Compassionate Support Training for Adults Impacted by Mental Illness: A Workshop Series Using Transformative Learning Pedagogy

This professional project aims to translate current compassion academic research and scholarship for my client partner, the Cache County chapter of NAMI, to aid the family members of loved ones who live with mental illness

The role of IKKalpha, IKKbeta and NF-kappaB in the progression of breast cancer

Breast cancer is the most common female cancer in the UK and, despite earlier detection and improved treatments, remains the second most common cause of cancer death in women. Although therapies exist for breast cancer, including endocrine therapy for oestrogen receptor (ER) positive tumours, resistance to current treatment remains a major problem. The molecular mechanisms of endocrine resistance have yet to be fully elucidated and in order to improve treatment for patients this needs to be addressed. Clinically breast cancer presents as several distinct diseases with different outcomes and molecular profiles. Over the past decade, through the use of molecular profiling, the number of different subtypes of breast cancer has grown and understanding the pathways driving each subtype may allow a stratified approach to therapy, allowing patients to receive the treatment which will be of most benefit. The Nuclear Factor kappa B (NF-κB) pathways regulate the transcription of a wide range of genes involved in the immune response, inflammation, proliferation and apoptosis. Many of these processes are hallmarks of cancer and NF-κB has been hypothesised to have a role in tumorigenesis. The aim of the current study was to investigate the role of both NF-κB pathways in the pathogenesis and recurrence of breast cancer. Immunohistochemistry was employed to assess key components of the canonical and non-canonical NF-κB pathways on a tissue microarray (TMA) of 544 patients with full clinical follow up and clinical information including ER status, subtype, necrosis, apoptosis and angiogenesis. Nuclear expression of p65 phosphorylated at serine 536 was associated with angiogenesis and shorter recurrence free interval. Cytoplasmic expression of IKKα was associated with cell death (apoptosis and necrosis) and a shorter recurrence free interval was also observed for those with high expression. These observations between phospho-p65/IKKα and recurrence free interval, when subdivided by ER status, remained significant in ER positive tumours but were negated in ER negative tumours. When split further into subtype, a diverging role for each was observed with phospho-p65 associating with recurrence in luminal B tumours and IKKα with luminal A tumours. Other members of the NF-κB pathways (p65, IKKβ, NIK and RelB) were not associated with recurrence free interval. When these results were tested in an independent cohort, IKKα remained significant on recurrence free interval and breast cancer specific survival in ER positive tumours however phospho-p65 was only marginally associated with breast cancer specific survival. Variability of phospho-p65 is a major issue in IHC studies and therefore an alternative marker of the canonical NF-κB pathway is required. Analysis of expression in this second cohort also revealed that high levels of IKKα in the cytoplasm were associated with recurrence on tamoxifen. This marker may therefore be able to be employed as a diagnostic tool to predict patients who are likely to display endocrine resistance and may represent a therapeutic strategy in combination with endocrine therapy, or for patients after endocrine resistance has occurred. Further examination of the pathways in breast cancer cell lines also demonstrated a difference between ER positive and ER negative breast cancer. In ER negative MDA-MB-231 cells phosphorylation of p65 (from the canonical NF-κB pathway) and phosphorylation of p100 (from the non-canonical NF-κB pathway) was apparent even in untreated control cells, suggesting constitutive activation. Expression was however found to be inducible in ER positive MCF7 cells. In order to investigate whether kinases involved in activation of each pathway, IKKβ in the canonical pathway and IKKα in the non-canonical NF-κB pathway, had potential as targets in breast cancer, we examined the phenotypic impact of silencing their expression in breast cancer cell lines. Silencing IKKβ induced apoptosis and decreased cell viability in both MCF7 and MDA-MB-231 cells but reduction in expression of IKKα only impacted on cell viability and apoptosis in ER positive MCF7 cells. This data, consistent with results from the clinical specimens, has therefore revealed that inhibitors of IKKα are likely to be most beneficial in the treatment of ER positive tumours. These results suggest that the NF-κB pathways are associated with recurrence in patients with ER positive tumours with each pathway possibly associating with recurrence in different subtypes. Additional studies in a larger cohort, including patients receiving aromatase inhibitors are required, accompanied by extensive mechanistic studies to further explore the roles of IKKα and IKKβ in breast cancer. These observations highlight that different subgroups of breast cancer may have different signalling pathways driving progression and therefore patients are likely to benefit from different therapeutic strategies

Additive Manufacturing Waste Management System - Plastic Extrusion Process

3D printing is a fast-growing market with its main source of waste being PLA and ABS plastics. In 2019, the global additive manufacturing market grew to over $10.4 billion, crossing the pivotal double-digit billion threshold for the first time in its nearly 40-year history. (SmarTech Analysis, 2020 Additive Manufacturing Market Outlook and Summary of Opportunities Report). The waste is generated from failed prints and rejected support structures which are common occurrences for personal use. Plastic recycling has become one of the leading discussions of environmental protection and waste management. The 3D market currently does not offer an effective and affordable solution for handling the waste generated. To help bring awareness to this need, companies such as Print Your Mind 3D has offered challenges to hobbyists and corporations to solve this problem. Due to the continuing growth of the additive manufacturing market, and the constant attention on plastics and how they harm the environment, a solution must be found to reduce or eliminate the waste stream of 3D printing. (GB

Mapping Circadian Output Pathways in Neurospora crassa

Circadian clocks are ubiquitous in eukaryotic organisms, providing the ability to anticipate regularly occurring stressful environmental changes. The molecular clock leads to a change in physiology of the organism such that it is prepared for predictable changes. While the external signals detected by the clock, as well as the molecular mechanism of clock components have been extensively characterized, less is known about how the clock manifests time of day information to the organism as a whole. Our lab has focused on identifying output pathways from the clock, using the model organism Neurospora crassa. We have previously demonstrated the circadian regulation of the conserved Mitogen Activated Protein Kinase (MAPK) OS-2 pathway, a homolog of the mammalian p38 pathway, and necessary for maintaining osmotic homeostasis in Neurospora. I present data indicating the circadian regulation of the 2 other MAPK pathways in Neurospora, the mammalian ERK1 and ERK2 like MAPKs, MAK-1 and MAK-2, and show that they are outputs of the clock. Furthermore, I identified around 500 genes that are mis-regulated when MAK-1 is deleted; greater than 25% of those genes are predicted to be clock-controlled. I demonstrated that the clock is signaling through the MAK-1 pathway to regulate 3 clock-controlled genes (ccgs) that encode proteins involved in several different biological processes including, stress response, cell wall formation, and mitochondrial phosphate transport. I established the circadian regulation of the transcript levels of 2 of the MAK-1 cascade components, mek-1 and mak-1. Additionally, I found that the accumulation of MEK-1 protein is clock-controlled, suggesting this is one mechanism by which the clock regulates the activity of MAK-1. Additional studies were carried out to elucidate the proteins that directly regulate the expression of mek-1 and mak-1; however, the mechanisms of direct clock control remain unclear and require further investigation. The finding that the circadian clock regulates all MAPK pathways in Neurospora, combined with the conservation of both the circadian clock and MAPK pathways in mammals provide compelling evidence that mammalian MAPK pathways are also regulated as clock output pathways to control circadian physiology. There is a strong link between aberrations in mammalian clocks, MAPKs, and disease, and therefore, an urgent need to further characterize the circadian regulation of the MAPK families, which will reveal new avenues for therapeutic treatments

Conformational Dynamics of metallo-β-lactamase CcrA during Catalysis Investigated by Using DEER Spectroscopy

Previous crystallographic and mutagenesis studies have implicated the role of a position-conserved hairpin loop in the metallo-β-lactamases in substrate binding and catalysis. In an effort to probe the motion of that loop during catalysis, rapid-freeze-quench double electron–electron resonance (RFQ-DEER) spectroscopy was used to interrogate metallo-β-lactamase CcrA, which had a spin label at position 49 on the loop and spin labels (at positions 82, 126, or 233) 20–35 Å away from residue 49, during catalysis. At 10 ms after mixing, the DEER spectra show distance increases of 7, 10, and 13 Å between the spin label at position 49 and the spin labels at positions 82, 126, and 233, respectively. In contrast to previous hypotheses, these data suggest that the loop moves nearly 10 Å away from the metal center during catalysis and that the loop does not clamp down on the substrate during catalysis. This study demonstrates that loop motion during catalysis can be interrogated on the millisecond time scale

Automated Core Removal System for the NASA/BHI Mars Drill

NASA is in the process of developing an autonomous drill for use in an unmanned mission to Mars. NASA along with Baker Hughes, Inc. (BHI) has designed a drill capable of drilling a core sample from as deep as twenty meters below the Martian surface. Since the mission is unmanned, an automated process for removal of drilled material from the drill is needed. The Trinity Tigernauts, T.N.G. design group is working on an automation system for the Mars drill that will assist in recovery of these core samples. These samples could be analyzed for signs of water and life. The team researched space automation, interplanetary automation, and automated interplanetary drilling, and visited related collaborators. Ultimately, the Tigernauts developed a prototype system on paper for this removal process. Next semester the team plans to construct and test this prototype design

Automatic Core Removal System for a Mars Drill

NASA has discovered the existence of water under the surface of Mars. To access this water, an automated drilling solution has been proposed. It has been the focus of the Trinity Tigernauts T.N.G. to automate the core removal system, one aspect of the overall automation. A mock drill was constructed based on the Johnson Space Center and Baker Hughes Inc. Mark II drill. The Tigernauts designed a robotic arm structure to remove the core sample from the drill bit of the mock drill. The automation process is controlled by a programmable logic controller (PLC) interfacing with motors, encoders, and limit switches. The PLC automation code is complete and has been tested through simulation. In addition, the circuitry has been integrated with the PLC. The current prototype also performs several of the mechanical operations manually; however, full integration of the mechanical and electrical components has not be accomplished

Relationship between tumour PTEN/Akt/COX-2 expression, inflammatory response and survival in patients with colorectal cancer

In patients with colorectal cancer (CRC), local and systemic inflammatory responses have been extensively reported to associate with cancer survival. However, the specific signalling pathways responsible for inflammatory responses are not clear. The PTEN/Akt pathway is a plausible candidate as it may play a role in mediating inflammation via COX-2, and has been associated with cancer progression. This study therefore examined the relationship between tumour PTEN/Akt/COX-2 expression, inflammatory responses and survival in CRC patients using a tissue microarray. In 201 CRC patients, activation of tumour-specific PTEN/Akt significantly associated with poorer CSS (12.0yrs v 7.3yrs, P=0.032), poorer differentiation (P=0.032), venous invasion (P=0.008) and peritoneal involvement (P=0.004). Patients were stratified for peri-nuclear expression of COX-2 to examine associations with inflammatory responses. In patients with absent peri-nuclear COX-2 expression, activation of tumour-specific PTEN/Akt significantly associated with poorer CSS (11.9yrs v 5.4yrs, P=0.001), poorer differentiation (P=0.018), venous invasion (P=0.003) and peritoneal involvement (P=0.001). However, no associations were seen with either the local or systemic inflammatory responses. In CRC patients, tumour-specific PTEN/Akt pathway activation was significantly associated with poorer CSS, particularly when peri-nuclear COX-2 expression was absent. However, activation of the PTEN/Akt pathway appears not to be responsible for the regulation of inflammatory responses

The microbiome associated with equine periodontitis and oral health

Equine periodontal disease is a common and painful condition and its severe form, periodontitis, can lead to tooth loss. Its aetiopathogenesis remains poorly understood despite recent increased awareness of this disorder amongst the veterinary profession. Bacteria have been found to be causative agents of the disease in other species, but current understanding of their role in equine periodontitis is extremely limited. The aim of this study was to use high-throughput sequencing to identify the microbiome associated with equine periodontitis and oral health. Subgingival plaque samples from 24 horses with periodontitis and gingival swabs from 24 orally healthy horses were collected. DNA was extracted from samples, the V3–V4 region of the bacterial 16S rRNA gene amplified by PCR and amplicons sequenced using Illumina MiSeq. Data processing was conducted using USEARCH and QIIME. Diversity analyses were performed with PAST v3.02. Linear discriminant analysis effect size (LEfSe) was used to determine differences between the groups. In total, 1308 OTUs were identified and classified into 356 genera or higher taxa. Microbial profiles at health differed significantly from periodontitis, both in their composition (p < 0.0001, F = 12.24; PERMANOVA) and in microbial diversity (p < 0.001; Mann–Whitney test). Samples from healthy horses were less diverse (1.78, SD 0.74; Shannon diversity index) and were dominated by the genera Gemella and Actinobacillus, while the periodontitis group samples showed higher diversity (3.16, SD 0.98) and were dominated by the genera Prevotella and Veillonella. It is concluded that the microbiomes associated with equine oral health and periodontitis are distinct, with the latter displaying greater microbial diversity