Submitted in partial compliance with the requirements of the Master’s Degree in Technology: Homoeopathy, Durban University of Technology, Durban, South Africa, 2016. / Aim The aim of this study was to compare the nuclear magnetic resonance spectra of Sulphur 12c samples produced by the traditional Hahnemannian method with Sulphur 12c samples produced using sonication as an alternative method of agitation. Sonication, while not widely employed as an agitating technique in the homoeopathic potentisation process, is a highly effective agitation process which produces effects on liquids that closely resemble the effect of traditional Hahnemannian hand succussion (Bhattacharyya et al. 2008). Thus, this study sought to reveal whether or not homoeopathic remedies produced by sonication bore a close enough physicochemical resemblance to traditional hand succussed remedies to be considered as a viable equivalent. Methodology Five sample groups were manufactured for analysis, all by means of serial dilution at the centesimal ratio (1:100) to the 12c potency, and with agitation between dilution levels where applicable. Three of the sample groups were experimental, namely the Sulphur 12c Hahnemannian, Sulphur 12c sonicated and Sulphur 12c both (succussion and sonication). The Sulphur 12c Hahnemannian samples were produced by hand according to the German Homoeopathic Pharmacopoeia (Benyunes 2005), which includes an agitation phase of 10 hand succussions. Sonicated samples were produced according to the Hahnemannian method as far as possible, however the agitation phase consisted of 30 seconds of sonication in a sonication bath at 40Hz in accordance with related studies (Sukul, Sinhabau, and Sukul 1999: 58-59; Sukul et al. 2001a: 187). Sulphur 12c both (succussion and sonication) samples underwent ten hand succussions and 30 seconds of sonication at 40Hz between dilution levels. Two of the sample groups were controls, namely Sulphur 12c unagitated and Lactose 12c unagitated, neither of which underwent agitation between dilution phases but were otherwise produced according to the German Homoeopathic Pharmacopoeia specification (Benyunes 2005). All samples were raised to the 12c potency level in 87% alcohol from a 3CH triturate. The Lactose 12c unagitated control was derived from a 3CH triturate of lactose, while the other samples were all derived from a 3CH triturate of Sulphur. The sample groups were sent for nuclear magnetic resonance (NMR) spectroscopy at the Department of Chemistry at Stellenbosch University. The NMR device used was the Varian UnityInova 600 NMR Spectrometer ®, with a Deuterated DMSO insert added as an instrument frequency lock. Samples were drawn and analysed by Dr D.J. Brand. One sample was drawn from each sample group. The chemical shift and relative integration values for the OH, H2O, CH2, and CH3 peaks of the NMR spectra were captured and tabulated using Microsoft Excel© 2013. The statistical analysis was performed with the aid of SPSS Version 22. The chemical shift and relative integration values for the H2O, OH, CH2 and CH3 peaks were used in the statistical analysis. The Kruskal-Wallis method was performed for the five sample groups to ascertain whether or not a statistically significant difference existed between the five sample groups. Comparisons between individual paired groups were conducted by means of the non-parametric Mann-Whitney test. The significance interval was set at α = Results The chemical shift values of the CH2 peaks of the samples showed a clear similarity between the samples produced by Hahnemannian hand succussion, sonication and both (succussion and sonication) as well as a clear difference between these three samples and the two controls. The relative integration values, however, showed no clear trends in support of or detracting from the hypotheses. Conclusion In terms of the CH2 peak chemical shift values it can be concluded that distinct similarities exist between 12c potency level of Sulphur produced by Hahnemannian hand succussion and sonication, and that the two methods of agitation produce similar structural properties in samples of the 12c potency level. Furthermore in terms of the chemical shift values, succussion and sonication develop remedies that are distinct from unagitated remedies of equivalent potency level. Thus, these findings support the use of sonication as a potentially viable alternative to hand succussion as a method of agitation in the potentisation process. Further studies need to be conducted however, with the inclusion of a greater variety of potency levels in order to possibly reveal more trends in terms of the relative integration values as these values were inconclusive in this study. / M

CoordenaÃÃo de AperfeiÃoamento de Pessoal de NÃvel Superior / Atualmente à crescente a busca por uma alimentaÃÃo mais saudÃvel e com isso surge a possibilidade da inclusÃo de frutos e hortaliÃas nos hÃbitos alimentares. A manga à um fruto bastante consumido e produzido no Brasil e os minimamente processados (MP) representam praticidade porque sÃo produtos que jà vem prontos para consumo, sem perda de tempo com operaÃÃes de descasque, corte, etc. Durante as etapas de processamento hà um estresse dos tecidos, que leva à geraÃÃo de radicais livres que podem causar a degradaÃÃo do fruto, por isso, se faz necessÃrio a utilizaÃÃo de tratamentos capazes de minimizar as perdas de qualidade. Nesse contexto surge o ultrassom (US) que à uma tecnologia emergente com potencial de exploraÃÃo e utilizaÃÃo. As ondas ultrassÃnicas provocam principalmente a compressÃo e expansÃo dos tecidos, cujos resultados vÃo depender da matriz alimentar. Este trabalho teve como objetivo avaliar o efeito do US na qualidade de mangas âTommy Atkinsâ MP, analisando o sistema de defesa antioxidante, componentes do metabolismo oxidativo e a atividade de enzimas hidrolÃticas da parede celular. Para tanto, as amostras foram expostas à sonicaÃÃo na frequÃncia de 25 kHz por 30 minutos em banho de US. Observouâse que o tratamento causou uma reduÃÃo na acidez titulÃvel (AT) e nos sÃlidos solÃveis (SS), porÃm apÃs sete dias de armazenamento a 4 oC, manteve elevada a relaÃÃo SS/AT (46,54). Os teores de aÃÃcares tambÃm foram reduzidos (15%), bem como Ãgua foi incorporada (4,6%), o que mostra uma modificaÃÃo na estrutura celular, que pode ser comprovada pelo decrÃscimo na firmeza apÃs sete dias (de 241,73 para 139,38 N) e aumento na atividade da pectinametilesterase. A cor foi degradada apÃs sete dias (∆L = 3,24 e ∆C = 4,42), resultando em escurecimento, contudo as amostras controle tambÃm escureceram. Em relaÃÃo ao sistema antioxidante, nÃo houve grandes modificaÃÃes, tendo em vista ser um fator benÃfico se comparado a outros mÃtodos de conservaÃÃo, com destaque apenas para a perda de vitamina C (de 328,10 para 190,18 mg Ãcido ascÃrbico/kg matÃria fresca (MF)). Houve ainda reduÃÃo na peroxidaÃÃo de lipÃdios de membrana com o tratamento (de 23,58 para 18 ηmol MDA/g MF). Concluiâse com base neste estudo, que a aplicaÃÃo de US em mangas âTommy Atkinsâ MP tem seus pontos negativos na firmeza e na cor, contudo, nÃo afetou drasticamente os sistemas antioxidante enzimÃtico e nÃo enzimÃtico, alÃm de levar a uma reduÃÃo no teor de aÃÃcar. Todavia, fica claro a necessidade de explorar esta tecnologia na fisiologia pÃsâcolheita de frutos, assim como, otimizar e combinar a tÃcnica com outros processos que diminuam seus efeitos negativos e favoreÃam os positivos. / Currently is growing to search for healthier eating with that comes the possibility for inclusion of fruits and vegetables in the diet. Mango fruit is widely consumed and produced in Brazil and fresh â cut (FC) represents convenience because them are products that comes ready for consumption without loss of time with operations peeling, cutting, etc., and these steps there is a stress tissue leading to generation of free radicals that can lead to a degradation fruit, so it is necessary a type of processing that minimize the loss of quality and yet ensures the safety of the final product. In this context arises the ultrasound (US) that is an emerging technology that has potential for exploitation and utilization. The ultrasonic waves cause primarily tissue compression and expansion, whose the results will depend on the food matrix. This study aimed to evaluate the effect of US in quality of FC 'Tommy Atkins' mangoes, analyzing the antioxidant defense system, metabolism oxidative compounds and the activity of cell wall hydrolytic enzymes. The samples were exposed to 25 kHz frequency sonication for 30 minutes in US bath. It was observed tha t the treatment caused a reduction in titratable acidity (TA) and soluble solids (SS), but after seven days of storage at 4 o C, maintaining the high SS/TA ratio (46.54). The sugars was also reduced (15%) and water incorporated (4.6%), which indicates a change in cell structure, that can be evidenced by decreased in firmness after seven days (from 241.73 to 139.38 N) and increased activity of pectinmethylesterase. The color wa s degraded after seven days (ΔL = 3.24 and ΔC = 4.42), resulting in darkening, but the control samples also darkened. In relation to the antioxidant system, there were no significant changes in order to be a beneficial factor compared to other preservation methods, particularly only to the loss of vitamin C (328.10 to 190.18 mg ascorbic acid/kg weight fresh (WF), still there was a reduction in lipid peroxidation of membrane (from 23.58 to 18 η mol MDA/g WF). It concluded to base on this study, FC 'Tommy Atkins' mango has its drawbacks on firmness and color, however, did not affect severely the enzymatic and non â enzymatic antioxidant x systems, and lead to a reduction in sugar content. However, it is clear the necessitate to explore the application of this technology in post harvest fruits physiology as well as to optimize and to combine the technique with other processes that reduce their negative effects and promote the positives.

Two issues hindering the clinical application of image-guided transcostal focused ultrasound surgery (FUS) are the organ motion caused by cardiac and respiratory movements and the presence of the ribcage. Intervening ribs absorb and reflect the majority of ultrasound energy excited by an acoustic source, resulting in insufficient energy delivered to the target organs of the liver, kidney, and pancreas. Localized hot spots also exist at the interfaces between the ribs and soft tissue and in highly absorptive regions such as the skin. The aim of this study is to assess the effects of transmitted beam distortion and frequency-dependent rib heating during trans-costal FUS, and to propose potential solutions to reduce the side effects of rib heating and increase ultrasound efficacy. Direct measurements of the transmitted beam propagation were performed on a porcine rib cage phantom, an epoxy rib cage phantom and an acoustic absorber rib cage phantom, in order of their similarities to the human rib cage. Finite element analysis was used to investigate the rib cage geometry, the position of the target tissue relative to the rib cage, and the geometry and operating frequency of the transducer. Of particular importance, frequency-dependent heating at the target and the intervening ribs were estimated along with experimental verification. The ratio of ultrasonic power density at the target and the ribs, the time-varying spatial distribution of temperature, and the ablated focus of each sonication are regarded as key indicators to determine the optimal frequency. Following that, geometric rib-sparing was evaluated by investigating the operation of 2D matrix arrays to optimize focused beam shape and intensity at target. Trans-costal FUS is most useful in treating tumours that are small and near the surface of the abdominal organs, such as the liver, kidney and pancreas. However, for targets deep inside these organs, severe attenuation of acoustic energy occurs, suggesting that pure ultrasound thermal ablation with different heating patterns will have limited effects in improving the treatment efficacy. Results also demonstrate that the optimal ultrasound frequency is around 0.8 MHz for the configurations considered, but that it may shift to higher frequencies with changes in the axial and lateral positions of the tumours. In this work, I aimed to reduce the side effects of rib heating and increase the ultrasound efficacy at the focal point in trans-costal treatment. However, potential advanced techniques need to be explored for further enhanced localized heating in trans-costal FUS.

Biosensor development has the potential to meet the need for rapid, sensitive, and specific detection of pathogenic bacteria from natural sources. An antibody-based fiber-optic biosensor assay to detect low levels of Vibrio vulnificus in estuarine waters following an enrichment step was developed. The principle of the sensor is based on an immuno-sandwich assay where an anti-V. vulnificus polyclonal capture antibody preparation was first immobilized on a polystyrene fiber-optic waveguide using a biotin-avidin association. The capture antibody is responsible for binding the target cells to the waveguide. Cyanine-5-conjugated anti-V. vulnificus polyclonal antibodies are subsequently allowed to bind to immobilized cells, and detection occurs when a photodetector collects emitted light (670-710 nm) from the fluorophore, which is excited with 635-nm laser light produced by the Analyte 2000 biosensor. Any detection signal greater than a pre-determined threshold signal is considered to be a positive detection event, while any signal lower than the threshold is considered no detection. This immunosensor assay proved highly specific when tested against whole cells and cell extracts from V. cholerae, V. parahaemolyticus, V. alginolyticus, and E. coli. isolates. Following a four hour enrichment in PNCC broth, and in a total of less than seven hours, the assay was able to detect cell extracts from as few as 100 V. vulnificus colony forming units suspended in sterile water. This method holds promise for detection of low numbers V. vulnificus and other autochthonous pathogens in estuarine waters.

The inability to transport molecules efficiently and easily into cells and across tissues is one of the major limitations of developing drug delivery systems. A novel approach to overcoming this problem could be the use of low-frequency ultrasound to make cell membranes and tissues more permeable. Previous studies show that normally impermeant molecules can be transported into cells exposed to ultrasound; however, the mechanism by which this occurs is not well understood. Our hypothesis is that low frequency ultrasound can reversibly disrupt membrane structure, thus allowing diffusion-driven intracellular delivery of molecules through a breach in the cell membrane. The effects of ultrasound are not limited to uptake of molecules; there can also be significant loss of cell viability after sonication. Therefore, the focus of this work is to determine the mechanisms by which molecular uptake and cell death occur from ultrasound exposure. The long-term goal of this work is to increase the number of viable cells that experience uptake by controlling the effects that cause cell death. Our data have show that large molecules (r ≤ 28 nm) can be taken into cells after exposure to 24 kHz (10% duty cycle for 2 s of exposure time at 0.1 pulse length over a range of pressures) ultrasound and that uptake of these molecules can occur even after sonication ended. In experiments developed to isolate the mechanism(s) of uptake, DU145 prostate cancer cells depleted of ATP energy and intracellular calcium showed no uptake of calcein, a small fluorescent molecule (MW = 623 Da), nor did sonicated lipid bilayers (red blood cell ghosts), suggesting that uptake is calcium mediated and requires active mechanisms in viable cells. Multiple types of microscopy, including electron and laser scanning confocal, showed evidence of large plasma membrane disruptions which support the hypothesis that transport of molecules into cells occurs through repairing wounds. Microscopy studies also indicated that much if the sonication-mediated death can occur by instantaneous cellular lysing and rapid cell death (within minutes post-exposure) due to wound-instigated necrosis; in addition, characteristics of rapidly induced controlled death modes were seen and found to be non-caspase-mediated within an hour after sonication ended.

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FundaÃÃo Cearense de Apoio ao Desenvolvimento Cientifico e TecnolÃgico / O trabalho objetivou o estudo dos efeitos do ultrassom, como prÃ-tratamento na secagem convectiva de maÃÃ Fuji, sobre a perda de Ãgua, perda de sÃlidos, difusividade efetiva da Ãgua, tempo de secagem, cor e atividade enzimÃtica de polifenoloxidase e peroxidase. A partir dos resultados obtidos, determinou-se a melhor condiÃÃo para substituiÃÃo dos aÃÃcares naturais da fruta por estÃvia (aÃÃcar nÃo calÃrico). Observou-se perda de Ãgua, perda de sÃlidos, aumento de 85% da difusividade efetiva da Ãgua, reduÃÃo de 45% no tempo de secagem, reduÃÃo de 39% das alteraÃÃes de cor e reduÃÃo da atividade enzimÃtica de polifenoloxidase e peroxidase da maÃÃ Fuji. Estabeleceu-se como melhor condiÃÃo para substituiÃÃo de aÃÃcares naturais por estÃvia aquela onde a fruta foi sonificada por 20 minutos na proporÃÃo entre Ãgua e fruta de 6:1 (p/p). O uso de estÃvia como aÃÃcar nÃo calÃrico substituto dos aÃucares naturais da maÃÃ Fuji, aliado ao processo de sonificaÃÃo, gerou um produto com valor energÃtico final 12% menor do que a fruta in natura, com tempo de recuperaÃÃo da doÃura inicial da fruta estimado em 13 segundos de sonificaÃÃo em soluÃÃo de estÃvia 10%. Dessa forma, a sonificaÃÃo figura como alternativa tecnolÃgica para a obtenÃÃo de maÃÃ Fuji desidratada, com tempo de secagem reduzido, baixo valor energÃtico, baixa alteraÃÃo de cor e menor atividade enzimÃtica de polifenoloxidase e peroxidase. / The work aimed to study the effects of ultrasound as a pretreatment on convective drying of Fuji apple, about the loss of water, loss of solid, effective diffusivity of water, drying time, color and enzymatic activity of polyphenol oxidase and peroxidase. From the results obtained, it was determined the best condition for replacement of the natural sugars of the fruit by stevia (no-calorie sugar). There was loss of water, solids loss, 85% increase of the effective diffusivity of water, 45% reduction in drying time, a 39% reduction of the color changes and reduction in enzymatic activity of peroxidase and apple PPO Fuji . Set up as best condition for replacement of natural sugars stevia by one where the fruit was sonicated for 20 minutes in water and fruit ratio of 6:1 (w / w). The use of stevia as a non-caloric sugar substitute for the natural sugars of Fuji apple, along with the sonication process, generated a product with the final energy value 12% lower than the fresh fruit, with a recovery time of the initial sweetness of the fruit estimated 13 second sonication in a solution of 10% stevia. Thus, the sonication figure as an alternative technology for obtaining dried Fuji apple with reduced drying time, low energy, low color shift and reduced enzymatic activity of polyphenol oxidase and peroxidase.

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