Digitized by Kansas Correctional Industries

Thesis (M.Sc.) -- University of Adelaide, Dept. of Microbiology, 1972.

Thesis (Ph. D.)--University of Missouri--Columbia, 1998. / "May 1998." Typescript. Vita. Includes bibliographical references (leaves 71-76). Also available on the Internet.

Thesis (Ph. D.)--University of Missouri--Columbia, 1998. / Typescript. Vita. Includes bibliographical references (leaves: 71-76). Also available on the Internet.

Background: Keratoconjunctivitis sicca (KCS) is a chronic ocular disease of both dogs and humans that can result in ocular discomfort, corneal opacification, and vision loss. Lacritin, a protein found in the tears of many species, has been shown to play a role in lacrimation and corneal health. Because of its role as a potential lacrimostimulant, assessment of endogenous lacritin levels could reveal a correlation between lacritin and tear production in the dog. Objectives: To determine if tear lacritin concentrations are decreased in canine eyes affected by KCS. Animals: 58 client-owned dogs (tear samples from 55 eyes with normal tear production and 55 eyes diagnosed with KCS). Methods: All eyes underwent an ophthalmic exam, including Schirmer Tear Testing (STT), anterior segment assessment, and tear sample collection. Tear samples were evaluated for their total protein concentrations via BCA assay and lacritin concentrations via ELISA. Results: Total protein of canine tears is increased in KCS-affected eyes as compared to normal eyes. Tear lacritin as a component of total tear protein is significantly decreased in tears from KCS-affected eyes. When measured as a concentration (mass per volume of aqueous tears), lacritin is not significantly different between KCS-affected eyes and normal eyes, nor were they strongly correlated to STT values. Conclusions and Clinical Importance: Total tear protein levels were significantly increased in canine KCS. When quantified as a proportion of total tear sample protein, tear lacritin levels are decreased in KCS-affected eyes. Relative to tear volume, tear lacritin levels are not significantly different between KCS-affected eyes and normal eyes. Assessment of lacritin supplementation in canine KCS is warranted to evaluate potential effects on lacrimation and ocular surface health. / Master of Science / Keratoconjunctivitis sicca describes a syndrome of inadequate pre-ocular tear film, due to decreased quantity or quality of tears on the ocular surface. A multitude of clinical signs and sequelae result, including ocular discomfort, ocular discharge, and corneal opacification that may lead to vision loss. Current therapies aim at improving endogenous tear production; standard therapy, however, may not significantly improve disease in up to 30% of canine patients and 40% of human patients. Development of additional therapies could improve the vision and comfort of patients with KCS. Lacritin, a protein found in the tears of many species, has been shown to play a role in lacrimation and corneal health. Because of its role as a potential stimulant of tear production, assessment of endogenous lacritin levels could reveal a correlation between lacritin and tear production in the dog. This project aims to compare tear lacritin concentrations in dogs with normal tear production versus dogs with KCS. Tear samples were collected from normal dogs (n=55) and those affected with KCS (n=55). Samples were analyzed for their total tear protein levels via BCA assay and for their lacritin levels via ELISA analysis. Total tear protein levels and tear lacritin levels were compared to Schirmer Tear Test results, a routine method of diagnosing KCS in dogs. Tear lacritin was significantly decreased in KCS, relative to the quantity of total tear proteins. As a portion of the total volume of tears, lacritin was not significantly different in KCS-affected eyes as compared to normal eyes. Further investigation is warranted to determine the effects lacritin supplementation may have on canine KCS, in regard to tear production and clinical signs of KCS.

No description available.

INTRODUCTION: There are two main procedures responsible for correcting refractive error: laser-assisted in situ keratomileusis (LASIK) and photorefractive keratectomy (PRK). The most common postoperative complication of all laser vision correction (LVC) surgeries is keratoconjunctivitis sicca, or dry eye. In addition to physical irritation, dry eye can cause poor visual acuity and reduced quality of life. There are inconsistencies in the literature about which procedure results in higher rates of dry eye thus we explored the likelihood of a dry eye intervention within the first year after surgery. METHODS: Performing a retrospective chart review of all patients who underwent LVC procedures between 2009-2019 at a private Boston ophthalmology clinic, we were able to quantify the proportion of patients that required postoperative dry eye interventions within the 12 months following surgery. At this clinic, a dry eye intervention was defined as one of two treatments: punctal plug insertion or prescription medication (Restasis or Xiidra) use. RESULTS: A total of 11,175 LASIK eyes from 5,920 individuals, 1,549 LASEK eyes from 880 individuals, and 2,006 PRK eyes from 1,165 individuals were included in the analysis. Comparing the proportion of LASIK dry eye interventions to the proportion of PRK/LASEK dry eye interventions via a 2-sample z-test at an alpha level of significance of 0.05, yielded a z-value= 5.7 and a p-value= <0.0001 at a 95% CI (0.0102 - 0.0208). CONCLUSION: Our results suggest a greater incidence of postoperative dry eye interventions for PRK/LASEK patients compared to LASIK patients in the 12 months following LVC surgery. This study was limited in regards to both objective and subjective indicators of dryness; therefore future studies should attempt to be more comprehensive in evaluating postoperative dry eye. However, this study should help surgeons in the decision making process of which refractive eye surgery to recommend in order to reduce postoperative dry eye incidence and improve overall patient quality of life and satisfaction. / 2022-06-08T00:00:00Z

A Ceratoconjuntivite Seca (KCS Keratoconjunctivitis Sicca) é uma desordem imunomediada e resulta de alterações do componente aquoso do filme lacrimal e da deficiência dos componentes lipídicos e mucoso.Seu diagnóstico é baseado no Teste Lacrimal de Schirmer (TLS) e no Teste de Ruptura do Filme Lacrimal (TRFL) e tem como sinais clínicos: secreção mucopurulenta, hiperemia conjuntival, blefaroespasmos, fotofobia, incômodo, dor, vascularização, opacidade corneana e pigmentação, além de cegueira em casos avançados. O tratamento convencional consiste em aplicações diárias de Ciclosporina 0,2% ou Tacrolimus 0,03% (pomada ou colírio oftálmicos), que apesar de controlar a doença, são custosos, não curativos e exigem alto comprometimento da interação paciente-proprietário. A terapia celular usando células-tronco (CT) traz uma nova esperança para doenças sem tratamento efetivo. Neste trabalho utilizamos CT mesenquimais (CTM) obtidas a partir de membrana amniótica (CTMA) de cães obtidas a partir do descarte destes tecidos em campanhas de castrações em diferentes tempos gestacionais, sem formação tumoral quando submetidas ao teste tumorigênico durante 60 dias. Dois animais com KCS crônica foram tratados com duas injeções de CTMA com intervalo de 30 dias, sendo a primeira de 0,5x106 células e a segunda de 1x106 células em cada glândula. Na segunda semana após a terapia foi observado aumento da TLS sugerindo um benéficio da terapia que foi diminuindo com o passar das semanas. O TRFL oscilou durante os testes e não apresentou diferenças significativas. A terapia celular utilizando CTMA de cães melhorou a condição ocular nos dois casos em momentos e parâmetros variados, com repercussão na melhoria da superfície, mas não houve regressão do quadro clínico. Investigações futuras em estágios menos avançados da doença podem ajudar a elucidar os mecanismos pelos quais esse efeito foi obtido / Keratoconjunctivitis Sicca (KCS) is an immune-mediated disorder and results from changes in the aqueous component of the tear film and also from the impairment of lipid and mucous components. Diagnosis is based on Lacrimal Schirmer Test (LST) and Tear Film Break Test (TFBT) and its clinical signs: purulent discharge, conjunctival hyperemia, blefaroespasmos, photophobia, discomfort, pain, vascularization, corneal opacity and pigmentation, addition of blindness in advanced cases. The standard treatment consists of daily applications of 0.2% cyclosporin or tacrolimus 0.03% (ophthalmic ointment or drops), that in spite of controlling the disease are expensive, require the patient does not curative and and require greater involvement of the patient-owner interaction. Cell therapy using stem cells (SC) brings new hope for diseases without effective treatment. In this work we used CT mesenchymal cells (MSCs) obtained from dogs amniotic membrane (AMSC) in the capanhas of castration at different times gestacionai without tumor formation tumorigenic when subjected to the test during 60 days. Two animals with chronic KCS were treated with two injections of CTMA with 30-day intervals, the first of which 0,5x106 cells and the second of 1x106 cells in each gland. In the second week after therapy was an increase in LST that has been decreasing over the weeks. The BTTF fluctuated during testing and no significant differences. Cell therapy using AMSC dogs improved the eye condition in both cases at times and different parameters, with consequent improvement in ocular surface but there was no regression of the clinical picture. Future research in less advanced stages of the disease may help to elucidate the mechanisms by which this effect was obtained

The combat against viral diseases has been, and still is, a major challenge in the field of drug development. Viruses are intracellular parasites that use the host cell ma-chinery for their replication and release. Therefore it is difficult to target and destroy the viral particle without disturbing the essential functions of the host cell. This thesis describes studies towards antiviral agents targeting adenovirus type 37 (Ad37), which causes the severe ocular infection epidemic keratoconjunctivitis (EKC). Cell surface oligosaccharides serve as cellular receptors for many pathogens, including viruses and bacteria. For EKC-causing adenoviruses, cell surface oligo-saccharides with terminal sialic acid have recently been shown to be critical for their attachment to and infection of host cells. The work in this thesis support these re-sults and identifies the minimal binding epitope for viral recognition. As carbo-hydrate–protein interactions in general, the sialic acid–Ad37 interaction is very weak. Nature overcomes this problem and vastly improves the binding affinity by presenting the carbohydrates in a multivalent fashion. Adenoviruses interact with their cellular receptors via multiple fiber proteins, whereby it is likely that the ideal inhibitor of adenoviral infections should be multivalent. This thesis includes design and synthesis of multivalent sialic acid glycoconjugates that mimic the structure of the cellular receptor in order to inhibit adenoviral attachment to and infection of human corneal epithelial (HCE) cells. Synthetic routes to three different classes of sialic acid conjugates, i.e. derivatives of sialic acid, 3’-sialyllactose and N-acyl modified sialic acids, and their multivalent counterparts on human serum albumine (HSA) have been developed. Evaluation of these conjugates in cell binding and cell infectivity assays revealed that they are effective as inhibitors. Moreover the results verify the hypothesis of the multivalency effect and clearly shows that the power of inhibition is significantly increased with higher orders of valency. Potential inhibi-tors could easily be transferred to the eye using a salve or eye drops, and thereby they would escape the metabolic processes of the body, a major drawback of using carbohydrates as drugs. The results herein could therefore be useful in efforts to develop an antiviral drug for treatment of EKC.

Το σύνδρομο Sjögren (ΣΣ) είναι μια αυτοάνοση νόσος που προσβάλλει κυρίως τους σιελογόνους και δακρυϊκούς αδένες οδηγώντας στην ξηρά επιπεφυκίτιδα και την ξηροστομία. Ιστοπαθολογικά χαρακτηρίζεται από λεμφοκυτταρική διήθηση Τ και Β κυττάρων, με επικράτηση των ενεργοποιημένων CD4 + Τ βοηθητικών κυττάρων. Οι ακριβείς παθογενετικοί μηχανισμοί που είναι υπεύθυνοι για το ΣΣ δεν είναι ακόμη πλήρως γνωστοί, ωστόσο φαίνεται ότι συμμετέχουν περιβαλλοντικοί, ορμονικοί και γενετικοί παράγοντες. Ακόμη, απορρύθμιση των κυτταροκινών που παράγονται από Th1, Th2 και Th17 κύτταρα εμπλέκονται στην ανάπτυξη του ΣΣ. Σκοπός: Σε αυτή τη μελέτη, αναλύσαμε την έκκριση κυτταροκινών από υποσύνολα Τ-κυττάρων από το περιφερικό αίμα ασθενών με πρωτοπαθές ΣΣ, χρησιμοποιώντας συνθετικά πεπτίδια τα οποία αντιστοιχούσαν σε επιτόπους Τ-κυττάρων των αυτοαντιγόνων La / SSB. Μέθοδοι: Μονοκύτταρα περιφερικού αίματος (PBMC) απομονώθηκαν από 23 ασθενείς με πρωτοπαθές σύνδρομο Sjogren (πΣΣ) και 14 από την ομάδα ελέγχου και καλλιεργήθηκαν παρουσία ή απουσία μιτογόνου διέγερσης (ΡΗΑ) και σε διάφορες συγκεντρώσεις των πεπτιδίων. Τo PBMC υποβλήθηκε σε κυτταρική ανοσοφαινοτύπιση και κυτταρομετρία ροής μετά από επώαση με το πεπτίδιο. ELISA χρησιμοποιήθηκε για τη μέτρηση της IL-2, IL-4, IL-5, IL-10, ΤGF-β και IFN-γ σε υπερκείμενα από καλλιέργειες κυττάρων των PBMC ή καθαρές ομάδες κυττάρων. Αποτελέσματα: Δεν παρατηρήθηκε σημαντική διαφορά στον κυτταρικό πολλαπλασιασμό στο περιφερικό αίμα των υγιών είτε παρουσία είτε απουσία PHA ενώ ανεστάλη ο κυτταρικός πολλαπλασιασμός σε ασθενείς με πΣΣ παρουσία PHA. Το ποσοστό των CD4 + CD25 + Τ λεμφοκυττάρων στο περιφερικό αίμα αυξήθηκε σε ασθενείς με πΣΣ σε σύγκριση με τους υγιείς. Οι IL-4, IL-2, ΙFΝ-γ και IL-5 δεν ανιχνεύθηκαν στα υπερκείμενα των υγιών και ασθενών, είτε τα κύτταρα καλλιεργήθηκαν μόνα ή υπό την παρουσία των πεπτιδίων. Η L-10 και ο TGF-β, έδειξαν μια διαφοροποίηση στην έκκριση τους σε σχέση με τους υγιείς. Συμπεράσματα: Το αυξημένο ποσοστό των CD4+CD25+ Τ κυττάρων δείχνει την ύπαρξη ανοσορυθμιστικού μηχανισμού που είναι παρόν στο ΣΣ, ενώ η αύξηση της IL-10 μπορεί τελικά να συμβάλλει στην παθογένεια της νόσου. / Sjögren’s syndrome (SS) is a common autoimmune disease mainly of salivary and lachrymal glands, leading το xerostomia and keratoconjunctivitis sicca. Histopathological findings in SS include focal lymphocytic infiltrates that contain T cells and B cells with a predominance of activated CD4+ helper T cells. The exact pathogenetic mechanisms responsible for SS are not yet fully known, however environmental, hormonal and genetic factors may be involved in SS pathogenesis. Dysregulation of several cytokines produced by the infiltrating Th1, Th2 and also Th17 cells have also been implicated in the development and progression of SS. Purpose: In this study, we analyzed the cytokine expression patterns of peripheral blood T-cell subsets in primary SS (pSS) patients using synthetic peptides that map to T-cell epitopes of the La/SSB autoantigen. Methods: Peripheral Blood Mononuclear Cells (PBMC) were isolated from 23 patients with pSS and 14 controls and were cultured in the presence or absence of mitogenic stimulation (PHA) and various concentrations of the peptides. PBMC were subjected to cell phenotyping and FACS analysis after incubation with peptide. ELISA was used to measure IL-2, IL-4, Il-5, Il-10, TGF-b and IFN-γ cytokine production in culture supernatants from cell culture of PBMC or purified cell groups. Results: No significant difference in cellular proliferation was observed in isolated PBMCs from healthy donors in the presence or absence PHA, whereas an inhibition of cellular proliferation was noticed in the presence of PHA in pSS patients. The percentage of CD4+CD25+ T lymphocytes in peripheral blood (PB) was increased in pSS patients compared with controls. IL-4, IL-2, IFN-γ and IL-5 were not detected in the supernatants of both controls and patients, whether the cells were cultured alone or in the presence of the peptides. IL-10 and TGF-b showed a differentiation in their secretion from controls. Conclusion: The increased percentage of the CD4+CD25+ T cells, shows that an enhanced immunoregulatory mechanism is present in SS and increased production of IL-10 in SS patients may contribute to the pathogenesis of the disease.