The crude skin secretion of the pepper frog leptodactylus labyrinthicus is rich in metallo and serine peptidases

Peptidases are ubiquitous enzymes involved in diverse biological processes. Fragments from bioactive peptides have been found in skin secretions from frogs, and their presence suggests processing by peptidases. Thus, the aim of this work was to characterize the peptidase activity present in the skin secretion of Leptodactylus labyrinthicus. Zymography revealed the presence of three bands of gelatinase activity of approximately 60 kDa, 66 kDa, and 80 kDa, which the first two were calcium-dependent. These three bands were inhibited either by ethylenediaminetetraacetic acid (EDTA) and phenathroline; thus, they were characterized as metallopeptidases. Furthermore, the proteolytic enzymes identified were active only at pH 6.0–10.0, and their activity increased in the presence of CHAPS or NaCl. Experiments with fluorogenic substrates incubated with skin secretions identified aminopeptidase activity, with cleavage after leucine, proline, and alanine residues. This activity was directly proportional to the protein concentration, and it was inhibited in the presence of metallo and serine peptidase inhibitors. Besides, the optimal pH for substrate cleavage was determined to be 7.0–8.0. The results of the in gel activity assay showed that all substrates were hydrolyzed by a 45 kDa peptidase. Gly-Pro-AMC was also cleaved by a peptidase greater than 97 kDa. The data suggest the presence of dipeptidyl peptidases (DPPs) and metallopeptidases; however, further research is necessary. In conclusion, our work will help to elucidate the implication of these enzymatic activities in the processing of the bioactive peptides present in frog venom, expanding the knowledge of amphibian biology.

Morphological and molecular diversity of Colletotrichum spp. causing pepper spot and anthracnose of lychee (Litchi chinensis) in Australia

Since the 1980s a new disease has been affecting Australian lychee. Pepper spot appears as small, black superficial lesions on fruit, leaves, petioles and pedicels and is caused by Colletotrichum gloeosporioides, the same fungus that causes postharvest anthracnose of lychee fruit. The aim of this study was to determine if a new genotype of C.gloeosporioides is responsible for the pepper spot symptom. Morphological assessments, arbitrarily-primed PCR (ap-PCR) and DNA sequencing studies did not differentiate isolates of C.gloeosporioides from anthracnose and pepper spot lesions. The ap-PCR identified 21 different genotypes of C.gloeosporioides, three of which were predominant. A specific genotype identified using ap-PCR was associated with the production of the teleomorph in culture. Analysis of sequence data of ITS and -tubulin regions of representative isolates did not group the lychee isolates into a monophyletic clade; however, given the majority of the isolates were from one of three genotypes found using ap-PCR, the possibility of a lychee specific group of C.gloeosporioides is discussed.

Novel ATPase activity of the polyprotein intermediate, Viral Protein genome-linked-Nuclear Inclusion-a protease, of Pepper vein banding potyvirus

Potyviruses temporally regulate their protein function by polyprotein processing. Previous studies have shown that VPg (Viral Protein genome-linked) of Pepper vein banding virus interacts with the NIa-Pro (Nuclear Inclusion-a protease) domain, and modulates the kinetics of the protease. In the present study, we report for the first time that VPg harbors the Walker motifs A and B, and the presence of NIa-Pro, especially in cis (cleavage site (E191A) VPg-Pro mutant), is essential for manifestation of the ATPase activity. Mutation of Lys47 (Walker motif A) and Asp88:Glu89 (Walker motif B) to alanine in E191A VPg-Pro lead to reduced ATPase activity, confirming that this activity was inherent to VPg. We propose that potyviral VPg, established as an intrinsically disordered domain, undergoes plausible structural alterations upon interaction with globular NIa-Pro which induces the ATPase activity. (C) 2012 Elsevier Inc. All rights reserved.

Metaphilosophy and educating: Stephen C. Pepper's selectivism as a basis for educational excellence

Atheory of educating is always derived from philosophical tenets. In Western society these tenets are concerned primarily with the provision, maintenance and evolution of knowledge for use by future generations. The provision of knowledge for future generations is for the purpose of ensuring cultural and biological survival. Essentially this provision involves two major criteria: first, that only that knowledge which has been judged to be exce11 ent shou1d be passed on and, second, in add it ion to providing knowledge claims, the evidence for knowledge claims must also be extended in order to fully enrich meaning for an individual involved in a learning experience. Embedded in such a theory of educating are a concept of educational excellence and a concept of the provision of evidence for knowledge claims. This thesis applied the contributions of metaphilosophy to the concepts of educational excellence and the provision of evidence. The metaphilosophy of Stephen C. Pepper was examined for its contributions to a theory of educating and a concept of educational excellence. Metaphilosophy is concerned with making knowledge meaningful. It is a subject matter which may be studied in and of itself and it is a method for acquiring meaning by interpreting knowledge. Historically people have interpreted the knowledge of the world from basically four adequate world views which Pepper termed formism, mechanism, contextual ism and organicism. He later proposed a fifth world view which he termed selectivism. In this thesis these world views were shown to contribute in a variety of ways to educational excellence, most particularly as they allow for interpretations and analysis of evidence about knowledge claims. Selecti vismwas examined in depth and was shown to contribute to educational excellence in two major ways; first, as a world hypothesis which offers an interpretation of the evidence for knowledge claims and, second, as a metahypothesis which provides knowledge about the nature of knowledge. Finally the importance of metaphilosophy in contributing to cultural survival was demonstrated in a discussion of the potential impact of selectivism on a theory of educating and educational excellence.

Problems and prospects of india's major spice-pepper

This study is directed to examine how far price fluctuations in pepper can be controlled in the Indian context so as to have a reasonable and stable income for the primary producers which will ensure an adequate ‘encouragement for higher production and better export earnings. In a study of the methods of controlling violent price fluctuations a important question is that whether the present system of management of supply is satisfactory or not. It is more so when the demand is likely to be sanimlatsd by the importers and wholesalers of the foreign countries. Though pepper is the most important of all the spices gross in India, little work has been done so far to study the problems and prospects of this commodity.

Processing, procurement and marketing of pepper with particular reference to co-operative sector

The present Study is designed to gather, record and analyse data on history of pepper, pepper production, procurement and marketing with particular reference to Kerala. The main emphasis is given to study the'role of cooperative sector with regard to procurement and export efforts and also the services rendered by cooperative sector agencies under MARKETFED and NAFED to pepper trade. The scope of the Study covers the botany, methods of cultivation, fertilizer application, pest control management and other related aspects of pepper. Taking into consideration Kerala's supremacy in pepper cultivation and production, detailed study of its production, procurement, internal and export marketing with reference to Kerala has been given importance. As Kerala accounts for 96 per cent1 of the pepper cultivation and 94 per cent of the pepper production, the present study is entirely confined to Kerala

Characterisation, immunolocalisation and antifungal activity of a lipid transfer protein from chili pepper (Capsicum annuum) seeds with novel alpha-amylase inhibitory properties

Lipid transfer proteins (LTPs) were thus named because they facilitate the transfer of lipids between membranes in vitro. This study was triggered by the characterization of a 9-kDa LTP from Capsicum annuum seeds that we call Ca-LTP(1). Ca-LTP(1) was repurified, and in the last chromatographic purification step, propanol was used as the solvent in place of acetonitrile to maintain the protein`s biological activity. Bidimensional electrophoresis of the 9-kDa band, which corresponds to the purified Ca-LTP(1), showed the presence of three isoforms with isoelectric points (pIs) of 6.0, 8.5 and 9.5. Circular dichroism (CD) analysis suggested a predominance of alpha-helices, as expected for the structure of an LTP family member. LTPs immunorelated to Ca-LTP(1) from C. annuum were also detected by western blotting in exudates released from C. annuum seeds and also in other Capsicum species. The tissue and subcellular localization of Ca-LTP(1) indicated that it was mainly localized within dense vesicles. In addition, isolated Ca-LTP(1) exhibited antifungal activity against Colletotrichum lindemunthianum, and especially against Candida tropicalis, causing several morphological changes to the cells including the formation of pseudohyphae. Ca-LTP(1) also caused the yeast plasma membrane to be permeable to the dye SYTOX green, as verified by fluorescence microscopy. We also found that Ca-LTP(1) is able to inhibit mammalian alpha-amylase activity in vitro.