Recent advances in the field of synthetic biology have enabled the use of macrophages as living cell diagnostics for early cancer detection. Presently, macrophages have been engineered to be macrophage sensors, which produce a reporter signal upon activation of the ARG1 promoter—an event which will occur in the tumor microenvironment (Aalipour et al., 2019). Incorporation of Boolean logic gates can help refine the specificity of this sensor, to ensure that it is only activated in the tumor microenvironment and not in inflammatory or wound healing environments. In order to do this, it is necessary to identify genes that can be used as markers that are differentially specific to tumor microenvironments compared to inflammatory microenvironments. Because tumor microenvironments are commonly infiltrated with M2-polarized macrophages and inflammatory environments are commonly infiltrated with M1- polarized macrophages, assessing relative gene expression from M1 and M2 type macrophages could be a useful avenue for identifying genes to be used in Boolean logic gates for enhancing the specificity of the previously developed macrophage sensor. First, macrophages must be polarized to the M1 and M2 phenotypes, and second, quantitative real time PCR must be used to identify genes that are upregulated in either M1s or M2s.

Osteoarthritis is a degenerative bone disease that affects over 27 million Americans (Neogi, 2013). It is characterized by the loss of functional cartilage that normally bears mechanical stresses, which is then transferred onto bone and leads to untreatable pain. The objective of this study was to histologically characterize the nature of subchondral bone cysts found in different anatomical sites of the femoral heads from patients who had undergone arthroplastic surgical replacement. Femoral heads were collected from sixteen patients (5 males and 11 females) aged 43-77 years who had all underwent a total hip arthroplasty. Heads were fixed for two weeks and computerized tomography (micro CT) was then carried out to identify subchondral bone cysts (regions within the head devoid of bone tissue). The head was segmented into three regions: The large cyst region, where the largest micro CT identified cyst was found, the primary compressive region, in accordance with the orientation of the trabeculae, and the anterior cartilage region which was where the healthiest articular cartilage is typically found. These regions were then dissected and slices were decalcified and embedded in paraffin for histological processing. The embedded tissues where stained with either Safranin-O and Fast green or hematoxylin and eosin to visualize the histological properties of each region. The subchondral bone cysts were then identified histologically within the sections and matched to the micro CT findings as an area of subchondral bone devoid of normal trabeculation. Three separate types of cysts were identified from the above criteria: fibrous cysts composed of dense fibrous tissues; cartilage intrusion cysts, containing cartilage tissues that had moved deep to the subchondral bone plate; and fatty cysts containing high amounts of unilocular adipocytes. Throughout the course of this study it was noted that the fibrous cysts tended to be more numerous and larger than the other two types. This finding is in accord with the bony contusion theory that subchondral cysts are created from excessive mechanical loading and can grow larger as the bone becomes increasingly ill equipped to handle stress. The cartilage intrusion cysts most likely arose from the process outlined in the synovial intrusion theory, where a small amount of synovial fluid breaks through the subchondral bone plate and creates a cyst. This would account for why the cartilaginous cysts were found so close to the plate and were much smaller in size than their fibrous counterparts. The fatty cysts ranged in size and location with the larger ones deep to the articulating surface while the smaller ones were superficial, suggesting a mixed mechanism of formation. The varying locations and identities of the cysts may provide evidence to both of the conflicting theories of subchondral bone cyst formation. Further research is indicated to help elucidate the pathogenesis of subchondral bone cysts.

Hypertensive disease in pregnancy, including preeclampsia, is the leading cause of perinatal morbidity and mortality in the United States. Preeclampsia, characterized by new onset hypertension and proteinuria, can cause seizures, cerebral hemorrhage, pulmonary edema, placental abruption, premature birth, and death. The only definitive treatment for preeclampsia is delivery of the fetus. However, studies have shown that daily low dose aspirin (LDA) can reduce the risk of preeclampsia without increasing negative outcomes, such as bleeding. Despite this evidence, LDA has yet to be implemented routinely in prenatal care in the United States. The aim of this study is to improve LDA implementation by increasing adherence to the medication once it is prescribed by the provider. While some studies have shown that providing patient education or increasing family support increases medication adherence, very few studies have assessed medication adherence in a pregnant population. Even fewer studies have assessed adherence to LDA to prevent preeclampsia. We are proposing a quality improvement project within the OBGYN clinic at Boston Medical Center to assess the barriers to medication adherence, to identify effective patient-directed implementation strategies to increase adherence, and to reach an adherence rate of 90% in twelve months. Identifying and implementing effective strategies to increase medication adherence to LDA has the potential to reduce the rate of preeclampsia and the associated negative perinatal outcomes.

BACKGROUND: Sirtuin-1 (SirT1), a nicotinamide adenine dinucleotide+ –dependent deacetylase, is a key enzyme in the cellular response to metabolic, inflammatory, and oxidative stresses; however, the role of endogenous SirT1 in the vasculature has not been fully elucidated. Our goal was to evaluate the role of vascular smooth muscle SirT1 in the physiological response of the aortic wall to angiotensin II, a potent hypertrophic, oxidant, and inflammatory stimulus. METHODS AND RESULTS: Mice lacking SirT1 in vascular smooth muscle (i.e., smooth muscle SirT1 knockout) had drastically high mortality (70%) caused by aortic dissection after angiotensin II infusion (1 mg/kg per day) but not after an equipotent dose of norepinephrine, despite comparable blood pressure increases. Smooth muscle SirT1 knockout mice did not show any abnormal aortic morphology or blood pressure compared with wild-type littermates. Nonetheless, in response to angiotensin II, aortas from smooth muscle SirT1 knockout mice had severely disorganized elastic lamellae with frequent elastin breaks, increased oxidant production, and aortic stiffness compared with angiotensin II–treated wild-type mice. Matrix metalloproteinase expression and activity were increased in the aortas of angiotensin II–treated smooth muscle SirT1 knockout mice and were prevented in mice overexpressing SirT1 in vascular smooth muscle or with use of the oxidant scavenger tempol. CONCLUSIONS: Endogenous SirT1 in aortic smooth muscle is required to maintain the structural integrity of the aortic wall in response to oxidant and inflammatory stimuli, at least in part, by suppressing oxidant-induced matrix metalloproteinase activity. SirT1 activators could potentially be a novel therapeutic approach to prevent aortic dissection and rupture in patients at risk, such as those with hypertension or genetic disorders, such as Marfan’s syndrome.

The homeless population is one of the most medically vulnerable patient populations in the world. They face numerous barriers to obtaining appropriate, timely health care and have many competing needs. As a consequence, they are more likely to present to hospitals and clinics with more advanced stages of diseases that could have been prevented. This poses a significant public health crisis and highlights a need to intervene and implement preventative tools in this population. Past studies have focused on the need for preventative resources for more immediately fatal disease processes. It is important to expand resources to include prevention for diseases that homelessness directly increases the risk of developing, such as skin cancer. The homeless population is at a much higher risk of developing skin cancer due to their chronic sun exposure and lack of access to appropriate sun protection. There is a significant gap in the medical literature regarding skin cancer prevention in the homeless. Existing research has collectively shown that the homeless are at high risk for developing skin cancer due to their increased number of risk factors. Additionally, the homeless have been shown to have a higher incidence of malignant/premalignant skin growths. All suggest the need to implement skin cancer prevention to reduce skin cancer risk factors among the homeless population. This proposal focuses on implementing prevention efforts in the homeless in the form of a dermatology intervention at a community health fair in order to reduce their risk factors for developing skin cancer in the future. The intervention will consist of education, free skin exams, and sun protective resources. The results of this study may provide a method to decrease skin cancer risk factors among the homeless that can be incorporated into local community health fairs. The dual public health crises of providing healthcare to the homeless and the rising rates of skin cancer are addressed by proposing an innovative intervention to decrease skin cancer risk factors in this highly susceptible population of people.

BACKGROUND: Pediatric food allergy is increasing in prevalence, and a number of potential immunotherapies are being developed in an attempt to address this health issue. However, there have been no studies investigating parental concerns and priorities regarding selecting a potential immunotherapy for their child. OBJECTIVES: To describe parental immunotherapy preferences and the factors influencing those preferences. METHODS: A survey was developed to understand parental food allergy therapy preferences. Cognitive interviews were performed with parents of food-allergic children (n = 6) to ensure the feasibility and comprehensibility of the survey. The online survey was then disseminated to parents of children with food allergies via social media venues from February 1, 2016 to March 7, 2016 (N = 246). Descriptive statistics were used to report and analyze the attitudes and perceptions of parents considering enrolling their child in a food allergy therapy. RESULTS: Among parents of food-allergic children, 50% (n = 123) reported that if food allergy therapies were made publicly available, they would enroll their children in a therapy. Survey data demonstrated that 69.5% (n = 171) of participants ranked epicutaneous immunotherapy (EPIT) as their first choice, (22.8%, n = 56) ranked oral immunotherapy (OIT) as their first choice, and 7.7% (n = 19) ranked sublingual immunotherapy (SLIT) as their first choice. The majority of parents (62.60%, n = 154) cited the safety profile of a specific therapy as the principal factor influencing their choice of preferred immunotherapy. CONCLUSION: This study suggests that many parents would choose to enroll their children in a food allergy therapy if given the opportunity. The majority of parents participating in this study preferred EPIT to OIT and SLIT. Though comfort and efficacy are important factors when choosing an immunotherapy, the principal concern of participants was the safety profile of the therapy.

The study of compulsive eating has been swiftly gaining attention in both preclinical and clinical research. Compulsive eating behaviors characterize obesity and several eating disorders and can be conceptualized as being composed of three main elements: 1. habitual overeating, 2. overeating to alleviate a negative emotional state, and 3. overeating despite negative consequences. At a preclinical level, developing appropriate and clinically-relevant animal models and tests has been a barrier to investigating the neurobiological substrates of compulsive eating with the purpose of refining pharmacological interventions for forms of obesity and eating disorders. Throughout this review, we will describe the strategies used to develop animal models, first detailing experimental manipulations that are most commonly used to facilitate development of compulsive eating behavior and then we will focus on the tests used to measure compulsive eating as defined by the three aforementioned elements. Retuning the methodological approach towards compulsive eating behavior is essential to understand the complex mechanisms underlying the maladaptive food intake in forms of obesity and eating disorders. / 2020-07-03T00:00:00Z

Reconstructive microvascular surgery is performed on over 5 million patients in the United States each year. Important sub-populations are patients with breast cancer and trauma to their lower extremities, which affects over 120,000 patients. While data has demonstrated that the failure rates for these free tissue transfers as a result of vascular clot formation is limited, there lacks a mechanism to predict the chances of such failures. Such complications increase hospital length of stay and health care costs. The aim of the study was to demonstrate the efficacy of using Thromboelastography (TEG) as a point-of-care test in order to identify hypercoagulable patients, monitor coagulability changes over the course of free flap reconstruction, and ultimately use this data to predict post-operative microvascular compromise. Following IRB approval, we attempted to recruit both lower extremity trauma patients (N=5) and patients with active breast cancer (N=5) undergoing free flap reconstruction. Blood was collected pre-operatively and intra-operatively, at the time of anastomosis. Standard and Heparinase TEG were performed on each sample within 2 hours of collection. We were unable to recruit patients from the trauma population but did identify 5 breast cancer patients. Hypercoagulability was defined as having low R and high α-angle values (p = 0.04 and 0.02, respectively), in the presence or absence of high MA and low R-values. Based on this definition, 80% of patients were found to become hypercoagulable over the course of the surgical procedure. While no patients experienced post-operative microvascular complications. As a result, we discovered that TEG has the ability to identify hypercoagulability and that over the course of a free flap reconstruction, breast cancer patients are likely to become more hypercoagulable when compared to baseline values. Following the recruitment of more patients, TEG could potentially be a useful supplemental tool in microvascular surgery in order to monitor coagulability changes and ultimately be used to predict post-operative thrombotic complications. / 2020-07-03T00:00:00Z

Clustered Regularly-Interspaced Short Palindromic Repeats (CRISPR) and its CRISPR-associated (Cas) proteins have revolutionized genetic engineering. Since its discovery, it has supplanted previous methods of gene editing with its versatility and ease-of-use. Furthermore, the possibility of developing CRISPR-Cas9-based treatment plans for genetic therapy has piqued the interest of scientists. Much research has been done to evaluate its potential for use in medicine as well as the obstacles that prevent it from clinical applications. Although CRISPR-Cas9 can be effective at inducing double strand breaks in DNA, its overall template insertion efficiency is often not sufficient. By default, somatic cells repair damage to their DNA by undergoing nonhomologous end joining (NHEJ), which is not compatible with precise gene insertion. For a desired sequence to be inserted at the break site, homology directed repair (HDR) is required. Several methods of NHEJ inhibition and HDR stimulation have been evaluated and were effective at increasing HDR efficiency in vitro, but these results cannot be generalized for in vivo environments. Accordingly, results from CRISPR-Cas9 research have shown more promise for hematological diseases than for others. In order to address this disparity, additional work on its efficacy in vivo must be done. CRISPR-Cas9 has the potential to become the basis of future treatments for genetic disorders. Prior to this, treatments will need to account for the complex nature of in vivo processes. This thesis examines the current literature to gauge how far CRISPR-Cas9 technology is from use on human patients.

Basal Cell Carcinoma (BCC) is one of the most commonly diagnosed nonmelanoma skin cancers (NMSC) in the world. While BCC is usually readily treatable with surgical excision of the tumor, its metastatic form (mBCC) is far deadlier, with an average patient survival of eight months following diagnosis. A majority of mBCC cases is tied to mutations causing dysfunction in the Hedgehog Pathway (HHP), a pathway that induces basal cells in the skin to divide rapidly and repeatedly. For this reason, there has been an emphasis over the last thirty years on developing novel drug-based therapeutics for the treatment of mBCC that specifically target the HHP. This paper will examine the body of published research investigating current and developing mBCC treatments, with a focus on the therapeutic modalities of the two current treatments for mBCC, Vismodegib and Sonidegib, and one potential treatment currently under investigation, Itraconazole/ATO. A review of the clinical research will detail the mechanism of action for each drug with regard to the HHP and its components, the efficacy of the treatment, and adverse effects seen in patients during treatment.