Biological activity of Pyrrole-Imidazole polyamides in vivo

This thesis focuses on biological activity of pyrrole-imidazole polyamides in vivo. The work presented includes experiments underlining sequence selectivity of these compounds in living cells and potential methods to improve it. A large fraction of this thesis is devoted to activity of Py-Im in murine models of cancer. We investigated the pharmacokinetics and biodistribution of two compounds – targeted to 5'-WGGWCW-3' and 5'-WTWCGW-3' sequences – and characterized their activity by measuring their effects on tumor growth, gene expression in vivo and in tissue culture, and their effects on physiology of tumors. The initial theoretical studies suggested that a large fraction of genomic sites are bound by Py-Im polyamides non-specifically and experimental data shows that the programmed binding sequence is not a sole determinant of the patterns of gene regulation. Despite the likely presence of non-specific effects of Py-Im polyamides in living cells, in vivo administration of Py-Im polyamides resulted in tolerable host toxicity and anti-tumor activity. Py-Im polyamide targeted to Estrogen Receptor Response Element showed downregulation of ER-driven gene expression in tumor cells, while the compound targeted to hypoxia response element reduced vascularization of tumors and their growth rate, induced apoptosis of cells in hypoxic areas and reduced expression of proangiogenic and prometastatic factors. Further studies, showed that polyamides distributed to many of the tested tissues and their FITC-conjugates showed nuclear uptake. The gene expression effects were also present in murine tissues, such as liver and kidneys, indicating a potential for use for Py-Im polyamides in non-cancerous diseases.

Antilipolytic and hypotriglyceridemic effects of dietary Salvia triloba Lf (Lamiaceae) in experimental rats

Purpose: Pancreatic triacylglycerol lipase (PL) is a noteworthy pharmacological target for the management of dyslipidemia, and diabetes and obesity. This study was aimed to evaluate the modulatory effects of Salvia triloba L.f. (Lamiaceae) leaves methanol extract (ME) on a high fat diet (HFD)-induced hypertriglyceridemia in rats, with complementary in vitro evaluation of sage PL-inhibitory potential. Methods: Pre-induction of HFD hypertriglyceridemia sage leaves ME (750 mg/kg) was orally supplemented (via gastric intubation) to overnight fasting rats (n = 5). Potential plant modulation of PL was also quantified in vitro by a colorimetric assay (n = 3). For comparison, the effect of Orlistat was similarly evaluated as reference standard. Results: Compared to Orlistat, supplementation of S. triloba at a dose of 750 mg/kg b.wt significantly reversed the HFD-induced postprandial hypertriglyceridemia in experimental overnight fasting rats (p < 0.001 vs. HFD rats). Dietary sage caused 66.4 % reduction in plasma triglycerides. Compared to Orlistat which exerted antilipolytic activity, with half-maximal inhibitory concentration (IC50) of 0.114 ± 0.004 μg/mL), sage inhibited PL activity in vitro in a dose-dependent manner IC50 of 100.80 ± 9.07 μg/mL) Conclusion: Sage has dual hypotriglyceridemic and antilipolytic properties which indicate that it can potentially be used to suppress body weight gain.

Thermodynamic Analysis of Rare Earth Chlorination and Bromination Processes

Research funded by the Army Research Laboratory (ARL), the Metallurgical and Materials Engineering Department at Montana Tech investigated various methods of extracting and refining rare earth elements (REEs) from mineral ores and concentrates. Extensive thermodynamic, thermogravimetric and differential thermal analyses were performed to evaluate the relative stabilities of various REE compounds in order to assess potential methods for selective separation and recovery of specific REEs. Conversion of rare earth oxides (REO) to rare earth chlorides or bromides is a possible initial step in pyrometallurgical and hydrometallurgical processing of REEs. REO can be converted to chlorides or bromides by roasting in the presence of a chloridizing or bromidizing reactant. (e.g. NH4Cl and NH4Br).

Evaluation of Indigenous Grains from the Peruvian Andean Region for Antidiabetes and Antihypertension Potential Using In Vitro Methods

The health-relevant functionality of 10 thermally processed Peruvian Andean grains (five cereals, three pseudocereals, and two legumes) was evaluated for potential type 2 diabetes-relevant antihyperglycemia and antihypertension activity using in vitro enzyme assays. Inhibition of enzymes relevant for managing early stages of type 2 diabetes such as hyperglycemia-relevant alpha-glucosidase and alpha-amylase and hypertension-relevant angiotensin I-converting enzyme (ACE) were assayed along with the total phenolic content, phenolic profiles, and antioxidant activity based on the 1,1-diphenyl-2-picrylhydrazyl radical assay. Purple corn (Zea mays L.) (cereal) exhibited high free radical scavenging-linked antioxidant activity (77%) and had the highest total phenolic content (8 +/- 1 mg of gallic acid equivalents/g of sample weight) and alpha-glucosidase inhibitory activity (51% at 5 mg of sample weight). The major phenolic compound in this cereal was protocatechuic acid (287 +/- 15 mu g/g of sample weight). Pseudocereals such as Quinoa (Chenopodium quinoa Willd) and Kaniwa (Chenopodium pallidicaule Aellen) were rich in quercetin derivatives (1,131 +/- 56 and 943 +/- 35 mu g [expressed as quercetin aglycone]/g of sample weight, respectively) and had the highest antioxidant activity (86% and 75%, respectively). Andean legumes (Lupinus mutabilis cultivars SLP-1 and H-6) inhibited significantly the hypertension-relevant ACE (52% at 5 mg of sample weight). No alpha-amylase inhibitory activity was found in any of the evaluated Andean grains. This in vitro study indicates the potential of combination of Andean whole grain cereals, pseudocereals, and legumes to develop effective dietary strategies for managing type 2 diabetes and associated hypertension and provides the rationale for animal and clinical studies.

Anemia screening in potential female blood donors: comparison of two different quantitative methods

Anemia screening before blood donation requires an accurate, quick, practical, and easy method with minimal discomfort for the donors. The aim of this study was to compare the accuracy of two quantitative methods of anemia screening: the HemoCue 201(+) (Aktiebolaget Leo Diagnostics) hemoglobin (Hb) and microhematocrit (micro-Hct) tests. Two blood samples of a single fingerstick were obtained from 969 unselected potential female donors to determine the Hb by HemoCue 201(+) and micro-Hct using HemataSTAT II (Separation Technology, Inc.), in alternating order. From each participant, a venous blood sample was drawn and run in an automatic hematology analyzer (ABX Pentra 60, ABX Diagnostics). Considering results of ABX Pentra 60 as true values, the sensitivity and specificity of HemoCue 201(+) and micro-Hct as screening methods were compared, using a venous Hb level of 12.0 g per dL as cutoff for anemia. The sensitivities of the HemoCue 201(+) and HemataSTAT II in detecting anemia were 56 percent (95% confidence interval [CI], 46.1%-65.5%) and 39.5 percent (95% CI, 30.2%-49.3%), respectively (p < 0.001). Analyzing only candidates with a venous Hb level lower than 11.0 g per dL, the deferral rate was 100 percent by HemoCue 201(+) and 77 percent by HemataSTAT II. The specificities of the methods were 93.5 and 93.2 percent, respectively. The HemoCue 201(+) showed greater discriminating power for detecting anemia in prospective blood donors than the micro-Hct method. Both presented equivalent deferral error rates of nonanemic potential donors. Compared to the micro-Hct, HemoCue 201(+) reduces the risk of anemic female donors giving blood, specially for those with lower Hb levels, without increasing the deferral of nonanemic potential donors.

Evaluation of rhBMP-2 and Natural Latex as Potential Osteogenic Proteins in Critical Size Defects by Histomorphometric Methods

This in vivo study evaluated the osteogenic potential of two proteins, recombinant human bone morphogenetic protein-2 (rhBMP-2) and a protein extracted from natural latex (Hevea brasiliensis, P-1), and compared their effects on bone defects when combined with a carrier or a collagen gelatin. Eighty-four (84) Wistar rats were divided into two groups, with and without the use of collagen gelatin, and each of these were divided into six treatment groups of seven animals each. The treatment groups were: (1) 5 mu g of pure rhBMP-2; (2) 5 mu g of rhBMP-2/monoolein gel; (3) pure monoolein gel; (4) 5 mu g of pure P-1; (5) 5 mu g of P-1/monoolein gel; (6) critical bone defect control. The animals were anesthetized and a 6 mm diameter critical bone defect was made in the left posterior region of the parietal bone. Animals were submitted to intracardiac perfusion after 4 weeks and the calvaria tissue was removed for histomorphometric analysis. In this experimental study, it was concluded that rhBMP-2 allowed greater new bone formation than P-1 protein and this process was more effective when the bone defect was covered with collagen gelatin (P < 0.05). Anat Rec, 293:794-801, 2010. (C) 2010 Wiley-Liss, Inc.

Modern scientific methods and their potential in wastewater science and technology

Application of novel analytical and investigative methods such as fluorescence in situ hybridization, confocal laser scanning microscopy (CLSM), microelectrodes and advanced numerical simulation has led to new insights into micro-and macroscopic processes in bioreactors. However, the question is still open whether or not these new findings and the subsequent gain of knowledge are of significant practical relevance and if so, where and how. To find suitable answers it is necessary for engineers to know what can be expected by applying these modern analytical tools. Similarly, scientists could benefit significantly from an intensive dialogue with engineers in order to find out about practical problems and conditions existing in wastewater treatment systems. In this paper, an attempt is made to help bridge the gap between science and engineering in biological wastewater treatment. We provide an overview of recently developed methods in microbiology and in mathematical modeling and numerical simulation. A questionnaire is presented which may help generate a platform from which further technical and scientific developments can be accomplished. Both the paper and the questionnaire are aimed at encouraging scientists and engineers to enter into an intensive, mutually beneficial dialogue. (C) 2002 Elsevier Science Ltd. All rights reserved.

Comparison of different methods in estimating potential evapotranspiration at Muda Irrigation Scheme of Malaysia

Evapotranspiration (ET) is a complex process in the hydrological cycle that influences the quantity of runoff and thus the irrigation water requirements. Numerous methods have been developed to estimate potential evapotranspiration (PET). Unfortunately, most of the reliable PET methods are parameter rich models and therefore, not feasible for application in data scarce regions. On the other hand, accuracy and reliability of simple PET models vary widely according to regional climate conditions. The objective of the present study was to evaluate the performance of three temperature-based and three radiation-based simple ET methods in estimating historical ET and projecting future ET at Muda Irrigation Scheme at Kedah, Malaysia. The performance was measured by comparing those methods with the parameter intensive Penman-Monteith Method. It was found that radiation based methods gave better performance compared to temperature-based methods in estimation of ET in the study area. Future ET simulated from projected climate data obtained through statistical downscaling technique also showed that radiation-based methods can project closer ET values to that projected by Penman-Monteith Method. It is expected that the study will guide in selecting suitable methods for estimating and projecting ET in accordance to availability of meteorological data.

Extracting Urban Food Potential: design-based methods for digital and bodily cartography

Sweden’s recent report on Urban Sustainable Development calls out a missing link between the urban design process and citizens. This paper investigates if engaging citizens as design agents by providing a platform for alternate participation can bridge this gap, through the transfer of spatial agency and new modes of critical cartography. To assess whether this is the case, the approaches are applied to Stockholm’s urban agriculture movement in a staged intervention. The aim of the intervention was to engage citizens in locating existing and potential places for growing food and in gathering information from these sites to inform design in urban agriculture. The design-based methodologies incorporated digital and bodily interfaces for this cartography to take place. The Urban CoMapper, a smartphone digital app, captured real-time perspectives through crowd-sourced mapping. In the bodily cartography, participant’s used their bodies to trace the site and reveal their sensorial perceptions. The data gathered from these approaches gave way to a mode of artistic research for exploring urban agriculture, along with inviting artists to be engaged in the dialogues. In sum, results showed that a combination of digital and bodily approaches was necessary for a critical cartography if we want to engage citizens holistically into the urban design process as spatial agents informing urban policy. Such methodologies formed a reflective interrogation and encouraged a new intimacy with nature, in this instance, one that can transform our urban conduct by questioning our eating habits: where we get our food from and how we eat it seasonally.

Participatory Farm Management methods for assessing the suitability of potential innovations. A case study on green manuring options for tomato producers in Ghana

This paper examines the potential of using Participatory Farm Management methods to examine the suitability of a technology with farmers prior to on-farm trials. A study examining the suitability of green manuring as a technology for use with wet season tomato producers in Ghana is described. Findings from this case-study demonstrate that Participatory Budgeting can be used by farmers and researchers to analyse current cultivation practices, identify the options for including green manures into the system and explore the direct and wider resource implications of the technology. Scored-Causal Diagrams can be used to identify farmers' perceptions of the relative importance of the problem that the technology seeks to address. The use of the methods in this examine evaluation process appears to have the potential to improve the effectiveness and efficiency of the adaptive research process. This ensures that technologies subsequently examined in trials ate relevant to farmers' interests, existing systems and resources, thereby increasing the chances of farmer adoption. It is concluded that this process has potential for use-with other technologies and in other farming systems. (C) 2002 Elsevier Science Ltd. All rights reserved.

Methods for evaluating the physiological potential of scarlet eggplant (Solanum aethiopicum) seeds

Fundação de Amparo à Pesquisa do Estado de São Paulo (FAPESP)

Methods for the evaluation of hydroelectric potential

Includes bibliography