Typescript (photocopy). / Digitized by Kansas Correctional Industries

Thesis (MSc)--University of Stellenbosch, 2003. / ENGLISH ABSTRACT: The effect of the bacteriocin-like peptide AS-48, produced by Enterococcus faecalis FAIRE 92, was tested against a mastitis isolate of Staphylococcus aureus in an in vivo and in vitro study. During initial tests peptide AS-48 showed no significant activity towards S. aureus, even with a ten-fold concentrated cell-free supernatant. Activity was obtained only after purification with Triton X-114 phase partitioning, followed by cation exchange chromatography. Titers for the purified peptide varied between 3200 and 12800 AU/ml. The purified peptide also exhibited activity towards Streptococcus agalactiae and Streptococcus dysgalactiae, but not against Escherichia coli. The size of peptide AS-48 was determined at 7150 Da, based on electronspray mass spectrometry and SDS-PAGE. Complete inhibition of cell growth was obtained by adding 1ml of the purified peptide (3200 AU/ml) to 100 ml of cells of S. aureus in the lag growth phase. When the same concentration of peptide AS-48 was added to a culture of S. aureus in mid-exponential growth, a slight decrease in viable cell numbers was recorded, which lasted for only 30 min. Cell growth commenced thereafter. In situ experiments in cows were done with purified peptide AS-48, encapsulated in liposomes. These in vivo studies were conducted by administering peptide AS-48 (6400 AU/ml) to different udder quarters. In a prevention trial, i.e. where quarters were pretreated with peptide AS-48, a reduction close to 90% in the viable cell numbers of S. aureus was recorded relative to the control quarters, which were not treated with the peptide. A 50% reduction in somatic cell count (SCC) was recorded. In the treatment trial, i.e. infected quarters treated with peptide AS-48, a reduction of up to 94% in viable cell numbers of S. aureus was recorded. In the same quarters, a reduction in SCC amounted to almost 80%. A recombinant strain was constructed by conjugating plasmid 92 (p92), encoding peptide AS-48, from Enterococcus faecalis FAIRE 92 to E. faecalis FA2/Ent, which produces enterocins 1071A and 1071B. Southern blot hybridization experiments revealed thepresence of plasmid p92 in the recipient strain without the loss of plasmid pEF1071, which encodes enterocins 1071A and 1071B. All three antimicrobial peptides, i.e. enterocin 1071A, enterocin 1071B and peptide AS-48, were produced in transconjugant FA2/Ent/AS-48. The spectrum of antimicrobial activity of the transconjugant was greater than that recorded for strains FA2/Ent and FAIRE 92, respectively and included E. faecalis, Bacillus cereus, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus curvatus, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus sakei, Leuconostoc cremoris, Leuconostoc pentosaceus, Staphylococcus carnosus and S. aureus. These organisms are not inhibited by strain FA2/Ent. However, low levels of peptide AS-48 was produced by strain FA2/Ent/AS-48. Further research in fermentation and gene expression will be needed before the transconjugant E. faecalis FA2/Ent/AS-48 may be used in the treatment of mastitis. / AFRIKAANSE OPSOMMING: Die effek van die bakteriosien-agtige, peptied AS-48, geproduseer deur Enterococcus faecalis FAIRE 92, is gedurende ‘n in vivo en in vitro studie teen ‘n mastitiese Staphylococcus aureus-isolaat getoets. Aanvanklike toetse met peptied AS-48, selfs tienvoudig gekonsentreerde selvrye supernatant, het geen beduidende aktiwiteit teen S. aureus getoon nie. Aktiwiteit is eers verkry na suiwering met Triton X-114 fase-skeiding gevolg deur katioon uitruilingschromatografie. Titers vir die gesuiwerde peptied het tussen 3200 en 12800 AE/ml gewissel. Die gesuiwerde peptied het ook aktiwiteit teen Streptococcus agalactiae en Streptococcus dysgalctiae getoon, maar nie teen Escherichia coli nie. Peptied AS-48 het ‘n molekulêre massa van 7150 Da, soos bepaal met elektronsproeimassa spektrometrie en SDS-PAGE. Totale inhibisie van selgroei is verkry deur 1 ml gesuiwerde peptied AS-48 (3200 AE/ml) by ‘n 100 ml kultuur van S. aureus in die sloerfase te voeg. Dieselfe konsentrasie peptied AS-48, toegevoeg tydens die mideksponensiële groeifase, het egter slegs ‘n klein vermindering in die aantal lewende selle teweeg gebring en het ook vir slegs ‘n 30 min geduur. Selgroei het hierna weer normaal voort gegaan. In situ eksperimente op koeie is uitgevoer met gesuiwerde peptied AS-48, geenkapsuleerd in liposome. Hierdie In vivo studies is onderneem deur peptied AS-48 (6400 AE/ml) in verskillende kwarte van die uier, kunsmatig of reeds geïnfekteerd met S. aureus, toe te dien. In ‘n voorkomings-eksperiment waar kwarte vooraf met peptied AS- 48 behandel is, is ‘n verlaging van byna 90% in die lewende seltelling van S. aureus relatief tot die kontrole kwarte, sonder behandeling met peptied AS-48, verkry. ‘n 50% verlaging in die somatiese seltelling (SST) is verkry. In die behandelings-eksperiment, waar geïnfekteerde kwarte met peptied AS-48 behandel is, is ‘n verlaging van byna 90% in lewende S. aureus selle gevind. In dieselfde kwarte is ‘n verlaging van byna 80% in die SST genoteer.‘n Rekombinante ras is gekonstrueer deur plasmied 92 (p92), wat kodeer vir peptied AS- 48, vanaf Enterococcus faecalis FAIRE 92 na E. faecalis FA2/Ent, wat enterosien 1071A en 1071B produseer, te konjugeer. Southern-klad hibridisasie het die teenwoordigheid van plasmied p92 in die ontvanger ras, sonder die verlies van plasmied pEF1071 wat enterosien 1071A en 1071B kodeer, getoon. Al drie antimikrobiese peptiede, nl. enterosien 1071A, enterosien 1071B en peptied AS-48, is deur die transkonjugant FA2/Ent/AS-48 geproduseer. Die spektum van antimikrobiese aktiwiteit van die transkonjugant vand die transkonjugant is breër as dié van rasse FA2/Ent en FAIRE 92, onderskeidelik en het ook E. faecalis, Bacillus cereus, Lactobacillus acidophilus, Lactobacillus casei, Lactobacillus curvatus, Lactobacillus fermentum, Lactobacillus plantarum, Lactobacillus reuteri, Lactobacillus sakei, Leuconostoc cremoris, Leuconostoc pentosaceus, Staphylococcus carnosus en S. aureus ingesluit. Hierdie organismes word nie deur ras FA2/Ent geïnhibeer nie. Lae vlakke van peptied AS-48 is egter deur ras FA2/Ent/AS-48 geproduseer. Verdere navorsing in fermentasie en geenuitdrukking is nodig voordat E. faecalis FA2/Ent/AS-48 in die behandeling van mastitis gebruik kan word.

Thesis (MCurr)--Stellenbosch University, 2014. / ENGLISH ABSTRACT: The aim of this study was to compare the impact of a four-day structured Basic Infection Prevention and Control course on the knowledge of, and adherence to, Standard Precautions in clinical nursing practice amongst nurses who had completed the course and those who did not. The specific precautionary measures of investigation included hand hygiene, personal protective equipment (PPE) and sharps management. The secondary aim of the study was to identify any personal and contextual factors that influenced the application of such Standard Precautions measures in public healthcare facilities within the Cape Winelands and Overberg District. Sixty eight students (those who had been trained) with a similar number of controls (who had not been trained) were enrolled in the study. Although both the participants and controls had the knowledge, their adherence to hand hygiene, PPE and sharps management in clinical nursing practice was poor. Staff attitude was found to be the main factor for non-adherence. The knowledge of the participants was good as they had answered most of the questions correctly. It seems as if there was retention of knowledge after the four-day Basic Infection Prevention and Control course. There were, however, no significant differences between the two groups. For both groups attitude and behavioural change must be addressed in order to improve adherence to hand hygiene, PPE and sharps management. The findings of the study will form recommendations towards improved infection prevention and control practices at public healthcare facility level in the Cape Winelands District. / AFRIKAANSE OPSOMMING: Die doel van die studie was om die impak van 'n 4-dag gestruktureerde Basiese Infeksiebeheerkursus op die kennis en toepassing van Standaard Voorsorgmaatreëls in kliniese praktyk in die Kaapse Wynland en Overberg Distrikte ondersoek, vergeleke met 'n groep wat nie die kursus bygewoon het nie. Die spesifieke Voorsorgmaatreëls wat ondersoek is, het handhigiëne, die gebruik van beskermende drag en die hantering en beheer van skerpvoorwerpe ingesluit. Die studie het ook gekyk na enige kontekstuele en persoonlike faktore wat die toepassing van Standaard Voorsorgmaatreëls in openbare gesondheidsorgfasiliteite beïnvloed. Agt en sestig verpleegkundiges het die 4-dag Basiese Infeksiebeheerkursus bygewoon en 'n gelyke aantal kontrole studente het nie die kursus bygewoon nie. Alhoewel beide groepe die kennis van handhigiëne, die dra van beskermende drag en die hantering van skerpvoorwerpe gehad het, was die toepassing van die Standaard Voorsorgmaatreëls in kliniese praktyk baie swak. Personeel se houding was die grootste faktor wat gelei het tot die nie-toepassing van Standaard Voorsorgmaatreëls. Die kennis van die kursusgangers was goed, want albei groepe het die meeste van die vrae korrek beantwoord. Die waarneming wat gemaak is, is dat die kursusgangers se kennis wel verbeter het na die bywoning van die 4-dag Basiese Infeksiebeheerkursus. Data weerspieël egter geen noemenswaardige verskille tussen die groepe nie. Beide groepe se houding en gedrag moet aangespreek word om die toepassing van handhigiëne, die dra van beskermende drag en die hantering van skerpvoorwerpe te verbeter. Die bevindinge van die studie sal gebruik word om aanbevelings te maak ten einde infeksiebeheerpraktyke in die Kaapse Wynland Distrik te verbeter.

Spores of foodborne pathogens such as Clostridium botulinum, Clostridium perfringens and Bacillus cereus are widely distributed in nature. Presence of those spores in food products, particularly C. botulinum spores in vacuum packed, ready-to-eat low-acid products, is a great safety concern. The research here described is a first effort towards understanding the role of the spore coat proteins in the inactivation of bacterial spore using high pressure processing. This study proposes a coat protein solubilization methodology using non-ionic detergents minimizing protein damage and compatible with spectroscopy methods. The methodology developed here was compared with approaches proposed in the literature with respect to protein yield, protein fractions identified, amino acid composition and suitability with spectroscopy techniques for the further analysis of coat proteins. Bacillus subtilis ATCC 6633 spore coat proteins were solubilized (n=3) using octyl-β-D-glucopyranoside (OGP) at room temperature and urea/sodium dodecyl sulphate (UDS) at 37C and 70C. Analysis of variance (ANOVA) showed no significant (95% confidence) differences between the three repetitions of the three spore coat protein solubilization methods. Protein yield was significantly larger (95% confidence) when using UDS at 70C as compared to UDS at 37C. OGP gave the lowest protein yield but allowed circular dichroism (CD) analysis of the spore coat protein solution with minimum blank signal. SDS-PAGE revealed that the UDS-70C coat protein solutions consisted of five major and six minor proteins ranging 6 to 65 kD while the OGP solution appear to consist of four major and nine minor bands in the same mw range. Amino acid analysis of the protein extracted by the OGP method was conducted using reverse phase HPLC (RP-HPLC) and compared with published information. The OGP spore coat protein solution showed a higher proportion of aspartate, glutamate, alanine and tyrosine. Pressure, heat and time effects were studied on spore coat proteins obtained from untreated and pressure-treated B. subtilis ATCC 6633 spores. Pressure treatments of spores, and of extracted spore coat protein solutions, at 50 kpsi (345 mPa) and 85 kpsi (586 mPa) for 10 and 30 min at constant 85C along with appropriate heat- and pressure-only controls and untreated sample, were used to study the effect of pressure, heat and time on spore coat proteins. Both spore coat protein solubilization procedures showed a significant reduction in protein yield for pressure-only, heat-only and pressure/heat treated spores when compared with untreated spores. When OGP-solubilized proteins from untreated spores were pressure treated, SDS-PAGE profile showed an increasing overall band intensity with increasing pressure and time. In the case of protein solution obtained from pressure-treated spores the electrophoretic pattern showed the loss of higher molecular weight proteins. The significance of this study is that for the first time we have observed extensive changes on spore coat proteins caused by pressure, as well as heat treatments. Future studies will examine what is the probable physiological role of the proteins damaged by these physical treatments. An advantage of the protein solubilization here developed will allow the application of spectroscopy techniques to characterize changes in spore coat proteins. / Graduation date: 2003

A quantitative competitive binding "triple sandwich" enzyme immunoassay was used to evaluate pathogen/class-specific antibody responses in Holstein-Friesian calves vaccinated against Clostridium perfringens B-toxin at various ages postpartum. Vaccination of dams at six weeks and again at two weeks prepartum increased pathogen-specific antibody levels in their colostrum and respective calf's serum. Calves initially vaccinated at three days produced both a primary and secondary pathogen-specific antibody response, whereas calves initially vaccinated at 12 and 21 days produced only secondary responses. Maternally-derived antibodies were found to suppress neonatal antibody production following primary immunization. They were also found to influence secondary humoral immune responses, although in a diminished capacity. Pathogen-specific IgG and IgM concentrations in dams' sera and colostra were found related to subsequent pathogen-specific IgG and IgM neonatal serum concentrations. Only pathogen-specific IgA in dams' colostra was correlated to neonatal levels, possibly owing to a different origin and role of this immunoglobulin class. All class-specific colostral immunoglobulin levels were related to subsequent neonatal concentrations. Based on results from this experiment, it is recommended that calves be vaccinated at three days postpartum with a booster administered at 63 days.

Thirty-one college age females enrolled in aerobic dance conditioning classes were subjects. The purposes were to determine if aerobic dance is sufficiently vigorous to elicit a heart rate capable of producing a training effect and to determine whether a training effect heart rate level can be maintained. The narco-biosystems telemetry was used to monitor heart rates. Each subject performed one aerobic dance at three trials. Twelve-minute run/walk pretests and post-tests were given to assign fitness levels. Results of the study indicate that aerobic dance is an effective cardiovascular training program. Resting heart rates decreased, fitness levels improved, and training intensity heart rates were elicited and maintained at each of the trials.

This dissertation discusses anti-HIV-1 microbicide research. In particular, it concentrates on microbicide formulation and delivery. Microbicides are anti-HIV-1 agents that when applied in the human vagina or rectum may prevent sexual HIV-1 transmission. Although most of the anti-HIV-1 agents being developed as microbicides are active in vitro, they have proved to be ineffective in vivo. A review of microbicide development over the last decade expounds the view that unsatisfactory microbicide failures may be a result of inefficient delivery systems employed. Thus, necessitating a thorough scientific qualitative and quantitative investigation of important aspects involved in HIV-1 transmission as a prerequisite for microbicide development. In this dissertation it is postulated that intravaginal targeting of HIV-1 increases the chances of microbicide success, wherein vaginal micro-environmental factors including pH would be maintained at HIV-1 prohibitive acidic levels to ward off other sexually transmitted diseases which compromise vaginal epithelial barrier properties. Furthermore, targeting early stages of the HIV-1 infection accompanied by computation and delivery of appropriate microbicide quantities could result in an effective microbicide formulation. In an effort to address microbicide formulation challenges, an intravaginal delivery system able to deliver anti-HIV-1 agents (zidovudine and BP36) over 28 days was formulated. This delivery system is a caplet-shaped composite system comprising zidovudine (AZT) and BP36-loaded pectin-mucin-polyethylene glycol submicrospheres embedded within a poly(D,L-lactide), magnesium stearate, polyvinyl acetate/polyvinylpyrolidone (Kollidon® SR) and poly(acrylic acid) based polymeric caplet matrix. The delivery system was tested in vitro and in vivo in the pig model. X-ray imaging illustrated the delivery system swelling and its matrix contrast fading over time as vaginal fluid permeated the matrix’s core. Plasma, vaginal fluid and tissue drug was detected and quantified using ultra performance liquid chromatography-tandem photodiode array detector. AZT plasma and vaginal fluid concentrations measured on days; 3, 7, 14, 21 and 28 decreased gradually with time. Vaginal tissue AZT concentrations (after 28 days) were higher than plasma AZT concentrations and were in the same range as vaginal fluid AZT concentrations. The herbal extract, BP36, was detected in plasma, vaginal fluid and tissue but was only qualitatively analysed due to its lack of standardization. Histopathological analysis of excised vaginal tissue revealed different scores of abnormalities comprising mild to moderate epithelial proliferation and exocytosis, subepithelial leukocyte influx, perivascular cell cuffing and isolated epithelial erosion, stromal fibrosis and isolated tissue necrosis.

Orientador: Lenice do Rosário de Souza / Coorientador: Hélio Amante Miot / Banca: Marli Terezinha Cassamassimo Duarte / Banca: Walter Belda Junior / Resumo: A infecção pelo HPV é a DST mais comum no mundo. Dos 630 milhões de casos novos anuais, trinta milhões são relacionados a verrugas anogenitais. Embora a forma subclínica seja mais comum, o aparecimento de condilomas também está associado à diminuição da imunidade causada pelo HIV. Tendo em vista as altas taxas de prevalência da coinfecção HPV/HIV, principalmente entre homens que fazem sexo com homens, os objetivos deste trabalho foram analisar a prevalência de verrugas anogenitais em homens portadores de HIV/aids e identificar fatores associados. Estudo transversal, descritivo e analítico com 159 homens atendidos em serviço de referência de Botucatu (SP). Dados sociodemográficos, comportamentais e clínicos foram relacionados ou não à presença de verrugas anogenitais. Após análise hierárquica dos dados, as variáveis com valor de p menor do que 0,2 foram incluídas no modelo logístico multivariado não condicional. Foram diagnosticados 49 (31,0%) pacientes HIV+ com verrugas anogenitais, cuja média de idade foi de 44,6 ± 9,6 anos. Os principais fatores associados às verrugas foram: escolaridade menor que ou igual a oito anos; maior tempo de diagnóstico do HIV; tratamento antirretroviral irregular; baixas contagens de linfócitos T CD4+. Verrugas anogenitais são prevalentes e relacionadas à imunossupressão da infecção pelo HIV. Ações como o cuidado integral do paciente e educação e prevenção em saúde colaboram para diagnóstico precoce e diminuição da vulnerabilidade a DSTs / Abstract: HPV infection is the most usual STD in the world responsible for 630 million new cases annually, of which 30 million are related with anogenital warts. Although subclinical form is the most common, warts are also associated with reduction of HV-related immunity. There are high rates involving prevalence of the association HIV/HPV around the world. Following this point of view the aim of this study was analyze the prevalence of wart in HIV-infected men and describe the associated factors. cared by a reference service in the Botucatu city - São Paulo/Brazil. Sociodemographic, behavioral and clinical data were collected by an individual interview and medical records. There were 49 (31,0%) HIV-infected men with anogenital warts with mean age of 44,6 ± 9,6 years. The main associated factors were: have up to 8 years of study; longer duration of HIV-infection; irregular treatment to HIV-infection; lower counts of lymphocytes T CD4+. Genital warts are prevalent and are associated with HIV- infection magnitude. Actions such as the complete care over the patients, increasing the education and prevention in health system could help to do a previously, diagnosis and decreasing the vulnerability to STD / Mestre

Thesis (MTech (Environmental Health))--Peninsula Technikon, Cape Town, 2004 / AIM &; OBJECTIVES: a) To quantify human vibration exposnres among the various similar occupational groups present on the mine, b) To determine the degree of vibration risk posed onto the mineworkers and c) To recommend and implement a sustainable human vibration management control progranune. METHODOLOGY: A cross-sectional descriptive study design was carried out on 135 mine workers employed in various similar occupational groups at Rossing Uranium mine, Namibia Data acquisition originated from a multi-stage proportionally stratified random sampling technique. An approved Human Vibration measuring instrument was utilized to measure Hand-arm and Wholebcdy vibration exposure levels [Aeq (m/s2 )] prevalent among the similar exposure groups. A structured questionnaire, developed specifically for the actual work environment enabled the collection of information such as work history, type of vibration exposure, exposure duration and vibration symptoms. Furthermore, group specific results [A(8)] were computed by means of the latest internationally accepted Health & Safety Executive Vibration calculators. In addition, statistical analyses were performed in order to establish the occupational groups that are at increased risk for the development of hand-arm and wholebody vibration induced health disorders.

The purpose of this study was to examine the effectiveness and exposure of an implementation strategy, which included a 4-day in-class training with two follow-up technical assistance calls, on mediating factors hypothesized to be positively associated with staff’s intention to use a five-session, couples-based HIV and other sexually transmitted prevention intervention. The Consolidated Framework for Implementation Research (CFIR) guided the study aims and analysis of the direct effect of exposure to the implementation strategy and 3 factors hypothesized to mediate the implementation strategies’ effect on intention to implement a couples-based intervention. Individual staff characteristics and an organizational process variable informed by Social Cognitive Theory (SCT), the Diffusion of Innovation Theory and Theory of Planned Action were examined. Two hundred and fifty-three staff, predominantly African American and Latina, from 80 organizations, were recruited from HIV service agencies, clinics and community-based organization from New York City and other regions of New York State. They were randomized by agency to either a multimedia condition or a traditional paper-based version of the couples-based intervention and received the implementation strategy 4-day, in-class intervention training followed by a technical assistance phone call at 3 and 6-months post training. Findings suggest that greater exposure to the implementation strategy in days and contacts was significantly associated with an increase in staff’s intention to implement the intervention at six months. Further, while a statistically significant effect of the implementation strategy dose on the mediators examined was not detected, the implementer’s experience of these mediators defined as self-efficacy for couples-based implementation, positive perception of the intervention’s characteristics and the perceived availability of an organizational intervention Champion was found to be significantly associated with the outcome variable intention to implement, and also was found to reduce the dosage effect of the implementation strategy on intention. Further examination of the implementation strategy’s content and dosage is needed to identify how increased intention to utilize an intervention at 6 months and 12 months following training and technical assistance may be enhanced through greater attention to and measurement of these mediators in addition to the implementation strategy dosage effect. Of note, the dosage effect on intention was found to diminish at the 12 month follow-up period suggesting the importance of timely support and planning prior to and post implementation strategies to increase utilization of an innovation. Implications for HIV prevention theory, and social work research, practice and policy are discussed.