An ethnopharmacological approach to multidrug -resistant Staphylococcus aureus: Evaluation of Italian plants used in the traditional healing of skin disease

One-third of botanical remedies from southern Italy are used to treat skin and soft tissue infections (SST's). Methicillin-resistant Staphylococcus aureus (MRSA), a common cause of SSTIs, is responsible for increased morbidity and mortality from infections. Therapeutic options are limited by antibiotic resistance. Many plants possess potent antimicrobial compounds for these disorders. Validation of traditional medical practices is important for the people who rely on medicinal plants. Moreover, identification of novel antibiotics and anti-pathogenic agents for MRSA is important to global healthcare.^ I took an ethnopharmacological approach to understand how Italian medicinal plants used for the treatment of SSTIs affect MRSA growth and virulence. My hypothesis was that plants used in folk remedies for SSTI would exhibit lower cytotoxicity and greater inhibition of bacterial growth, biofilm formation and toxin production in MRSA than plants used for remedies unrelated to the skin or for plants with no ethnomedical application. The field portion of my research was conducted in the Vulture-Alto Bradano area of southern Italy. I collected 104 plant species and created 168 crude extracts. In the lab, I screened samples for activity against MRSA in a battery of bioassays. Growth inhibition was analyzed using broth microtiter assays for determination of the minimum inhibitory concentration. Interference with quorum-sensing (QS) processes, which mediate pathogenicity, was quantified through RP-HPLC of δ-toxin production. Interference with biofilm formation and adherence was assessed using staining methods. The mammalian cytotoxicity of natural products was analyzed using MTT cell proliferation assay techniques.^ Although bacteriostatic activity was limited, extracts from six plants used in Italian folk medicine (Arundo donax, Ballota nigra, Juglans regia, Leopoldia comosa, Marrubium vulgare, and Rubus ulmifolius ) significantly inhibited biofilm formation and adherence. Moreover, plants used to treat SSTI demonstrated significantly greater anti-biofilm activity when compared to plants with no ethnomedical application. QSI activity was evident in 90% of the extracts tested and extracts from four plants ( Ballota nigra, Castanea saliva, Rosmarinus officinalis, and Sambucus ebulus) exhibited a significant dose-dependent response. Some of the plant remedies for SSTI identified in this study can be validated due to anti-MRSA activity.^

Simulating Forest Shade to Study the Developmental Ecology of Tropical Plants: Juvenile Growth in Three Vines in India

Both light quantity and quality affect the development and autoecology of plants under shade conditions, as in the understorey of tropical forests. However, little research has been directed towards the relative contributions of lowered photosynthetic photon flux density (PPFD) versus altered spectral distributions (as indicated by quantum ratios of 660 to 730 nm, or R:FR) of radiation underneath vegetation canopies. A method for constructing shade enclosures to study the contribution of these two variables is described. Three tropical leguminous vine species (Abrus precatorius L., Caesalpinia bondicela Fleming and Mucuna pruriens (L.) DC.) were grown in two shade enclosures with 3-4% of solar PPFD with either the R:FR of sunlight (1.10) or foliage shade (0.33), and compared to plants grown in sunlight. Most species treated with low R:FR differed from those treated with high R:FR in (1) percent allocation to dry leaf weight, (2) internode length, (3) dry stem weight/length, (4) specific leaf weight, (5) leaf size, and (6) chlorophyll a/b ratios. However, these plants did not differ in chlorophyll content per leaf dry weight or area. In most cases the effects of low R:FR and PPFD were additional to those of high R:FR and low PPFD. Growth patterns varied among the three species, but both low PPFD and diminished R:FR were important cues in their developmental responses to light environments. This shadehouse system should be useful in studying the effects of light on the developmental ecology of other tropical forest plants.

Age Differences among Female Sex Workers in the Philippines: Sexual Risk Negotiations and Perceived Manager Advice

Consistent condom use among high risk groups such as female sex workers (FSWs) remains low. Adolescent female sex workers are especially at higher risk for HIV/STI infections. However, few published studies have compared the sexual risk negotiations among adolescent, emerging adult, and older age groups or the extent a manager’s advice about condom use is associated with an FSW’s age. Of 1,388 female bar/spa workers surveyed in the southern Philippines, 791 FSW who traded sex in the past 6 months were included in multivariable logistic regression models. The oldest FSWs (aged 36–48) compared to adolescent FSWs (aged 14–17) were 3.3 times more likely to negotiate condoms when clients refused condom use. However, adolescent FSWs received more advice from their managers to convince clients to use condoms or else to refuse sex, compared to older FSWs. Both adolescent and the oldest FSWs had elevated sexually transmitted infections (STIs) and inconsistent condom use compared to other groups. Having a condom rule at the establishment was positively associated with condom negotiation. Factors such as age, the advice managers give to their workers, and the influence of a condom use rule at the establishment need to be considered when delivering HIV/STI prevention interventions.

Shade promotes thorn development in a tropical liana, Artabotrys hexapatalus (Annonaceae)

The liana Artabotrys hexapetalus (L.f.) Bhand., which is widely planted in the Tropics and native to African rain forests, produced new reiterations (new leader shoots) normally and after damage induced by Hurricane Andrew (August 24, 1992). In each new orthotropic shoot, there is a gradient in lateral branch structures from basal thorns, to vegetative leafy branches, to distal leafy flowering branches. We noted that reiterations developing in shade had more thorns than similar reiterations developing in full sun. Tents with clear (66% photosynthetically active radiation [PAR]) and shaded plastic film (12%–14% PAR) were placed over nodes when the axillary buds began to expand to produce reiteration shoots. After 2 mo of growth inside the tents and in the open, the types of lateral outgrowths (thorn vs. branch) were recorded. Shoots in spectrally neutral shade (red to far red of full sun) and spectrally altered shade (red to far red of canopy shade) produced significantly more thorns at the lower nodes of the shoots as compared to those in full sun. Shoots in control clear plastic tents were the same as those in full sun. We conclude that the fate of lateral bud development is controlled by irradiance (light level) but not by light quality. Increased thorn production in shade could be advantageous to plants growing in the deep shade of rain forests. Thorns in the self-shaded regions of the plant, and well below the forest canopy, could aid in protection from herbivory and in climbing by acting as hooks.

The effects of sex of teacher, sex of scenario principal, and leadership style on the performance ratings of scenario principals by teachers

This study was designed to address questions regarding the effects of sex and leadership style on teacher perceptions of principal effectiveness. On a researcher-designed instrument, middle school teachers rated the effectiveness of a scenario principal's response in several situations. The responses reflected varying levels of Task and Relationship Behavior.^ The design incorporated two between subjects factors (Teacher Sex and Principal Sex) and one within subjects factor (Leadership Style) which was treated as a repeated measure. An analysis of variance revealed no significant effects except for Leadership Style. Overall, High Task/High Relationship behavior rated significantly higher and Low Task/Low Relationship rated significantly lower than the others. The null hypothesis concerning differences could not be rejected and the stated research hypotheses were not supported.^ Additional analyses of variance were conducted substituting subject demographic variables for Teacher Sex in the research design. No significant interactions or main effects other than Leadership Style were noted when either Age or Ethnicity were substituted.^ A significant two-way interaction was noted for Teacher Experience and Leadership Style (p =.0316). Less experienced teachers rated principal's performance lower when exhibiting High Task/Low Relationship style than did more experienced teachers. A significant three-way interaction was noted for Administrative Aspiration x Principal Sex x Leadership Style (p =.0294). Teachers who indicated an intent to enter administration differed more on their ratings between male and female principals exhibiting mixed styles of High Task/Low Relationship and Low Task/High Relationship than did teachers who indicated no or undecided.^ Sex of the teacher appears less important than sex of the principal on performance ratings. Results suggest further study of the effects of teacher experience and teacher administrative aspiration on perceptions of principal effectiveness. ^

Biogeochemical factors affecting the fate and transport of mercury in the terrestrial environment at Oak Ridge, Tennessee

Mercury (Hg) contamination problem in the United Sates has been an important issue due to its potential threat to human and ecological health. This thesis presents a study of two Hg-contaminated sites along the East Fork Poplar Creek (EFPC) at Oak Ridge. The top soils from the terrestrial areas, along with the soils from three vertical soil horizons at the EFPC bank were sampled and analyzed for total-Hg (THg), methyl-Hg, total-organic-carbon (TOC), and pH. The stream bank soils were also analyzed for the stable-Hg-isotopes (198Hg, 199Hg, 200Hg, 201Hg, and 202Hg). Furthermore, some of the soil samples (n=7) from the same study sites were investigated for phytoavailability of mercury as measured by degree of Hg translocation in aboveground biomass of Impatiens walleriana plants grown in the soils.^ The results showed a significant difference (p<0.001) in THg concentrations for the forest soils (42.40±4.98 mg/kg) and the grassland soils (8.71±2.30 mg/kg). The higher THg and methyl-Hg concentrations were commensurate with the higher TOC content in the soils (p<0.001). Also, the THg concentrations for the upstream site was higher (129.08±34.14 mg/kg) than the downstream site (24.31±3.47 mg/kg). The two sites also differed in their stable Hg isotope compositions (p<0.001 for δ199Hg). The stable isotope analysis indicated the increased level of mass dependent isotopic fractionation with increasing depths along the EFPC bank. The difference between the two study sites was also prominent in case of the Hg uptake by the plants, with higher Hg uptake from the upstream soils compared to that from the downstream soils. A significant correlation, r=0.93 p<0.01, was observed between the Hg uptake and the soil-THg concentrations. THg was higher in the leaves (1161.87±310.01 μg/kg) than in the flowers (206.13±55.23 μg/kg) or the stems (634.54±403.35μg/kg). ^ The level of Hg contamination increased with decreasing distance from the point source and was highly influenced by plants/microbes, soil-organic-content, and Hg-speciation. The isotopic study indicated the existence of an additional Hg source in the EFPC watershed, possibly atmospheric Hg-deposition. These findings are worth taking into account while planning any Hg remediation effort and developing Hg loading criteria as per the National Pollutant Discharge Elimination System (NPDES) Program.^

Shoot architecture and growth in the south Florida wet prairie macrophyte, Eleocharis cellulosa

Eleocharis cellulosa is a dominant macrophyte in Everglades wet prairie communities. The development of the shoot system in the genus has been described as sympodial but with an unusual adnation of the horizontal and vertical shoots. The growth pattern of E. cellulosa was studied from field collected plants and plants grown in the greenhouse. Plants were extracted and measurements of horizontal and vertical shoot were taken. Dissections, paraffin sectioning and SEM's were used to examine shoot structure in early developmental stages. E. cellulosa was transplanted from the field to the greenhouse and different levels of Nitrogen and Phosphorus were added to determine how it responded phenotypically. Dissections and microscopy showed that growth of the vertical shoots of E. cellulosa is sympodial, while growth of the horizontal shoots is mixed, beginning monopodially then transforming to sympodial growth. Additions of nutrients did not have any effect on the morphology of E. cellulosa.

The effects of sex of teacher, sex of scenario principal and leadership style on the performance ratings of scenario principals by teachers

This study was designed to address questions regarding the effects of sex and leadership style on teacher perceptions of principal effectiveness. On a researcher-designed instrument, middle school teachers rated the effectiveness of a scenario principal's response in several situations. The responses reflected varying levels of Task and Relationship Behavior. The design incorporated two between subjects factors (Teacher Sex and Principal Sex) and one within subjects factor (Leadership Style) which was treated as a repeated measure. An analysis of variance revealed no significant effects except for Leadership Style. Overall, High Task/High Relationship behavior rated significantly higher and Low Task/Low Relationship rated significantly lower than the others. The null hypothesis concerning differences could not be rejected and the stated research hypotheses were not supported. Additional analyses of variance were conducted substituting subject demographic variables for Teacher Sex in the research design. No significant interactions or main effects other than Leadership Style were noted when either Age or Ethnicity were substituted. A significant two-way interaction was noted for Teacher Experience and Leadership Style (p = .0316). Less experienced teachers rated principal's performance lower when exhibiting High Task/Low Relationship style than did more experienced teachers. A significant three-way interaction was noted for Administrative Aspiration x Principal Sex x Leadership Style (p = .0294). Teachers who indicated an intent to enter administration differed more on their ratings between male and female principals exhibiting mixed styles of High Task/Low Relationship and Low Task/High Relationship than did teachers who indicated no or undecided. Sex of the teacher appears less important than sex of the principal on performance ratings. Results suggest further study of the effects of teacher experience and teacher administrative aspiration on perceptions of principal effectiveness.

The developmental responses of papaya leaves to simulated canopy shade

The developmental responses of plants to shade underneath foliage are influenced by reductions in irradiance and shifts in spectral quality (characterized by reductions in the quantum ratio of red to far-red wavelengths, R:FR). Previous research on the influence of shadelight on leaf development has neglected the reductions in R:FR characteristic of foliage shade, and these studies have almost certainly underestimated the extent and array of developmental responses to foliage shade. We have studied the effects of reduced irradiance and R:FR on the leaf development of papaya (Carica papaya L., Caricaceae). Using experimental shadehouses, replicates of plants grown in high light conditions (0.20 of sunlight and R:FR = 0.90) were compared to low light conditions (0.02 of sunlight) with either the spectral quality of sunlight (R:FR = 0.99) or of foliage shade (F:FR = 0.26). Although many characteristics, such as leaf thickness, specific leaf weight, stomatal density, palisade parenchyma cell shape, and the ratio of mesophyll air surface/leaf surface were affected by reductions in irradiance, reduced R:FR contributed to further changes. Some characters, such as reduced chlorophyll a/b ratios, reduced lobing, and greater internode length, were affected primarily by low R:FR. The reduced R:FR of foliage shade, presumably affecting phytochrome equilibrium, strongly influences the morphology and anatomy of papaya leaves.

Pollination, Herbivory, and Habitat Fragmentation: Their Effects on the Reproductive Fitness of Angadenia berteroi, a Native Perennial Plant of the South Florida Pine Rocklands

Angadenia berteroi is a tropical perennial subshrub of the pine rocklands with large yellow flowers that set very few fruits. My dissertation seeks to elucidate the factors that affect the reproductive fitness of Angadenia berteroi a native species of the south Florida pine rocklands. I provide novel information on the pollination biology of this native species. I also assess the effects of herbivory on growth and the reproductive success of A. berteroi. Finally, I elucidate how habitat fragmentation and quality are correlated with reproductive fitness of this native perennial plant. Using a novel experimental approach, I determined the most effective pollinator group. I used nylon fishing line of widths corresponding to proboscis diameter of the major groups of visitors to examine pollen removal and deposition. In the field, I estimated visitation frequency and efficacy of each pollinator type. Using potted plants, I exposed flowers to single visit from different types of pollinators to measure fruit set. I performed artificial defoliation with scissors on plants growing in the greenhouse to assess the effects of defoliation before flowering as well as during flowering. Additionally, I used structural equation modelling (SEM) to elucidate how A. berteroi reproductive fitness was affected by habitat fragmentation and quality. My experiments provide evidence that Angadenia berteroi is specialized for bee pollination; though butterflies, skippers and others also visit its flowers, A. berteroi is exclusively pollinated by two native bees of the South Florida pine rocklands . This research also demonstrated that herbivory by the oleander moth may have direct and indirect effects on Angadenia berteroi growth and reproductive success. The SEM results suggested that habitat quality (litter depth and subcanopy cover) may favor reproduction in native species of the South Florida pine rocklands that are properly maintained by periodic fires and exotic control. Insights from this threatened and charismatic species may provide impetus to properly manage remaining pine rocklands in South Florida for this and other endemic understory species.