Metabolic dysregulation accompanying SARS-CoV-2 infection is a key determinant of disease severity. In this study, we performed extensive data mining of multiple existing single-cell RNA seq datasets of COVID-19 BALFs, in combination with high-dimensional immune cell profiling of PBMCs from COVID-19-infected patients, to get a comprehensive, systemic profile of the immunometabolic regulation of adaptive immunity during severe COVID-19. Hypoxia, a hallmark of COVID-19 ARDS, was found to elicit a global metabolic reprogramming in effector lymphocytes. In response to oxygen and nutrient-deprived microenvironments, these cells shift from aerobic respiration to increase their dependence on anaerobic processes including glycolysis, mitophagy, and glutaminolysis to fulfill their bioenergetic demands. We find that these metabolically reprogrammed CD8 and NK cells, under persistent antigen stimulation, become exhausted, displaying impaired cytotoxic function and anti-viral efficacy. We demonstrate that dysregulated metabolism significantly impairs memory lymphocyte differentiation, including the formation of memory NK and tissue resident CD8 memory cells. Unsupervised clustering techniques revealed multiple distinct, differentially abundant CD8 and NK memory cell states that are marked by high glycolytic flux, mitochondrial dysfunction, and cellular exhaustion, further highlighting the connection between disrupted metabolism and impaired memory cell function in COVID-19. Overall, our findings provide novel insight on how SARS-CoV-2 infection affects host immunometabolism and anti-viral response during COVID-19.

<p>Localization and subtype distribution of the receptors for neurotensin and opiods in canine small intestine was studied by the radioligand binding technique.</p> <p>Extensive dissection procedure was developed to separate a) longitudinal muscle (LM) layer containing myenteric plexus (MP), b) the circular muscle (CM) layer containing the deep muscular plexus (DMP) and c) the submucosa containing the submucous plexus (SMP).</p> <p>Purified membranes were prepared from the LM, MP, CM, DMP and SMP by differential and the density gradient centrifugation and using the markers 5'-nucleotidase (for smooth muscle plasma membranes), NADPH cytochome C reductase (for endoplasmic reticulum), cytochrome C oxidase (for mitochondrial membranes), specific binding of [^3H]saxitoxin for the neuronal membranes and the content of the vasoactiev intestinal polypeptide immunoreactive material as a measure of intact synaptosomes.</p> <p>The fractions enriched in the membranes from LM, CM, DMP, MP and SMP were used to study the distribution and properties of neurotensin receptors using [^125]Tyr^3-neurotensin and of opioid receptors using [^3H]diprenorphine, [^3H]etorphine and [^3H]ethylketocyclazocine.</p> <p>Neurotensin receptors were confined to the CM, DMP and SMP. These receptors has the similar affinity and recognition properties at their high affinity sites (Kd 0.1 - 0.2 nM). The low affinity receptors on the DMP were of lower affinity than their CM counterparts (Kd 40 nM vs 3nM). The receptors on the CM differed from those on the DMP in their radiation target size (mw - 190, 000 da on the CM and - 120, 000 da. on the DMP). Reduced disulfide bridges were required for the binding to both the CM and DMP neurotensin receptors. However, in vitro, the excitation, but not the inhibition, of the circular muscle strips to added neurotensin was abolished on reduction of the disulfide bonds.</p> <p>Opioid receptors were present on the DMP, MP and SMP but not on any smooth muscle. The receptors on all the three plexuses had similar affinity for the non-selective opioid ligand ([3H]diprenorphine (Kd - 0.1 - 0.2 nM). Both the DMP and MP contained -40-45% 5-subtypes of opioid receptors as assessed in competition studies. All three plexuses contained similar porportion of k-subtype (10-15%), assessed by competition studies with dynorphin [1-13] and U-50488H, and confirmed by saturation experiments with [^3H]ehtylketocyclazocine with or without sheilded 5-receptors. Ionic regulation of these receptors was similar to those observed for the opioid receptors in other systems.</p> <p>Therefore the action of neurotensin on the motility of the canine small intestine may be the combined result of the its action on the smooth muscle and on the modulation of the release of other mediators presynaptically at the DMP level. The action of opioids on the motility is probably through the modulation of the release of other mediatorset the DMP and MP level: direct action of opioids on the smooth muscle is not supported by the present studies.</p> / Thesis / Doctor of Philosophy (PhD)

Although the importance of breastfeeding is almost universally accepted, the rates of breastfeeding are improving very slowly. The current literature provides much insight as to how medical professionals such as obstetricians and lactation specialists could improve the rates of breastfeeding. The pediatrician could encourage breastfeeding at a prime opportunity: during post-natal visits. Unfortunately, there is limited research on what role the pediatrician plays in breastfeeding success. This study used in-depth, qualitative interviews from four pediatricians to gather data on the true role of the pediatrician in breastfeeding. These pediatricians were selected based on the diverse patient population they serve, considering factors such as race, socioeconomic status, and geographic density. The hypothesis stated that pediatricians have a strong positive impact on the culture around breastfeeding but may not be fulfilling their duties to promote breastfeeding to the fullest extent, as compared to the June 2022 recommendation set forth by the American Academy of Pediatrics. Through this study, we can see that pediatricians follow the AAP recommendations in some areas and some recommendations are simply unrealistic for their patients. We also understand how some disparities, specifically the socioeconomic disparity, manifests in mothers and how that affects pediatric practice. This information can be used to improve these AAP guidelines for pediatricians and help improve pediatric practice.

The intent of this thesis is to determine whether the deubiquitinating enzyme ubiquitin specific protease 30 (USP30) is cleaved by Omi/HtrA2 (hereafter referred to as Omi) protease during mitochondrial stress. USP30 is a mitochondrial protein that is anchored in the outer mitochondrial membrane and has components in the intermembrane space (IMS) as well as in the cytoplasm. USP30's IMS component has a six-amino-acid sequence that is very similar to Omi's consensus cleavage sites. Under normal conditions, Omi resides exclusively within the IMS; therefore, if Omi were to cleave USP30, it would target the part of the protein located in the IMS component. Omi is known to play a crucial role in a variety of diseases including cancers, neurodegenerative, and metabolic disorders. Since Omi is a serine protease, it is assumed to carry its normal function through the direct cleavage and degradation of specific substrates. If USP30 deubiquitinase is a bona fide substrate of Omi, this will provide new and important information on the mechanism by which Omi regulates the polyubiquitination process during mitochondrial stress.

Lung cancer is the leading cause of cancer death in the United States and worldwide. The higher mortality among patients with lung cancer is related to cases being diagnosed in late stage where treatment is limited. Urban and rural health outcomes are potentially influenced by differences in accessibility to health care services. We are unaware of existing research examining geographic differences in or factors related to lung cancer mortality in Florida. Therefore, this study aims to examine lung cancer mortality differences between urban and rural counties in Florida. We examined all 67 counties in Florida to investigate if a rural disparity exists in lung cancer mortality. We collected data from the National Cancer Institute (NCI) State Cancer Profile. We assigned urban and rural designation using the United States Census Bureau definition based on population density. We then used a t-test for unequal variances to compare the mean lung cancer mortality rate for the urban counties versus rural counties using Microsoft Excel. We used ArcGIS Pro software to create three maps: one showing urban and rural county designations, one showing lung cancer mortality, and a third bivariate map of urban and rural designations combined with age-adjusted lung cancer mortality rates together to examine the geographic distributions in Florida. We observed a mean age-adjusted lung cancer mortality of 62.7 per 100,000 for rural counties in Florida and a mean of 44.2 per 100,000 for urban counties which was significantly different using a t-test for unequal variances (p < 0.0001). The counties with the highest lung cancer mortality were grouped geographically in Florida’s Big Bend area. Overall, rural counties had a significantly higher average lung cancer mortality than the urban counties in Florida. Future research should examine factors affecting higher lung cancer mortality in rural counties such as screening locations and access to care.

Although many studies have found a substantial racial disparity in infant feeding habits, the factors contributing to this unique disparity and potential solutions remain unknown. The general importance of social interactions and medical interventions in successful breastfeeding has been studied, but little research addresses the specific experiences of breastfeeding Black mothers interacting with their medical providers. This paper examines the perspectives of Black women with breastfed infants on their encounters with healthcare providers during prenatal, perinatal, and postnatal periods. Using qualitative data from 22 in-depth interviews addressing infant feeding decisions and experiences, this paper analyzes the link between healthcare interactions and the breastfeeding experience among Black women.

Receptor-interacting protein 2 (RIP2) is a kinase which mediates signaling downstream of the bacterial peptidoglycan sensors Nucleotide-binding Oligomerization Domain (NOD) 1 and 2. Genetic loss or pharmaceutical inhibition of RIP2 has been shown to be beneficial in multiple inflammatory disease models with the effects largely attributed to reducing pro-inflammatory signaling downstream of peptidoglycan recognition. However, given the widespread expression of this kinase and its reported interactions with numerous other proteins, it is possible that RIP2 has many other unappreciated roles. In this work, we report the involvement of RIP2 in two novel pathways. First, we demonstrate a NOD2 dependent role for RIP2 in mediating ROS production downstream of FcƴR engagement. In these studies, we show that different members of the Src family kinases (SFKs) can promote RIP2 tyrosine phosphorylation and activation, providing a potential mechanism for such involvement. Second, we discover a novel role for RIP2 in the production of specialized pro-resolving lipid mediators (SPMs). SPMs are biologically active mediators which promote resolution by dampening the inflammatory immune response and enhancing the return to homeostasis. We show that NOD2 agonism leads to RIP2-dependent production of different SPMs in an in vivo murine peritonitis model. Using overexpression studies, we demonstrate that RIP2 promotes the modification and relocalization of various enzymes involved in SPM biosynthesis. Overall, our findings shed light on novel functional roles for NOD2 and RIP2 outside of peptidoglycan sensing. These data will have important implications for newly developing RIP2 targeted therapies by potentially determining the molecular basis for in vivo efficacy (or lack thereof) as well as elucidating a potential optimal therapeutic window for administration of such therapies for treatment of various inflammatory diseases.

This research provides ultrastructural information regarding pulmonary surfactant secretion from stimulation of adrenergic pathways to the lung via the stellate ganglion. This information will help clarify the important role of the sympathetic nervous system in surfactant disorders. It also provides insight into sympathetic mechanisms causing surfactant derangement in adult and infant respiratory distress syndromes (RDS). Basic cellular mechanisms of surfactant metabolism, including synthesis and packaging in lamellar bodies, were studied. Results shed light on these basic processes of lung cell function. The results of this study provide new avenues for studying a problem in health care that has long resisted advances, that of surfactant replacement therapy. It is expected that the results of this research, taken with information from other researches, will ultimately reduce the 50% mortality rate of the 150,000 patients annually suffering from RDS (Andreadis and Petty 1985).

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