Group A streptococcal disorders can result from autoantibodies generated against M proteins. These autoantibodies cross react with the basal ganglia resulting in movement disorders. Previously, we demonstrated binding of streptococcal mAb10F5, with CPu and phospholipids. To determine if mAb10F5 binding to basal ganglia and phospholipids is due to virulence of the antibody or antibody subtype, rats were injected with control IgG2 antibodies and euthanized after 24, 48, or 72 hours. Brains were harvested and immunofluorescence was used to analyze brain slices. Control IgG2 rats showed significantly less fluorescence in the CPu than mAb10F5 injected rats at every time point. These findings reaffirm 10F5 is an anti-basal ganglia antibody. To evaluate mechanism of antibody entry, mAb10F5 was examined for anti-phospholipid activity. MAb10F5 displayed greater affinity to phospholipids when compared to IgG2 controls. Our findings support mAb10F5 is an anti-basal ganglia and anti-phospholipid antibody due to its own virulence. / Access to thesis permanently restricted to Ball State community only / Department of Physiology and Health Science

Autoantibodies generated against M proteins can cause post-streptococcal disorders such as Rheumatic Fever. A severe complication of rheumatic fever is rheumatic heart disease which may involve both cardiomyopathy and valvulitis. Rheumatic fever has been associated with the class I M protein epitope of Group A streptococcus (GAS). This epitope can be recognized by monoclonal antibodies (mAbs) 10B6 and 10F5. Previously, we demonstrated binding of streptococcal mAb10F5 in the heart tissue (apex, atria, and valves) of Lewis rats as compared to anti-myosin binding. To determine if mAb10F5 binding in the heart is due to virulence of the antibody or antibody subtype, rats were injected with control IgG2 antibodies and euthanized after 24, 48, or 72 hrs. Hearts were harvested and immunofluorescence was used to analyze the hearts. The immunofluorescence intensities for IgG2b were compared to mAb10F5 using previously acquired data. Control IgG2b rats showed significantly less immunofluorescence intensities in the heart regions than mAb10F5 injected rats at the 48 and 72 hr time points. These findings reaffirm mAb10F5 as an anti-cardiac antibody thatbinds heart tissue due its own virulence. To differentiate between the two IgG subtypes, binding intensities of IgG2a were compared to the binding intensities of IgG2b. The binding intensities of IgG2a increased with time. This finding was supported by previous work in our laboratory suggesting IgG2a remained in the bloodstream longer than the IgG2b. / Access to thesis permanently restricted to Ball State community only. / Department of Physiology and Health Science

Thesis (Ph.D.)--University of Wollongong, School of Biological Sciences. / Typescript. Includes bibliographical references: leaf 269-331.

Abstract Recurrent pharyngitis causes doctor visits, antibiotics use and absences from school or work and thus worsens patients’ quality of life (QOL). Even though tonsillectomy is often performed for recurrent pharyngitis, there is limited evidence of the tonsillectomy benefit concerning both researcher- and patient-recorded outcomes. The intent of this work was to find out if tonsillectomy reduces numbers of pharyngitis episodes or symptom days, if tonsillectomy improves patients’ QOL and if there are any clinical factors predicting QOL benefit after tonsillectomy. Seventy adult patients with recurrent streptococcal pharyngitis (2001–2005) and 86 patients with recurrent pharyngitis of any origin (2007–2010) were enrolled for two randomised controlled trials. Patients with recurrent pharyngitis of any origin were followed up either before (control group, n=40) or after (tonsillectomy group, n=46) tonsillectomy. At five months of follow-up, 17 (43%) patients in the control group and 2 (4%) patients in the tonsillectomy group consulted a physician for pharyngitis. Thirty-two (80%) patients in the control group and 18 (39%) patients in the tonsillectomy group experienced any kind of pharyngitis episode. Only one episode was considered severe. The numbers of days with throat pain and fever were significantly lower in the tonsillectomy group. QOL of 142 responders measured by Glasgow Benefit Inventory (GBI) six months after tonsillectomy showed improvement: median GBI total score was +27. However, GBI total scores varied considerably between the patients (range −19 to +69). Only one patient reported declined QOL. The number of prior pharyngitis episodes, frequent throat pain, untreated dental caries and chronically infected tonsils were the best clinical factors predicting QOL improvement. The precision of these predictions was still quite low. The results of this work suggest that tonsillectomy reduces numbers of acute pharyngitis episodes and symptoms. Although most of the episodes are not severe, tonsillectomy still generally improves patients’ QOL. The distribution of QOL benefit is broad, however. Throat-related morbidity before tonsillectomy is the only clinical factor that was associated with patient satisfaction. / Tiivistelmä Toistuvat nielutulehdukset aiheuttavat paljon lääkärikäyntejä, antibioottihoitoja sekä poissaoloja töistä tai opinnoista ja huonontavat potilaiden elämänlaatua. Toistuvien nielutulehdusten vuoksi päädytään usein nielurisaleikkaukseen, vaikka tutkimusnäyttö leikkauksen hyödystä on vähäistä. Tämän väitöskirjatyön tavoitteena oli tutkia, vähentääkö nielurisaleikkaus nielutulehdusten määrää tai oireita sekä selvittää leikkauksenjälkeistä elämänlaatua ja siihen liittyviä ennustekijöitä. Tutkimusaineisto koostui kahta eri satunnaistettua kliinistä koetta varten rekrytoiduista potilaista: 70 potilasta, joiden toistuvien nielutulehdusten aiheuttaja oli A-ryhmän streptokokki (2001–2005) ja 86 potilasta, joiden toistuvien nielutulehdusten etiologialle ei asetettu vaatimuksia (2007–2010). Potilaat, joilla nielutulehdusten etiologia oli avoin, satunnaistettiin kahteen ryhmään: kontrolliryhmää (n=40) seurattiin ennen nielurisaleikkausta ja leikkausryhmää (n=46) sen jälkeen, molempia 5 kuukauden ajan. Seurannassa 17 (43 %) kontrolliryhmän potilasta ja 2 (4 %) leikkausryhmän potilasta hakeutui lääkäriin nielutulehduksen vuoksi. Kontrolliryhmän potilaista 32 (80 %) ja leikkausryhmän potilaista 18 (39 %) sairasti nielutulehduksen vähintään kerran. Vain yksi episodi luokiteltiin vaikeaksi. Nielukipu- ja kuumepäiviä oli merkittävästi vähemmän leikkausryhmässä. Nielurisaleikkauksen vaikutusta elämänlaatuun tutkittiin Glasgow Benefit Inventory (GBI) -kyselyllä kuusi kuukautta leikkauksen jälkeen. Yhteensä 142 potilasta vastasi kyselyyn. GBI:n mediaanitulos +27 osoitti leikkauksen parantavan elämänlaatua. GBI-tulokset kuitenkin vaihtelivat huomattavasti potilaiden välillä (−19 – +67), vaikkakin vain yksi potilas raportoi elämänlaatunsa heikentyneen. Aiempien nielutulehdusten määrä, usein toistuva nielukipu, hoitamaton karies ja kroonisesti tulehtuneet nielurisat ennustivat parhaiten potilastyytyväisyyttä leikkauksen jälkeen, mutta näidenkin tekijöiden ennustearvo oli melko heikko. Tulosten perusteella nielurisaleikkaus vähentää akuutteja nielutulehduksia sekä oirepäiviä. Vaikka sairastamisjaksot ovat harvoin vaikeaoireisia, leikkaus parantaa useimmiten elämänlaatua, mutta hyödyn määrä vaihtelee merkittävästi potilaiden välillä. Ainoastaan leikkausta edeltävä nielun oireilun määrä ennustaa leikkaushyötyä jossain määrin.

Pneumonia is considered to be one of the leading causes of death worldwide. The outcome depends on both, proper antibiotic treatment and the effectivity of the immune response of the host. However, due to the complexity of the immunologic cascade initiated during infection, the latter cannot be predicted easily. We construct a biomathematical model of the murine immune response during infection with pneumococcus aiming at predicting the outcome of antibiotic treatment. The model consists of a number of non-linear ordinary differential equations describing dynamics of pneumococcal population, the inflammatory cytokine IL-6, neutrophils and macrophages fighting the infection and destruction of alveolar tissue due to pneumococcus. Equations were derived by translating known biological mechanisms and assuming certain response kinetics. Antibiotic therapy is modelled by a transient depletion of bacteria. Unknown model parameters were determined by fitting the predictions of the model to data sets derived from mice experiments of pneumococcal lung infection with and without antibiotic treatment. Time series of pneumococcal population, debris, neutrophils, activated epithelial cells, macrophages, monocytes and IL-6 serum concentrations were available for this purpose. The antibiotics Ampicillin and Moxifloxacin were considered. Parameter fittings resulted in a good agreement of model and data for all experimental scenarios. Identifiability of parameters is also estimated. The model can be used to predict the performance of alternative schedules of antibiotic treatment. We conclude that we established a biomathematical model of pneumococcal lung infection in mice allowing predictions regarding the outcome of different schedules of antibiotic treatment. We aim at translating the model to the human situation in the near future.

In an attempt to more fully understand the generation of antibody diversity to carbohydrate antigens, we produced and characterized a panel of hybridoma cell lines specific for group A streptococcal carbohydrate from mice injected with the intact bacteria (minus the hyaluronic acid capsule and cell wall protein antigens). We have analyzed the use of heavy and light chain variable region genes in the early (day 7) and late response (hyperimmune) and have determined the nucleotide sequence of the dominant VH gene used in several of our hybridomas. Our data allowed us to assess the extent to which the recombination of various V, D, and J gene segments and somatic mutation contribute to antibody diversification in this system. In this report we confirm that a minimum of two VH and four VK gene segments are used to encode this response. We extend this analysis to show that multiple D and J gene segments are used and that a significant amount of junctional variability is tolerated in CDR 3. Our results also suggest that there is a positive selection for somatic mutation in CDR 1 during the hyperimmune response to group A streptococcal carbohydrate.

Thesis (Ph.D.)--University of Melbourne, The School of Population Health and Department of Medicine, 2007. / Typescript. Includes bibliographical references (leaves 174-186).

Garcinia kola is one of the plants used in folklore remedies for the treatment of microbial infections. Bacterial resistance to commonly used antibiotics has necessitated the search for newer and alternative compounds for the treatment of drug resistant microbial infections. This study focuses on the bioactivity of G. kola seeds on Streptococcus pyogenes (ATCC 49399), Staphylococcus aureus (NCTC 6571), Plesiomonas Shigelloides (ATCC 51903) and Salmonella typhimurium (ATCC 13311), organisms which can cause illnesses from mild to severe with potentially fatal outcomes. The crude ethyl acetate, ethanol, methanol, acetone and aqueous extracts were screened by agar-well diffusion method and the activities of the extract were further determined by Minimum Inhibitory Concentration (MIC) and Minimum Bactericidal Concentration (MBC) assays. The inhibition zones ranged from 0 - 24 mm, while MIC and MBC of the extract ranged between 0.04 - 1.25 mg/mL and 0.081 - 2.5 mg/mL respectively. Chloroform/ Ethyl Acetate/ Formic acid (CEF) solvent system separated more active compounds followed by Ethyl Acetate/ Methanol/ Water (EMW) and Benzene/ Ethanol/ Ammonium Hydroxide (BEA). The extracts were fractionated by Thin Layer Chromatography (TLC). Bioautography was used to assess the activity of the possible classes of compounds present in the more active extracts. Column chromatography was used to purify the active compounds from the mixture while Gas Chromatography-Mass Spectrometry (GC-MS) was used to identify the phyto components of the fractions. The MIC of the fractions ranged between 0.0006 - 2.5 mg/mL. CEF 3 (F3), CEF 11 (F11) and CEF 12 (F12) revealed the presence of high levels fatty acids Linoleic acid, 1, 2-Benzenedicarboxylic acid and 2, 3-Dihydro-3, 5-dihydroxy-6-methyl, respectively. The results obtained from this study justify the use of this plant in traditional medicine and provide leads which could be further exploited for the development of new and potent antimicrobials.

INTRODUÇÃO: A Faringotonsilite é doença comum nos consultórios e prontosocorros de pediatria. OBJETIVOS: Avaliar o impacto da realização rotineira da prova rápida para pesquisa de estreptococo do grupo A (PRE) no diagnóstico e tratamento da faringotonsilite aguda em crianças e adolescentes atendidos em um Hospital Geral. MÉTODOS: Trata-se de um estudo prospectivo, observacional, de protocolo de atendimento, instituído no Pronto-Socorro do Hospital Universitário da Universidade de São Paulo para o atendimento de crianças e adolescentes com diagnóstico de faringotonsilite aguda. RESULTADOS: Foram estudadas 1039 crianças e adolescentes. Com base no quadro clínico, antibiótico seria prescrito em 530 pacientes (51%), e com o uso da PRE e/ou cultura de orofaringe foi prescrito em 268 (25,8%) pacientes. Das 509 crianças que não receberiam antibiótico pelo quadro clínico, 157 tiveram PRE e/ou cultura de orofaringe positiva. O diagnóstico baseado no quadro clínico apresentou sensibilidade de 63,06% (IC-95%:62,95-63,17%); especificidade de 57,33% (IC-95%:57,25-57,41%); valor preditivo positivo de 50,57% (IC-95%:50,47-50,66%) e valor preditivo negativo de 69,16% (IC-95%: 50,47-50,66%). CONCLUSÕES: Neste estudo o diagnóstico clínico da faringotonsilite estreptocócica mostrou baixa sensibilidade e especificidade. O uso rotineiro da prova rápida para pesquisa de estreptococo permitiu uma redução do uso de antibiótico e a identificação de crianças e adolescentes com faringotonsilite estreptocócicas que não receberiam antibiótico e estariam sob o risco das complicações da infecção estreptocócica / BACKGROUND: Sore throat is a common disease in the pediatric emergency room. OBJECTIVES: The objective of this study was to evaluate the impact of routine performance of rapid antigen detection test (RADT) in the diagnosis and treatment of acute pharyngitis in children treated at an academic hospital. METHODS: This is a prospective, observational, protocol compliance, established at the Emergency of Hospital Universitário - Universidade de São Paulo for the care of children and adolescents diagnosed with acute pharyngitis. RESULTS: We studied 1039 children and adolescents. Based on clinical findings, antibiotic would be prescribed in 530 patients (51%) and using the RADT or sore throat culture was prescribed in 268 patients. Of the 509 children who did not receive antibiotics for the clinical, 157 had positive RADT or sore throat culture. The diagnosis based on clinical sensitivity was 63,06% (IC 95% 62,95- 63,17%), specificity 57,3% (IC 95% 57,25-57,41%), positive predictive value of 50,57% (IC 95% 50,47-50,66%) and negative predictive value of 69,16% (IC 95% 50,47-50,66%). CONCLUSIONS: In this study the clinical diagnosis of streptococcal pharyngitis had low sensitivity and specificity. The routine use of rapid test for streptococcal research led to a reduction of antibiotic use and the identification of a risk group for complication of streptococcal infection

INTRODUÇÃO: A Faringotonsilite é doença comum nos consultórios e prontosocorros de pediatria. OBJETIVOS: Avaliar o impacto da realização rotineira da prova rápida para pesquisa de estreptococo do grupo A (PRE) no diagnóstico e tratamento da faringotonsilite aguda em crianças e adolescentes atendidos em um Hospital Geral. MÉTODOS: Trata-se de um estudo prospectivo, observacional, de protocolo de atendimento, instituído no Pronto-Socorro do Hospital Universitário da Universidade de São Paulo para o atendimento de crianças e adolescentes com diagnóstico de faringotonsilite aguda. RESULTADOS: Foram estudadas 1039 crianças e adolescentes. Com base no quadro clínico, antibiótico seria prescrito em 530 pacientes (51%), e com o uso da PRE e/ou cultura de orofaringe foi prescrito em 268 (25,8%) pacientes. Das 509 crianças que não receberiam antibiótico pelo quadro clínico, 157 tiveram PRE e/ou cultura de orofaringe positiva. O diagnóstico baseado no quadro clínico apresentou sensibilidade de 63,06% (IC-95%:62,95-63,17%); especificidade de 57,33% (IC-95%:57,25-57,41%); valor preditivo positivo de 50,57% (IC-95%:50,47-50,66%) e valor preditivo negativo de 69,16% (IC-95%: 50,47-50,66%). CONCLUSÕES: Neste estudo o diagnóstico clínico da faringotonsilite estreptocócica mostrou baixa sensibilidade e especificidade. O uso rotineiro da prova rápida para pesquisa de estreptococo permitiu uma redução do uso de antibiótico e a identificação de crianças e adolescentes com faringotonsilite estreptocócicas que não receberiam antibiótico e estariam sob o risco das complicações da infecção estreptocócica / BACKGROUND: Sore throat is a common disease in the pediatric emergency room. OBJECTIVES: The objective of this study was to evaluate the impact of routine performance of rapid antigen detection test (RADT) in the diagnosis and treatment of acute pharyngitis in children treated at an academic hospital. METHODS: This is a prospective, observational, protocol compliance, established at the Emergency of Hospital Universitário - Universidade de São Paulo for the care of children and adolescents diagnosed with acute pharyngitis. RESULTS: We studied 1039 children and adolescents. Based on clinical findings, antibiotic would be prescribed in 530 patients (51%) and using the RADT or sore throat culture was prescribed in 268 patients. Of the 509 children who did not receive antibiotics for the clinical, 157 had positive RADT or sore throat culture. The diagnosis based on clinical sensitivity was 63,06% (IC 95% 62,95- 63,17%), specificity 57,3% (IC 95% 57,25-57,41%), positive predictive value of 50,57% (IC 95% 50,47-50,66%) and negative predictive value of 69,16% (IC 95% 50,47-50,66%). CONCLUSIONS: In this study the clinical diagnosis of streptococcal pharyngitis had low sensitivity and specificity. The routine use of rapid test for streptococcal research led to a reduction of antibiotic use and the identification of a risk group for complication of streptococcal infection