The major objective of the research was to improve lychee postharvest management, through a greater understanding of mechanical injury and moisture loss. Mechanical injury is a known cause of postharvest loss in lychee, but previously published information has been limited to broad observations. In this study, the symptoms of mechanical damage in lychee were defined, including quantitative measurement of colour changes. Impact injury caused protuberance tip darkening, cracking of the pericarp and significant changes in skin colour. Compression also typically caused tip darkening, and severe loads were capable of puncture, shape distortion and skin cracking. Abrasion and vibration injuries were characterised by strong yellowing of pericarp colour, possibly due to the leakage of cell contents onto the fruit surface. Vibration also caused significant darkening and loss of colour saturation. Vibration has not previously been mentioned as an issue in lychee postharvest management, but appeared to be as important a problem as desiccation browning at the wholesale level, both in incidence and severity. Mechanically damaged fruit consistently showed increased ethylene and carbon dioxide synthesis, and moisture loss was increased by up to 30%. Some significant changes in skin biochemistry and cuticle properties were also detected. The study of damaged tissue by SEM revealed distinctive patterns of surface tissue disruption. Open pericarp cracking was a particularly detrimental injury, causing significantly increased electrolyte leakage and rapid pathogen development. The effects of load characteristics, such as magnitude, method of application, site, repetition and cushioning, on the extent of damage were defined. Fruit characteristics such as cultivar, gross morphology, temperature, hydration and surface wetness were shown to significantly affect damage levels. Small seed size was correlated with increased cracking susceptibility. Fruit surface wetness exacerbated vibration or abrasion damage. Turgid fruit were less susceptible to vibration and abrasion damage, but showed increased susceptibility to impact cracking. Previously neglected aspects of desiccation browning research were studied, including cultivar and maturity effects, sites of moisture loss and the role of air currents. Cultivar effects on moisture loss were obscured by pre-harvest factors, but consistent cultivar differences were detected in desiccation browning, possibly related to skin thickness. In contrast, maturity levels over a marketable range had little effect on weight loss or browning. Moisture was lost fairly evenly over the fruit surface, but poor postharvest handling appeared to massively increase loss from the protuberance tips. Moisture loss was shown to substantially increase ethylene synthesis. The crucial role of air currents in exacerbating lychee moisture loss was emphasised, and the relationship between air speed and weight loss was defined. The research contributed to a greater understanding of the processes of mechanical damage and moisture loss in lychee, leading to improved protocols for the postharvest management of the fruit. Improved management of mechanical damage and moisture loss will ultimately improve fruit quality and reduce postharvest losses, hence increasing returns to industry.

MUC4 is a heterodimeric mucin glycoprotein expressed in the epithelia of tissues. Previous studies in our laboratory demonstrated that MUC4 protein expression is regulated by exogenous growth factors and that MUC4 is found in complex with the receptor tyrosine kinase ErbB2. MUC4 protein expression in airway epithelia was evaluated using molecular biology techniques. The impact of the protein on ErbB2 activation was evaluated post mechanical wounding of airway epithelia, and upon MUC4 RNA silencing. MUC4 levels were increased with exposure to the differentiating agent retinoic acid and decreased upon exposure to epidermal growth factor, a proliferative agent. In the absence of MUC4, ErbB2 phosphorylation was diminished. These results support the hypothesis that MUC4 expression is enhanced during differentiation of epithelia. Furthermore these findings provide evidence for an additional level of ErbB regulation in airway injury and subsequent epithelial wound healing.

The major objective of the research was to improve lychee postharvest management, through a greater understanding of mechanical injury and moisture loss. Mechanical injury is a known cause of postharvest loss in lychee, but previously published information has been limited to broad observations. In this study, the symptoms of mechanical damage in lychee were defined, including quantitative measurement of colour changes. Impact injury caused protuberance tip darkening, cracking of the pericarp and significant changes in skin colour. Compression also typically caused tip darkening, and severe loads were capable of puncture, shape distortion and skin cracking. Abrasion and vibration injuries were characterised by strong yellowing of pericarp colour, possibly due to the leakage of cell contents onto the fruit surface. Vibration also caused significant darkening and loss of colour saturation. Vibration has not previously been mentioned as an issue in lychee postharvest management, but appeared to be as important a problem as desiccation browning at the wholesale level, both in incidence and severity. Mechanically damaged fruit consistently showed increased ethylene and carbon dioxide synthesis, and moisture loss was increased by up to 30%. Some significant changes in skin biochemistry and cuticle properties were also detected. The study of damaged tissue by SEM revealed distinctive patterns of surface tissue disruption. Open pericarp cracking was a particularly detrimental injury, causing significantly increased electrolyte leakage and rapid pathogen development. The effects of load characteristics, such as magnitude, method of application, site, repetition and cushioning, on the extent of damage were defined. Fruit characteristics such as cultivar, gross morphology, temperature, hydration and surface wetness were shown to significantly affect damage levels. Small seed size was correlated with increased cracking susceptibility. Fruit surface wetness exacerbated vibration or abrasion damage. Turgid fruit were less susceptible to vibration and abrasion damage, but showed increased susceptibility to impact cracking. Previously neglected aspects of desiccation browning research were studied, including cultivar and maturity effects, sites of moisture loss and the role of air currents. Cultivar effects on moisture loss were obscured by pre-harvest factors, but consistent cultivar differences were detected in desiccation browning, possibly related to skin thickness. In contrast, maturity levels over a marketable range had little effect on weight loss or browning. Moisture was lost fairly evenly over the fruit surface, but poor postharvest handling appeared to massively increase loss from the protuberance tips. Moisture loss was shown to substantially increase ethylene synthesis. The crucial role of air currents in exacerbating lychee moisture loss was emphasised, and the relationship between air speed and weight loss was defined. The research contributed to a greater understanding of the processes of mechanical damage and moisture loss in lychee, leading to improved protocols for the postharvest management of the fruit. Improved management of mechanical damage and moisture loss will ultimately improve fruit quality and reduce postharvest losses, hence increasing returns to industry.

Orientador: José Fernando Durigan / Banca: Ben Hur Mattiuz / Banca: Paulo Ademar Martins Leal / Banca: Josalba Vidigal de Castro / Banca: José Flávio Diniz Nantes / Resumo: O objetivo deste trabalho foi avaliar os efeitos de injúrias mecânicas, por impacto, compressão e corte, na fisiologia de abacates, através de análises físicas, químicas, bioquímicas e sensoriais, tomografia de ressonância magnética e microscopia eletrônica de varredura. O impacto foi aplicado nos lados opostos do fruto e provocados por queda livre de 2,00 m; a compressão, submetendo-os a pesos de 117,6 N, por 24 horas; e o corte, com quatro incisões longitudinais, com 80 mm de comprimento e 4 mm de profundidade, nos lados opostos do fruto. As diferentes lesões aplicadas aos abacates ocasionaram danos irreparáveis à qualidade dos frutos, onde os efeitos mais nocivos foram encontrados com os cortes. As imagens tomográficas não demonstraram a ocorrência de lesionamento interno em abacates 'Hass', enquanto que os frutos da 'Quintal', submetidos à compressão e ao impacto, apesar de não mostrarem lesões externas, as imagens indicaram a ocorrência de lesionamento interno e a evolução do mesmo durante o amadurecimento. A microscopia eletrônica de varredura mostrou que os tecidos da polpa de abacates 'Quintal' injuriados apresentaram desordem celular em suas estruturas e que eles podem ser protegidos por diferentes processos de cicatrização. / Abstract: This work aimed to evaluate the effects of mechanical injuries, for impact, compression and cut, in the physiology of avocados, by physical, chemical, biochemical and sensorial analysis, and by use of magnetic resonance imaging and scanning electron microscopy. The impact was applied in the opposed sides of the fruit and provoked by free fall from 2.00 m; compression, submitting them to weights of 117.6 N, for 24 hours; and cut, with four longitudinal incisions, with 80 mm of length and 4 mm of depth, in the opposed sides of the fruit. The different injuries applied to the avocados caused irreparable damages to the quality of these fruits, where the most injurious effects were found for the cuts. The tomography images did not indicate the occurrence of internal injuries in 'Hass' avocados, while the fruits of the 'Quintal', submitted to the compression and the impact, although did not show external lesions, the images indicated the occurrence of internal injuries and evolution of them during the ripening. The scanning electron microscopy showed that 'Quintal' avocados tissues mechanically injured presented cellular disorder in their structures and that they can be protected by different cicatrization processes. / Doutor

Made available in DSpace on 2017-07-24T19:22:10Z (GMT). No. of bitstreams: 1 Ricardo_Kern.pdf: 242426 bytes, checksum: a174b7b70f1d6dd81756070f120c5387 (MD5) Previous issue date: 2006-02-23 / O constante aperfeiçoamento dos testes genéticos, como o Teste de Micronúcleos (MN), os tornaram importantes auxiliares na prevenção do câncer. Com o objetivo de melhor compreender a etiologia do desenvolvimento de neoplasias bucais, o presente trabalho se propôs a estudar a influência do fumo, álcool, trauma mecânico e de substância contidas em colutórios sobre a Freqüência de Micronúcleos (FMN) em células epiteliais bucais de alunos do curso de Odontologia. Para tanto, 40 alunos foram divididos em 4 grupos assim caracterizados G1 – Abstêmios (controle); G2 Alcoolistas; G3 – Usuários de aparelho ortodôntico; G4 – Fumantes alcoolistas, e submetidos ao TMN. Posteriormente estes mesmos alunos receberam de forma aleatória, 4 diferentes tratamentos, por 10 dias, a base de colutórios: T1 (óleos essenciais), T2 (álcool 11%), T3 (álcool, 11% + clorexidina 0,12%), T4 (clorexidina 0,12%), sendo então novamente submetidos ao TMN. Os resultados da FMN demonstraram diferenças significativas entre os 4 grupos G (p = 0,043 Kruskal-Wallis), sendo indicado diferença entre G1 e G3 (Mann-Whitney p ? 0,01) e entre G1 e G4 (Mann-Whitney p ? 0,05), contudo, em relação aos tratamentos com colutórios, todos os tratamentos T não alcançaram resultados significativos (Wilcoxon p > 0,05). Os resultados sugerem que o trauma mecânico causado por aparelho ortodôntico assim como a associação de fumo com bebidas alcoólicas favorecem um aumento na prevalência de MN quando comparados ao controle. A utilização de colutórios bucais, em curto prazo, não foi capaz de causar aumento na freqüência de Micronúcleos

Made available in DSpace on 2014-06-11T19:33:40Z (GMT). No. of bitstreams: 0 Previous issue date: 2006-04-19Bitstream added on 2014-06-13T19:23:56Z : No. of bitstreams: 1 sanches_j_dr_jabo.pdf: 1095068 bytes, checksum: ecb21f712a2b32ecca0a02942022c438 (MD5) / Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) / Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP) / O objetivo deste trabalho foi avaliar os efeitos de injúrias mecânicas, por impacto, compressão e corte, na fisiologia de abacates, através de análises físicas, químicas, bioquímicas e sensoriais, tomografia de ressonância magnética e microscopia eletrônica de varredura. O impacto foi aplicado nos lados opostos do fruto e provocados por queda livre de 2,00 m; a compressão, submetendo-os a pesos de 117,6 N, por 24 horas; e o corte, com quatro incisões longitudinais, com 80 mm de comprimento e 4 mm de profundidade, nos lados opostos do fruto. As diferentes lesões aplicadas aos abacates ocasionaram danos irreparáveis à qualidade dos frutos, onde os efeitos mais nocivos foram encontrados com os cortes. As imagens tomográficas não demonstraram a ocorrência de lesionamento interno em abacates 'Hass', enquanto que os frutos da 'Quintal', submetidos à compressão e ao impacto, apesar de não mostrarem lesões externas, as imagens indicaram a ocorrência de lesionamento interno e a evolução do mesmo durante o amadurecimento. A microscopia eletrônica de varredura mostrou que os tecidos da polpa de abacates 'Quintal' injuriados apresentaram desordem celular em suas estruturas e que eles podem ser protegidos por diferentes processos de cicatrização. / This work aimed to evaluate the effects of mechanical injuries, for impact, compression and cut, in the physiology of avocados, by physical, chemical, biochemical and sensorial analysis, and by use of magnetic resonance imaging and scanning electron microscopy. The impact was applied in the opposed sides of the fruit and provoked by free fall from 2.00 m; compression, submitting them to weights of 117.6 N, for 24 hours; and cut, with four longitudinal incisions, with 80 mm of length and 4 mm of depth, in the opposed sides of the fruit. The different injuries applied to the avocados caused irreparable damages to the quality of these fruits, where the most injurious effects were found for the cuts. The tomography images did not indicate the occurrence of internal injuries in 'Hass' avocados, while the fruits of the 'Quintal', submitted to the compression and the impact, although did not show external lesions, the images indicated the occurrence of internal injuries and evolution of them during the ripening. The scanning electron microscopy showed that 'Quintal' avocados tissues mechanically injured presented cellular disorder in their structures and that they can be protected by different cicatrization processes.

ATP is a (co)transmitter and signaling molecule in the CNS. It acts at a multitude of ligand-gated cationic channels termed P2X to induce rapid depolarization of the cell membrane. Within this receptor-channel family, the P2X7 receptor (R) allows the transmembrane fluxes of Na+, Ca2+, and K+, but also allows the slow permeation of larger organic molecules. This is supposed to cause necrosis by excessive Ca2+ influx, as well as depletion of intracellular ions and metabolites. Cell death may also occur by apoptosis due to the activation of the caspase enzymatic cascade. Because P2X7Rs are localized in the CNS preferentially on microglia, but also at a lower density on neuroglia (astrocytes, oligodendrocytes) the stimulation of this receptor leads to the release of neurodegeneration-inducing bioactive molecules such as pro-inflammatory cytokines, chemokines, proteases, reactive oxygen and nitrogen molecules, and the excitotoxic glutamate/ATP. Various neurodegenerative reactions of the brain/spinal cord following acute harmful events (mechanical CNS damage, ischemia, status epilepticus) or chronic neurodegenerative diseases (neuropathic pain, Alzheimer’s disease, Parkinson’s disease, multiple sclerosis, amyotrophic lateral sclerosis) lead to a massive release of ATP via the leaky plasma membrane of neural tissue. This causes cellular damage superimposed on the original consequences of neurodegeneration. Hence, blood-brain-barrier permeable pharmacological antagonists of P2X7Rs with excellent bioavailability are possible therapeutic agents for these diseases. The aim of this review article is to summarize our present state of knowledge on the involvement of P2X7R-mediated events in neurodegenerative illnesses endangering especially the life quality and duration of the aged human population.

O raquitismo das soqueiras (RSD), causado pela bactéria Leifsonia xyli subsp. xyli (Lxx) é uma das principais doenças na cana-de-açúcar, contudo, as informações sobre a biologia do patógeno, bem como sobre sua interação com cana são limitadas. Desta forma, três estudos foram conduzidos com o objetivo de verificar a interferência de fatores do hospedeiro e do ambiente na cinética populacional da Leifsonia xyli subsp. xyli (Lxx) in planta e in vitro. Num primeiro, foi quantificada, através de PCR em tempo real (qPCR), a colonização da bactéria de plantas de duas variedades submetidas a estresse por restrição hídrica e por injúria mecânica infligida através de corte da planta. Num segundo, o desenvolvimento in vitro da bactéria foi avaliado, por 10 dias, em resposta à adição ao meio de cultivo de fluido vascular de plantas de cana. Já o terceiro estudo avaliou o efeito do hormônio ácido abscísico (ABA) na cinética populacional de Lxx em cana. No primeiro estudo, os resultados apontaram efeito positivo tanto do estresse hídrico como da injúria no título bacteriano em plantas da variedade CB 49-260. Já na variedade RB83 5486, não houve efeito significativo destes agentes de estresse no título bacteriano. No segundo estudo, o maior desenvolvimento da população de Lxx ocorreu em meio de cultivo sem adição de fluido vascular, ao passo que o desenvolvimento da população em presença de fluido da variedade resistente não inoculada com Lxx foi maior, quando em presença de fluido de plantas da mesma variedade inoculadas com a bactéria. A adição de fluido vascular da variedade suscetível, por outro lado, inibiu substancialmente o desenvolvimento das culturas bacterianas. A análise cromatográfica do fluido vascular indicou diferenças na concentração de compostos do fluido das duas variedades, bem como entre plantas da mesma variedade inoculadas ou não com a bactéria. No terceiro estudo, apesar das variações nos títulos da Lxx detectados ao longo do tempo, tanto nas plantas testemunhas como nas plantas tratadas com ABA, não houve diferenças significativas (p>0,05) entre os tratamentos dentro de um período de avaliação de 4 meses. Em seu conjunto, os resultados, além de fornecerem informações inéditas sobre a interação entre Lxx e cana, ainda permitiram direcionar futuros estudos a fim de aprimorar o conhecimento sobre os mecanismos de colonização do hospedeiro desta bactéria. / Ratoon Stunting Disease (RSD) caused by the bacterium Leifsonia xyli subsp. xyli (Lxx) is a major disease in sugarcane; however, information on the biology of this pathogen and on its interaction with sugarcane is limited. Three studies were conducted to analyze effects of the host and environment on the population kinetics of Leifsonia xyli subsp. xyli (Lxx) in plant and in vitro. In the first treatment, we used real time PCR (qPCR) to quantify bacterial colonization in two sugarcane varieties subjected to water stress and mechanical injury. In the second treatment, we evaluated the in vitro growth of the bacteria for 10 days after the addition of 15% of sugarcane vascular fluid to the culture medium. The third treatment evaluated the effect of the application of abscisic acid (ABA) on Lxx colonization in sugarcane. In the first treatment, the results showed an increase in Lxx colonization in the variety CB 49-260 exposed to water stress and mechanical injury. For the variety RB83 5486, there was no significant effect of these stresses on the bacterial colonization. In the second treatment, the greatest development of Lxx population occurred in the culture medium without the addition of vascular fluid (control), while bacterial population in the presence of fluid from resistant variety non-inoculated with Lxx was greater, compared to the growth in the presence of plant fluids of the same variety inoculated with the bacteria. The addition of vascular fluid of the susceptible variety, on the other hand, significantly inhibited bacterial growth. The chromatographic analysis of the vascular fluid indicated differences in the compounds concentration in the fluid of the two varieties, as well as in plants of the same variety inoculated or not with the bacteria. In the third study, despite variations in Lxx colonization detected over time, the control plants and plants treated with ABA showed no significant differences (p> 0.05) among treatments during the trial period of four months. The results, in addition to providing new information about the interaction between Lxx and sugarcane, allowed to direct further studies on mechanisms of host colonization of this bacterium.

O raquitismo das soqueiras (RSD), causado pela bactéria Leifsonia xyli subsp. xyli (Lxx) é uma das principais doenças na cana-de-açúcar, contudo, as informações sobre a biologia do patógeno, bem como sobre sua interação com cana são limitadas. Desta forma, três estudos foram conduzidos com o objetivo de verificar a interferência de fatores do hospedeiro e do ambiente na cinética populacional da Leifsonia xyli subsp. xyli (Lxx) in planta e in vitro. Num primeiro, foi quantificada, através de PCR em tempo real (qPCR), a colonização da bactéria de plantas de duas variedades submetidas a estresse por restrição hídrica e por injúria mecânica infligida através de corte da planta. Num segundo, o desenvolvimento in vitro da bactéria foi avaliado, por 10 dias, em resposta à adição ao meio de cultivo de fluido vascular de plantas de cana. Já o terceiro estudo avaliou o efeito do hormônio ácido abscísico (ABA) na cinética populacional de Lxx em cana. No primeiro estudo, os resultados apontaram efeito positivo tanto do estresse hídrico como da injúria no título bacteriano em plantas da variedade CB 49-260. Já na variedade RB83 5486, não houve efeito significativo destes agentes de estresse no título bacteriano. No segundo estudo, o maior desenvolvimento da população de Lxx ocorreu em meio de cultivo sem adição de fluido vascular, ao passo que o desenvolvimento da população em presença de fluido da variedade resistente não inoculada com Lxx foi maior, quando em presença de fluido de plantas da mesma variedade inoculadas com a bactéria. A adição de fluido vascular da variedade suscetível, por outro lado, inibiu substancialmente o desenvolvimento das culturas bacterianas. A análise cromatográfica do fluido vascular indicou diferenças na concentração de compostos do fluido das duas variedades, bem como entre plantas da mesma variedade inoculadas ou não com a bactéria. No terceiro estudo, apesar das variações nos títulos da Lxx detectados ao longo do tempo, tanto nas plantas testemunhas como nas plantas tratadas com ABA, não houve diferenças significativas (p>0,05) entre os tratamentos dentro de um período de avaliação de 4 meses. Em seu conjunto, os resultados, além de fornecerem informações inéditas sobre a interação entre Lxx e cana, ainda permitiram direcionar futuros estudos a fim de aprimorar o conhecimento sobre os mecanismos de colonização do hospedeiro desta bactéria. / Ratoon Stunting Disease (RSD) caused by the bacterium Leifsonia xyli subsp. xyli (Lxx) is a major disease in sugarcane; however, information on the biology of this pathogen and on its interaction with sugarcane is limited. Three studies were conducted to analyze effects of the host and environment on the population kinetics of Leifsonia xyli subsp. xyli (Lxx) in plant and in vitro. In the first treatment, we used real time PCR (qPCR) to quantify bacterial colonization in two sugarcane varieties subjected to water stress and mechanical injury. In the second treatment, we evaluated the in vitro growth of the bacteria for 10 days after the addition of 15% of sugarcane vascular fluid to the culture medium. The third treatment evaluated the effect of the application of abscisic acid (ABA) on Lxx colonization in sugarcane. In the first treatment, the results showed an increase in Lxx colonization in the variety CB 49-260 exposed to water stress and mechanical injury. For the variety RB83 5486, there was no significant effect of these stresses on the bacterial colonization. In the second treatment, the greatest development of Lxx population occurred in the culture medium without the addition of vascular fluid (control), while bacterial population in the presence of fluid from resistant variety non-inoculated with Lxx was greater, compared to the growth in the presence of plant fluids of the same variety inoculated with the bacteria. The addition of vascular fluid of the susceptible variety, on the other hand, significantly inhibited bacterial growth. The chromatographic analysis of the vascular fluid indicated differences in the compounds concentration in the fluid of the two varieties, as well as in plants of the same variety inoculated or not with the bacteria. In the third study, despite variations in Lxx colonization detected over time, the control plants and plants treated with ABA showed no significant differences (p> 0.05) among treatments during the trial period of four months. The results, in addition to providing new information about the interaction between Lxx and sugarcane, allowed to direct further studies on mechanisms of host colonization of this bacterium.