TRPV1 is an ion channel of great importance for pain and heat sensing. The EC50 (threshold dose) of the ion channel TRPV1 is highly variable depending on the cell type in which it is expressed. For example, the EC50 of TRPV1 in HEK293T/17 (a cultured mamalian cell line) is ~1 µM whereas the EC50 of TRPV1 in isolated DRG neurons is ~ 100 nM. Numerous explanations have been proposed for this phenomenon, with one possible explanation being the varying lipid contents of the plasma membrane regulating the EC50 of TRPV1. The cholesterol content of the plasma membrane is a candidate for regulating TRPV1, as it is known to vary among types of cells. We measured TRPV1 function with patch-clamp electrophysiology to determine the effect of removing cholesterol from the plasma membrane on channel function. Inside-out excise patches remain stable for tens of minutes, allowing for cholesterol removal and replacement by flowing a Methylated-β-Cyclodextrin, a ring of sugar molecules that bind cholesterol, across the patch. In intact HEK293T/17 cells, we estimated the cholesterol content to be about 30%. Thus, the Methylated-β-Cyclodextrin is expected to produce a very large change in plasma membrane properties as removes cholesterol. Recordings from these patches showed that both the EC50 for activation of TRPV1 by capsaicin and the capsaicin-induced maximum current were unaffected by patch treatment with Methylated-β-Cyclodextrin. These results are significant as they shows that TRPV1 function is unaffected by a large reduction in plasma membrane cholesterol. Because removal of such an abundant component of the plasma membrane likely produces meaningful changes in the bulk properties of the bilayer, these data also indicate that lipid regulation of TRPV1 is caused by lipids which have specific, chemical interactions with the channel, rather than the bulk properties of the plasma membrane.

Transient receptor potential vanilloid receptor 1(TRPV1) is a nonselective cation channel whose activity can be regulated by different kinds of factors including vanilloids such as capsaicin. Nerve growth factor (NGF) has been shown to cause the rapid sensitization of nociceptors neurons expressing TRPV1 under inflammatory hyperalgesia. In other words, TRPV1 is involved the sensation of pain caused by inflammation. It’s been suggested that tropomyosin receptor kinase A (trkA) and its downstream effectors including phosphoinositide 3-kinase (PI3K) play a key role in TRPV1 potentiation and we want to further examine the molecular signaling mechanism. For the purpose of our study, we utilize a system based on a light-sensitive molecule phytochrome B (PhyB) to mimic inflammatory responses in cells. Under 650nm light, PhyB undergoes conformational change and binds to phytochrome interaction factor (PIF). By fusing proteins of interest to PIF, PhyB-PIF interaction controlled by light can be used to manipulate signaling pathways including PI3K pathway. To study the potentiating effect of PI3K on TRPV1, we examine NIH3T3 cells transfected with proteins of interest using calcium imaging. We compare cell responses to capsaicin before and after PI3K translocation activated by 650 nm light. Our experimental data suggest a significant increase of calcium signalling after translocating PI3K to the plasma membrane. In the future, we are hoping to learn more about how the downstream effectors can affect TRPV1 trafficking. We will perform similar procedures as described using dominant-negative Akt and Rac1 cells and investigate the specific mechanisms. Eventually, we hope to utilize what we know about TRPV1 in pain treatment development.

 Approximately 500,000 Americans undergo surgery for lower back problems each year. Despite dramatic improvements over the last decade in perioperative care, the outcome of spine surgery is associated with prolonged hospitalization and poor pain management. In April 2014, Harborview Medical Center introduced an enhanced recovery protocol, called the Perioperative Enhanced Recovery from Major Spine Surgery (PERMSS), which aims to reduce significant physiological stress and shorten hospitalization by implementing pre-operative nutrition supplement, standardized analgesic administration, and early mobilization. The purpose of this study is to investigate the efficacy of PERMSS on post-operative pain relief. This project takes a quantitative approach and follows the descriptive correlational design. From June 2015 to Feb 2016, fifty PERMSS and fifty non-PERMSS patients were recruited and interviewed in their hospital rooms on their first postoperative day. The validated International Pain Outcome survey was used to capture comprehensive data including patient demographics, comorbidity, surgical procedure, analgesic regimen, and patient-reported outcomes. In April 2016, data analysis will be conducted through Stata MP V13.1. to compare the difference between PERMSS and non-PERMSS patient outcomes. We expect a significant improvement in pain management, reflected by less time spent in severe pain, decreased interference in recovery activities, and improved patient satisfaction. The result of this study leads to two potential benefits: first, to introduce enhanced recovery protocol to other similar complex surgeries; and secondly, to reduce reliance on opioids especially in spinal patients who have a 15-30% likelihood to develop failed back surgery syndrome (FBSS) and become frequent victims of opioid use disorders. In contrast to traditional practice that relies heavily on opioids, PERMSS may help address this problem with its emphasis on multi-modal analgesic, which is a combination of opioid and non-opioid prescription.

The first member of the vanilloid sensing sub-family of transient receptor potential cation channels (TRPV1) is an ion channel located primarily in the nociceptive (pain-sensing) neurons of the peripheral nervous system. It is most strongly activated by high temperatures, protons, and capsaicin, the molecule responsible for the spice of hot chili peppers. From these examples of TRPV1 activators, it has been deduced that this ion channel plays a key role in the sensory transduction of pain that results from thermal burns, acid burns, and ingestion of noxious chemicals. Thus, the TRPV1 ion channel has potential to be a drug target for innovative pain medications and therapies. Before this is feasible, physiologically relevant details of TRPV1 structure and function must be elucidated. Expanding the understanding of how TRPV1 interacts and changes in various environments is also crucial. My research aims to determine if the lipid environment surrounding TRPV1 has significant impacts on its structural conformation. Electron resonance techniques such as EPR and DEER can be used to detect structural information of proteins; however, a number of steps precede this goal. First, I expressed and purifyied TRPV1 in large quantities. Next, I reconstituted the purified channel into vesicles. Vesicles are spheres of phospholipid bilayer and more accurately resemble the plasma membrane environment in which TRPV1 naturally occurs. Then electron resonance techniques were used to compare the structure of detergent-stabilized TRPV1 and TRPV1 reconstituted into vesicles. The comparison of TRPV1 structure in different lipid environments has a lot of implications in the applicability of structural studies conducted outside of physiologically relevant systems and for the future study of TRPV1.

The postpartum period can be a challenging experience for many women as they adjust to the physical and social changes of new motherhood. Breastfeeding is one of the postpartum tasks that may be more difficult than expected. Additionally, many women may feel that their postpartum body fails to meet an idealized image, leading to body dissatisfaction. Mindfulness-based interventions have been developed for stress reduction in a variety of health contexts, including pregnancy. Mindfulness-based interventions may provide strategies for new mothers to handle the challenges of breastfeeding the infant and to improve their body image. The purpose of this study is to explore whether participants in a mindfulness based childbirth and parenting class (MBCP) during pregnancy found mindfulness skills beneficial to their breastfeeding experiences and postpartum body image. Women who participated in a ten week MBCP course during pregnancy were interviewed within the first year postpartum to discuss their experiences. The semi-structured interview guide included questions on how participants may have used mindfulness to approach a variety of positive and negative experiences. Findings demonstrate that women used mindfulness to appreciate the positive aspects of their postpartum experiences as well as handle the challenges they faced. During breastfeeding sessions participants deliberately focused on mindful attention on their infants. Women also used mindfulness to appreciate their postpartum bodies for the achievement of giving birth and to show self-compassion for the physical challenges of recovery. Findings have implications for future research on the postpartum experience and intervention design.

The Hidden Valley is a new theory that extends the Standard Model, our current best description of particles and forces that make up our universe. The Hidden Valley theory, along with others, extends the Standard Model in an attempt describe phenomena like dark matter not yet included in the Standard Model. Hidden Valley particles do not interact with Standard Model particles, and are therefore impossible to directly detect. But by using their predicted decay products, we may be able to prove their existence. We are currently running simulations to try to give us an idea of what we should be looking for when we use data from the Large Hadron Collider at CERN. Features of the model and the search strategy will be presented.