Populations of an unknown species of Mimulus, herein described as Mimulus hymenophyllus Meinke, were located and studied near Horse Creek, a tributary of the Snake River in eastern Wallowa County, Oregon. The new species is differentiated from its apparent closest relative, Mimulus jungermannioides Suksd., by an annual habit, long petioles, corollas that are three to four times the length of the calyx, rounded capsules, and seeds up to 0.85 mm long. Mimulus hymenophyllus and M. iungermannioides are members of the section Paradanthus, and are morphologically allied with Mimulus moschatus Dougl. in Lindl. and several related taxa. The Mimulus moschatus complex of species is distinguished within the genus by having firmly adherent placentae, glandular-sticky foliage and stems, prismatic calyces with equal to sub-equal teeth, elongate pedicels, and deciduous, funnelform corollas ranging in color from canary to lemon yellow. Mimulus hvmenophyllus is endemic to the steep canyons of Horse and Cow Creeks in Wallowa County, at altitudes ranging from 850 to 1300 m. The species is restricted to the damp crevices of vertical basalt cliffs, often growing in the shade under overhangs. The surrounding plant community is dominated by several coniferous species, primarily Pseudotsuga menziesii, pinus ponderosa, and Abies arandis. The habitat of M. hymenophyllus is isolated, and is presently not in danger from human disturbance. However, the species is currently maintained as a state and federal candidate for listing as threatened or endangered, based on limited abundance and narrow geographic range. Studies of the seed and seedling biology of Mimulus hymenophyllus and its Pacific Northwest relatives (i.e., M. moschatus, M. floribundus, M. jungermannioides, M. patulus, M. pulsiferae, M. washingtonensis, and M. breviflorus) were also conducted. Germination trials indicated that seeds of M. moschatus, M. jungermannioides, and M. floribundus are capable of germination immediately or soon after capsules dehisce, and do not become dormant upon later exposure to warm or cold temperatures. Seed lots of M. hymenophyllus and M. breviflorus germinate rapidly when first mature, but subsequently develop complete or partial dormancy after prolonged exposure to late summer temperatures. Dormant seeds of these species, as well as those of M. patulus, M. Dulsiferae, and M. washingtonensis which are dormant when capsules dehisce, will germinate readily after several weeks of cold-wet stratification. Four of the five species lacking innate seed dormancy develop inflated fruiting calyces that temporarily trap seeds as capsules dehisce. This prolongs the dispersal rate of seed crops, thereby minimizing the potential for catastrophic seedling mortality due to mass germination of cohorts during unfavorable environmental conditions. These species retain seed on the parent plant for significantly longer periods than species with unmodified calyces. Seed dispersal from parent plants is accomplished by wind and flowing water, except in M. hvmenophvllus, where negatively phototropic pedicels orient ripe capsules towards the darkened cliff substrate for dispersal directly into crevices. Two of the rarest species studied, M. hymenophyllus and M. patulus, were found to co-occur in nature with an indigenous and rather weedy, unrelated species of Mimulus, i.e., M. nasutus Greene. Experimental plantings showed that seedling lots of M. nasutus emerge sooner and over a shorter period than those of the two uncommon species, suggesting that M. nasutus may outcompete them and thereby contribute to their rarity. Seedlings of M. breviflorus and M. floribundus, which typically occur in habitats subject to sudden fluctuations in soil moisture, exhibit adaptations for accelerated sexual reproduction under experimental drought conditions. This trait, when coupled with inflated fruiting calyces, enables these species to exploit unpredictably arid environments despite the potential disadvantage of nondormant seeds. Mimulus jungermannioides and M. moschatus are not adapted for precocious sexual maturation, and offset their lack of seed dormancy by having a perennial life cycle and by occurring in more predictably mesic habitats. Significant differences between the annual and perennial species were observed in the timing and amount of seed production during droughtstress. The information provided in this thesis will be valuable to systematists and conservation biologists. Several of the species studied have been promoted for protection under endangered species laws. The formal naming of Mimulus hymenophyllus legitimizes efforts for the management of this unique species, while seed and seedling biology data will be important to germplasm studies, phylogenetic assessments, and possible reintroduction of species or populations should extirpation occur. / Graduation date: 1991

Thesis (Ph. D.)--Ohio State University, 2003. / Title from first page of PDF file. Document formatted into pages; contains xiv, 119 p.; also includes graphics (some col.). Includes abstract and vita. Advisor: Andrea D. Wolfe, Dept. of Evolution, Ecology, and Organismal Biology. Includes bibliographical references (p. 109-119).

Inaug.-Diss.--Zürich. / Includes bibliographical references.

No description available.

Thesis (Ph. D.)--University of Oklahoma, 1995. / Includes bibliographical references (212-216).

Thesis (Ph. D.)--University of Wisconsin--Madison, 1971. / Typescript. Vita. Includes bibliography.

Graduation date: 2000

Thesis (doctoral)--Universität Zürich, 1906. / Includes bibliographical references (p. [59]-61).

A biflorina é uma o-naftoquinona que apresenta uma variedade de atividades biológicas, entre as quais pode-se destacar a atividade antitumoral. Esta molécula é obtida a partir das raízes da planta Capraria biflora L.(Schrophulariaceae) originária das Antilhas e América do Sul e habita zonas temperada e tropical. No continente europeu é muito usada como ornamento. No Brasil, pode ser encontrada nos Estados de Goiás, Minas Gerais e na faixa litorânea entre o Piauí até o Espírito Santo. O presente trabalho traz informações referentes aos primeiros estudos de caracterização estrutural até os mais recentes relatos sobre a atividade biológica desta molécula. Neste trabalho avaliamos os efeitos citotóxicos e detecção de apoptose tardia por análise in situ de células tratadas com a biflorina. Para isso utilizamos a linhagem tumoral HeLa e a não tumoral HEK-293 tratadas com biflorina nas concentrações de 5 - 50 μg/mL por 24h, 48h e 72h. A citotoxicidade foi avaliada exclusivamente para o tratamento de 48h em seis linhagens celulares diferentes sendo elas: Hep-2, HeLa, HT-29, A-375, A-549 e HEK-293. Os resultados mostraram citotoxicidade seletiva da biflorina contra a linhagem não tumoral HEK-293 (IC50 = 56,01 ± 1,17 μg/mL) comparada com todas as linhagens de células tumorais analisadas, com IC50 variando de 29,44 ± 1,32 μg/mL até 47,37 ± 3,21 μg/mL. Modificações morfológicas em células HeLa foram observadas após o tratamento de 48 horas com biflorina de acordo com o aumento da concentração (5-50 μg/mL). Além disso, na maior parte dos casos, observou-se um incremento do estágio de apoptose tardia, na análise in situ da imunocoloração de anexina V de todas as linhagens (Hep-2, HeLa, HT-29, A-375, A-549 e HEK-293) após tratamento com a biflorina. A apoptose tardia para HEK-293 (77,69 ± 6,68%) foi mais evidente em concentrações mais elevadas de biflorina em comparação com as linhagens tumorais testadas. Os resultados indicam que a biflorina mostrou uma importante citotoxicidade contra linhagens de células tumorais. No entanto, mais estudos são necessários para entender melhor os mecanismos envolvidos na citotoxicidade e morte celular programada. / Submitted by Marcelo Teixeira (mvteixeira@ucs.br) on 2014-06-16T12:51:14Z No. of bitstreams: 1 Dissertacao Gabrielle Gianna N. S. Wisintainer.pdf: 1578186 bytes, checksum: 77fe847912ace71147729d4182942bf2 (MD5) / Made available in DSpace on 2014-06-16T12:51:14Z (GMT). No. of bitstreams: 1 Dissertacao Gabrielle Gianna N. S. Wisintainer.pdf: 1578186 bytes, checksum: 77fe847912ace71147729d4182942bf2 (MD5) / Biflorin is an o-naphthoquinone with proven cytotoxic effects on tumor cells with antimicrobial, antitumor and antimutagenic activities. Biflorin is an isolated compound taken from the roots of the plant Capraria biflora L. (Schrophulariaceae), indigenous of the West Indies and South America, that inhabits temperate and tropical areas. In the European continent, this plant is used as an ornament. In Brazil, it can be found in the states of Goiás, Minas Gerais and the coastal strip between Piauí in the Northeast, and it extends to the states of Ceará, Pernambuco, Pará and Espírito Santo. In this work, we report the information related to the first structural characterization studies, as well as the latest reports concerning the biological activity of this molecule. It was work verified the cytotoxic effects of biflorin and detection of late apoptosis by in situ analysis. Initially, tumor HeLa and non-tumor HEK-293 cells were treated with biflorin for 24h, 48h and 72h at a range of 5-50 μg/mL. The cytotoxicity was further evaluated exclusively for 48h treatment on six different cell lines Hep-2, HeLa, HT-29, A-375, A-549 and HEK-293. The results indicate that biflorin showed selective cytotoxicity against non-tumor line HEK-293 (IC50 = 56,01 ± 1,17 μg/mL) compared to all tumor cells analyzed in a concentration dependent manner, with IC50 ranging from 29.44 ± 1.32 μg/mL to 47,37 ± 3,21 μg/mL. Substantial morphological changes in HeLa cells were observed after 48h treatment with biflorin with increased concentrations (5- 50 μg/mL) of extract. In addition, in situ immunostaining of annexin V showed that all lines were majority seen at late apoptotic stages in a dose-dependent manner. Late apoptosis for HEK-293 was more evident (77.69 ± 6.68%) at higher extract concentrations compared to all tumor lines tested. The data here presented indicate that biflorin showed an important cytotoxicity against tumor cell lines. However, more studies are needed to better understand the pathways involved in programmed cell death.

A biflorina é uma o-naftoquinona que apresenta uma variedade de atividades biológicas, entre as quais pode-se destacar a atividade antitumoral. Esta molécula é obtida a partir das raízes da planta Capraria biflora L.(Schrophulariaceae) originária das Antilhas e América do Sul e habita zonas temperada e tropical. No continente europeu é muito usada como ornamento. No Brasil, pode ser encontrada nos Estados de Goiás, Minas Gerais e na faixa litorânea entre o Piauí até o Espírito Santo. O presente trabalho traz informações referentes aos primeiros estudos de caracterização estrutural até os mais recentes relatos sobre a atividade biológica desta molécula. Neste trabalho avaliamos os efeitos citotóxicos e detecção de apoptose tardia por análise in situ de células tratadas com a biflorina. Para isso utilizamos a linhagem tumoral HeLa e a não tumoral HEK-293 tratadas com biflorina nas concentrações de 5 - 50 μg/mL por 24h, 48h e 72h. A citotoxicidade foi avaliada exclusivamente para o tratamento de 48h em seis linhagens celulares diferentes sendo elas: Hep-2, HeLa, HT-29, A-375, A-549 e HEK-293. Os resultados mostraram citotoxicidade seletiva da biflorina contra a linhagem não tumoral HEK-293 (IC50 = 56,01 ± 1,17 μg/mL) comparada com todas as linhagens de células tumorais analisadas, com IC50 variando de 29,44 ± 1,32 μg/mL até 47,37 ± 3,21 μg/mL. Modificações morfológicas em células HeLa foram observadas após o tratamento de 48 horas com biflorina de acordo com o aumento da concentração (5-50 μg/mL). Além disso, na maior parte dos casos, observou-se um incremento do estágio de apoptose tardia, na análise in situ da imunocoloração de anexina V de todas as linhagens (Hep-2, HeLa, HT-29, A-375, A-549 e HEK-293) após tratamento com a biflorina. A apoptose tardia para HEK-293 (77,69 ± 6,68%) foi mais evidente em concentrações mais elevadas de biflorina em comparação com as linhagens tumorais testadas. Os resultados indicam que a biflorina mostrou uma importante citotoxicidade contra linhagens de células tumorais. No entanto, mais estudos são necessários para entender melhor os mecanismos envolvidos na citotoxicidade e morte celular programada. / Biflorin is an o-naphthoquinone with proven cytotoxic effects on tumor cells with antimicrobial, antitumor and antimutagenic activities. Biflorin is an isolated compound taken from the roots of the plant Capraria biflora L. (Schrophulariaceae), indigenous of the West Indies and South America, that inhabits temperate and tropical areas. In the European continent, this plant is used as an ornament. In Brazil, it can be found in the states of Goiás, Minas Gerais and the coastal strip between Piauí in the Northeast, and it extends to the states of Ceará, Pernambuco, Pará and Espírito Santo. In this work, we report the information related to the first structural characterization studies, as well as the latest reports concerning the biological activity of this molecule. It was work verified the cytotoxic effects of biflorin and detection of late apoptosis by in situ analysis. Initially, tumor HeLa and non-tumor HEK-293 cells were treated with biflorin for 24h, 48h and 72h at a range of 5-50 μg/mL. The cytotoxicity was further evaluated exclusively for 48h treatment on six different cell lines Hep-2, HeLa, HT-29, A-375, A-549 and HEK-293. The results indicate that biflorin showed selective cytotoxicity against non-tumor line HEK-293 (IC50 = 56,01 ± 1,17 μg/mL) compared to all tumor cells analyzed in a concentration dependent manner, with IC50 ranging from 29.44 ± 1.32 μg/mL to 47,37 ± 3,21 μg/mL. Substantial morphological changes in HeLa cells were observed after 48h treatment with biflorin with increased concentrations (5- 50 μg/mL) of extract. In addition, in situ immunostaining of annexin V showed that all lines were majority seen at late apoptotic stages in a dose-dependent manner. Late apoptosis for HEK-293 was more evident (77.69 ± 6.68%) at higher extract concentrations compared to all tumor lines tested. The data here presented indicate that biflorin showed an important cytotoxicity against tumor cell lines. However, more studies are needed to better understand the pathways involved in programmed cell death.