Purine nucleoside phosphorylase: A potential target for the development of drugs to treat T-cell- and apicomplexan parasite-mediated diseases

Purine nucleoside phosphorylase (PNP) catalyzes the reversible phosphorolysis of nucleosides and deoxynucleosides, generating ribose 1-phosphate and the purine base, which is an important step of purine catabolism pathway. The lack of such an activity in humans, owing to a genetic disorder, causes T-cell impairment, and thus drugs that inhibit human PNP activity have the potential of being utilized as modulators of the immunological system to treat leukemia, autoimmune diseases, and rejection in organ transplantation. Besides, the purine salvage pathway is the only possible way for apicomplexan parasites to obtain the building blocks for RNA and DNA synthesis, which makes PNP from these parasites an attractive target for drug development against diseases such as malaria. Hence, a number of research groups have made efforts to elucidate the mechanism of action of PNP based on structural and kinetic studies. It is conceivable that the mechanism may be different for PNPs from diverse sources, and influenced by the oligomeric state of the enzyme in solution. Furthermore, distinct transition state structures can make possible the rational design of specific inhibitors for human and apicomplexan enzymes. Here, we review the current status of these research efforts to elucidate the mechanism of PNP-catalyzed chemical reaction, focusing on the mammalian and Plamodium falciparum enzymes, targets for drug development against, respectively, T-Cell and Apicomplexan parasites-mediated diseases.

Effect of foliar and substrate fertilization on lady palm seedling growth and development

Lady palm, [Rhapis excelsa (Thunberg) Henry ex. Rehder] is one of the most cultivated ornamental palms in the world, for use as a vase plant or in shaded landscapes. Because limited information exists on lady palm response to fertilizers, the objective of this study was to evaluate the effect of different types of fertilization and substrates on lady palm seedling growth and development. Three year old lady palms were planted in 8-L pots, filled with a mix of soil, manure, and sand 1:1:1 (v:v:v), placed under a 50% shade, and irrigated with microspray. Treatments were substrate fertilization with 500 g P(2)O(5) and 100 g K(2)O per m(3); fertilization with 1.8 kg of P(2)O(5) (simple superphosphate) per m3; 50 g of nitrogen (N), P(2)O(5), and K(2)O of a granulated fertilizer (10:10:10) per m(3), control (without fertilization), and a foliar fertilization in addition to these treatments using the commercial product Biofert (8:9:9). Treatments were replicated four times in a randomized block design. Each treatment plot consisted of four plants. Data were collected at 140, 170, 200, 230, 260, and 290 days after transplanting (DAT) for plant heights, stem diameter at substrate level, number of leaves, shoots, and canopy, roots fresh and dry matter samples were harvest at 290 days. Foliar fertilization resulted in significantly greater plant height in a 140, 120, 200, and 230 DAT and plant diameter on the 140, 260, and 290 DAT. There was interaction among factors for number of leaves with fertilization based on P(2)O(5) and K(2)O when leaf fertilizer was added that resulted in a greater number of leaves.

Effect of water stress on seed germination and seedling growth in Oryza sativa L.

The effect of water stress induced by application of polyethylene glycol 6000 during seed germination and seedling growth of Oryza sativa L. cv. IAC 165 was analysed. The seed germination was inhibited by the decrease in the water potential of the medium, the inhibition being greater under white light than under continuous darkness. When the seedling was submitted to water stress (-0.51 MPa) white light inhibited growth of root, coleoptile-and leaf, while under no stress conditions white light caused increase in growth of root and leaf and only inhibition of coleoptile growth. © 1990 Kluwer Academic Publishers.

Effect of five substrates and three seed types on Calendula officinalis (Asteraceae) germination and seedling development

The investigation was carried out during May - June 1996 in Agronomical Sciences College - UNESP, Campus de Botucatu. The objective was to study the effect of seed types (carenate, alate and orbicular) and different substrates (vermiculite, sand, soil, commercial mixture and producer's mixture) on Calendula officinalis germination and seedling development. The design was in randomized blocks, with 4 replications. The germination index of the seeds was evaluated, as well as the development of aerial part and root system of the seedlings. Results showed that seed types have no relation with seedling development, therefore, germination was better on alate and orbicular seeds in C. officinalis. With reference to the substrates, the producer's mixture and the commercial mixture gave better germination and seedling development.

Chorismate synthase: An attractive target for drug development against orphan diseases

The increase in incidence of infectious diseases worldwide, particularly in developing countries, is worrying. Each year, 14 million people are killed by infectious diseases, mainly HIV/AIDS, respiratory infections, malaria and tuberculosis. Despite the great burden in the poor countries, drug discovery to treat tropical diseases has come to a standstill. There is no interest by the pharmaceutical industry in drug development against the major diseases of the poor countries, since the financial return cannot be guaranteed. This has created an urgent need for new therapeutics to neglected diseases. A possible approach has been the exploitation of the inhibition of unique targets, vital to the pathogen such as the shikimate pathway enzymes, which are present in bacteria, fungi and apicomplexan parasites but are absent in mammals. The chorismate synthase (CS) catalyses the seventh step in this pathway, the conversion of 5-enolpyruvylshikimate-3-phosphate to chorismate. The strict requirement for a reduced flavin mononucleotide and the anti 1,4 elimination are both unusual aspects which make CS reaction unique among flavin-dependent enzymes, representing an important target for the chemotherapeutic agents development. In this review we present the main biochemical features of CS from bacterial and fungal sources and their difference from the apicomplexan CS. The CS mechanisms proposed are discussed and compared with structural data. The CS structures of some organisms are compared and their distinct features analyzed. Some known CS inhibitors are presented and the main characteristics are discussed. The structural and kinetics data reviewed here can be useful for the design of inhibitors. © 2007 Bentham Science Publishers Ltd.

miRNApath: A database of miRNAs, target genes and metabolic pathways

MicroRNAs (miRNAs) are small non-coding RNAs that regulate target gene expression and hence play important roles in metabolic pathways. Recent studies have evidenced the interrelation of miRNAs with cell proliferation, differentiation, development, and diseases. Since they are involved in gene regulation, they are intrinsically related to metabolic pathways. This leads to questions that are particularly interesting for investigating medical and laboratorial applications. We developed an miRNApath online database that uses miRNA target genes to link miRNAs to metabolic pathways. Currently, databases about miRNA target genes (DIANA miRGen), genomic maps (miRNAMap) and sequences (miRBase) do not provide such correlations. Additionally, miRNApath offers five search services and a download area. For each search, there is a specific type of input, which can be a list of target genes, miRNAs, or metabolic pathways, which results in different views, depending upon the input data, concerning relationships between the target genes, miRNAs and metabolic pathways. There are also internal links that lead to a deeper analysis and cross-links to other databases with more detailed information. miRNApath is being continually updated and is available at http://lgmb.fmrp.usp.br/mirnapath. ©FUNPEC-RP.

Video target tracking by using competitive neural networks

A target tracking algorithm able to identify the position and to pursuit moving targets in video digital sequences is proposed in this paper. The proposed approach aims to track moving targets inside the vision field of a digital camera. The position and trajectory of the target are identified by using a neural network presenting competitive learning technique. The winning neuron is trained to approximate to the target and, then, pursuit it. A digital camera provides a sequence of images and the algorithm process those frames in real time tracking the moving target. The algorithm is performed both with black and white and multi-colored images to simulate real world situations. Results show the effectiveness of the proposed algorithm, since the neurons tracked the moving targets even if there is no pre-processing image analysis. Single and multiple moving targets are followed in real time.

Seedling growth of Schizolobium parahyba on different substrates and irrigation levels

The present study aimed to assess the behavior of seedlings of Schizolobium parahyba (Vell.) Blake growing on different substrates and irrigation levels, and to study the use of urban waste compost in substrates. The experiment was carried out in the College of Agricultural Sciences, Universidade Estadual Paulista (UNESP), Department of Agricultural Engineering, Jaboticabal Campus, state of São Paulo, Brazil. The experimental design used was completely randomized, consisting of 30 treatments in a factorial design with 15 substrates and two irrigation levels in four replicates. The substrates were composed of different materials: urban waste, Plantmax®, dry cattle manure, vermiculite and soil. For the study of seedling growth, the following characteristics were evaluated: height (H), root-collar diameter (D), number of leaves, shoot dry weight, root dry weight, total dry weight, H/D ratio, Dickson's quality index, and the ratio between height and shoot dry weight (H/SDW). Assessments of the first three characteristics were carried out 20, 35, 50 and 65 days after sowing. Results showed that urban waste compost increased the growth of Schizolobium parahyba. There were significant differences for the irrigation levels tested; with better results for 150% ET compared to 100% ET irrigation level.

Wavelets and decision trees for target detection over sea surface using cosmo-skymed SAR data

We are investigating the combination of wavelets and decision trees to detect ships and other maritime surveillance targets from medium resolution SAR images. Wavelets have inherent advantages to extract image descriptors while decision trees are able to handle different data sources. In addition, our work aims to consider oceanic features such as ship wakes and ocean spills. In this incipient work, Haar and Cohen-Daubechies-Feauveau 9/7 wavelets obtain detailed descriptors from targets and ocean features and are inserted with other statistical parameters and wavelets into an oblique decision tree. © 2011 Springer-Verlag.

Seed and seedling surface-sterilization for in vitro culture of tillandsia gardneri (Bromeliaceae)

Tillandsia gardneri is a bromeliad with ornamental value and a wide geographical distribution over Brazil. However, due to habitat loss and illegal overcollection in the wild it is included as a vulnerable species in the official list of endangered plants of the State of Rio Grande do Sul, Brazil. The development of a protocol for T. gardneri seed propagation in vitro may be useful for reintroducing plants in their natural habitats, and for germplasm conservation. A difficult problem encountered during the establishment of an in vitro culture is explants disinfection, especially when working with endangered species, from which explant availability is restricted. Thus, the establishment of a sterilization protocol is crucial for the initiation and success of a micropropagation system for T. gardneri. The objective of this study was to evaluate the effect of sodium hypochlorite concentration and exposure time in seed and seedling surface disinfection, tissue sensitivity and development. Sodium hypochlorite solutions (10 or 20%/5, 10 or 15 min; 25%/5 or 10 min; and 50%/5 min) were effective in eliminating seed superficial contaminants. There was no significant difference among the effective sterilization treatments in relation to seed germination (%), and seedling length and number of leaves, after 120 days in vitro. Also, no damage to seed and seedling tissues were observed. Surface sterilization of seedlings, for initiation of an in vitro culture, required higher concentrations of sodium hypochlorite (25%/15 min; 20 or 50%/5, 10 or 15 min; and 40%/5 and 10 min) for controlling fungal and yeast contamination, compared to seed sterilization. No significant differences among these treatments were found in relation to seedling length and number of leaves, after 60 days in vitro.

Seedling morphology and development in the epiphytic cactus Epiphyllum phyllanthus (L.) Haw. (Cactaceae: Hylocereeae)

Seedling morphology is relevant in classification, taxonomy, and vegetation studies to understand plant life cycles, germination succession and requirements, and developmental progression. However, most morphological studies of seedlings lack analysis of organ anatomy, impeding the comprehension of series of development and establishment in a particular environment. Here, we have taken a traditional anatomical approach to examine the stages of seedling development in Epiphyllum phyllanthus, a holo-epiphytic cactus of tribe Hylocereeae. The goals were 1) to offer a comprehensive description of growth series in E. phyllanthus seedlings based on morphological and anatomical analysis and 2) to examine the initial growth phases in the life cycle of this species to identify organ development and understand their adaptive significance in relation to seedling establishment. Our results include descriptions of seed morphology, embryonic features, and seedling vascularization pattern in the root, hypocotyl, cotyledons, and epicotyl. The morphological and developmental patterns in E. phyllanthus seedlings have potential phylogenetic and ontogenetic implications in the Cactaceae. Characters such as the presence of mucilage on the seed coat, the lack of seed operculum, and large cotyledons in E. phyllanthus are comparable to basal cacti, but the root anatomy is more similar to columnar relatives. At the familial level, there is an apparent trend in decreasing number of phloem and xylem poles in the stele of primary root, correlated with degree of specialization and advanced phylogenetic position: tetrarch to septarch-octarch in basal lineages, tetrarch Cereus-type in columnar species, to the diarch vascular system in Rhipsalideae and some species with cylindric/globose stem. © Torrey Botanical Club.

Effects of substrate and temperature on the emergence of Phacelia sp. (Boraginaceae) seedling

This study aimed to define the best substrate and temperature for the emergence of Phacelia sp. seedling, annual ornamental flower gardens. The experimental design was entirely randomized in a factorial scheme 37 (three different types of substrates: vermiculite, sand and sphagnum combined with seven temperature conditions: room temperature, constant at 20, 25, 30 and 35C, and alternated at 20-30C and 25-35C) with 4 replications of 100 seeds each. Emergence (%E) and Emergence Rate (ER) were evaluated. The means of the resulting values were then compared by the Tukey test at 5% confidence level. There was a significant interaction amongst substrates and temperatures for all analyzed variables. For seeds sown in vermiculite and sand, the seedlings had higher %E and emerged fastest at 20C, room temperature and 20-30C that did not present statistically significant data. In sphagnum, seedlings showed greater %E in the alternated at 20-30C, room temperature and 20C and emerged quickly at room temperature and 20C that did not present statistically significant data. At room temperature and 20C, the seedlings had higher %E and emerged faster in vermiculite and sand that did not present statistically significant data. At 25, 30 and 25-35C, the seedlings showed better %E and emerged fastest in sand. At 35C, the seedlings showed either in vermiculite and sand were not significantly different in their emergence, but emerged faster in sand. There was not significantly different among substrates in their emergence at 20-30C, but the seedlings emerged faster in vermiculite and sand. It was concluded that the Phacelia sp. seedlings in all substrates showed greater %E at room temperature, 20C and 20-30C. For all temperatures, the seedlings growth parameters were superior when seeds were sown in sand.

Cryopreservation, early seedling development, and genetic stability of Oncidium flexuosum Sims

An efficient cryopreservation protocol was developed for mature seeds of Oncidium flexuosum Sims. Seed morphology, protocorm formation, and early seedling development were also assessed. The effects of phloroglucinol and Supercool X-1000® as cryoprotectant additives in the vitrification solution were investigated. Dehydration using the plant vitrification solution 2 (PVS2) for 60 and 120 min prior to immersion in liquid nitrogen promoted the highest frequency of in vitro seed germination 6 weeks following culture on half-strength Murashige and Skoog (1/2 MS) medium. Mature seeds submitted to vitrification for 120 min in PVS2 and 1 % phloroglucinol at 0 °C enhanced germination by 68 %, whereas in PVS2 and 1 % Supercool X-1000® germination was just moderately enhanced (26 %). In vitro-germinating seedlings developed healthy shoots and roots without the use of plant growth regulators. After 6 months of growth, there were no differences between in vitro- and ex vitro-grown seedlings for various phenotypic characteristics, including shoot length, number of leaves, number and length of roots, and fresh and dry weight. Seedlings were transferred to greenhouse conditions and successfully acclimatized, further developing into normal plants with over 90 % survival. Comparative analysis of seedlings from control and vitrified seeds using flow cytometry indicated that no change in ploidy levels occurred as a result of cryopreservation, therefore maintaining seedlings genetic stability. In this study, vitrification with PVS2 for 120 min with the addition of 1 % phloroglucinol offers a simple, safe, and feasible protocol for cryopreservation of O. flexuosum mature seeds. © 2013 Springer Science+Business Media Dordrecht.

No changes in seedling recruitment when terrestrial mammals are excluded in a partially defaunated Atlantic rainforest

One of the most intriguing questions in ecology is how to identify which and how many species will be able to inhabit human-modified landscapes. Large-bodied mammals structure plant communities by trampling, herbivory, seed dispersal and predation, and their local extinction may have pervasive consequences in plant communities due to the breakdown of key interactions. Although much attention has been given to understanding the effects of defaunation on plant communities, information on the potential impacts on plant functional groups (seed dispersal, seed size and seedling leaves defense) inhabiting continuous forests after defaunation is scarce. We conducted mammal surveys (line transects and camera trapping) to determine the defaunation status of a continuous Atlantic forest in Brazil. Then, we evaluated the effects of defaunation on seedling diversity, richness and abundance of functional groups using 15 plot-pairs (each pair with one open and one exclusion plot) monitored over 36. months. We found that the studied area is partially defaunated because it exhibits high abundance of primates, while terrestrial mammals, such as large rodents and ungulates, are rare. We found no significant changes in either seedling richness and diversity or in the seedling composition of plant functional groups in response to mammal exclosure. Seedling mortality and recruitment were similar between plot types. Our findings suggest that at semi-defaunated areas, where arboreal species are still present, terrestrial mammals have low impacts on the plant community reassembly. © 2013 Elsevier Ltd.