBF-2 localizes to P granules in germ cells of the C. elegans while FBF-1 does not. We propose that the different localization patterns exhibited between the FBF-1 and 2 are due to these VR’s. Analysis of which VR or combination of VR’s is responsible for this difference in localization was undertaken through chimeric protein assembly and insertion into the C. elegans genome. Assembly of DNA encoding chimeric proteins with various VR’s and a tagging Fluorescent Protein (GFP) present was achieved through fusion Polymerase Chain Reaction (PCR) and BP/LR clonase plasmid assembly. Introduction of chimeric DNA constructs is in progress, and done through Crispr-CAS-9 genome editing. Expression of the modified proteins and assessment of localization patterns will be carried out using GFP visualization. The poster will discuss our observations and preliminary conclusions.