O cloreto de trifeniltetrazólio (TTC) foi empregado neste trabalho visando padronização de método alternativo para o teste de limite microbiano em medicamentos e cosméticos, com redução no tempo de análise, além de possibilitar a aplicação da técnica de semeadura em profundidade em amostras hidroinsolúveis com opacidade e carga contaminante menor que 102 UFC/g (mL).Na técnica de tubos múltiplos, a aplicação do corante não possibilitou a redução do tempo de incubação, pois a sensibilidade para detecção de suspensão bacteriana por meio de coloração do trifenilformazano foi de 108 UFC/mL após 1 hora de reação e 107 UFC/mL após 4 horas, suspensões estas visivelmente turvas. A adição de glicose e ácido glutâmico no meio de caseina-sola, nas concentrações de 0,5% e 0,1%, respectivamente, não aumentou a sensibilidade da reação. O polissorbato 20, adicionado ao meio de cultura, líquido ou sólido, promoveu a difusão do trifenilformazano das células para o meio, fator que não favorece a visualização da massa celular no meio líquido e nem das colônias. A concentração mínima de TTC que resultou em coloração máxima no meio líquido foi de 500 e 1000 µg/mL, dependendo da espécie bacteriana. Entretanto, suspensões bacterianas com 102, 10 e 1 UFC/mL testadas, pela técnica de tubos multiplos, com adição do corante na forma de solução após 20 horas em meio de caseina-soja, forneceram resultados equivalentes, quanto ao número de tubos positivos e negativos frente às duas concentrações. A comparação desta técnica alternativa com a da subcultura demonstrou ser o tempo de 20 horas de incubação insuficientes para detecção pelo TTC, quando inoculadas alíquotas de 1 mL de suspensão com número da ordem de 1 UFC/mL Na técnica de semeadura em profundidade, após incubação de 48 horas, a coloração das colônias bacterianas foi efetuada cobrindo-se a superfície do meio de cultura com 5 mL de ágar a 1,0% contendo TTC. A concentração de 0,025% do corante promoveu a coloração das colônias após 1 hora. Entretanto, na presença de suspensão de hidróxido de alumínio a 6% a concentração mínima eficaz de TTC foi de 0,1%. Para leveduras em meio líquido essa concentração foi de 2.500 µ/mL no meio neutralizado. Em meio sólido, na presença de material opalescente, a concentração eficaz foi de 2,0%, com o TTC adicionado na dispersão de ágar em tampão pH 7,5. Amostras de produtos comercializados com característica opalescente foram submetidas à análise pelo método de tubos múltiplos com sub-cultura e pela revelação com TTC. Simultaneamente as mesmas foram testadas pelo método de semeadura em profundidade, inoculando-se 2 mL das diluições 1:4 e 1:10 com revelação das colônias com o corante. O tempo para obtenção dos resultados foi de 72 horas pelo método oficial e de 50 horas pelo método com TTC, tanto em meio líquido como em sólido. A comparação dos dados obtidos pelos 3 métodos não resultou em diferença significativa para a maioria das amostras testadas. Entretanto, os coeficientes de variação foram sempre menores para o método de semeadura em profundidade com a amostra diluída de 1:4, indicando maior precisão deste sobre os outros, principalmente em relação ao número mais provável. / The conventional methods for bioburden assessment are restrictive when applied to evaluate microbiological quality of pharmaceuticals and cosmetics. Triphenyltetrazolium chloride (TTC) was applied in this standartization study as alternative method for microbial limit test of pharmaceuticals and cosmetics. This method aimed at to reduce the assay time by conciliating the pour-plate technique analysis low bioburden of samples containing opacifiers. The application of this dye in the multiple tubes technique did not reduce incubation time, because the bacterial suspension sensitivity to triphenylformazan colour was 108 CUF/mL to 1 hour reaction and 107 CUF/mL to 4 hours. Suspensions with these bioburden are clearly turbid. The addition of 0.5% glicose and 0.1% glutamic acid to soybean-casein digest medium did not increase the reaction sensitivity. The addition of polissorbate 20 to solid ar liquid media promotted the difusion of triphenylformazan trom the cells to media; it rendered difficult of cellular mass detection in liquid medium and as much as colonies. The minimum concentration of TTC that produced maximum colour in liquid medium was 500 and 1,000 µg/mL, depending on the bacteria. However, bacterial suspensions with 102 CUF/mL tested by multiple tubes technique with 20 hours of incubation and by microbial growth detection after TTC addition provided equivalent numbers of positive and negative tubes with both concentrations. The comparison of this alternative method with that of sub-culture proved that 20 hours of microbial growth is not enough to be detected by TTC, when 1 mL of 1 CUF/mL suspension was planted. The dying of bacterial colonies was performed by covering the medium surface with 5 mL of 1% agar with TTC, molten at 48 C. The concentration of TTC that colored colonies in 1 hour was 0.025%. However, in presence of 6% aluminium hidroxide suspension the minimum effective concentration was 0.1%. The concentration required to detect yeasts in liquid media was 2,500 µg/mL in neutral pH condition. To detect yeast colonies in opalescent solid medium effective TTC concentration was 2%; in this case the dye was added to agar dispersed in 7.5 phosfate buffer. Samples of marketed produtcts were analysed through multiple tubes method with sub-culture and by TTC addition. Same samples were tested through pour-plate technique by inoculating 2 mL of 1:4 and 1:10 dilutions. The time required for resutts was 72 hours by the official method and 50 hours by alternative proposed method in liquid or solid medium. When the 3 method results were compared no significant differences were observed in almost all of the samples. However, the coefficient of variation related to pour-plate method results from 1:4 sample dilution was always lower than the others, mainly when compared with those obtained trom multiple tubes method. Therefore, the alternative pour-plate method is the most accurate one.

Nesta pesquisa, calculou-se, por meio da modelagem molecular, parâmetros físico-químicos importantes à capacidade anti-radicalar de compostos fenólicos extraídos de plantas da flora nacional, Chimarrhis turbinata e Arrabidaea samydoides. As propriedades eletrônicas também podem ser analisadas por meio de superfícies representadas por legendas de cores no campo 3D. Mapa de potencial eletrostático, distribuição orbitalar de HOMO e de LUMO e densidade de spin foram superfícies avaliadas neste trabalho. Em adição, estudos de QSAR (Quantitative Structure-Activity Relationships), cálculos de descritores moleculares holísticos por meio dos programas DRAGON e VOLSURF, cálculos estatísticos incluindo algoritmo genético e PLS (Partial Least Squares), demonstraram a influência de determinadas características moleculares como fundamentais à atividade biológica. A pesquisa concluiu que o grupo farmacofórico favorável à atividade antioxidante é estrutura que apresenta predominantemente características hidrofílicas, grupos hidroxila como substituintes, características eletrônicas favoráveis à doação de elétron e à estabilização do radical fenóxi formado, além de reduzido comprometimento estérico. Consideramos que os métodos empregados no trabalho podem ser considerados como abordagem inovadora para a Ciência Cosmética, indicando potencial ação antioxidante, que poderá ser utilizada em formulações antienvelhecimento. / In this research, the calculated physico-chemical parameters, by molecular modelling, have been reported in the literature for supplying important information about the antiradicalar behavior of phenolic compounds, as the studied herein from Chimarrhis turbinata sp. and Arrabidaea samydoides sp. The electronic properties also can be analyzed by means of surfaces represented by legends of colors in the 3D field. Map of electrostatic potential, HOMO and LUMO distribution orbitalar and spin density have been used in this work. In addition, QSAR studies (Quantitative Structure-Activity Relationships), calculations of holistic molecular descriptors by softwares DRAGON and VOLSURF, statistical analysis including genetic algorithm and PLS (Partial Least Squares), demonstrate the influence of the molecular structure in the biological activity. Therefore, pharmacofor favorable to the antioxidant activity structure that presents predominantly characteristic hydrophilic, groups hydroxyl as substituintes, electronic characteristics favorable to the donation of electron and the stabilization of the radical formed, besides reduced inibition esteric. These recent methods can be considered as an innovative approach for Cosmetic Science toward antioxidant action that could be used in antiaging products.

Submitted in partial compliance with the requirements for the Master’s Degree in Technology: Chiropractic, Department of Chiropractic and Somatology, Faculty of Health Sciences, Durban University of Technology, Durban, South Africa. 2017. / Background The growing popularity and competitive nature of the beauty industry has resulted in beauty therapists performing an extensive range of treatments to meet the demand, often at the jeopardy of their own well-being. Treatments include massage, waxing, pedicures and facials. These techniques involve vigorous and repetitive movements that may lead to overuse and strain on the musculoskeletal system, resulting in injury. Work-related activities together with factors such as socio-demographic, psychosocial, environmental and anthropometric can influence the prevalence of work-related musculoskeletal disorders (WRMDs). There has been little investigation into the WRMDs of beauty therapists both locally and internationally. Aim The aim of this study was to determine the prevalence, selected risk factors and impact of self-reported work related musculoskeletal disorders (WRMDs) in beauty therapists working within the hotel spa industry in the eThekwini municipality. Method A cross sectional epidemiological design was used to survey 254 beauty therapists working in the hotel spa industry in the eThekwini municipality of Durban. A pre-validated questionnaire was hand delivered to beauty therapists working at all hotel spas that agreed to participate. Informed consent was obtained. The survey contained questions related to musculoskeletal pain, socio-demographic, psycho-social, lifestyle and occupational factors. Results A response rate of 70% was obtained (n=178). The majority of the respondents were female (94.3%; n=165), the mean age of the respondents was 27.74 (SD±4.83) years. The respondents had been working as beauty therapists for an average of 5.91 (SD±4.19; n=176) years and performed on average 27.89 (SD±13.33; n=170) treatments per week, working 47.38 (SD±13.36; n=175) hours per week. Those working overtime worked on average 5.43 (SD±2.94; n=59) hours of overtime per week. There was a high rate of reported WRMDs with the lifetime, current and 12 month period prevalence being 86% (n=153), 85% (n=151) and 83% (n=148), respectively. The area most affected were the low back followed by the neck, the hand and wrist. Ninety percent of the respondents indicated that their pain was mild to moderate in nature, with half expressing an inability to cope with the pain, and 78% reporting that it interfered with their ability to work. Mental exhaustion (p=0.032), suffering from a concomitant co-morbidity (p=0.031), years worked as a beauty therapist (p˂0.001) and treating clients after hours (p=0.007) were significantly associated with the presence of musculoskeletal pain. Performing manual massage (p=0.043) and reporting feelings of exhaustion whilst performing specialised massage techniques (p=0.014) and applying makeup/eyelashes (p=0.022) were associated with the presence of WRMDs. Those who reported having co-worker (p=0.006) and managerial support (p˂0.001) were less likely to experience musculoskeletal pain. In contrast, feeling frustrated by work (p=0.007), being under pressure (p=0.005) and over worked (p=0.009) increased the chances of experiencing WRMDs. Working with the back in an awkward position (p˂0.001), standing for prolonged periods (p˂0.001), lifting heavy loads (p=0.019) and working in a hot and humid environment (p<0.001) increased the risk of WRMDs. Conclusion WRMDs have a significant impact on beauty therapists’ ability to work. Effective strategies to prevent and manage WRMDs in hotel spa beauty therapists is required, along with governmental regulation. / M