Fibroblast Growth Factor 8 and its Receptors in The Growth and Angiogenesis of Experimental Breast Cancer . With Special Reference to Thrombospondin-1 as a Novel Target Gene for FGF-8

The growth of breast cancer is regulated by hormones and growth factors. Recently, aberrant fibroblast growth factor (FGF) signalling has been strongly implicated in promoting the progression of breast cancer and is thought to have a role in the development of endocrine resistant disease. FGFs mediate their auto- and paracrine signals through binding to FGF receptors 1-4 (FGFR1-4) and their isoforms. Specific targets of FGFs in breast cancer cells and the differential role of FGFRs, however, are poorly described. FGF-8 is expressed at elevated levels in breast cancer, and it has been shown to act as an angiogenic, growth promoting factor in experimental models of breast cancer. Furthermore, it plays an important role in mediating androgen effects in prostate cancer and in some breast cancer cell lines. We aimed to study testosterone (Te) and FGF-8 regulated genes in Shionogi 115 (S115) breast cancer cells, characterise FGF-8 activated intracellular signalling pathways and clarify the role of FGFR1, -2 and -3 in these cells. Thrombospondin-1 (TSP-1), an endogenous inhibitor of angiogenesis, was recognised as a Te and FGF-8 regulated gene. Te repression of TSP-1 was androgen receptor (AR)-dependent. It required de novo protein synthesis, but it was independent of FGF-8 expression. FGF-8, in turn, downregulated TSP-1 transcription by activating the ERK and PI3K pathways, and the effect could be reversed by specific kinase inhibitors. Differential FGFR1-3 action was studied by silencing each receptor by shRNA expression in S115 cells. FGFR1 expression was a prerequisite for the growth of S115 tumours, whereas FGFR2 expression alone was not able to promote tumour growth. High FGFR1 expression led to a growth advantage that was associated with strong ERK activation, increased angiogenesis and reduced apoptosis, and all of these effects could be reversed by an FGFR inhibitor. Taken together, the results of this thesis show that FGF-8 and FGFRs contribute strongly to the regulation of the growth and angiogenesis of experimental breast cancer and support the evidence for FGF-FGFR signalling as one of the major players in breast cancers.

Net melon resistance to Didymella bryoniae according to grafting and potassium levels

Chemical control of the fungus Didymella bryoniae, the causal agent of the disease gummy stem blight in melon, is frequently inefficient; thus, alternatives such as grafting and nutrition must be studied. Rootstocks and potassium levels were tested aimed at controlling this disease in net melon under protected environment. The melon hybrid 'Bônus II', ungrafted and grafted onto 'Dinero' melon and 'Strong Tosa' pumpkin rootstocks, was cultivated and inoculated by using the toothpick insertion method with 7-mm mycelial disks from the isolate D. bryoniae Dbr 37; for control, only toothpick insertion was used. The plants were subjected to the following potassium levels: 0, 62.5, 125, 187.5, 250 mg L-1. Grafted 'Bônus II' melon plants were resistant to the fungus, whereas ungrafted ones were susceptible. The adopted potassium levels did not influence the stem lesion size or the survival of plants.

Effects of different light levels on the initial growth and photosynthesis of Croton urucurana Baill. in southeastern Brazil

Four levels of shading (full sunlight (0%), 30, 50, 70% of solar radiation interception on growth, chlorophyll concentration and photosynthetic rate were studied in Croton urucurana Baill., a pioneer plant species. This species seedlings are of potential interest for reforestation projects and recovery of degraded areas. The seedlings were grown in pots containing soil and sand (2:1) and later transferred to plastic bags of 3 dm³ and submitted to different levels of shading (30, 50, 70%) of solar radiation and full sunlight, as control. The experimental design was completely randomized with five replicates and each experimental unit was composed of five plants. The results suggest that plants submitted to 70% shading showed higher dry weight accumulation in leaf and root, and higher plant height and leaf area. However, the seedlings root system showed higher dry biomass under full sunlight. It was observed a tendency to increase chlorophyll concentration and to decrease photosynthetic rate with the increase of the shading level.

Chemical Biology Screen for Prostate Cancer Therapeutics

Prostate cancer initially responds to hormone-based therapeutics such as anti-androgen treatment or chemotherapeutics but eventually becomes resistant. Novel treatment options are therefore urgently needed. This thesis study applied a high-throughput screen of 4910 known drugs and drug-like small molecules to identify compounds that selectively inhibit growth of prostate cancer cells. In addition, the mechanisms underlying the cellular sensitivity to potent cancer selective compounds were addressed. Surprisingly, many of the compounds currently used in the clinics or studied in clinical trials were not cancer-selective. Only four drugs, aldehyde dehydrogenase inhibitor disulfiram (Antabus), antibiotic ionophore monensin, histone deacetylase inhibitor tricostatin A and fungicide thiram inhibited prostate cancer cell growth at nanomolar concentrations without major effects on non-malignant prostate epithelial cells. Disulfiram, monensin and a structurally similar compound to monensin, salinomycin, induced oxidative stress and inhibited aldehyde dehydrogenase activity. Moreover, monensin and salinomycin reduced androgen receptor signalling and steroidogenesis, enforced cell differentiation and reduced the overall levels of cancer stem cells. Taken together, novel and potentially prostate cancer-selective therapeutic agents were identified in this study, including the description of a multitude of intoxicating mechanisms such as those relating to oxidative stress. The results provide novel insights into prostate cancer biology and exemplify useful means of considering novel approaches to cancer treatment.

Effects of thermal environment and supplementation levels on the physiological parameters of moxotó goats in confined and semi-confined rising systems

The objective of this work was to determine the effect of environmental variables and supplementation levels on physiological parameters of Moxotó goats in confined and semi-confined rising systems, in the Brazilian semi-arid region. The semi-confined individuals were kept on a grass based diet during the day and arrested in the end of the afternoon. The confined animals were kept in a management center, receiving two diets composed by forage cactus and maniçoba hay into two different levels (0.5 and 1.5% of the body weight). Inside the management center and in the external environment the environmental comfort parameters were set high during the afternoon period characterizing a situation of thermal discomfort for the animals. During the morning the semi-confined animals presented an average respiratory frequency (69.5 mov min-1) and rectal temperature (39.5 ºC) higher than the confined ones (62.6 mov min-1 and 39.0 ºC, respectively). The confined and semi-confined animals were able to maintain their rectal temperature within normal limits, with increase in the cardiac beatings rate and respiratory frequency. The greater percentage of the used supplementations (1.5%) seemed to increase rectal temperature in the two studied rising systems.

Response of young 'Tahiti' lime trees to different irrigation levels

The aim of this study was to evaluate the effect of different irrigation levels on canopy and root growth, productivity, and fruit quality of young ‘Tahiti’ acid lime trees. The experiment was installed in Piracicaba, Brazil in a 1.0-ha orchard plot with ‘Tahiti’ acid lime trees, grafted on ‘Swingle’ citrumelo rootstock and carried out from August of 2002 to May 2005. Each treatment was assigned to a drip irrigation level, based on ETc as follows: T1) non-irrigated, T2) 25%, T3) 50%, T4) 75% and T5) 100% of ETc determined by weighing lysimeter presented in the orchard plot. Trunk diameter and tree height were evaluated monthly. The roots were evaluated when the trees were 30 and 48 months old. The yield and fruit quality was evaluated in 2004 and 2005. The results showed that irrigation did not influence root distribution in depth, and trees irrigated with 75% and 100% ETc showed horizontal root distribution concentrated until 0.6 m from the trunk. Irrigation did not improve the quality of fruit. Yield increased in all irrigated treatment, but the most efficient yield mean per unit of water applied was the 25% ETc treatment.

Establishment of Xaraés and Marandu grasses under levels of soil compaction

In Cerrado soils under grazing, changes occur in physical attributes, such as increased density, decreasing on the size of water stable aggregates, and macroporosity reduction. Thus, the aim of this study was to study the effect of compaction on the establishment of two forages. It was adopted a completely randomized design with three replications, in 2 x 4 factorial design, and two forages (Xaraés grass and Marandu grass), and four levels of compaction (soil densities of 1.0, 1.2, 1.4, and 1.6 Mg m-3). The following variables were evaluated 48 days after sowing: tiller population, plant height, dry matter production of shoots and components, leaf and stem, as well as the root dry mass. The stem dry mass decreased with soil density in a similar manner for both forages. It was observed that the leaf dry mass and shoots dry mass of Xaraés grass remained constant in the levels of soil compaction, not adjusting to any regression model. The establishment of Xaraés grass has not been negatively affected by compaction, which may be suitable for situations where there may be layers that restrict the growth of different forages.

Jatropha seed oil content and yield under different irrigation and potassium fertilization levels

The aim of this research was to evaluate the effects of different irrigation depths and potassium dosages, of Jatropha seed oil content and yield. The experimental design used was randomized blocks, in split-plots, with four replicates. The treatments were four water depths (plots) and four potassium dosages (subplots) applied through irrigation water. The water depths were applied based on the percentage of accumulated evaporation of a Class A (ECA) tank and of rainfall, as following: L0 = without irrigation, L40, L80 and L120, representing 40, 80 and 120% of the balance, respectively. The potassium dosages were K30, K60, K90 and K120 (30, 60, 90 and 120kg ha-1 of potassium, respectively). The oil extraction of samples was done through chemical extraction by organic solvent. The seeds used in this test were from the sampling of two seed productions from 2009, second year of crop production. It was possible to observe that irrigation use increased oil yield and decreased the oil content of Jatropha seed. Potassium fertilization did not influence oil content and yield. There was a relative increase of efficiency in water use producing oil until certain water depth, and after that there was a decrease.

Energetic analysis of landrace varieties and hybrids of corn produced in different technological levels of management

The aim of this study was to analyze, under the energetic point of view, the cultivation of corn in three management systems (low, medium and high-tech), using two landrace varieties ('Argentino' and 'BR da Várzea'), a double hybrid cultivar (SHS 4080) and a simple hybrid (IAC 8333). Five performance indicators were used: energy efficiency, liquid cultural energy, cultural efficiency, energy balance and productive energy efficiency. From the perspective of family farming, it was verified the largest social importance of the systems under low and medium levels of technology, due to the increase employment capacity of rural labor. The liquid cultural energy and energy balance were more favorable for the system under high technological level, unlike cultural efficiency and productive energy efficiency, which were significantly higher for medium and low technological levels. The variety 'Argentino' showed lower productive energy efficiency. The variety 'BR da Várzea', on the other hand, presented the potential to generate energy as much as the hybrids. In general, the biggest sustainability in the corn crop was achieved when the management system under medium and lower levels of technology were used.

Economic analysis of cashew early dwarf Crop BRS - 189 depending on water levels and doses of potassium fertilization

Water and fertilizer among the production factors are the elements that most restrict the production of cashew. The precise amount of these factors is essential to the success of the crop yield. This research aimed to determine the best factor-product ratio and analyze technical and economic indicators, of productivity of the cashew clone BRS 189 (Anacardium occidentale) to production factors water and potassium. The experiment was conducted from May 2009 to December 2009 in an experimental area of 56.0 m x 112.0 m in the irrigated Curu - Pentecoste, located in the municipality of Pentecoste, Ceará, Brazil. Production factors water (W) and potassium (K) were the independent variables and productivity (Y), the dependent variable. Ten statistical models that have proven satisfactory for obtaining production function were tested. The marginal rate of substitution was obtained through the ratio of the potassium marginal physical product and the water marginal physical product. The most suited model to the conditions of the experiment was the quadratic polynomial without intercept and interaction. Considering that the price of the water was 0.10 R$ mm -1, the price of the potassium 2.19 R$ kg -1 and the price of the cashew 0.60 R$ kg-1, the amounts of water and K2O to obtain the maximum net income were 6,349.1 L plant-1 of water and 128.7 g plant -1year, -1 respectively. Substituting the values obtained in the production function, the maximum net income was achieved with a yield of 7,496.8 kg ha-1 of cashew.

Germination and early growth of physic nut submitted to levels of salinity

Two experiments were conducted to evaluate the effect of salinity on early physic nut plant development. In the first trial, physic nut seeds were exposed to seven levels of salinity (0, 2, 4, 6, 8, 10 and 12dS m-1) with eight repetitions, using a substrate of paper soaked with solutions of CaCl2 and KCl. The treatments were evaluated based on the initial germination, total percentage of germination, and time necessary to germination of 50% of the seeds. Increased salinity reduced the first germination count and delayed the time to 50% germination. From 10dS m-1, there was a reduction in germination percentage. The second trial evaluated the effect of salinity on the growth of physic nut seeds. This trial, carried out inside a greenhouse, with a completely randomized design, was composed of five salinity treatments (0.02, 2, 4, 6 and 8dS m-1) with 5 replications. It was observed that salinity levels above 2dS m-1 affected plant development. The current study suggests that salinity management is an important factor to be considered to achieve the potential productivity of physic nut.

Drought resistance of sugar-cane crop for different levels of water availability in the soil

Soil water availability is the main cause of reduced productivity, and the early development period most sensitive to water deficit. This study aimed to evaluate the drought resistance of the varieties of sugar-cane RB867515 and SP81-3250 during the early development using different levels of water deficit on four soil depths. The experiment was conducted at the Department of Biosystems at Escola Superior de Agricultura "Luiz de Queiroz" (ESALQ/USP) in a greenhouse in soil classified as Oxisol, sandy loam texture (Series "Sertãozinho"). Once exhausted the level of available water in the soil, the dry strength of the studied strains are relatively low. Water balance with values less than -13 mm cause a significant decrease in the final population of plants, regardless of the variety, and values below -35 mm, leads to the death of all plants.

Stochastic variability of output levels in a pulp mill

Quite often, in the construction of a pulp mill involves establishing the size of tanks which will accommodate the material from the various processes in which case estimating the right tank size a priori would be vital. Hence, simulation of the whole production process would be worthwhile. Therefore, there is need to develop mathematical models that would mimic the behavior of the output from the various production units of the pulp mill to work as simulators. Markov chain models, Autoregressive moving average (ARMA) model, Mean reversion models with ensemble interaction together with Markov regime switching models are proposed for that purpose.

Hypothalamic-pituitary-gonadal function in men with liver cirrhosis before and after liver transplantation

Objective: To evaluate the influence of end-stage liver disease and orthotopic liver transplantation in the pituitary function and hormone metabolism before and after liver transplantation.Methods: In a prospective study, serum levels of follicle stimulating hormone (FSH), luteinizing hormone (LH), estradiol (E2) and prolactin (PRL) of 30 male patients with cirrhosis were determined two to four hours before and six months after liver transplantation. The results were compared according to the Model for End-stage Liver Disease (MELD).Results: male patients with liver cirrhosis have hypogonadism. FSH was normal, but inappropriately low due to androgen failure; E2 and PRL, on their turn, were high. After liver transplantation, FSH and LH levels increased (p < 0.05), whereas E2 and PRL normalized (p < 0.05). The MELD score did not influence changes in FSH, PRL and LH, however, the more severe the cirrhosis was, the more significant was the normalization of E2 (p = 0.01).Conclusion: Patients with cirrhosis and male hypogonadism have inappropriately normal levels of FSH and LH, associated with an increase in E2 and LRP. After liver transplantation, FSH and LH increased, while E2 and PRL returned to normal. Changes in E2 levels were most pronounced in patients with MELD > 18. The severity of cirrhosis had no influence on FSH, PRL and LH.

Salivary and serum cortisol levels, salivary alpha-amylase and unstimulated whole saliva flow rate in pregnant and non-pregnant

PURPOSE: To compare salivary and serum cortisol levels, salivary alpha-amylase (sAA), and unstimulated whole saliva (UWS) flow rate in pregnant and non-pregnant women. METHOD: A longitudinal study was conducted at a health promotion center of a university hospital. Nine pregnant and 12 non-pregnant women participated in the study. Serum and UWS were collected and analyzed every trimester and twice a month during the menstrual cycle. The salivary and serum cortisol levels were determined by chemiluminescence assay and the sAA was processed in an automated biochemistry analyzer. RESULTS: Significant differences between the pregnant and non-pregnant groups were found in median [interquartile range] levels of serum cortisol (23.8 µL/dL [19.4-29.4] versus 12.3 [9.6-16.8], p<0.001) and sAA (56.7 U/L [30.9-82.2] versus 31.8 [18.1-53.2], p<0.001). Differences in salivary and serum cortisol (µL/dL) and sAA levels in the follicular versus luteal phase were observed (p<0.001). Median UWS flow rates were similar in pregnant (0.26 [0.15-0.30] mL/min) and non-pregnant subjects (0.23 [0.20-0.32] mL/min). Significant correlations were found between salivary and serum cortisol (p=0.02) and between salivary cortisol and sAA (p=0.01). CONCLUSIONS: Serum cortisol and sAA levels are increased during pregnancy. During the luteal phase of the ovarian cycle, salivary cortisol levels increase, whereas serum cortisol and sAA levels decline.