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Control of Foodborne Pathogenic Bacteria Using Natural Plant AntimicrobialsReyna-Granados, Javier Rolando January 2012 (has links)
Foodborne pathogens are a threat to public health worldwide. Because many consumers prefer natural compounds to synthetic additives, research on safe plant-derived compounds with antimicrobial activity against foodborne pathogens is vital. The aim of this investigation was to evaluate the antimicrobial activities of plant essential oils (oregano, cinnamon, lemongrass), their active components (carvacrol, trans-cinnamaldehyde, citral) and plant-extracts such as green tea, apple skin extract, black and decaffeinated black tea, grapes seed and pomace extracts against foodborne bacteria. Salmonella enterica serotype Typhimurium DT104, and serotype Newport, were selected conducting an antibiotic screening on 23 Salmonella isolates using seven antibiotics to determine antibiotic resistance. Listeria monocytogenes (strain 101M; beef and pork sausage isolate; resistant to antimicrobials in past investigations) was included to represent gram-positive bacteria. Escherichia coli O157:H7 virulent isolates (932- apple juice isolate; ATCC 35150- human isolate; F4637- sprouts isolate; used as a cocktail) were selected after conducting a Multiplex PCR over nine E. coli O157:H7 isolates to detect shiga-toxin 1 and 2 genes. All antimicrobials were evaluated in vitro in phosphate buffered saline. In general, all pathogens were more susceptible to essential oils and their active components, than powder extracts. The most active antimicrobials from each category were directly applied on foods. The activity of oregano oil (0.5%) and green tea (3%) was evaluated against S. Typhimurium on chicken and S. Newport on tomatoes and sprouts, and the results showed that oregano oil was more effective. In addition, baby spinach leaf samples inoculated with green fluorescent protein labeled S. Newport were examined under confocal scanning laser microscope before and after antimicrobial treatments. Antimicrobial experiments against L. monocytogenes on sprouts, ham and bologna, carvacrol at 0.5% and grape seed extract at 3% were used and carvacrol showed better activity. Antimicrobial activity against E. coli O157:H7 was tested on romaine lettuce, spinach and ground beef using oregano oil at 0.5% and green tea at 3%. Both compounds were effective showing no recovery of E. coli O157:H7 from lettuce and spinach; however, was not reduced in ground beef. Antimicrobial plant compounds have the potential for reducing foodborne pathogenic bacteria on/in various foods.
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Potato seed tuber physiological age and tolerance of attack by the potato cyst nematode Globodera pallidaHaydock, Patrick Peter John January 1990 (has links)
Seed tubers conditioned to 0, 200 or 400 day-degrees above 4t were grown in land infested with Q. pallida. Plants from 400 day-degree seed emerged earlier, had larger canopies and greater dry weights early in the growing season than plants from 0 day-degree seed. However, physiological ageing reduced peak percentage ground cover and advanced crop senescence so that similar quantities of solar radiation were intercepted over the whole growth period. Overall, total and ware yields were not affected much by seed tuber physiological age. The partially resistant cultivars tested were more tolerant than the non resistant cultivars but tolerance was not usually affected by physiological age of seed tubers. The effects of nematicide treatment, initial Q. pallida population density, cultivar maturity class, cultivar resistance status and planting date on nematode multiplication, plant growth and tolerance of attack by Q. pallida are discussed. Using data from a variable temperature water bath experiment, probability and regression analysis estimated mean basal temperatures for the development of Q. pallida and Q. rostochiensis at 2.5 and S.rCi least variance analysis estimates were 3.5 and 4.rC respectively. Approximately 200 day-degrees above 3.5 and 4.rC were required from the inoculation of JJ2 of Q. pallida and G. rostochiensis to the peak numbers of JJ5 found in potato roots. From a range of chemicals tested for their ability to release antigen from Q. pallida cysts, sodium hypochlorite was found to be the most effective. Released antigen was detected using polyclonal antisera and monoclonal antibodies in an ELISA test. The potential for the development of an ELISA based diagnosis test for PCN using species-specific antibodies is discussed.
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Analysis of Reciprocal Inhibition Between Candida albicans and Opportunistic Pathogens Enterobacter aerogenes and Enterobacter cloacaeHall, Amanda, Pribanich, Steven, Fox, Sean 07 May 2020 (has links)
The fungal pathogen Candida albicans and the opportunistic bacterial pathogens Enterobacter aerogenes and Enterobacter cloacae are common sources of human disease. The colonization of proximal anatomical locations by these pathogens suggests that interspecies polymicrobial interactions between Candida albicans and Enterobacter species occur. In order to understand mechanisms of diseases caused by these pathogens and to further the study of disease prevention, analyzation of their combined activities was conducted in this study. Changes in fungal morphology, cellular viability, and colony density were investigated using fungal and bacterial co-cultures. The effects of the Candida secreted quorum sensing molecule farnesol on Enterobacter aerogenes and Enterobacter cloacae was studied to observe changes in Enterobacter viability and colony density. The effects of the presence of Enterobacter species on Candida albicans was studied by observing changes in Candida morphology and colony density. The mutant strain of Candida albicans AlS6-/- was also cultured with Enterobacter to determine if the presence of the ALS6 surface glycoprotein gene affected Candida viability and colony density in the presence of Enterobacter species. Statistically significant decreases were observed in all studied metrics between experimental and control groups. This indicated that the interactions observed between Candida albicans and Enterobacter species represent reciprocal inhibitions of cellular functionalities. As Candida albicans is the primary cause of human fungal infections and Enterobacter species are common causes of opportunistic infections, the study of polymicrobial interactions between Candida and Enterobacter species as conducted in this study is important to furthering efforts of human disease inhibition.
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Pathology and Osteological Observations of Early Pliocene Rhinoceros, Teleoceras aepysoma (Perissodactyla, Rhinocerotidae) from Gray Fossil Site, TennesseeScaife, Thomas 01 May 2024 (has links) (PDF)
Rhinoceroses were an important part of North America’s Paleogene and Neogene ecosystems, with Teleoceras aepysoma being one of the last representatives of this family. Specimens of T. aepysoma from the Gray Fossil Site (GFS) possess distinct/peculiar pathologies: including a pair of fused ribs and ankylosed phalanges. A qualitative description of the pathologies in the GFS T. aepysoma, including new material, was conducted to accurately identify pathologies and make interpretations about the life history of the GFS rhinos. Analysis suggests that rheumatoid arthritis is common in the lower limb bones of GFS rhinos. Additionally, the rib and toe pathologies are more severe than anticipated, with the ribs showing multiple stages of healing indicating repeated trauma, likely being the first direct evidence of agonistic behavior in Teleoceras. This study provides a glimpse of what pathological conditions rhinocerotids may have been vulnerable to through time, as well as a baseline for future studies.
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DIFFERENTIAL GENE EXPRESSION IN EQUINE CARTILAGINOUS TISSUES AND INDUCED CHONDROCYTESAdam, Emma N. 01 January 2016 (has links)
Degenerative joint disease, or osteoarthritis, is a major cause of lameness and morbidity in horses, humans, and dogs. There are no truly satisfactory cures for this widespread problem and current treatments all have limitations or unwanted side effects.
New cell-based strategies to repair joint surface lesions have generated a high level of interest, but have yet to achieve the full restoration of articular cartilage structure and function. Currently used therapy cells include autologous chondrocytes and adult mesenchymal cells such as bone marrow derived cells and adipose derived cells. Unfortunately, the resultant repair tissue is biomechanically inferior fibrocartilage. A critical gap in knowledge in this regard is a limited understanding of the specific cellular phenotype of normal, robust articular chondrocytes.
This thesis examines the global mRNA transcriptome of equine articular cartilage to test the hypothesis that adult articular chondrocytes have a unique gene expression profile. In the first part of the study, RNA-sequencing was used to compare the mRNA transcriptome of normal adult articular cartilage with five other cartilaginous tissues. From these comparisons, locus level gene expression and alternative splicing patterns have been identified that clearly distinguish articular cartilage. In the second part of the study, fetal (interzone, cartilage anlagen chondrocytes, dermal fibroblasts) and adult (bone marrow derived, adipose derived, articular chondrocytes, dermal fibroblasts) primary cells were grown in culture and stimulated to differentiate into chondrocytes. The chondrogenic differentiation potential as assessed by matrix proteoglycan and the expression of cartilage biomarker genes was highly variable among cell types. Together, these results advance our understanding of the specific phenotype of articular chondrocytes and the potential of prospective therapeutic progenitor cells to differentiate into articular chondrocytes. This new knowledge will improve efforts to optimize cell-based therapies for osteoarthritis and the repair of joint cartilage lesions.
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Diffuse Brain Injury Triggers Ultra-Rapid Perisomatic Traumatic Axonal Injury, Wallerian Change, and Non-Specific Inflammatory ResponsesKelley, Brian Joseph 01 January 2006 (has links)
A significant component of diffuse brain injury (DBI) is diffuse axonal injury (DAI) which is responsible for the morbidity and mortality associated with this condition. DAI and its experimental counterpart traumatic axonal injury (TAI) result in scattered microscopic pathology characterized by focal impairment of axonal transport leading to progressive swelling and delayed axotomy. DBI-mediated perisomatic axotomy does not result in acute neuronal death suggesting that delayed axotomy was responsible for this unanticipated response. To evaluate this hypothesis, we examined the spatiotemporal progression of DBI-mediated perisomatic TAI. LM / TEM identified impaired axonal transport within 15 - 30 min post-injury. Perisomatic TAI revealed somata and related proximal / distal axonal segments with normal ultrastructural detail continuous with axonal swellings. In other cases, axotomy was confirmed by loss of axonal continuity distal to the swelling. By 60 - 180 min post-injury, somatic, proximal segment, and swelling ultrastructure were comparable to earlier time points although swelling diameter increased. Distal segment ultrastructure revealed the initial stages of Wallerian degeneration. Axotomy sites did not internalize pre-injury administered dextran suggesting pathogenesis independent of altered axolemmal permeability. Given the rapidity of perisomatic axotomy, absence of axolemmal permeability may constitute the more significant finding in terms of somatic protection.DBI-mediated neuroinflammatory reactions were then examined to see if this non-lethal neuronal pathology evoked responses comparable to those following focal injury. Microglia / macrophage responses within diffusely injured loci uncomplicated by focal pathology were explored using LM, TEM, and confocal evaluations as was albumin immunoreactivity to assess injury-induced blood-brain barrier (BBB) alterations. Initially, microglial activation was observed within injured loci while microglia within adjoining regions maintained resting phenotypes. Scattered activated microglia were observed among injured axons though no clear associations were seen. Later, activated microglia contained myelin debris while only limited microglial aggregations were recognized. Macrophages also localized to injured loci with select cells approximating somata of axotomized neurons. Immune cell observations correlated with altered BBB permeability. These data indicated rapid, yet initially uncoordinated, and persistent immune cell reactivity to DBI pathology. Taken together, these responses suggest that histopathological evaluation following DBI may include non-lethal neuronal injury with unique neuroinflammatory findings.
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ROLE OF IL-17 AND TH17 CELLS IN HSV INDUCED OCULAR IMMUNOPATHOLOGYSuryawanshi, Amol Sahebrao 01 August 2011 (has links)
Herpes simplex virus (HSV) infection of the cornea leads to a blinding immuno-inflammatory condition of the eye also called stromal keratitis (SK). SK immunopathology is characterized by the infiltration of CD4+ T cells of Th1 phenotype as well as the development of new blood vessels into the normally avascular cornea. Studies in mouse models of SK have firmly established the role of CD4+ T cells, and particularly of Th1 phenotype, as the principal mediators of SK immunopathology. However, with the recent discovery of IL-17A and Th17 cells, the role of this cytokine as well as Th17 cells remains to be further defined. Recently it was shown that the normal cornea expresses VEGF-A, however its biological activity is impeded by its binding to a soluble form of VEGF-A receptor-1 (sVEGFR-1). Past studies have implicated the role of vascular endothelial growth factor-A (VEGF-A) in HSV induced corneal angiogenesis, however the source of VEGF-A as well as molecular mechanisms, particularly in the context of VEGF-A/sVEGFR-1 balance during HSV infection, are poorly understood.
The first part of this dissertation (I) reviews past literature on HSV induced corneal SK immunopathology. It focuses on the understanding of HSV-1 induced events that particularly results in corneal angiogenesis as well as tissue damage mediated by different type of cells as well as their secreted products. The next three parts (II-IV) focus on the mechanisms of HSV induced corneal angiogenesis as well as the relative role of Th1 and Th17 cells in SK immunopathology. Results in part II focuses on the relative role of IFN-γ/IL-17 as well as Th1/Th17 cells in HSV induced corneal immunopathology. The third section evaluate the significance of VEGF-A/sVEGFR-1 balance in HSV induced corneal neovascularization. Results in part IV focus on the role of IL-17A in altering the balance between VEGF-A and sVEGFR-1 post ocular HSV infection and subsequent corneal angiogenesis.
Collectively these studies identified novel mechanisms by which HSV infection of the cornea leads to the development of angiogenesis as well as corneal tissue damage and subsequent SK immunopathology, the most common cause of infectious blindness in the Western World.
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The Human Intruder Test: An Anxiety Assessment in Rhesus Macaques (Macaca Mulatta)Peterson, Emily J 23 November 2015 (has links)
The human intruder test (HIT) is a noninvasive tool widely used for assessing anxiety in rhesus macaques (Macaca mulatta). This thesis explores the HIT procedure and applies it to a population of monkeys with a self-injurious behavioral pathology. Individual variation on this test can be used to assess anxiety and temperament. The first experiment of this thesis applied two different procedures of the HIT to 17 monkeys at UMass. Monkeys displayed little response to the intruder, and no significant differences were detected for the two procedures. To determine whether these responses were unique to the UMass monkeys, their behavior was then compared to the behavior of monkeys at three other primate facilities. UMass monkeys showed less of a reaction compared to monkeys at other facilities. They came to the front of the cage when the intruder entered the room whereas the monkeys at other facilities moved to the back and showed virtually no threats to the intruder. One possible explanation is the increased exposure to humans that UMass monkeys experience. Even though the human running the HIT was a stranger, monkeys at UMass may not perceive a new human in front of their cage to be a threat. The second experiment tested the hypothesis that monkeys with a record of self-injurious behavior (SIB) would be more anxious in response to the HIT. The cage-side version of the HIT was applied to 41 monkeys with a record of self-injurious behavior and 36 matched controls. In contrast to our prediction, SIB subjects spent significantly less time showing anxious behavior and aggressive behavior toward the intruder as well as spent more time in the front of the cage. SIB subjects showed the same range of behaviors as controls, but significantly less behavioral change overall. These data add to the evidence from experiment one that the HIT may not be a sufficient novelty test to elicit a response in monkeys who are more often exposed to different people. An alternative explanation is that SIB is associated with a depressive like syndrome based on reduced overall activity and possibly lowered affect during the stare phase.
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Increased coding potential of Bovine Herpesvirus 1Jefferson, Victoria 08 December 2023 (has links) (PDF)
Bovine respiratory disease (BRD) costs the cattle industry millions of dollars in costs in treatment and loss every year in the United States. A significant pathogen often contributes to BRD is Bovine Herpesvirus 1 (BoHV-1), a double stranded DNA virus with the ability to establish latency in the trigeminal ganglia and neurons. Primary infection with BoHV-1 results in immunosuppression that increases the risk of secondary bacterial infection and pneumonia. Because herpesviruses infect their hosts for life and can be reactivated in times of stress, BoHV-1 can present a recurring risk of BRD. The following research aims to expand the knowledge of the genome of this costly agricultural pathogen and provide evidence of viral features that can be further explored to increase the efficiency of its control.
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CELLULAR AND MOLECULAR BASIS OF EQUINE ARTERITIS VIRUS PERSISTENT INFECTION IN THE STALLION REPRODUCTIVE TRACT: CHARACTERIZATION OF LOCAL HOST-PATHOGEN INTERACTIONS MEDIATING LONG-TERM VIRAL PERSISTENCECarossino, Mariano 01 January 2018 (has links)
Equine arteritis virus (EAV) has a global impact on the equine industry being the causative agent of equine viral arteritis (EVA), a reproductive, respiratory, and systemic disease of equids. A distinctive feature of EAV infection is that it establishes long-term persistent infection in the reproductive tract of stallions and is continuously shed in the semen (carrier state). Recent studies showed that long-term persistence is associated with a specific allele of the CXCL16 gene (CXCL16S). However, the cellular and molecular mechanisms underlying the establishment and maintenance of persistent infection are yet to be determined. The studies were undertaken herein unequivocally demonstrated that the ampulla is the main EAV tissue reservoir rather than immunologically privileged tissues (i.e., testes) and that EAV has specific tropism for stromal cells and CD8+ T and CD21+ B lymphocytes but not glandular epithelium in the reproductive tract. Furthermore, persistent EAV infection is associated with a significant humoral, mucosal antibody and inflammatory response at the site of persistence, characterized by induction of high levels of neutralizing antibodies (IgG1), mucosal anti-EAV-specific IgA, IgG1, IgG3/5, and IgG4/7 with variable neutralizing efficacy; and moderate, multifocal lymphoplasmacytic ampullitis, with significant infiltration of T lymphocytes (mainly CD8+ and low numbers of FOXP3+ lymphocytes), CD21+ B lymphocytes, diverse Ig-secreting plasma cells, and Iba-1+ and CD83+ tissue macrophages/dendritic cells. Moreover, EAV long-term persistent infection is associated with a CD8+ T lymphocyte transcriptional profile with upregulation of T-cell exhaustion-related transcripts and homing chemokines/chemokine receptors (CXCL9-11/CXCR3 and CXCL16/CXCR6), orchestrated by a specific subset of transcription factors (EOMES, PRDM1, BATF, NFATC2, STAT1, IRF1, TBX21), which are associated with the presence of the susceptibility allele (CXCL16S). Finally, these studies have determined that long-term EAV persistence is associated with the downregulation of a specific seminal exosome-associated miRNA (eca-mir-128) along with an enhanced expression of CXCL16 in the reproductive tract, a putative target of eca-mir-128. These findings provide evidence that this miRNA plays a crucial role in the regulation of the CXCL16/CXCR6 axis in the reproductive tract of persistently infected stallions, a chemokine axis strongly implicated in EAV persistence. The findings presented herein suggest that complex host-pathogen interactions shape the outcome of EAV infection in the stallion and that EAV employs complex immune evasion mechanisms favoring persistence in the reproductive tract. Further studies to identify specific mechanisms mediating the modulation of the CXCL16/CXCR6 axis and viral immune evasion in the reproductive tract of the EAV long-term carrier stallion are warranted.
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