Haematological stem cell transplantation (HSCT) is a revolutionary treatment for haematological malignancies. Although HSCT is potentially curative, patients usually develop stomatitis which is a common and debilitating complication after the transplantation. Furthermore, stomatitis may predispose patients to various complications which are associated with significantly increased morbidity and mortality. In some studies, ice water mouthwash has been shown to be an effective method for the prevention of stomatitis. However, a high-level evidence-based protocol on the prevention of stomatitis has not been fully developed and it is not commonly practiced in most HSCT centers at present. A well established protocol can help to minimize the patients’ suffering and avoid prolonged hospitalization. The nurses who are involved in patient education, assessment, care for, and coping with stomatitis, play an important role to bring these innovations into practice. In this regard, this translational research aims at developing an evidence-based protocol on using ice water mouthwash in the prevention of stomatitis for patients undergoing autologous HSCT. A systematic search for relevant literatures was performed with the use of five electronic databases. Six relevant studies were found. Critical appraisal on the relevant studies was conducted. The level of evidence extracted from the studies was graded according to the Scottish Intercollegiate Guidelines Network (SIGN) and were synthesized to establish the protocol for patients in the proposed setting. The implementation potential of the protocol was assessed in terms of the transferability, feasibility, and cost benefit ratio. An implementation and evaluation plan was established for comprehensive evidence-based protocol development. The successful implementation of the protocol will be beneficial for the patients undergoing HSCT as it may hasten their recovery, shorten their hospital stay, and minimize their distressing experience and suffering. / published_or_final_version / Nursing Studies / Master / Master of Nursing

published_or_final_version / Medicine / Master / Doctor of Medicine

M. Tech. / Stem cells are defined as undifferentiated cells that can proliferate indefinitely and have the capacity of both self-renewal and differentiation to one or more types of specialised cells. Traumatic tissue injury and age-related degenerative diseases are a major problem in South Africa and worldwide. Stem cells could be used for tissue engineering and reconstructive surgery. In treating these conditions, the main principle of stem cell therapy is the replacement of damaged and dead cells in injured tissues and organs with new healthy ones expanded in vitro from stem cells (Orlic et al., 2002). These cells can be isolated from adipose tissue in significant numbers and exhibit stable growth and proliferation kinetics in culture and could be differentiated into bone, fat, cartilage and muscle when treated with established lineage-specific factors (Zuk et al., 2002). Low Level Laser Therapy (LLLT) is currently applied in the treatment of numerous diseases and pathological conditions (Gasparyan et al., 2004). LLLT produces positive effects on irradiated cells and tissues such as proliferation of cells, capillary growth and adenosine triphosphate (ATP) activation (Schindl et al., 1998). Low level laser radiation at different intensities has been shown to stimulate as well as to inhibit cellular processes (Moore et al., 2005). Epidermal growth factor (EGF) is a growth factor that plays important roles in the regulation of cell growth, proliferation and differentiation. This study investigated the effect of low level laser radiation alone as well as in combination with EGF on adult adipose derived stem cells (ADSCs) isolated from human adipose tissue. ADSCs were isolated from human adipose tissue through collagenase digestion and cultured in DMEM-F12 containing 10% FBS and antibiotics and incubated at 37°C in a humidified atmosphere of 5% CO2 (Zuk et al., 2001). iii Semi-confluent monolayers of ADSCs were exposed to low level laser at 5 J/cm2 using 636 nm diode laser with a power density of 12.1 mW/cm2 at room temperature in the dark. Cell morphology was monitored at 0, 24 and 48 h using an inverted light inverted microscope. Cell viability was evaluated at 0, 24 and 48 h using the Trypan Blue exclusion test and an adenosine triphosphate (ATP) luminescence assay. bFGF (basic fibroblast growth factor) indirect ELISA and optical density assays were used to monitor cell proliferation at 0, 24 and 48 h post irradiation. In addition the expressions of stem cell markers, β1-integrin and Thy-1, were monitored by immunocytochemical live cell surface labelling and Western blot analysis. Cells were incubated with EGF to enhance proliferation and differentiation and the cell morphology, viability and proliferation were monitored as well as the expressions of stem cell markers, β1-integrin and Thy-1. Morphology of the cells was not altered by irradiating them with 5 J/cm2 using diode laser at 0, 24 and 48 h. Cell viability and proliferation showed an increase at 24 and 48 h post irradiation. At 0 h, there was no significant difference between irradiated and non-irradiated cells in cell viability and proliferation. There was an increase in the expression of β1-integrin and Thy-1 after irradiation as shown by Western blot analysis and immunocytochemical live cell surface labelling. Cell viability and proliferation showed a significant increase at all time points post irradiation with the addition of EGF. There was no noticeable change in cellular morphology at any time point. Low level laser irradiation of human ADSC’s at 636 nm with 5 J/cm2 and 12.1 mW/cm2 increased the viability and proliferation of these cells in vitro. Furthermore, low level laser irradiation appeared to increase the expression of stem cell markers, β1-integrin and Thy-1. In addition, laser irradiation did not alter the morphology of the cultured cells. The addition of EGF to the cells also increased their viability and proliferation as well the expression of the markers, β1-integrin and Thy-1. The study showed that laser irradiation stimulates two important cellular responses namely cell viability and proliferation which indicates that ADSCs may be suitable for tissue engineering and future cell differentiation studies.

M.Tech. / Stem cells possess self-renewal capacity, long-term viability, and multilineage potential. Stem cells play important roles in normal physiological and disease processes, they also have great therapeutic potential. However, there have been controversies surrounding stem cells in political, religious and ethical arenas. Although the use of certain stem cells (i.e. embryonic stem cells) and the means by which they are obtained contravene certain basic ethical laws, researchers have developed methods with which to ethically obtain and create stem cell lines. Stem cells can be classified as either: totipotent, pluripotent, multipotent, oligopotent and unipotent (Moore, 2007). Totipotent cells have the ability to differentiate into all cell types of an embryo, including the extra-embryonic and post embryonic tissues and organs. Pluripotent cells have the potential to differentiate into almost all tissues found in an embryo (including germ cells), but are not capable of giving rise to supporting cells and tissues. Multipotent stem cells have progeny of several differentiated cell types - but all within a particular tissue, organ, or physiological system. A good example of multipotent cells, are the haematopoietic stem cells that produce blood cell-restricted progenitors, as well as all cell types and elements, such as platelets, that are normal components of blood. Oligopotent stem cells produce two or more lineages within a specific tissue, such as neural stem cells that are able to produce subsets of neurons in the brain. Unipotent cells self-renew, as well as give rise to a single mature cell type, a prime example being the spermatogonial stem cells, that give rise to spermatozoa (Moore, 2007). Adult human subcutaneous adipose tissue contains cells with multilineage developmental plasticity like marrow-derived mesenchymal stem cells (Strem et al., 2005, Tong et al., 2000). Adipose derived stem cells can be obtained in abundance and can differentiate into osteogenic, adipogenic, myogenic and chondrogenic lineages when treated with appropriate growth factors.

Thesis (DScMedSc (Medical Sciences)--University of Stellenbosch, 2010. / Bibliography / ENGLISH ABSTRACT: See full text for abstract / AFRIKAANSE OPSOMMING: Geen opsomming was ingehandig met tesis

published_or_final_version / Anatomy / Doctoral / Doctor of Philosophy

published_or_final_version / Medicine / Master / Master of Philosophy

Orientador: Sigmar de Mello Rode / Coorientador: Lucio Murilo dos Santos / Banca: Antonio Olavo Cardoso Jorge / Banca: Fernando Callera / Resumo: Micro-organismos como fungos do gênero Candida, são habitantes comensais da cavidade bucal; em condições normais, co-existem com a microbiota normal sem provocar doenças. Entretanto, alterações locais ou sistêmicas como imunossupressão, desequilíbrio da microbiota oral, hipossalivação e mucosite, secundárias ao tratamento quimioterápico, predispõem pacientes com câncer a um alto risco de infecções fúngicas orais e sistêmicas. O objetivo desse trabalho foi avaliar a presença de leveduras do gênero Candida na cavidade bucal de pacientes onco-hematológicos submetidos a transplante autólogo de células-tronco hematopoiéticas (TACTH). Foram avaliados 27 pacientes, nos períodos pré, e pós-transplante. As amostras da cavidade bucal foram obtidas pela técnica de enxágüe bucal e semeadas em Chromagar Candida para triagem das cepas isoladas. Após o crescimento, foi extraído o DNA e submetido a identificação molecular (PCR) utilizando iniciadores para os genes ribossomais dessa levedura. Após a amplificação do fragmento esperado, as cepas foram sequenciadas utilizando sequenciador automático.Para análise descritiva e estatística dos resultados obtidos, os dados foram submetidos ao teste de Shapiro-Wilk para avaliação da normalidade. Em seguida, o teste de variância Wilcoxon foi utilizado. A significância adotada foi de 5%. Candida spp foi encontrada em 40,74% (11 pacientes), sendo que 2 (18,18%) possuíam mais de uma espécie, dos 9 pacientes colonizados por apenas uma espécie, 7 eram port... (Resumo completo, clicar acesso eletrônico abaixo) / Abstract: Microorganisms such as fungi of the genus Candida, are commensal inhabitants of the oral cavity; under normal conditions co-exist with normal microflora without causing disease. However, local or systemic changes such as immunosuppression, imbalance in the oral microbiota, hyposalivation and mucositis, secondary to chemotherapy, predispose cancer patients at high risk of oral and systemic fungal infections. The aim of this project is to evaluate the presence of Candida species in the oral cavity of hematological malignances patients undergoing autologous transplantation of hematopoietic stem cells (HSCT). We evaluated 27 patients in preand post-transplant. Samples of the oral cavity were obtained by oral rinse technique and plated on Chromagar Candida for screening of the strains. After growth, the DNA was extracted and subjected to molecular identification (PCR) using primers for the ribosomal genes in this yeast. After amplification of the expected fragment, the strains were sequenced using the sequencer automático. For descriptive and statistical analysis of the results, the data will be submitted to the Shapiro-Wilk test for evaluation of normality. Then the variance Wilcoxon test was used. The significance of 5% was adopted. Candida spp was found in 40.74% (11pacientes), and 2 (18.18%) had more than one species, of the 9 patients colonized by only one species, 7 patients had Candida albicans (77.77%) and 1 Candida dubliniensis (11.11%) and 1 C. krusei (11.11%). Mixed settlements, in the other two patients were composed of C. albicans + C. glabrata; C. albicans + C. dubliniensis. In the second collection (C2), 9 (81.81%) of Candida carriers remained colonized, including by non albicans species. The homology identification of species standardized strains was 85 99%. Within the conditions of this study it was possible to determine with ...(Resumo completo, clicar acesso eletrônico abaixo) / Mestre

published_or_final_version / abstract / Medicine / Doctoral / Doctor of Philosophy

published_or_final_version / Orthopaedics and Traumatology / Master / Master of Philosophy