Hematopoietic stem cell transplantation (HSCT) is an important form of therapy for hematological genetic disorders and malignancies, particularly hematological cancers. However, common usage of this procedure is obstructed by graft-versus-host disease (GvHD), in which transplanted donor T cells wage an attack on recipient antigens, causing severe tissue damage and mortality. GvHD prognosis remains poor, and current treatment methods continue to be insufficient, especially for patients with more advanced and severe GvHD. T cells have been identified as the fundamental force behind GvHD, and their cellular metabolism is deemed vital to their fate and function, especially in pathogenic environments. A hallmark of T cell metabolism in GvHD microenvironments is aerobic glycolysis, which maximizes biomass accumulation and supports growth and proliferation. Lactate dehydrogenase A (LDHA) is an essential enzyme that sustains this pathway and may be a potential therapeutic target. Using murine and in-vitro GvHD models, this study investigates the ameliorative impacts of LDHA inhibition on the fate and function of T cells following HSCT. The results reveal that LDHA depletion leads to an immunosuppressive donor T cell characterization that minimizes recipient harm induced by GvHD. Future studies should focus on investigating LDHA inhibition in in-vivo models to introduce a paradigm shift in the development of clinically relevant therapeutics.

The F-strain of Mycoplasma gallisepticum (FMG) is commonly used in vaccination programs to displace infections by more virulent natural or wild type Mycoplasma gallisepticum strains. However, a better understanding of the mechanisms responsible for altered egg production (EP) and egg quality in commercial layers infected with FMG is important, as these alterations can cause economic loss to the United States layer industry. This study was designed to examine potential mechanism(s) responsible for alterations in EP and egg quality by FMG-inoculation. The effects of FMG on production parameters and physiological characteristics of commercial laying hens were evaluated. In isolation units, 12 wk FMG inoculation delayed onset of lay approximately one wk, decreased overall EP, and decreased EP 34 wk post-inoculation. A 12 wk FMG inoculation also resulted in a higher incidence of fatty liver hemorrhagic syndrome, ovarian follicular regression, and decreased isthmal and vaginal proportions of the reproductive tract. Ovarian regression may be related to retarded production (liver), transport (blood), and/or uptake (ovary) of yolk particles. Changes in blood characteristics (i.e. lipid composition) with FMG colonization of the liver may become manifest through changes in egg constituents. As evidenced through changes in the relative weights of various reproductive organs, colonization of these organs by FMG, in addition to the liver, may also be a cause of the effects observed on EP. Increases in hematocrit, serum triglycerides, and plasma protein between 8 and 10 wk post FMG-inoculation, suggest that the initial weeks of EP are stressful. Post-peak decreases in these same variables suggest a more chronic inhibition on lipid and protein synthesis in the liver. Decreased blood lipid concentration may be directly responsible for the reductions in yolk lipid, cholesterol, and fatty acid deposition in 12 wk FMG-inoculated hens. Dual adverse effects in the caged layer facility on feed conversion and egg mass were realized in 22 wk FMG-inoculated birds. In contrast, a 12 wk FMG inoculation delayed onset of lay without a loss in total EP or egg mass. Therefore, inoculation with FMG at 12 wk is more practical and cost effective. Higher degrees of physiological stress experienced by hens in a caged layer facility may exacerbate the effects of FMG inoculation seen in the isolation units. These data demonstrate that alterations in performance and egg characteristics of commercial layers inoculated with FMG at either 12 or 22 wk of age and housed in either isolation units or caged layer facilities are related to mutual functional disturbances in the blood, liver, ovary, and oviduct without concomitant intestinal changes.

<p>Unfractionated heparin (UFH) is used as an adjunct during thrombolytic therapy. However, its use is associated with many clinical limitations, such as the inability to inhibit fibrin-bound coagulation factors, increasing the potential for sustained procoagulant activity. We have developed a covalent conjugate of antithrombin (AT) and heparin (ATH) with superior anticoagulant properties to those of UFH. Some advantages of ATH include enhanced inhibition of surface-bound enzymes and its ability to reduce the overall size and mass of clots <em>in vivo</em>. However, the potential interactions of UFH or ATH with the components of the fibrinolytic pathway are not well understood. Therefore, our study utilized discontinuous second order rate constant (<em>k₂</em>) assays to determine rates of inhibition of plasmin (Pn) in the presence or absence of fibrin by AT+UFH <em>vs.</em> ATH. In addition, we monitored the rates of Pn generation in a system comprised of preformed fibrin clots with the aim of evaluating the inhibitory effect of AT+UFH or ATH in this more native system. The <em>k₂</em>values for the inhibition of Pn without fibrin were 5.74x10⁶±0.278x10⁶ and 6.39x10⁶±0.588x10⁶ for AT+UFH and ATH, respectively (p=0.36). In the presence of fibrin, the <em>k₂</em>values decreased to 1.45x10⁶±0.0971x10⁶ for AT+UFH and 3.07x10⁶±0.192x10⁶for ATH (<em>p</em></p> / Master of Science (MS)

Introduction: Immune thrombocytopenia (ITP) is an autoimmune bleeding disorder characterized by a platelet count less than 100 x 109/L. Platelet-bound autoantibodies are detected in the peripheral blood of only 40-50% of ITP patients. The subset of ITP patients who do not have detectable autoantibodies may truly be seropositive, but autoantibodies may not be detected due to limitations of the assays. Studies have also suggested that autoantibodies might be sequestered in the bone marrow where autoantibodies may impair platelet production. In addition, detecting autoreactive antibody-secreting B cells using the Enzyme-linked Immunospot (ELISPOT) assay was shown to be highly sensitive. In this study, the bone marrow and the peripheral blood compartments of ITP patients were tested for the presence of anti-GPIIbIIIa and anti-GPIbIX IgG autoantibodies and autoreactive B cells. Methods: Bone marrow aspirate and peripheral blood samples were collected from ITP patients (n=12), non-immune thrombocytopenic patient controls (n=3), and healthy controls (n=5). Mononuclear cells were isolated and tested for the presence of anti-GPIIbIIIa and anti-GPIbIX IgG autoreactive B cells before stimulation and after stimulation with R848 and IL-2 using the ELISPOT assay. Anti-GPIIbIIIa and anti-GPIbIX IgG autoantibodies were detected in the bone marrow and the peripheral blood using the direct antigen capture assay. Results: In our study, we detected autoantibodies and autoreactive B cells of known specificity in the bone marrow of a subset of ITP patients. Detecting anti-GPIIbIIIa and anti-GPIbIX autoantibodies in the bone marrow or the peripheral blood had a sensitivity of 42% and testing both compartments increased the sensitivity to 58%, while maintaining 100% specificity. Autoreactive B cells were detected at low frequencies with low specificity in the bone marrow and the peripheral blood of a subset of ITP patients. The majority of the ITP patients without detectable autoantibodies in the peripheral blood did not have autoantibodies in the bone marrow, and autoreactive B cells were not detected in either compartment. Conclusion: Examining both the bone marrow and the peripheral blood compartments for autoantibodies or autoreactive B cells increased the sensitivity. Furthermore, detecting autoantibodies using the antigen capture assay is more sensitive and specific than detecting autoreactive B cells using the ELISPOT assay. / Thesis / Master of Science (MSc)

Bakgrund: Trots att stora framsteg har gjorts senaste åren kring behandling av hematologiska cancersjukdomar, avlider fortfarande en stor del av patienterna till följd av sjukdomen eller komplikationer relaterade till sjukdom och behandling. Patienter med hematologisk cancer tillbringar ofta långa perioder på sjukhus på grund av långtgående behov av intravenöstantibiotika, transfusioner och cytostatikabehandlingar. De erbjuds inte palliativ vård i samma utsträckning som patienter med solida tumörsjukdomar, detta trots att de visar sig ha liknande symtombörda som patienter med andra cancerformer. Flera studier har publicerats senaste åren som visar att tidig integrering av palliativ vård vid solida tumörsjukdomar förbättrar fysisk och psykisk hälsa, minskar symtombörda och medför positiva effekter på övergripande livskvalitet. Effekt av tidig integrering av palliativ vård vid hematologisk cancer anses därför angeläget att undersöka. Syfte: Att undersöka hur tidig integrering av palliativ vård för patienter med hematologisk cancer påverkar patientens livskvalitet. Metod: Litteraturöversikt med induktiv ansats har genomförts med litteratursökningar i databaserna CINAHL, Pubmed och PsycINFO. Totalt fjorton kvantitativa artiklar om tidig palliativ vård vid hematologisk cancer analyserades med begreppet personcentrerad vård som teoretiskt ramverk. Resultat: Flera aspekter av livskvalitet förbättrades vid tidig integrerad palliativ vård vid hematologisk cancer. Smärta och psykiska symtom minskade, patientdelaktighet ökade genom fler genomförda samtal och information om sjukdom och prognos. Färre dödsfall på intensivvårdsavdelning sågs och färre aggressiva behandlingar i livets slutskede. Genom specifika skattningsinstrument för hälsorelaterad eller sjukdomsspecifik livskvalitet påvisades lägre minskning av livskvalitet under sjukdomsförloppet jämfört med enbart sedvanlig vård. Slutsats: Palliativ vård tillsammans med sedvanligt hematologisk vård är till gagn för patienter med hematologisk cancer och kan implementeras tidigt i sjukdomsförloppet för att främja livskvaliteten. Fler randomiserade kontrollerade kliniska studier behövs för att kunna fortsätta utveckla en palliativ vård som kan tillgodose de specifika behov en patient med hematologisk cancer kan ha. / Background: Although great progress has been made in recent years in the treatment of hematological cancers, a large proportion of patients still die as a result of the disease, or complications related to disease and treatment. Patients with hematological cancer often spend long periods of time in hospital due to far-reaching needs for intravenous antibiotics, transfusions, and chemotherapy treatments. They are not offered palliative care to the same extent as patients with solid tumor diseases, despite the fact that they are found to have a similar symptom burden as patients with other cancers. Several studies have been published in recent years showing that early integration of palliative care in solid tumor diseases improves physical and mental health, reduces symptom burden, and has positive effects on overall quality of life. The effect of early integration of palliative care in hematological cancer is therefore considered important to investigate. Aim: The aim of this review was to explore how early integrated palliative care for patients with hematologic cancer affects the patients’ quality of life. Method: A literature review with an inductive approach has been conducted with literature searches in the databases CINAHL, Pubmed and PsycINFO. A total of fourteen quantitative studies on early palliative care in hematological cancer were analyzed using the concept of person-centered care as a theoretical framework. Results: Several aspects of quality of life were improved in early integrated palliative care in hematological cancer. Pain and psychological symptoms decreased, patient participation increased through more completed conversations and information about illness and prognosis. There were fewer deaths in intensive care units and fewer aggressive treatments at the end of life. Through specific assessment instruments for health-related or disease-specific quality of life, lower reductions in quality of life during the course of the disease were demonstrated compared to usual care alone. Conclusion: Palliative care together with usual hematological care is beneficial for patients with hematological cancer and can be implemented early in the course of the disease to promote quality of life. More randomized controlled clinical trials are needed to continue to develop palliative care that can meet the specific needs of a patient with hematological cancer.

Bakgrund: Trots att stora framsteg har gjorts senaste åren kring behandling av hematologiska cancersjukdomar, avlider fortfarande en stor del av patienterna till följd av sjukdomen eller komplikationer relaterade till sjukdom och behandling. Patienter med hematologisk cancer tillbringar ofta långa perioder på sjukhus på grund av långtgående behov av intravenöst antibiotika, transfusioner och cytostatikabehandlingar. De erbjuds inte palliativ vård i samma utsträckning som patienter med solida tumörsjukdomar, detta trots att de visar sig ha liknande symtombörda som patienter med andra cancerformer. Flera studier har publicerats senaste åren som visar att tidig integrering av palliativ vård vid solida tumörsjukdomar förbättrar fysisk och psykisk hälsa, minskar symtombörda och medför positiva effekter på övergripande livskvalitet. Effekt av tidig integrering av palliativ vård vid hematologisk cancer anses därför angeläget att undersöka. Syfte: Att undersöka hur tidig integrering av palliativ vård för patienter med hematologisk cancer påverkar patientens livskvalitet. Metod: Litteraturöversikt med induktiv ansats har genomförts med litteratursökningar i databaserna CINAHL, Pubmed och PsycINFO. Totalt fjorton kvantitativa artiklar om tidig palliativ vård vid hematologisk cancer analyserades med begreppet personcentrerad vård som teoretiskt ramverk. Resultat: Flera aspekter av livskvalitet förbättrades vid tidig integrerad palliativ vård vid hematologisk cancer. Smärta och psykiska symtom minskade, patientdelaktighet ökade genom fler genomförda samtal och information om sjukdom och prognos. Färre dödsfall på intensivvårdsavdelning sågs och färre aggressiva behandlingar i livets slutskede. Genom specifika skattningsinstrument för hälsorelaterad eller sjukdomsspecifik livskvalitet påvisades lägre minskning av livskvalitet under sjukdomsförloppet jämfört med enbart sedvanlig vård. Slutsats: Palliativ vård tillsammans med sedvanligt hematologisk vård är till gagn för patienter med hematologisk cancer och kan implementeras tidigt i sjukdomsförloppet för att främja livskvaliteten. Fler randomiserade kontrollerade kliniska studier behövs för att kunna fortsätta utveckla en palliativ vård som kan tillgodose de specifika behov en patient med hematologisk cancer kan ha. / Background: Although great progress has been made in recent years in the treatment of hematological cancers, a large proportion of patients still die as a result of the disease, or complications related to disease and treatment. Patients with hematological cancer often spend long periods of time in hospital due to far-reaching needs for intravenous antibiotics, transfusions, and chemotherapy treatments. They are not offered palliative care to the same extent as patients with solid tumor diseases, despite the fact that they are found to have a similar symptom burden as patients with other cancers. Several studies have been published in recent years showing that early integration of palliative care in solid tumor diseases improves physical and mental health, reduces symptom burden, and has positive effects on overall quality of life. The effect of early integration of palliative care in hematological cancer is therefore considered important to investigate. Aim: The aim of this review was to explore how early integrated palliative care for patients with hematologic cancer affects the patients’ quality of life. Method: A literature review with an inductive approach has been conducted with literature searches in the databases CINAHL, Pubmed and PsycINFO. A total of fourteen quantitative studies on early palliative care in hematological cancer were analyzed using the concept of person-centered care as a theoretical framework. Results: Several aspects of quality of life were improved in early integrated palliative care in hematological cancer. Pain and psychological symptoms decreased, patient participation increased through more completed conversations and information about illness and prognosis. There were fewer deaths in intensive care units and fewer aggressive treatments at the end of life. Through specific assessment instruments for health-related or disease-specific quality of life, lower reductions in quality of life during the course of the disease were demonstrated compared to usual care alone. Conclusion: Palliative care together with usual hematological care is beneficial for patients with hematological cancer and can be implemented early in the course of the disease to promote quality of life. More randomized controlled clinical trials are needed to continue to develop palliative care that can meet the specific needs of a patient with hematological cancer.

Aquacultured products assist the human demands for seafood so that foodfish supplies can remain sustainable and consistent. Although the fish-farming industry has seen dramatic growth and intensification in recent years, the latter has led to an increase in bacterial diseases and fish health management problems, resulting in major economic losses around the world. In addition to the lack of understanding of fish physiology, these complications are exacerbated by the inappropriate and controversial use of antibiotics. This work addressed these issues in striped catfish (Pangasius hypophthalmus) and Nile tilapia (Oreochromis niloticus), two economically important foodfish, by investigating alternative, more cost-effective options to promote fish health. The first two studies established reference intervals for immunology, hematology and plasma chemistry analytes in striped catfish in a recirculating aquaculture system (RAS). In a third study, the immunomodulatory effects after directly feeding probiotic strains of Bacillus subtilis NZ86 and O14VRQ in Nile tilapia were ascertained. This last study revealed that supplementation with both of the probiotic strains for 51 days stimulated several local and systemic innate immune responses of tilapia. When these transient probiotic bacteria were present in the gut, a pro-inflammatory environment was developed as evidenced by the localized higher expression of the cytokines tumor necrosis factor (TNF) – α and interleukin (IL) – 1β. Significant increases (p < 0.05) were noted differentially by both probiotic strains throughout the trial in plasma lysozyme content, alternative complement activity, and in the peripheral blood leukocyte profiles. Additionally, there were trends for increased levels of phagocytosis and respiratory burst in leukocytes of the anterior kidney and spleen at the end of the trial, suggesting the potential use of these probiotic strains for improved immune-competence. These findings help to understand and clarify the potential mechanism of action associated with the increased disease resistance recorded in preliminary studies with the same probiotic strains. Implementation of the tools established and validated in this work could be useful in evaluating fish welfare situations involving striped catfish grown in RAS conditions, and also show promise for a healthier foodfish supply where antibiotic applications practices could be minimized. / Ph. D. / Aquaculture, or fish farming, is one of the most prosperous production sectors of animal-derived food. Despite the success story of aquaculture, the fish industry is heavily plagued by bacterial diseases, which cause losses in billions of dollars annually around the world, and directly contribute to increases in human food insecurity. Since the options to cost-effectively address diseases are limited, I explored alternative ways to more safely monitor and also ensure optimal health in striped catfish and tilapia, two globally important aquaculture fishes. I investigated the values of different cellular and chemical components of the blood to monitor the health of striped catfish when grown in indoor recirculating conditions, in order to understand normal catfish physiology. The values of these blood components were comparable to those of other freshwater fishes. As part of another study, I supplemented probiotics in the diet of the tilapia for 51 days, and assessed the effects of these on the immune system of the fish. Dietary supplementation of the probiotics resulted in the presence of the probiotics in the gut of the fish. Furthermore, the presence of these microbes was tightly linked to elevated values of numerous functions of the immune system. These functions included levels of lysozyme, alternative complement, and percentage of neutrophils, which are all related with a state of heightened immunity in the animal host. The tools that I established and validated in this study are promising alternatives to optimize the health of these two important foodfish. Moreover, they could be useful for the fish farmer because of their greater cost-effectiveness, and can potentially lead to a safer foodfish supply by decreasing the reliance on antibiotics.

High-dose methotrexate (HD MTX) is critical for treating pediatric malignancies such as acute lymphoblastic leukemia and neuro-carcinoma. However, its significant toxicity due to drug accumulation poses substantial risks. This retrospective study assesses the impact of demographic factors on MTX toxicity and clearance in pediatric oncology patients. Patient records from Saint Mary Hospital were analyzed, focusing on two MTX administration protocols: a 24-hour infusion followed by alkaline hydration and a 4-hour infusion followed by alkaline hydration. We hypothesize that factors such as age, body surface area (BSA), and body mass index (BMI) are associated with MTX clearance and toxicity. The study found no significant difference in clearance between genders, but females exhibited higher toxicity rates. Ethnicity comparisons showed Caucasians had the fastest clearance, followed by Hispanics, African Americans, and others, with Hispanics experiencing the highest toxicity rates. Patients with a BSA of less than one had a lower risk of delayed clearance, although toxicity levels were similar across BSA groups. BMI analysis indicated that patients with a BMI over 25 were at a higher risk of toxicity. Taken together, these findings suggest the need for personalized treatment plans in pediatric oncology to enhance therapeutic efficacy and reduce adverse effects. Future research should expand the sample size and develop a risk stratification guideline to identify patients suitable for outpatient treatment.

This dissertation consists of five chapters, each of which focuses on various aspects of the forensic discipline of Firearms and Toolmarks. This dissertation for the most part attempts to highlight the exposure of projectiles to blood and the degradation over time of the fine detail, which is necessary for microscopic examination. This study should be of interest to students and qualified role-players in forensic science, the criminal justice system, the law community and the general population globally. Chapter 1 identifies the research problem and the necessary steps that were taken to ensure that the research methodol.ogy applied is relevant and reliable. Chapter 2 focuses on various factors that have to be considered in damage to bullets and investigation procedures that should be followed to ensure that physical evidence is preserved for submitting to a forensic science laboratory. Chapter 3 investigates the degradation effects of fired bullets exposed to various materials in a laboratory environment and the timelines associated with the degradation effects. Chapter 4 evaluates the examination procedures for fired bullets and the contributing factors that may influence the striation marks on bullets needed for microscopic examination. It also examines the scientific method used for firearm identification, and explores the admissibility of physical evidence in a court of law. The final chapter, Chapter 5 discusses the findings and recommendation of this research study. / Criminology / M. Tech. (Forensic Investigation)

In recent years, the genetic landscape of B-cell derived lymphoid malignancies, including chronic lymphocytic leukemia (CLL), has been rapidly unraveled, identifying recurrent genetic mutations with potential clinical impact. Interestingly, ~30% of all CLL patients can be assigned to more homogeneous subsets based on the expression of a similar or “stereotyped” B-cell receptor (BcR). Considering that biased distribution of genetic mutations was recently indicated in specific stereotyped subsets, in paper I, we screened 565 subset cases, preferentially assigned to clinically aggressive subsets, and confirm the SF3B1 mutational bias in subset #2 (45%), but also report on similarly marked enrichment in subset #3 (46%). In contrast, NOTCH1 mutations were predominantly detected in subsets #1, #8, #59 and #99 (22-34%). This data further highlights a subset-biased acquisition of genetic mutations in the pathogenesis of at least certain subsets. Aberrant NF-κB signaling due to a deletion within the NFKBIE gene previously reported in CLL warranted extended investigation in other lymphoid malignancies. Therefore, in paper II, we screened 1460 patients with various lymphoid malignancies for NFKBIE deletions and reported enrichment in classical Hodgkin lymphoma (27%) and primary mediastinal B-cell lymphoma (PMBL) (23%). NFKBIE-deleted PMBL cases had higher rates of chemorefractoriness and inferior overall survival (OS). NFKBIE-deletion status remained an independent prognostic marker in multivariate analysis. EGR2 mutations were recently reported in advanced stage CLL patients; thus, in paper III we screened 2403 CLL patients for mutations in EGR2. An overall mutational frequency of 3.8% was reported and EGR2 mutations were associated with younger age, advanced stage and del(11q). EGR2 mutational status remained an independent marker of poor outcome in multivariate analysis, both in the screening and validation cohorts. Whole-genome sequencing (WGS) of 70 CLL cases, assigned to poor-prognostic subsets #1 and #2 and indolent subset #4, were investigated in Paper IV and revealed a similar skewing of SF3B1 mutations in subset #2 and NOTCH1 mutations in subset #1 to that reported in Paper I. Additionally, an increased frequency of the recently proposed CLL driver gene RPS15 was observed in subset #1. Finally, novel non-coding mutational biases were detected in both subset #1 and #2 that warrant further investigation.