Identification and Functional Analysis of a Novel Tryptophyllin Peptide from the Skin of the Red-eye Leaf Frog, Agalychnis callidryas

Amphibian skin has proved repeatedly to be a largely untapped source of bioactive peptides and this is especially true of members of the Phyllomedusinae subfamily of frogs native to South and Central America. Tryptophyllins are a group of peptides mainly found in the skin of members of this genus. In this study, a novel tryptophyllin (TPH) type 3 peptide, named AcT-3, has been isolated and structurally-characterised from the skin secretion and lyophilised skin extract of the red-eye leaf frog, Agalychnis callidryas. The peptide was identified in and purified from the skin secretion by reverse-phase HPLC. MALDI-TOF mass spectrometry and MS/MS fragmentation sequencing established its primary structure as: pGlu-Gly-Lys-Pro-Tyr-Trp-Pro-Pro-Pro-Phe-Leu-Pro-Glu, with a non-protonated molecular mass of 1538.19Da. The mature peptide possessed the canonical N-terminal pGlu residue that arises from post-translational modification of a Gln residue. The deduced open-reading frame consisted of 63 amino acid residues encoding a highly-conserved signal peptide of approximately 22 amino acid residues, an intervening acidic spacer peptide domain, a single AcT-3 encoding domain and a C terminal processing site. A synthetic replicate of AcT-3 was found to antagonise the effect of BK on rat tail artery smooth muscle and to contract the intestinal smooth muscle preparations. It was also found that AcT-3 could dose-dependently inhibit the proliferation of human prostate cancer cell lines after 72h incubation.

Global transcript analysis of rice leaf and seed using SAGE technology.

We have compiled two comprehensive gene expression profiles from mature leaf and immature seed tissue of rice (Oryza sativa ssp. japonica cultivar Nipponbare) using Serial Analysis of Gene Expression (SAGE) technology. Analysis revealed a total of 50 519 SAGE tags, corresponding to 15 131 unique transcripts. Of these, the large majority (approximately 70%) occur only once in both libraries. Unexpectedly, the most abundant transcript (approximately 3% of the total) in the leaf library was derived from a type 3 metallothionein gene. The overall frequency profiles of the abundant tag species from both tissues differ greatly and reveal seed tissue as exhibiting a non-typical pattern of gene expression characterized by an over abundance of a small number of transcripts coding for storage proteins. A high proportion ( approximately 80%) of the abundant tags (> or = 9) matched entries in our reference rice EST database, with many fewer matches for low abundant tags. Singleton transcripts that are common to both tissues were collated to generate a summary of low abundant transcripts that are expressed constitutively in rice tissues. Finally and most surprisingly, a significant number of tags were found to code for antisense transcripts, a finding that suggests a novel mechanism of gene regulation, and may have implications for the use of antisense constructs in transgenic technology.

Artichoke leaf extract reduces symptoms of irritable bowel syndrome and improves quality of life in otherwise healthy volunteers suffering from concomitant dyspepsia: a subset analysis

Objectives: Does artichoke leaf extract (ALE) ameliorate symptoms of Irritable bowel syndrome (IBS) in otherwise healthy volunteers suffering concomitant dyspepsia? Methods: A subset analysis of a previous dose-ranging, open, postal study, in adults suffering dyspepsia. Two hundred and eight (208) adults were identified post hoc as suffering with IBS. IBS incidence, self-reported usual bowel pattern, and the Nepean Dyspepsia Index (NDI) were compared before and after a 2-month intervention period. Results: There was a significant fall in IBS incidence of 26.4% (p<0.001) after treatment. A significant shift in self-reported usual bowel pattern away from "alternating constipation/diarrhea" toward "normal" (p<0.001) was observed. NDI total symptom score significantly decreased by 41% (p<0.001) after treatment. Similarly, there was a significant 20% improvement in the NDI total quality-of-life (QOL) score in the subset after treatment. Conclusion: This report supports previous findings that ALE ameliorates symptoms of IBS, plus improves health-related QOL.

Analysis of spectroscopic measurements of leaf water content at terahertz frequencies using linear transforms

We provide a unified framework for a range of linear transforms that can be used for the analysis of terahertz spectroscopic data, with particular emphasis on their application to the measurement of leaf water content. The use of linear transforms for filtering, regression, and classification is discussed. For illustration, a classification problem involving leaves at three stages of drought and a prediction problem involving simulated spectra are presented. Issues resulting from scaling the data set are discussed. Using Lagrange multipliers, we arrive at the transform that yields the maximum separation between the spectra and show that this optimal transform is equivalent to computing the Euclidean distance between the samples. The optimal linear transform is compared with the average for all the spectra as well as with the Karhunen–Loève transform to discriminate a wet leaf from a dry leaf. We show that taking several principal components into account is equivalent to defining new axes in which data are to be analyzed. The procedure shows that the coefficients of the Karhunen–Loève transform are well suited to the process of classification of spectra. This is in line with expectations, as these coefficients are built from the statistical properties of the data set analyzed.

Proteomic analysis of the medicinal plant Artemisia annua: data from leaf and trichome extracts

This article contains raw and processed data related to research published by Bryant et al. [1]. Data was obtained by MS-based proteomics, analysing trichome-enriched, trichome-depleted and whole leaf samples taken from the medicinal plant Artemisia annua and searching the acquired MS/MS data against a recently published contig database [2] and other genomic and proteomic sequence databases for comparison. The processed data shows that an order-of-magnitude more proteins have been identified from trichome-enriched Artemisia annua samples in comparison to previously published data. Proteins known to have a role in the biosynthesis of artemisinin and other highly abundant proteins were found which imply additional enzymatically driven processes occurring within the trichomes that are significant for the biosynthesis of artemisinin.

An improved estimate of leaf area index based on the histogram analysis of hemispherical photographs

Leaf area index (LAI) is a key parameter that affects the surface fluxes of energy, mass, and momentum over vegetated lands, but observational measurements are scarce, especially in remote areas with complex canopy structure. In this paper we present an indirect method to calculate the LAI based on the analyses of histograms of hemispherical photographs. The optimal threshold value (OTV), the gray-level required to separate the background (sky) and the foreground (leaves), was analytically calculated using the entropy crossover method (Sahoo, P.K., Slaaf, D.W., Albert, T.A., 1997. Threshold selection using a minimal histogram entropy difference. Optical Engineering 36(7) 1976-1981). The OTV was used to calculate the LAI using the well-known gap fraction method. This methodology was tested in two different ecosystems, including Amazon forest and pasturelands in Brazil. In general, the error between observed and calculated LAI was similar to 6%. The methodology presented is suitable for the calculation of LAI since it is responsive to sky conditions, automatic, easy to implement, faster than commercially available software, and requires less data storage. (C) 2008 Elsevier B.V. All rights reserved.

Fractal analysis of leaf-texture properties as a tool for taxonomic and identification purposes: a case study with species from Neotropical Melastomataceae (Miconieae tribe)

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

RAPD analysis of the sexual state and sterile mycelium of the fungus cultivated by the leaf-cutting ant Acromyrmex hispidus fallax

That the symbiotic fungus of leaf-cutting ants only occasionally produces the sexual phase makes their identification confusing. This has occurred so rarely, either in laboratory nests, or in unbalanced field nests. that the possibility of contamination of the fungal garden by other fungi cannot be disregarded. In this paper we describe the formation of several basidiomata in a healthy and free-living nest of the leaf-cutting ant Acromyrmex hispidus fallax. The cultivation in vitro of the sterile mycelia (isolated from the fungal garden) with their typical inflated tips, and the similarity of both forms confirmed by RAPD analysis of their genomic DNA. The fungus was identified as Leucoagaricus gongylophorus.

Application of on-line C30 RP-HPLC-NMR for the analysis of flavonoids from leaf extract of Maytenus aquifolium

The application of on-line C30-reversed-phase high-pressure liquid chromatography-nuclear magnetic resonance spectroscopy is described for the analysis of tetraglycosylated flavonoids in aqueous and hydroalcoholic extracts of the leaves of Maytenus aquifolium (Celastraceae). Triacontyl stationary phases showed adequate separation for on-line 1H-NMR measurements at 600 MHz and allowed the characterisation of these flavonoids by detection of both aromatic and anomeric proton signals. Copyright (C) 2000 John Wiley and Sons, Ltd.

Stresses analysis in semi-elliptic leaf spring and comparison with the SAE J788 (1982) norm

In this work are presented the values found with the experimental testing, in the semi-elliptic leaf spring, utilizing 24 strain gages, distributed in five leaves of springs; these values have been compared to the calculated values found with the application of Norm SAE J788 (1982). The results showed discrepancy between the values measured and calculated and that the Norm is not indicated to determine the actuating stress in any point of any leaf of the leaf spring, but due to its simplicity and quickness of the process it presents good precision for the pre-development of the product. Copyright © 2002 Society of Automotive Engineers, Inc.

Biochemical Analysis of a Castor Bean Leaf Extract and its Insecticidal Effects Against Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae)

Plant extracts represent a great source of molecules, with insecticidal activity, which are used for pest control in several crop production systems. This work aimed to evaluate the toxicity of an aqueous extract of leaves of castor bean against larvae of Spodoptera frugiperda (Smith) (Lepidoptera: Noctuidae) in search for different classes of molecules with insecticidal activities by using in vitro assays. The effects of the castor bean leaf extract on the food utilization, development, and survival of S. frugiperda larvae was evaluated by feeding the larvae an artificial diet supplemented with different concentrations of the extract (0%, 1%, 2.5%, 5%, and 10% w/v). The effects observed were dose-dependent, and the highest concentration evaluated (10% w/v) was the one the most affected food utilization by altering the nutritional indices, as well as larval weight gain, development time, and survivorship. In vitro assays to detect saponins, lectins, and trypsin inhibitors in the castor bean leaf extract were performed, but only trypsin inhibitors were detected. No preference for the diet source was detected in S. frugiperda by feeding the larvae in choice experiments with diets containing different concentrations of the castor bean extract tested. The data obtained indicate the existence of a potential molecule in the tested extract of castor bean to be used as an alternative insecticide to be integrated in the management of S. frugiperda.

Microsurgical and laser ablation analysis of leaf positioning and dorsoventral patterning in tomato

Leaves are arranged according to regular patterns, a phenomenon referred to as phyllotaxis. Important determinants of phyllotaxis are the divergence angle between successive leaves, and the size of the leaves relative to the shoot axis. Young leaf primordia are thought to provide positional information to the meristem, thereby influencing the positioning of new primordia and hence the divergence angle. On the contrary, the meristem signals to the primordia to establish their dorsoventral polarity, which is a prerequisite for the formation of a leaf blade. These concepts originate from classical microsurgical studies carried out between the 1920s and the 1970s. Even though these techniques have been abandoned in favor of genetic analysis, the resulting insights remain a cornerstone of plant developmental biology. Here, we employ new microsurgical techniques to reassess and extend the classical studies on phyllotaxis and leaf polarity. Previous experiments have indicated that the isolation of an incipient primordium by a tangential incision caused a change of divergence angle between the two subsequent primordia, indicating that pre-existing primordia influence further phyllotaxis. Here.. we repeat these experiments and compare them with the results of laser ablation of incipient primordia. Furthermore. we explore to what extent the different pre-existing primordia influence the size and position of new organs. and hence phyllotaxis. We propose that the two youngest primordia (P-1 and P-2) are sufficient for the approximate positioning of the incipient primordium (I-1), and therefore for the perpetuation of the generative spiral, whereas the direct contact neighbours of I-1 (P-2 and P-3) control its delimitation and hence its exact size and position. Finally. we report L I specific cell ablation experiments suggesting that the meristem L-1 layer is essential for the dorsoventral patterning of leaf primordia.

Chemical and structural analysis of Eucalyptus globulus and E. camaldulensis leaf cuticles: a lipidized cell wall region

The plant cuticle has traditionally been conceived as an independent hydrophobic layer that covers the external epidermal cell wall. Due to its complexity, the existing relationship between cuticle chemical composition and ultra-structure remains unclear to date. This study aimed to examine the link between chemical composition and structure of isolated, adaxial leaf cuticles of Eucalyptus camaldulensis and E. globulus by the gradual extraction and identification of lipid constituents (cutin and soluble lipids), coupled to spectroscopic and microscopic analyses. The soluble compounds and cutin monomers identified could not be assigned to a concrete internal cuticle ultra-structure. After cutin depolymerization, a cellulose network resembling the cell wall was observed, with different structural patterns in the regions ascribed to the cuticle proper and cuticular layer, respectively. Our results suggest that the current cuticle model should be revised, stressing the presence and major role of cell wall polysaccharides. It is concluded that the cuticle may be interpreted as a modified cell wall region which contains additional lipids. The major heterogeneity of the plant cuticle makes it difficult to establish a direct link between cuticle chemistry and structure with the existing methodologies.

Does Leaf Position within a Canopy Affect Acclimation of Photosynthesis to Elevated CO2?1 : Analysis of a Wheat Crop under Free-Air CO2 Enrichment

Previous studies of photosynthetic acclimation to elevated CO2 have focused on the most recently expanded, sunlit leaves in the canopy. We examined acclimation in a vertical profile of leaves through a canopy of wheat (Triticum aestivum L.). The crop was grown at an elevated CO2 partial pressure of 55 Pa within a replicated field experiment using free-air CO2 enrichment. Gas exchange was used to estimate in vivo carboxylation capacity and the maximum rate of ribulose-1,5-bisphosphate-limited photosynthesis. Net photosynthetic CO2 uptake was measured for leaves in situ within the canopy. Leaf contents of ribulose-1,5-bisphosphate carboxylase/oxygenase (Rubisco), light-harvesting-complex (LHC) proteins, and total N were determined. Elevated CO2 did not affect carboxylation capacity in the most recently expanded leaves but led to a decrease in lower, shaded leaves during grain development. Despite this acclimation, in situ photosynthetic CO2 uptake remained higher under elevated CO2. Acclimation at elevated CO2 was accompanied by decreases in both Rubisco and total leaf N contents and an increase in LHC content. Elevated CO2 led to a larger increase in LHC/Rubisco in lower canopy leaves than in the uppermost leaf. Acclimation of leaf photosynthesis to elevated CO2 therefore depended on both vertical position within the canopy and the developmental stage.