Peptide DV-28 amide: Prototype of a novel class of bradykinin receptor antagonist isolated from the defensive skin secretion of bombinid toads

Kinestatin, isolated from the skin of the Chinese toad, Bombina maxima, was the first bradykinin B2 receptor antagonist identified in amphibians. Molecular cloning established that it is co-encoded with the bradykinin-related peptide, maximakinin, within one of several skin kininogens. To examine other species within the genus Bombina for the presence of structural homologues of kinestatin, we subjected skin secretion of the toad, Bombina orientalis, to HPLC fractionation with subsequent bioassay of fractions for antagonism of bradykinin activity using an isolated rat tail artery smooth muscle preparation. A single fraction was located that inhibited bradykinin-induced relaxation of rat arterial smooth muscle and MALDI-TOF analysis of this fraction revealed that it contained a single peptide of molecular mass 3198.5 Da. Further primary structural analysis of this peptide showed that it was a 28-mer with an N-terminal Asp (D) residue and a C-terminal Val (V) residue that was amidated. The peptide was named DV-28 amide in accordance with these primary structural attributes. Synthetic DV-28 amide replicated the observed bradykinin antagonistic effect within the smooth muscle bioassay in a dose-dependent manner. In addition, it was observed to inhibit the proliferation of human microvessel endothelial cells (HMECs) as assessed by MTT assay. Bioinformatic analysis revealed that DV-28 amide was, like kinestatin, co-encoded with a bradykinin receptor agonist on one of two skin kininogens identified in B. orientalis. DV-28 amide thus represents a novel class of bradykinin antagonist from skin secretions of bombinid toads that appear to be a rich source of such novel peptides.

The biology of habitat dominance; can microbes behave as weeds?

Competition between microbial species is a product of, yet can lead to a reduction in, the microbial diversity of specific habitats. Microbial habitats can resemble ecological battlefields where microbial cells struggle to dominate and/or annihilate each other and we explore the hypothesis that (like plant weeds) some microbes are genetically hard-wired to behave in a vigorous and ecologically aggressive manner. These 'microbial weeds' are able to dominate the communities that develop in fertile but uncolonized - or at least partially vacant - habitats via traits enabling them to out-grow competitors; robust tolerances to habitat-relevant stress parameters and highly efficient energy-generation systems; avoidance of or resistance to viral infection, predation and grazers; potent antimicrobial systems; and exceptional abilities to sequester and store resources. In addition, those associated with nutritionally complex habitats are extraordinarily versatile in their utilization of diverse substrates. Weed species typically deploy multiple types of antimicrobial including toxins; volatile organic compounds that act as either hydrophobic or highly chaotropic stressors; biosurfactants; organic acids; and moderately chaotropic solutes that are produced in bulk quantities (e.g. acetone, ethanol). Whereas ability to dominate communities is habitat-specific we suggest that some microbial species are archetypal weeds including generalists such as: Pichia anomala, Acinetobacter spp. and Pseudomonas putida; specialists such as Dunaliella salina, Saccharomyces cerevisiae, Lactobacillus spp. and other lactic acid bacteria; freshwater autotrophs Gonyostomum semen and Microcystis aeruginosa; obligate anaerobes such as Clostridium acetobutylicum; facultative pathogens such as Rhodotorula mucilaginosa, Pantoea ananatis and Pseudomonas aeruginosa; and other extremotolerant and extremophilic microbes such as Aspergillus spp., Salinibacter ruber and Haloquadratum walsbyi. Some microbes, such as Escherichia coli, Mycobacterium smegmatis and Pseudoxylaria spp., exhibit characteristics of both weed and non-weed species. We propose that the concept of nonweeds represents a 'dustbin' group that includes species such as Synodropsis spp., Polypaecilum pisce, Metschnikowia orientalis, Salmonella spp., and Caulobacter crescentus. We show that microbial weeds are conceptually distinct from plant weeds, microbial copiotrophs, r-strategists, and other ecophysiological groups of microorganism. Microbial weed species are unlikely to emerge from stationary-phase or other types of closed communities; it is open habitats that select for weed phenotypes. Specific characteristics that are common to diverse types of open habitat are identified, and implications of weed biology and open-habitat ecology are discussed in the context of further studies needed in the fields of environmental and applied microbiology.

Chroniques françaises, Histoire d'Alexandre, etc.

Contient : Chronique de Normandie, depuis le duc Aubert jusqu'à Henri III, roi d'Angleterre ; Chronique française abrégée de GUILLAUME DE NANGIS, jusqu'à Philippe le Bel ; Chronologie des rois de France jusqu'à Charles VI ; suivie de notes latines : « De Anglia » (fol. 111) ; — « Le serment que le Chancelier de France doit faire, » etc. (fol. 112) ; Chronique des quatre premiers Valois (1327-1393), publiée par S. Luce (Paris, 1862, In 8°) ; Histoire d'Alexandre le Grand (Historia de præliis du Pseudo-Callisthènes), en français ; « Le livre des meurs du gouvernement des seigneurs, appelé les Secrés des secrés d'ARISTOTE » ; « Proposicion faicte de par l'Université de Paris devant nos seigneurs de France et tout le conseil assemblez pour la reformacion du royaume, l'an 1405, le samedi VIIe jour de novembre, par Me Jehan Jarson..., chancellier en l'eglise Nostre-Dame de Paris » ; Recueil de pièces concernant la Normandie. — « Carta Normannorum. » (19 mars 1314 [1315]) ; Vidimus du texte latin du traité de Troyes entre Henri V et Charles VI (21 mai 1420) ; « Pour remonstrer en brief que les habitans du duchié de Normandie et mesmement les bourgois, manans et habitans de la ville de Rouen, ne doivent estre trais en France, ne sont tenus de y sortir jur[i]diction ne y respondre... » — « Raciones précédentes fuerunt composite per mag. Johannem Doule, licenciatum In legibus, condam advocatum In parlamento et magistrum requestarum regis dominum de Neufville » ; Texte français du traité de Troyes (21 mai 1420) ; « Arrest ou declaraciun que ung prelat ou eglise peult acquérir en ses liefs », en faveur de l'abaye de St-Ouen de Rouen (1373 et 1375), etc ; « Confirmatio Cartæ Normannorum » (16 nov. 1423) ; « Decretum unionis orientalis Ecclesiæ cum occidentali, publicalum... In concilio Florentino... 1439 » ; « Copia bulle [Eugenii IV] conservatorie studii Cadomensis » (18 mai 1439) ; Au bas de ce feuillet on lit cette note, de première main : « Cronicas Normannie et Francie, ceterasque alias scripturassuperius scriptas varias materias continentes scribi fecit Robertus, abbas Sancti Michaelis In periculo maris, apud Rothomagum, In pluribus annis, et finaliter perfecit usque hic, anno Domini 1436 » ; Ex-libris : « Messire Joachin de Dinteville, chevalier de l'ordre du Roy et gentilhomme de sa chambre, m'a donné ce livre à Troyes, en febvrier 1578. » (Signature grattée)

A Validated Methodology for Genetic Identification of Tuna Species (Genus Thunnus)

Tuna species of the genus Thunnus, such as the bluefin tunas, are some of the most important and yet most endangered trade fish in the world. Identification of these species in traded forms, however, may be difficult depending on the presentation of the products, which may hamper conservation efforts on trade control. In this paper, we validated a genetic methodology that can fully distinguish between the eight Thunnus species from any kind of processed tissue. Methodology: After testing several genetic markers, a complete discrimination of the eight tuna species was achieved using Forensically Informative Nucleotide Sequencing based primarily on the sequence variability of the hypervariable genetic marker mitochondrial DNA control region (mtDNA CR), followed, in some specific cases, by a second validation by a nuclear marker rDNA first internal transcribed spacer (ITS1). This methodology was able to distinguish all tuna species, including those belonging to the subgenus Neothunnus that are very closely related, and in consequence can not be differentiated with other genetic markers of lower variability. This methodology also took into consideration the presence of introgression that has been reported in past studies between T. thynnus, T. orientalis and T. alalunga. Finally, we applied the methodology to cross-check the species identity of 26 processed tuna samples. Conclusions: Using the combination of two genetic markers, one mitochondrial and another nuclear, allows a full discrimination between all eight tuna species. Unexpectedly, the genetic marker traditionally used for DNA barcoding, cytochrome oxidase 1, could not differentiate all species, thus its use as a genetic marker for tuna species identification is questioned

Grasshoppers and crickets (Insecta : Orthoptera) from the lower cretaceous of southern England

This is a synoptic monograph of fossil Orthoptera from the English Lower Cretaceous (Purbeck and Wealden groups). The previously described taxa of these insects are revised on the basis of type specimens and examination of extensive new material. Eight new genera and 30 new species are proposed: Probaisselcana cretacea sp. nov., Minelcana membranacea gen. et sp. nov., Panorpidium proximum sp. nov., P. bimacillatum sp. nov., ?P. parvum sp. nov. (Elcanidae); ?Cyrtophyllites cretaceus sp. nov. (Haglidae); Aenigmodus minutus gen. et sp. nov., Pseudaboilus wealdensis gen. et sp. nov., P. purbeckensis sp. nov., Tettigoilus sonorus gen. et sp. nov., ?Agrionidium obscurum sp. nov. (Prophalangopsidae); Notocearagryllus britannicus sp. nov., N. grandispeculum sp. nov., N. cordispeculum sp. nov., Anglogryllus lyristes gen. et sp. nov., A. rotundispeculum sp. nov.. Speculogryllus acutispeculum gen. et sp. nov., Sharategia davisi sp. nov., S. batchelorae sp. nov., S. baldocki sp. nov. (Baissogryllidae); ?Araripegryllus orientalis sp. nov. (Gryllidae); Deinovitimia occidentalis sp. nov. (Ensifera: infraorder incertae sedis); Cretoxya rasnitsyni gen. et sp. nov. (Tridactylidae); Locustopsis posterior sp. nov., Zeunerella prior sp. nov., Zessinia borealis sp. nov., Mesolocustopsis anglica sp. nov., M. angusta sp. nov., M. problematica sp. nov., and Britannacrida distincta gen. et sp. nov (Locustopsidae). The subfamily Baisselcaninae is synonymized with Elcaninae, and a new subfamily (Archelcaninae subfam. nov.) is proposed for a segregate of Elcaninae. A preliminary comparison of the Purbeck/Wealden with other Early Cretaceous orthopteran faunas is given. (c) 2006 Published by Elsevier Ltd.

INSECT CHYMOTRYPSINS: CHLOROMETHYL KETONE INACTIVATION AND SUBSTRATE SPECIFICITY RELATIVE TO POSSIBLE COEVOLUTIONAL ADAPTATION OF INSECTS AND PLANTS

Insect digestive chymotrypsins are present in a large variety of insect orders but their substrate specificity still remains unclear. Ewer insect chymotrypsins from 3 different insect orders (Dictyoptera, Coleoptera and two Lepidoptera) were isolated using affinity chromatography. Enzymes presented molecular masses in the range of 20 to 31 kDa and pH optima in the range of 7.5 to 10.0. Kinetic characterization. using different, colorimetric and fluorescent substrates indicated that insect chymotrypsins differ from, bovine chymotrypsin in their primary specificity toward small substrates (like N-benzoyl-L-Tyr p-nitroanilide) rather than on their preference for large substrates (exemplified by Succynil-Ala-Ala-Pro-Phe P-nitroanilide). Chloromethyl ketones (TPCK, N-alpha-tosyl-L-Phe chloromethyl ketone and Z-GGF-CK, N-carbobenzoxy-Gly-Gly-phe-CK) inactivated all chymotrypsins legated. Inactivation rates follow apparent first-order kinetics with variable second order rates (TPCK, 42 to 130 M(-1)s(-1); Z-GGF-CK, 150 to 450 M(-1)s(-1) that may be remarkably low for S. frugiperda chymotrypsin (TPCK, 6 M(-1)s(-1); Z-GGF-CK, 6.1 M(-1) s(-1)). Homology modelling and sequence alignment showed that. in lepidopteran chymotrypsins, differences in the amino acid residues in the neighborhood of the catalytic His 57 may affect its pKa, value. This is Proposed as the cause of the decrease in His 57 reactivity toward chloromethyl ketones. Such amino acid replacement in the active site is proposed. to be an adaptation to the presence of dietary ketones. (C) 2009 Wiley Periodicals, Inc.

Isolation and molecular identification of wine yeasts from a Brazilian vineyard

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

Antifungal activity of propolis on different species of Candida

Propolis is a resinous material collected by bees from the buds or other parts of plants. It is known for its biological properties, having antibacterial, antifungal and healing properties. The antifungal activity of propolis was studied in sensitivity tests on 80 strains of Candida yeasts: 20 strains of Candida albicans, 20 strains of Candida tropicalis, 20 strains of Candida krusei and 15 strains of Candida guilliermondii. The yeasts showed a clear antifungal activity with the following order of sensitivity: C. albicans > C. tropicalis > C. krusei > C. guilliermondii. Patients with full dentures who used a hydroalcoholic propolis extract showed a decrease in the number of Candida.

Yeast diversity isolated from grape musts during spontaneous fermentation from a brazilian winery

Saccharomyces and non-Saccharomyces yeast species from a winery located in Brazil were identified by ribosomal gene-sequencing analysis. A total of 130 yeast strains were isolated from grape surfaces and musts during alcoholic fermentation from Isabel, Bordeaux, and Cabernet Sauvignon varieties. Samples were submitted to PCR-RFLP analysis and genomic sequencing. Thirteen species were identified: Candida quercitrusa, Candida stellata, Cryptococcus flavescens, Cryptococcus laurentii, Hanseniaspora uvarum, Issatchenkia occidentalis, Issatchenkia orientalis, Issatchenkia terricola, Pichia kluyveri, Pichia guilliermondii, Pichia sp., Saccharomyces cerevisiae, and Sporidiobolus pararoseus. A sequential substitution of species during the different stages of fermentation, with a dominance of non-Saccharomyces yeasts at the beginning, and a successive replacement of species by S. cerevisiae strains at the final steps were observed. This is the first report about the yeast distribution present throughout the alcoholic fermentation in a Brazilian winery, providing supportive information for future studies on their contribution to wine quality. © 2013 Springer Science+Business Media New York.

Seleção de leveduras isoladas de uvas e mostos com atividade enzimáticas para melhoramento de vinhos

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

Sazonalidade, sinantropia e preferência por iscas de dípteros necrófagos da região de Rio Claro, SP

Pós-graduação em Ciências Biológicas (Zoologia) - IBRC

La comunicazione chimica in Blattaria: contributo al miglioramento della lotta

CHEMICAL COMMUNICATION IN BLATTARIA: CONTRIBUTION TO THE IMPROVEMENT OF THE CONTROL TECNIQUES The management of cockroach infestations in urban environment has undergone some changes in recent years by moving to the predominant use of baits, thanks to the awareness of the risks connected with the use of spray insecticides. The effectiveness of a bait is determined by the collective performance of its components, including active and inert ingredients, the food attractant and any other attractive odour. This research has focused on the behavioral responses of Italian sinanthropic cockroaches to semiochemicals and food attractants, for the purpose of evaluating a possible practical application and of contributing to the improvement of control techniques. Behavioral and morphological studies have been carried out on Supella longipalpa (F.), a small cockroach that is spreading in Italy. Behavioral assays showed that the fourth and fifth tergites of females are significantly more attractive than other region of the body. Cuticular pores and ducts ending in glandular structures (observed with a S.E.M. = Scanning Electron Microscope) are present in large number on these tergites, suggesting that they could be involved in the production and the release of sexual pheromone. Cockroaches produce an aggregation pheromone that is excreted along with their frass and that consists of volatile and non-volatile compounds, mainly amines and steroidal glycosides. The effectiveness of faecal extracts obtained from excrements of Blattella germanica (L.), Blatta orientalis L., Periplaneta americana (L.) and S. longipalpa was evaluated, both at intraspecific and interspecific level, using a "Y" tube olfactometer. Bioassays showed that faecal extracts obtained with methanol have a good intraspecific attractiveness and, in some cases, they showed also interspecific behavioral responses. A gel was prepared, having physical characteristics that could give a good resistance to dehydration, as a potential basis for a new bait; the gel was then added faecal extracts, obtained with methanol from B. germanica and S. longipalpa frass. Arena-tests showed that the new gel containing faecal extracts is more attractive than some commercial gel formulations used as comparison: it was the only product that could attract 100% of insects placed in the arenas in 4-5 days. In conclusion, the substances involved in chemical communication of Blattaria may be able to effectively increase the attractiveness of products for monitoring and controlling cockroaches.

Per un'archeologia del lavoro in eta' protobizantina. Organizzazione e committenza del cantiere edilizio attraverso i marchi dei marmorari

La ricerca riguarda lo studio del cantiere edilizio protobizantino, con particolare riferimento al ciclo della lavorazione del marmo. Quest’ultimo viene analizzato sotto il profilo amministrativo, tecnico, sociale ed artigianale. L’elemento guida della ricerca sono i marchi dei marmorari, sigle apposte da funzionari e maestranze durante il processo produttivo. Dapprima, fonti letterarie ed epigrafiche, tra cui le sigle di cava e officina su marmo, vengono esaminate per ricostruire il sistema alto-imperiale di amministrazione delle cave e di gestione dei flussi marmorei, nonché l’iter tecnico-artigianale adottato per la produzione dei manufatti. Il confronto con i dati disponibili per la tarda antichità, con particolare riferimento alle cave di Proconneso, evidenzia una sostanziale continuità della prassi burocratico-amministrativa, mentre alcuni cambiamenti si riscontrano nell’ambito produttivo-artigianale. Il funzionamento degli atelier marmorari viene approfondito attraverso lo studio dei marchi dei marmorari. Si tratta di caratteri greci singoli, multipli o monogrammi. Una ricognizione sistematica delle sigle dalla pars Orientalis dell’impero, reperite in bibliografia o da ricognizioni autoptiche, ha portato alla raccolta di circa 2360 attestazioni. Per esse si propone una classificazione tipologica tra sigle di cava, stoccaggio, officina. Tra le sigle di cava si annoverano sigle di controllo, destinazione/committenza, assemblaggio/posizionamento. Una particolare attenzione è riservata alle sigle di officina, riferibili ad un nome proprio di persona, ovvero al πρωτομαΐστωρ, il capo-bottega che supervisionava il lavoro dei propri artigiani e fungeva da garante del prodotto consegnato alla committenza. Attraverso lo studio comparato delle sigle reperite a Costantinopoli e in altri contesti si mette in luce la prassi operativa adottata dagli atelier nei processi di manifattura, affrontando anche il problema delle maestranze itineranti. Infine, sono analizzate fonti scritte di varia natura per poter collocare il fenomeno del marmo in un contesto socio-economico più ampio, con particolare riferimento alle figure professionali ed artigianali coinvolte nei cantieri e al problema della committenza.