The cranberry scare of 1959 was the first food scare in the United States involving food additives to have a national impact. It was also the first event to test the Delaney clause, part of a 1958 amendment to the 1938 Food, Drug and Cosmetics Act prohibiting cancer-causing chemicals in food. Although lasting only a few weeks, the scare significantly affected the cranberry industry and brought the regulation of chemical residues in food to the national stage. Generated by a complex interaction of legislation, technology, media, and science, the scare had far-reaching effects in all areas of the cranberry industry, food legislation, and the perception of the public toward additives and residues in their food. The ripples caused by the scare permanently altered the cranberry industry and, after numerous subsequent scares and challenges to the law, eventually resulted in the repeal of the Delaney clause. The goal of this investigation was to demonstrate how the social, scientific, and political climates in the United States interacted and led to such an event. It shows how science, politics, and contemporary social anxiety combined, with technology as a catalyst, and how the resulting scare left significant marks on the development of both legislation and industry. It also improves our understanding of this seminal event in American social history by exploring the events surrounding the scare, as well as by comparing the perspectives and reactions of the public, the Eisenhower administration, the cranberry industry, and other industries affected by the scare and its aftermath.

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The impact of cranberry juice was investigated with respect to the initial adhesion of three isogenic Burkholderia cepacia bacteria with different extracellular polymeric substance (EPS) producing capacities: a wild-type cepacian EPS producer PC184, mutant bacteria PC184rml with reduced EPS production, and PC184bceK with a deficiency in EPS production. Adhesion experiments demonstrated that in the presence of cranberry juice, the adhesive capacity of PC184 was largely reduced, while cranberry juice had little impact on the adhesion of either mutant. Thermodynamic modeling supported results from adhesion experiments. For PC184, the surface free energy change Gadh switched from negative in the absence of cranberry juice to positive when cranberry juice was added. Surface force apparatus (SFA) and scanning electron microscopy (SEM) studies demonstrated strong adsorption of cranberry juice components to bacterial EPS. It was concluded that cranberry juice components could impact bacterial initial adhesion by adhering to EPS and impairing bacterial adhesive capacity. / Environmental Engineering

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No. of bitstreams: 1 silva_ibl_dr_bot_int.pdf: 1148926 bytes, checksum: d627f33c13d713fb2ae92530fb8ecde7 (MD5) Previous issue date: 2018-02-27 / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / Introdução: Embora a LTU seja conhecida pela humanidade desde a antiguidade, o real mecanismo envolvido na formação do cálculo renal ainda não foi completamente esclarecido. Trata-se de um processo complexo, envolvendo uma doença multifatorial resultante de interações entre fatores ambientais, predisposição genética e distúrbios metabólicos. Alterações nos fatores dietéticos têm sido propostas para prevenir ou minimizar o risco da litíase recorrente. Dentre estes fatores, o cranberry tem sido sugerido como potencial agente terapêutico capaz de atenuar os efeitos da hiperoxalúria, porém com resultados ainda controversos. Objetivo: Avaliar as eventuais repercussões metabólicas e histológicas decorrentes da suplementação oral da dieta com cranberry, sobre as alterações morfológicas e urinárias secundárias à hiperoxalúria provocada por agentes indutores, em um modelo experimental em ratos. Métodos: Foram utilizados 40 ratos machos da raça Sprague-Dawley, randomicamente distribuídos em quatro grupos: GRUPO 1 (G1: n=7) controle clínico; GRUPO 2 [G2: Etileno glicol (EG) a 0,5%+vitamina D3 (VD3), n=11] no qual a hiperoxalúria foi induzida a partir da administração de EG diluído em água e ofertado em associação com a VD3 (Colecalciferol) na dose de 0,5 μM; GRUPO 3 [G3: EG 0,5%+VD3+Cranberry); n=11] no qual os animais receberam as mesmas drogas ofertadas ao G2 acrescido de xarope de cranberry na dose de 800mg/kg peso/dia; GRUPO 4 (G4, n=11) animais suplementados apenas com a cranberry na mesma dose do G3. Foi coletada urina de 24 horas após 28 dias de intervenção para estudo metabólico. Neste dia os animais foram eutanasiados e os rins removidos para análise histopatológica/morfométrica, análise do estresse oxidativo no parênquima renal, bem como dosagem do cálcio no parênquima. Resultados: Dentre os parâmetros urinários avaliados, observou-se significativa redução do citrato em todos os grupos em relação ao controle, enquanto que o oxalato mostrou-se significativamente aumentado nos G2 e G3 quando comparado ao G1 (85,8, 93,6 e 51,7 mg/24h, respectivamente). O cálcio urinário foi significativamente menor nos grupos induzidos (G2: 0,84, G3: 1,07 e G1: 1,76mg/dL). A análise histomorfométrica revelou que apenas os animais dos G2 e G3 desenvolveram nefrolitíase sem, no entanto, apresentar diferença significativa entre si na contagem dos cristais intratubulares. Da mesma forma, considerando-se a análise histopatológica, apenas os animais induzidos (G2 e G3) exibiram atrofia, extravasamento estromal e infiltrado inflamatório no parênquima renal, em um padrão bastante semelhante entre os dois grupos. Com relação a análise do estresse oxidativo, os níveis de hidroperóxido de lipídio mostraram-se aumentados no grupo induzido (G2), quando comparado aos demais grupos. Como esperado, a quantificação do cálcio no parênquima renal foi significativamente maior em G2 e G3 quando comparado aos grupos sem indução. Conclusão: Embora a suplementação alimentar com cranberry tenha apresentado alguns efeitos positivos sobre a análise do estresse oxidativo, tais benefícios não foram suficientemente relevantes para impactar positivamente na redução da deposição dos cristais de OxCa e/ou na prevenção dos danos histológicos ao parênquima renal dos animais submetidos ao modelo de indução de hiperoxalúria. / Introduction: Although the importance of UTL, the actual mechanism involved in kidney stone formation has not yet been fully elucidated. It is a complex process, involving a multifactorial disease resulting from interactions between environmental factors, genetic predisposition and metabolic disorders. Changes in dietary factors have been proposed to prevent or minimize the risk of recurrent lithiasis. Among them, cranberry has been suggested as a potential therapeutic agent that can minimize the hyperoxaluria effects. However, the results are controversial. Objective: Evaluate the possible metabolic and histological repercussions resulting from oral supplementation with cranberry, on the morphological and urinary alterations secondary to hyperoxaluria, in an experimental model in rats. Methods: It was used 40 Sprague Dawley male rats and were randomized into four groups: Group 1 [(G1: n = 7) clinical control]; Group 2 [G2: Ethylene glycol (EG) 0.5% + Vitamin D3 (VD3), n = 11], which hyperoxaluria was induced by the administration of EG diluted in water and offered in association with VD3 (Cholecalciferol) at a dose of 0.5 uM; Group 3 [G3: 0.5% EG + VD3 + cranberry; n = 11], which the animals received the same drugs offered to the G2 plus cranberry at a dose of 800mg / kg body weight / day; Group 4 (G4, n = 11) which the animals are supplemented only with the same dose of cranberry in G3. 24 hours urine was collected after 28 days of intervention for metabolic study. On this day the animals were euthanized and the kidneys removed for histopathological / morphometric analysis; oxidative stress analysis; and measurement of renal parenchymal calcium. Results: Among the urinary parameters evaluated, there was significant reduction in the citrate in all the groups compared to the control group, while the oxalate was significantly increased in G2 and G3 compared to G1 (85.8, 93.6 and 51.7mg/24h, respectively). The urinary calcium was significantly lower in the induced groups (G2: 0.84, G3: 1.07 and G1: 1.76 mg/dL). Histomorphometric analysis revealed that only the animals of G2 and G3 developed nephrolithiasis without, however, no substantially differences from each other in the counting of intratubular crystals were found. Similarly, considering the histopathologic analysis, only the induced animals (G2 and G3) exhibited atrophy, stromal extravasation and inflammatory infiltrate in the renal parenchyma in a similar pattern between the two groups. Regarding to the analysis of oxidative stress, an increase of lipid hydroperoxide levels was found in induced group (G2), while compared with all others. As expected, the quantification of calcium in the renal parenchyma was significantly higher in G2 and G3 as compared to groups without induction. Conclusion: Although dietary supplementation with cranberry had some positive effects on the oxidative stress analysis, these benefits were not sufficiently relevant to reduce the deposition of CaOx crystals and / or the prevention of histological damage to the renal parenchyma in rats with secondary hyperoxaluria obtained from the administration of induced agents. / FAPESP: 2015/1006-1

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Escherichia coli et Enterococcus faecalis sont deux principaux agents pathogènes impliqués dans les infections du tractus urinaire (ITU) en médecine de ville et à l’hôpital. Ces espèces bactériennes sont responsables d’ITU aigües avec des phénomènes de récurrence et dans des ITU chroniques. La consommation d'antibiotiques est directement corrélée à la résistance des bactéries uropathogènes ce qui montre l'importance de contrôler l'utilisation des antibiotiques et de développer des traitements préventifs et curatifs alternatifs pour les infections urinaires.La consommation alimentaire de cranberry et de leurs extraits est traditionnellement associée avec le maintien en bonne santé des voies urinaires. Par ailleurs, certaines études cliniques semblent montrer un effet préventif des ITU associé à la consommation alimentaire de cranberry. In vitro et ex vivo, la consommation de ces extraits par l’homme réduit l’adhérence de certaines souches d’E. coli aux cellules épithéliales urinaires et la formation de biofilm de différentes espèces. L’hypothèse de travail est que la consommation alimentaire d’extraits de cranberry conduise à la formation de métabolites urinaires qui diminuent l'adhérence des bactéries uropathogènes à l’épithélium urinaire. Ce mécanisme serait à la base de la prévention des ITU par consommation d’extraits de cranberry. Cependant, les métabolites bioactifs restent largement méconnus. / Escherichia coli and Enterococcus faecalis are two main pathogens involved in urinary tract infections (ITU) in town medicine and in the hospital. These bacterial species are responsible for acute UTIs with recurrence phenomena and in chronic ITUs. The consumption of antibiotics is directly correlated with the resistance of uropathogenic bacteria, which shows the importance of controlling the use of antibiotics and of developing alternative preventive and curative treatments for urinary infections.Cranberry consumption of their extracts is traditionally associated with the maintenance of healthy urinary tract. In addition, some clinical studies seem to show a preventive effect of ITUs associated with cranberry consumption. In vitro and ex vivo, the consumption of these extracts by humans reduces the adhesion of certain E. coli strains to urinary epithelial cells and biofilm formation of different species. The working hypothesis is that the consumption of cranberry extracts leads to the formation of urinary metabolites that decrease the adhesion of uropathogenic bacteria to the urinary epithelium. This mechanism would be the basis for the prevention of ITU by consumption of cranberry extracts. However, bioactive metabolites remain largely unknown.

As a species domesticated only in the last century, agricultural cranberry plants (Vaccinium macrocarpon) remain little removed from their wild relatives. Thus, it is a potential model species for studies of the earliest stages of domestication; however, there is little available quantitative information on its wild population biology and ecology. As such information is vital to studies of the ecological changes occurring during domestication, the purpose of this study was to consolidate the relevant knowledge available and conduct a preliminary search for patterns in the invertebrate communities of wild bogs. The alpha diversity was found to be greater than the overall (gamma diversity), which is likely a result of the metric used and the fact that there was minimal overlap in rare species between bogs. In addition, alpha diversity was found to be significantly negatively correlated with bog age. Two pairs of species were found to be correlated with each other, Blunt-nosed (Limotettix vaccinia (Van Duzee)) and Sharp-nosed leafhopper (Scaphytopius sp.), along with fleabeetle (Sysena frontalis (F.)) and firebeetle (Cryptocephalus incertus (Oliv.). In addition, it was found that spiders are significantly more common in western bogs, while firebeetle is significantly more common in eastern bogs. However, this may be indicative of a correlation with bog age rather than a true correlation with geographic location. More work is needed to determine the true dynamics driving these findings, so that the information could eventually be used to improve the efficacy and decrease the environmental impact of pest management on agricultural bogs.

"Bacterial infections persist as a public threat due to the ease by which bacteria adapt to commonly used antibiotics. In addition, bacteria on surfaces develop protective communities called biofilms that hinder the ability of antibiotics to completely eliminate the pathogens. The rapid development of bacterial resistance to antibiotics has made pharmaceutical companies reluctant to fund new antibiotics research. Hence, novel approaches to prevent and treat infections are needed. The development of infections can be divided into three steps: adhesion, invasion and multiplication. Antibiotics target at the latter two step and are prone to bacterial resistance as passive strategies. Bacterial adhesion to host cells/implanted medical devices is the first step leading to following invasion and multiplication. However, fundamental understanding of bacterial adhesion process is still lacking. The current studies are aimed to systematically investigate biological interactions between pathogenic bacteria and host cell, proteins and biomaterials with both macro and micro scale approaches. The macro scale methods include bacterial adhesion assay, viability studies, and thermodynamic modeling. The micro scale methods include direct adhesion force measurements, ultra surface visualization via atomic force microscopy (AFM) and surface structure modeling. Our work combines experiments and modeling aimed at understanding the initial steps of the bacterial adhesion process, focusing on two case studies: 1) Mechanisms by which cranberry can prevent urinary tract infections through interfering with bacterial adhesion; and 2) Design of anti-adhesive and antimicrobial coatings for biomaterials. We make direct adhesion force measurements between bacteria and substrates with an atomic force microscope (AFM), and combine such experiments with thermodynamic calculations, in order to develop a set of tools that allows for the prediction of whether bacteria will attach to a given surface. These fundamental investigations of the bacterial adhesion process help elucidate the underlying mechanisms behind bacterial adhesion, thus leading to improved clinical outcomes for a number of biomedical applications. "

L’infection urinaire (IU) est un problème majeur de Santé publique. La cystite aiguë touchant principalement les femmes est la plus fréquente des IU. La bactérie la plus fréquemment isolée au cours de ces IU est Escherichia coli. Une des particularités de la cystite est sa propension à récidiver. Le traitement préconisé pour ces infections est la prise d’antibiotiques, qui peut être fréquente en cas de cystites récidivantes. C’est dans ce contexte que de nouvelles stratégies doivent être développées afin de prévenir et traiter les IU récidivantes. Parmi ces différentes stratégies, l’utilisation de produits naturels tels que la canneberge (Vaccinium macrocarpon) apparaît comme prometteuse. En effet, des études précédentes ont montré que la canneberge a un effet négatif sur l’adhésion des bactéries aux cellules superficielles de l’épithélium vésical facilitant l’élimination des bactéries par le flux urinaire. Cette activité est portée par la proanthocyanidine de type A (PAC-A). D’autre part, une étude menée par notre équipe a montré que l’effet de la canneberge sur l’adhésion et la virulence de souches d’E. coli uropathogènes pouvait être potentialisé par l’ajout d’un autre composé naturel : la propolis. Depuis l’Antiquité ses propriétés anti-bactériennes sont reconnues et des études plus récentes ont démontré son impact sur des bactéries à Gram positif mais également sur deux bactéries à Gram négatif : E. coli et Pseudomonas aeruginosa. Ce travail de thèse a permis : i) de décrire l’impact de la canneberge, de la propolis et de leur association sur le transcriptome d’une souche clinique d’E. coli uropathogène (G50). Cette analyse transcriptomique a montré que la canneberge entrainait une sous-expression de gènes liés à l’adhésion, mais également de gènes liés à la mobilité et à la formation de biofilm. En revanche, la canneberge augmentait l’expression des gènes liés au métabolisme du fer ainsi qu’à la réponse au stress. Ces effets étaient potentialisés par l’ajout de la propolis. En parallèle, des tests phénotypiques menés sur une collection de souches d’E. coli uropathogènes sur la mobilité et la formation de biofilm ont confirmé les résultats précédents ; ii) de développer un test, basé sur les précédents travaux de transcriptomique, permettant une évaluation standardisée de l’effet de la PAC-A sur E. coli, indépendamment de sa concentration car il n’existe pas de techniques standardisées pour doser cette molécule. C’est ainsi que 4 gènes (tsr, ftnA, fecB, feoB) ont été sélectionnés, le suivi de leur expression permettant une mesure de l’activité anti-bactérienne de la canneberge; iii) de mesurer l’effet potentialisateur de la propolis sur l’activité des antibiotiques utilisés dans le traitement des IU. C’est ainsi qu’il a été montré que l’ajout de la propolis permettait d’augmenter l’activité bactéricide des antibiotiques testés et de diminuer les concentrations minimales inhibitrices de ces antibiotiques. / Urinary Tract Infection (UTI) is a major problem of public health. Acute cystitis which touches mostly women is the most common form of UTI. The bacteria which is mostly isolated in an UTI is Escherichia coli. A particularity of cystitis is to come back. In this context news strategies have to be developed to prevent and cure recurrent UTI. One of these strategies is the utilization of natural products like the cranberry (Vaccinium macrocarpon) which is promising. Indeed, previous studies showed the negative impact of cranberry on the adhesion of bacteria on the superficial cells of bladder which help the elimination of bacteria by the urinary flux. This activity is carried by the type A proanthocyanidin (PAC-A). Moreover, a study lead by our team has demonstrated an improvement of the activity of cranberry on the adhesion and the virulence of uropathogenic E. coli (UPEC) by another natural product: the propolis. Since Antiquity its antibacterial activities have been recognize and more recent studies have demonstrated its impact of Gram positive bacteria and also on two Gram negative bacteria: E. coli and Pseudomonas aeruginosa. This thesis has allowed for: i) the description of the impact of cranberry, propolis and its association on the transcriptome of a clinical strain of UPEC (G50). This transcriptomic analyze have shown that the cranberry down regulated genes linked to the adhesion and also genes linked to the motility and biofilm formation. However the cranberry up regulated genes linked to the iron metabolism and the stress response. These effects are improve by the addition of propolis. Concurrently phenotypics tests have been conducted on a collection of UPEC on the motility and the biofilm formation and they confirmed the previous results; ii) the development of a test, based on our transcriptomic results, enable to performed a standardized evaluation of the impact of PAC-A on E. coli, independently of its concentration. Indeed, this molecule cannot be measure in a standard way. Four genes have been selected (tsr, ftnA, fecB, feoB), the monitoring of their expression allow us to measure the anti-bacterial efficiency of the cranberry; iii) the measurement of the potential effect of the propolis of the antibiotic’s activities used to treat UTIs. Thus it have been observed that the addition of propolis improve the bactericide activity of the antibiotics tested and reduces the minimal inhibitory concentration of these antibiotics.