Master of Science (MS), Bowling Green State University, 2021, Forensic Science

Conventional forensic detection of saliva relies on the enzymatic activity or presence of α-amylase. α-amylase based detection methods face a variety of problems: α-amylase is not specific to saliva, varies in concentration between individuals and over time, and is found in nonhuman sources. Previous studies have investigated various species of bacteria native to the human oral cavity as an answer to the problems presented by α-amylase. This study aimed to further understand of how four of these oral bacteria (Streptococcus salivarius, Streptococcus oralis, Veillonella atypica and Prevotella maculosa) persist through time and temperature degradation studies. Saliva samples from a single donor were subjected to five temperature conditions, ranging from -15°C to 57°C, and five time conditions, ranging from 0 to 28 days. The concentration of bacterial DNA present at each condition was measured by quantitative polymerase chain reaction. In this individual, S. oralis was by far the most prominent species of the four examined and due to this dominance appears most robust. Each of the bacterial species studied displayed marked DNA loss at very high temperatures, but V. atypica had the lowest starting value and was most susceptible to the conditions. At 4°C and -15°C, each bacterial species showed no significant DNA loss through 28 days. This analysis of the bacterial species present did not influence the amount of amplifiable human DNA present.

Committee: Crystal Oechsle PhD (Advisor); Jon Sprague PhD (Committee Member); Travus Worst PhD (Committee Member) Subjects: Biology

Master of Science, The Ohio State University, 1965, Graduate School

Committee: Not Provided (Other) Subjects:

Master of Science, The Ohio State University, 2007, Graduate School

Committee: Not Provided (Other) Subjects:

Doctor of Philosophy, The Ohio State University, 2024, Biomedical Sciences

Severe acute respiratory syndrome associated coronavirus 2 (SARS-CoV-2) infected, asymptomatic individuals are an important contributor to Coronavirus Disease 2019 (COVID-19) transmission. SARS-CoV-2-specific immunoglobulin (Ig)—as generated by the immune system following infection or vaccination—has helped limit SARS-CoV-2 transmission from asymptomatic individuals to susceptible populations (e.g. elderly). Here, we describe the relationships between COVID-19 incidence and SARS-CoV-2 lineage, viral load, saliva Ig levels (SARS-CoV-2-specific IgM, IgA and IgG), and ACE2 binding inhibition capacity in asymptomatic individuals between January 2021 and May 2022. These data were generated as part of a large university COVID-19 monitoring program in Ohio, United States of America, and demonstrate that COVID-19 incidence among asymptomatic individuals occurred in waves which mirrored those in surrounding regions, with saliva SARS-CoV-2 viral loads becoming progressively higher in our community until vaccine mandates were established. Among the unvaccinated, infection with each SARS-CoV-2 lineage (pre-Omicron) resulted in saliva Spike-specific IgM, IgA, and IgG responses, the latter increasing significantly post-infection and being more pronounced than Nucleocapsid-specific IgG responses. Vaccination resulted in significantly higher Spike-specific IgG levels compared to unvaccinated infected individuals, and uninfected vaccinees' saliva was more capable of inhibiting Spike function. Vaccinees with breakthrough Delta infections had Spike-specific IgG levels comparable to those of uninfected vaccinees; however, their ability to inhibit Spike binding was diminished. These data are consistent with COVID-19 vaccines having achieved hoped-for effects in our community, including the generation of mucosal antibodies that inhibit Spike and lower community viral loads, and suggest breakthrough Delta infections were not due to an absence of vaccine-elicited Ig, but instead limited Sp (open full item for complete abstract)

Committee: Richard Robinson (Advisor); Fernanda Novais (Advisor); Amy Lovett-Racke (Committee Chair); Ian Davis (Committee Member); Adriana Forero (Committee Member) Subjects: Biomedical Research; Immunology; Microbiology

Master of Science, The Ohio State University, 2022, Dentistry

Background: The gingival sulcus hosts a diverse microbial community that exists in a state of dynamic equilibrium with the immuno-inflammatory system. When this equilibrium is disrupted, it can result in gingivitis and periodontitis, two diseases affecting the periodontal tissues. Although both diseases are primarily caused by plaque, other factors can increase risk for disease, and the extent and severity can be modified by host systemic factors1 . This is also may be the reason that the susceptibility of the disease varies among different individuals. The different individual response to plaque has been documented and has been assigned to just plaque quantity or quality. Finding from previous papers helps us to understand that there are high and low responders to the dental plaque. Since both host and bacterial variables can affect the severity of the immuno-inflammatory response to oral biofilms, it is important to employ a comprehensive methodology to investigate differences in both host and bacterial factors between normal, low and high responders. Therefore, we aimed to investigate clinical differences in host response to bacterial plaque with metabolomic signatures in saliva in a group of orally and systemically healthy individuals. Methods: 60 subjects were recruited following informed consent and divided into groups based on gingival index (GI), plaque index (PI) and GI/PI ratio. Metabolomic analysis was performed on saliva samples using MALDI_TOF. The data normalized by root mean square transformation and the m/z points were annotated. Results: The study subjects were stratified as Normal responders (NR), Low responders (LR) and High responders (HR). The mean ± standard error (median, range) for PI in NR was 0.463 ± 0.064 (0, 0.48 – 0.74), for LR was 1.059 ± 0.073 (1, 0.48 – 1.74) and for HR was 0.699 ± 0.081 (0.5, 0.48 – 1.36). The mean plaque score was significantly different between NR and LR as well as LR and HR. The mean± standard error (median, (open full item for complete abstract)

Committee: Purnima Kumar (Advisor) Subjects: Dentistry

Master of Science, The Ohio State University, 2022, Dentistry

Background and Objectives: The etiopathogenesis of peri-implant diseases is not well understood. Although dental plaque biofilm related bacteria may be the major player, the longterm integrity of implant devices made from titanium alloys and its effect on peri-implant pathogenesis have been recently questioned. Our group previously reported the presence of titanium ion (Ti) within peri-implant and adjacent gingival crevicular fluid (PICF and GCF) not only at sites diagnosed with peri-implant diseases but also with peri-implant health. We also published on peri-implant tissue response to surgical trauma following implant placement surgery by studying PICF and adjacent GCF cytokine content. The current study aims to evaluate inflammatory response and possible Ti contamination during early phases of peri-implant wound healing following surgical placement of a dental implant fixture via two different surgical protocols (one versus two stage). Materials and Methods: Patients scheduled to receive their first dental implant placement surgery at an edentulous site were recruited. Demographic information and clinical measurements of patient's periodontal status were documented at initial visit. Saliva, GCF, PICF and gingival biopsy samples were collected prior to (saliva, GCF, PICF) and during (gingival biopsy) surgery, 1 week, 4 weeks (saliva, GCF, PICF) and at 4 months (saliva, GCF, PICF from all sites, gingival biopsy only from implant sites placed with two stage protocol) post-operatively. The GCF and PICF samples were analyzed for inflammatory mediator content using V- plex assays and GCF, PICF, saliva samples for titanium release using inductively coupled plasma-mass spectrometry [ICP-MS]. Gingival biopsies were processed for H&E and IHC staining to study inflammatory infiltrates and macrophage phenotypes. Multivariable regression analysis was conducted to investigate possible associations between cytokine content, Ti contamination and possible clinical determinant (open full item for complete abstract)

Committee: Binnaz Leblebicioglu (Advisor); Dimitris Tatakis (Committee Member); Damian Lee (Committee Member) Subjects: Dentistry

Master of Science (MS), Bowling Green State University, 2020, Forensic Science

Oral fluid (OF) is rapidly becoming a new media for assisting law enforcement in determining if a subject is driving under the influence of drugs (DUID). Preliminary research shows that drugs can be identified in OF in conjunction with blood, and drug concentrations in OF and blood correlate. Despite availability of several roadside devices to test for drugs in OF, the roadside devices are considered a presumptive test. The results from these roadside tests must be confirmed with a validated liquid chromatography- mass spectrometer (LC-MS) instrumentation. A validated instrument for confirmation of OF results is important, and this study validates BGSU's Shimadzu 8050 LC-MS. For validation, the instrument must pass various guidelines set by Scientific Working Group for Forensic Toxicology (SWGTOX) Standard Practices for Method Validation in Forensic Toxicology and others for accuracy, precision, linearity, Limit of Detection (LOD) and Limit of Quantitation (LOQ), carryover, interference, stability, and matrix effects. Due to the Covid-19 global pandemic, only accuracy, precision, linearity, and LOD and LOQ were accessed. The validation studies were conducted over five days (not consecutive) with two runs being conducted during each 24-hour period for a total of 10 runs. A total of 83 different analytes were accessed. The 83 analytes covered a broad range of drugs with abuse potential. The results of the validation study showed that the instrument is highly precise for the vast majority of analytes, but the cannabinoids, particularly delta-9-tetrahydrocannabinol (THC), were troublesome. Linearity for all analytes were accessed using the R^2 of the calibration curve, and all analytes were above the 0.95 limit. The LOD and LOQ study proved that the cutoff for each analyte is correctly higher than the factor of 2 limit for cutoff/LOQ.

Committee: Jon Sprague RPh, PhD (Advisor); Travis Worst PhD (Committee Member); Phillip Gibbs PhD (Committee Member) Subjects: Biology; Chemistry; Molecular Chemistry; Organic Chemistry; Pharmaceuticals; Pharmacology; Pharmacy Sciences; Physical Chemistry; Statistics; Toxicology

Doctor of Philosophy, University of Toledo, 2019, Biomedical Engineering

The development of detection methods for the novel biomarkers can have a significant impact on the research and clinical applications such as drug discovery, disease diagnosis, and treatment monitoring. Histatin 3 (H3) is an antimicrobial salivary peptide that possesses the capability of being a therapeutic agent against oral candidiasis and has recently been linked to acute stress as a potential novel biomarker. Stress biomarkers reflect the physical and cognitive performance of an individual, and their monitoring in real-time is of vital importance for the high-risk jobs, including military, pilot, and surgeon, where higher vigilance is required for an extended period. The salivary levels of H3 also have been correlated with the HIV-infection and associated oral candidiasis. Therefore, monitoring H3 levels in human saliva can provide essential information about an individual's health status, including HIV-infection, oral candidiasis, and acute stress. Additionally, H3 detection could serve as therapeutic drug monitoring if H3 can be established as an alternative therapeutic agent. The currently available detection techniques for H3 are gel chromatography, high-performance liquid chromatography (HPLC), mass spectrometry (MS), and antibody-based immunoassays. The Chromatographic and mass-spectroscopic methods are laborious, utilize expensive instrumentation, require trained personnel, and time-consuming. Whereas antibody-based immunoassays are not widely validated, expensive, sensitive to temperature, and have a short lifespan. This void in analytical methods is not just for H3 but also applies to several other biomarkers in saliva. Even though saliva is considered as an optimal biofluid, several limitations are impeding its use in diagnostic and research. The major hurdles include the deficient concentration of biomarkers, need of laboratory-based preprocessing to remove mucin and interfering particulate matters, and lack of standard sample collection methods. A (open full item for complete abstract)

Committee: Cameron Brent PhD (Advisor); Giovannucci David PhD (Committee Member); Kim Dong-Shik PhD (Committee Member); Pappada Scott PhD (Committee Member); Pirnstill Casey PhD (Committee Member) Subjects: Analytical Chemistry; Biochemistry; Biomedical Engineering; Biomedical Research; Chemical Engineering; Chemistry; Health Care; Health Sciences; Molecular Biology; Nanotechnology

Doctor of Philosophy (PhD), University of Toledo, 2019, Biomedical Sciences (Neurosciences and Neurological Disorders)

The parotid gland produces saliva components critical for oral health. Secretory output is controlled by autonomic noradrenergic and cholinergic activity on parotid acini, the basic units of saliva production. The discovery of gamma-aminobutyric acid (GABA) and its receptors in the salivary glands hints at an unexplored mechanism of autonomic salivation. The purpose of this study was to elucidate this peripheral GABAergic system in the mouse parotid gland and define its effect on acinar regulation. Immunofluorescent imaging of the autonomic ganglia innervating the parotid revealed GABA-positive axons and neuronal soma. GABA labeling was most prominent in the sympathetic ganglia versus parasympathetic. Patch clamp of neuronal cultures showed that parotid-supplying autonomic neurons respond to GABA and exhibit reduced excitability upon GABA application. In the parotid gland, RT-PCR detection and immunofluorescent imaging of GABA receptor subunits indicated GABA may act on tissue components. However, both live cell calcium imaging and whole-cell recordings of dispersed acinar cells revealed no direct effect on acinar function. In contrast, synaptic preservation via tissue slice preparation revealed that nerve ending depolarization caused an acinar calcium response that was diminished with GABA application. Our data suggested GABA had no direct effect on the parotid acini but may downregulate acinar function by inhibiting autonomic neural activity at the ganglia as well as neurotransmitter release in the salivary tissue.

Committee: David Giovannucci (Committee Chair); Joseph Margiotta (Committee Member); Joshua Park (Committee Member); Guillermo Vazquez (Committee Member); Bruce Bamber (Committee Member) Subjects: Neurosciences

Doctor of Philosophy, The Ohio State University, 1972, Graduate School

Committee: Not Provided (Other) Subjects: Health Sciences

Doctor of Philosophy, The Ohio State University, 1966, Graduate School

Committee: Not Provided (Other) Subjects: Chemistry

Bachelor of Science, Ashland University, 2016, Biology/Toxicology

Iron deficiency anemia is the most common nutritional disorder in the world and is a condition in which the concentration of iron in the blood is significantly lowered. Iron plays an important role in the delivery of oxygen to tissues for cellular respiration. One consequence of iron deficiency anemia is a shift toward carbohydrate metabolism, which can be measured through the respiratory exchange ratio (RER), or the ratio of carbon dioxide production to oxygen consumption. This project's objective was to determine the metabolic implications of diets with low iron and high fat levels as measured through RER. Five populations containing three mice each were provided diets with varying levels of fat and iron: a high-fat high-iron diet, a low-fat low-iron diet, a high-fat low-iron diet, a positive control diet, and a negative control diet. Mice were evaluated biweekly using an iWorx metabolic chamber and a GA-200 gas analyzer to assess changes in the ratio of carbon dioxide exhalation to oxygen inspiration. Data was analyzed with correlation tests, ANOVAs, and MANOVAs using R v. 3.2.3 software. Results indicate that iron plays a significant role in allowing the body to continue carbohydrate metabolism under respiratory stress. A side project seeking to quantify iron in saliva as a means of verifying the success of the research diet was completed but iron concentrations were found to be below detection limits of the procedure. The results of this study suggest some key physiological differences in iron deficient mice, which significantly impact the understanding and treatment of iron deficiency anemia.

Committee: Dolly Crawford Ph.D. (Advisor); Jeffrey Weidenhamer Ph.D. (Committee Member); Mason Posner Ph.D. (Committee Member); Kristin Simokat Ph.D. (Committee Member) Subjects: Biology; Chemistry; Physiology

MA, University of Cincinnati, 2009, Allied Health Sciences : Speech-Language Pathology

Dysphagia is among the most serious and common disorders experienced by elderly adults and adults who have undergone treatment for oral-pharyngeal cancer. Research has demonstrated that adequate lingual pressure is one of the most significant factors in one's ability to swallow safely, assisting in the avoidance of penetration and aspiration of food and liquid. Lingual pressures decrease, however, with age and with disease-related processes. This study examined the lingual pressure measurements during saliva swallows of a healthy adult female (participant one) and an adult female who underwent a partial glossectomy with free flap reconstruction and radiation treatment (participant two). The KayPENTAX Swallowing Signals Lab (Kay Elemetrics, Lincoln Park, NJ) was used to measure peak lingual pressure during saliva swallows, using three-bulb tongue arrays, so that comparisons between the two participants could be made. The results revealed that the mean lingual pressure measurements during saliva swallows between the two participants varied greatly. For participant one, mean anterior lingual pressures were 106.4 and 108.4 (time period 1 and 2), while the mean posterior lingual pressures were 323 and 373.6 (time period 1 and 2). Participant two's mean anterior lingual pressures were 30.4 and 11.6 (time period 1 and 2), and her mean posterior lingual pressures were 58.6 and 66.2 (time period 1 and 2). The peak lingual pressures for participant one and participant two are similar to the peak lingual pressures of healthy adults and adults who have undergone head and neck cancer treatment. The posterior tongue pressures were greater than the anterior tongue positions in both participants, as the posterior tongue's role is to propel the bolus posteriorly into the esophagus with significant force against the posterior pharyngeal wall. Despite participant two's reduction in lingual pressure measurements in comparison to participant one, the trend for the posterior lingual pres (open full item for complete abstract)

Committee: Lisa Kelchner (Committee Chair); Aimee Dietz (Committee Member) Subjects: Speech Therapy

Doctor of Philosophy, The Ohio State University, 2012, Nutrition Program, The Ohio State University

Anthocyanins (ACN), natural pigment flavonoids, exhibit chemopreventive, chemotherapeutic, and anti-inflammatory activities in the oral cavity. ACN have recently been included into products for the promotion of oral health. However, it is unclear which ACN-rich foods could be most efficacious. Since fruits and vegetables contain unique mixtures of ACNs, an understanding of effect of ACN structure on metabolism, bioavailability, and bioactivity is needed for providing sound dietary recommendations and development of products containing ACN for promotion of oral health. ACN extracts from blueberry, chokeberry, black raspberry, red grape, and strawberry were incubated ex vivo for 60 min in human saliva (Chapter 2). Degradation of chokeberry ACN was primarily enzymatic and mediated by bacteria. Glycosides of delphinidin (Dp) and petunidin (Pt) were more susceptible to chemical degradation than those of cyanidin (Cy), pelargonidin (Pg), peonidin (Pn) and malvidin (Mv) in both intact and artificial saliva. Stability of di- and tri-saccharide conjugates of anthocyanidins was slightly greater than that of mono-saccharide derivatives. The ex vivo degradation was significantly decreased in saliva obtained after volunteers rinsed their mouths with anti-bacterial chlorhexidine. I next examined whether the ex vivo finds could be recapitulated in vivo by having subject retain for 5 minutes either chokeberry or red grape juice in their mouths (Chapter 3). Retained juice, oral washings and buccal scrapings were collected and analyzed for stability of ACN in the oral cavity, binding to buccal mucus and uptake into buccal cells. Cy-3-glucoside preferentially accumulated with the buccal cells compared to other Cy glycosides. Anti-bacterial treatment resulted in greater stability of ACN in retained juice. Loss of Dp glucosides exceeded that of Pt, Cy, Pn and Mv glucosides during oral retention of red grape juice. Lesser relative amounts of Dp and Pt glucosides were associated with mu (open full item for complete abstract)

Committee: Mark Failla PhD (Committee Chair); M. Monica Giusti PhD (Committee Co-Chair); Purnima Kumar DDS PhD (Committee Member); Steven Schwartz PhD (Committee Member); Yusen Liu PhD (Other) Subjects: Alternative Medicine; Dental Care; Dentistry; Food Science; Immunology; Nutrition

Doctor of Philosophy, The Ohio State University, 2004, Nursing

Many studies document the negative effects associated with providing care for an ill relative. The overall purpose of this research was to increase the understanding of factors affecting health in young caregivers of cancer patients. The specific aims were to: identify competing conceptualizations of the effects of caregiving, to present a multidimensional framework with which to examine the caregiver experience from a biopsychosocial perspective, and to test the feasibility of a methodology for studying relationships among variables related to stress and health outcomes in spouses of patients with cancer. Literature was reviewed to form the basis of a concept analysis and theory construction. These were used as the basis for the design of the pilot study. The framework for this study combines aspects of the Transactional Stress Model with Psychoneuroimmunology. A descriptive, cross-sectional design was used. The convenience sample consisted of spouses of cancer patients diagnosed within the previous 12 months. The couples were all parents with at least one dependent child. Measures included self-report and physiological measures of stress, health, and immune function. Physiologic stress was measured by salivary cortisol assay, and immune function by salivary secretory IgA. Correlational analyses were used to examine relationships among variables. The sample included 13 subjects, and was 61.5% female Results revealed a mean morning cortisol level of 0.9036 ug/dL (SD= .1825 ug/dL), and mean sIgA level of 299.97 ug/ml (SD= 320.66 ug/ml). Potential problems with including salivary assays in research are discussed. Women had lower general health scores than men. Individuals with larger family incomes reported better general health, had higher sIgA and cortisol levels. Lower income individuals reported higher perceived stress, and high perceived stress was associated with lower general health scores. Occupational status was a strong predictor of the physiologic stress an (open full item for complete abstract)

Committee: Carol Kennedy (Advisor) Subjects:

Doctor of Philosophy (PhD), University of Toledo, 2012, College of Medicine

There is emerging consensus that P2X4 and P2X7 ionotropic purinoceptors (P2X4R and P2X7R) are critical players in regulating [Ca2+]i dynamics and fluid secretion in the salivary gland. In contrast, details regarding their compartmentalization and selective activation, contributions to the spatiotemporal properties of intracellular signals and roles in regulating protein exocytosis and ion channel activity have remained largely undefined. Moreover, information regarding modulation of P2X class of receptors by cAMP, the major purinergic cell signaling pathway for protein secretion has been limited. To address these gaps in our understanding, we profiled mouse parotid acinar cells using live-cell imaging to follow the spatial and temporal features of ATP-evoked Ca2+ dynamics and exocytotic activity. Selective activation of P2X7Rs revealed an apical-to-basal [Ca2+]i signal that initiated at the sub-luminal border and propagated with a wave speed estimated at 17.3 ± 4.3 μm/s (n = 6). The evoked Ca2+ spike consisted of Ca2+ influx and Ca2+-induced Ca2+ release from intracellular Ca2+ channels. In contrast, selective activation of P2X4Rs induced a Ca2+ signal that initiated basally and propagated toward the lumen with a wave speed of 4.3 ± 0.2 μm/s (n = 8) that was largely independent of intracellular Ca2+ channel blockade. Consistent with these observations, P2X7R expression was enriched in the sub-luminal regions of acinar cells while P2X4R appeared localized to basal areas. We also predict that P2X7R enriching the apical regions of acinar cells are activated by luminal ATP. We also showed that P2X4R and P2X7R activation evokes exocytosis in parotid acinar cells. Our studies also demonstrate that the P2X4R-mediated [Ca2+]i rise and subsequent protein exocytosis was enhanced by ivermectin (IVR) thereby making it a potential target treatment of salivary hypofunction diseases. Additionally, experiments designed to assess crosstalk between purinoceptors (P2X4R and P2X7R) and (open full item for complete abstract)

Committee: David Giovannucci PhD (Advisor); Joseph Margiotta PhD (Committee Member); Kathryn Eisenmann PhD (Committee Member); Joshua Park PhD (Committee Member); Guillermo Vazquez PhD (Committee Member) Subjects: Biology; Neurobiology; Neurology; Neurosciences; Pharmacology; Physiology