Identification and molecular cloning of a novel neuromedin U analog from the skin secretions of toad Bombina maxima

Amphibian skin contains rich neuropeptides. In the present study, a novel neuromedin U (NmU) analog was isolated from skin secretions of Chinese red belly Load Bombina maxima. Being 17-amino acids long, its primary structure was established as DSSGIVGRPFFLFRPRN-NH2, in which the C-terminal 8-residue segment (FFLFRPRN) is the same as that of rat NmU, while the N-terminal part DSSGIVGRP shows a great sequence variation compared with those of NmU peptides from different resources. The peptide, named Bm-NmU-17, was found to elicit concentration-dependent contractile effects on smooth muscle of rat uterus horns. The cDNA Structure of the peptide, as obtained by a 3'-RACE strategy and subsequently cloning from a skin cDNA library, was found to contain a coding region of 438 nucleotides. The encoded precursor is composed of 145 amino acids with a single copy of Bm-NmU-17 located towards the C-terminus. The sequence of the peptide is preceded by a dibasic site (Lys-Arg) and followed by the sequence of Gly-Arg-Lys, providing the sites of cleavage and releasing of the mature peptide. (c) 2005 Elsevier B.V. All rights reserved.

Pachymedusa dacnicolor tryptophyllin-1 (PdT-1): structural characterization, pharmacological activity and cloning of precursor cDNA.

Tryptophyllins are a heterogenous group of amphibian skin peptides originally identified in skin extracts of Neotropical leaf frogs, Phyllomedusa sp., by chemical means. Until now, biosynthetic precursor structure and biological activity remain unreported. Here we describe the isolation of a novel, post-translationally modified tryptophyllin, Lys-Pro-Hyp-Ala-Trp-Val-Pro.amide (PdT-1), from the skin secretion of the Mexican leaf frog, Pachymedusa dacnicolor. Using a 3'- and 5'-RACE strategy and an in vitro skin cDNA library, the PdT-1-encoding precursor was cloned and found to consist of an open-reading frame of 62 amino acids with a single copy of PdT-1 located towards the C-terminus. A synthetic replicate of PdT-1 was found to be a potent myoactive agent, relaxing mammalian arterial smooth muscle and contracting small intestinal smooth muscle at nanomolar concentrations. PdT-1 is thus the first amphibian skin tryptophyllin to be pharmacologically characterized and the first whose precursor cDNA has been cloned.

Bradykinins and their precursor Cdnas from the skin of the Fire-Bellied Toad (Bombina orientalis)

Bradykinin and (Thr6)-bradykinin have been identified in the defensive skin secretion of the fire-bellied toad, Bombina orientalis. The homologous cDNAs for both peptides were cloned from a skin library using a 3'- and 5'-RACE strategy. Kininogen-1 (BOK-1) contained an open-reading frame of 167 amino acid residues encoding four repeats of bradykinin, and kininogen-2 (BOK-2) contained an open-reading frame of 161 amino acid residues encoding two repeats of (Thr6)-bradykinin. Alignment of both precursor nucleotide and amino acid sequences revealed a high degree of structural similarity. These amphibian skin kininogens/preprobradykinins are not biologically analogous to mammalian kininogens.

Purification, characterization and molecular cloning of chymotrypsin inhibitor peptides from the venom of Burmese Daboia russelii siamensis

One novel Kunitz BPTI-like peptide designated as BBPTI-1, with chymotrypsin inhibitory activity was identified from the venom of Burmese Daboia russelii siamensis. It was purified by three steps of chromatography including gel filtration, cation exchange and reversed phase. A partial N-terminal sequence of BBPTI-1, HDRPKFCYLPADPGECLAHMRSF was obtained by automated Edman degradation and a Ki value of 4.77. nM determined. Cloning of BBPTI-1 including the open reading frame and 3' untranslated region was achieved from cDNA libraries derived from lyophilized venom using a 3' RACE strategy. In addition a cDNA sequence, designated as BBPTI-5, was also obtained. Alignment of cDNA sequences showed that BBPTI-5 exhibited an identical sequence to BBPTI-1 cDNA except for an eight nucleotide deletion in the open reading frame. Gene variations that represented deletions in the BBPTI-5 cDNA resulted in a novel protease inhibitor analog. Amino acid sequence alignment revealed that deduced peptides derived from cloning of their respective precursor cDNAs from libraries showed high similarity and homology with other Kunitz BPTI proteinase inhibitors. BBPTI-1 and BBPTI-5 consist of 60 and 66 amino acid residues respectively, including six conserved cysteine residues. As these peptides have been reported to have influence on the processes of coagulation, fibrinolysis and inflammation, their potential application in biomedical contexts warrants further investigation. © 2013 Elsevier Inc.

Performance analysis and prediction in triathlon

Performance in triathlon is dependent upon factors that include somatotype, physiological capacity, technical proficiency and race strategy. Given the multidisciplinary nature of triathlon and the interaction between each of the three race components, the identification of target split times that can be used to inform the design of training plans and race pacing strategies is a complex task. The present study uses machine learning techniques to analyse a large database of performances in Olympic distance triathlons (2008–2012). The analysis reveals patterns of performance in five components of triathlon (three race “legs” and two transitions) and the complex relationships between performance in each component and overall performance in a race. The results provide three perspectives on the relationship between performance in each component of triathlon and the final placing in a race. These perspectives allow the identification of target split times that are required to achieve a certain final place in a race and the opportunity to make evidence-based decisions about race tactics in order to optimise performance.

Influence of Different Performance Levels on Pacing Strategy During the Women's World Championship Marathon Race.

Purpose To analyse pacing strategies displayed by athletes achieving differing levels of performance during an elite level marathon race. Methods Competitors in the 2009 IAAF Women’s Marathon Championship were split into Groups 1, 2, 3, and 4 comprising the first, second, third, and fourth 25% of finishers respectively. Final, intermediate, and personal best (PB) times of finishers were converted to mean speeds, and relative speed (% of PB speed) was calculated for intermediate segments. Results Mean PB speed decreased from Group 1 to 4 and speed maintained in the race was 98.5 + 1.8%, 97.4 + 3.2%, 95.0 + 3.1% and 92.4 + 4.4% of PB speed for Groups 1-4 respectively. Group 1 was fastest in all segments and differences in speed between groups increased throughout the race. Group 1 ran at lower relative speeds than other groups for the first two 5 km segments, but higher relative speeds after 35km. Significant differences (P<0.01) in the percentage of PB speed maintained were observed between Groups 1 and 4, and 2 and 4 in all segments after 20 km, and Groups 3 and 4 from 20-25 km and 30-35 km. Conclusions Group 1 athletes achieved superior finishing times relative to their PB than athletes in other Groups who selected unsustainable initial speeds resulting in subsequent significant losses of speed. It is suggested that psychological factors specific to a major competitive event influenced decision making by athletes and poor decisions resulted in final performances inferior to those expected based on PB times.

Race as political strategy by US presidential candidates: a case study

Race has played an important part in US presidential politics in contemporary history. Different political parties and candidates have followed covert strategies playing on the prejudices of white voters both cognitively and emotionally by linking racerelated issues to the majority's individual and group interests. This elite discourse carried to the public by the mainstream media, along with media's practices of stereotyping, priming, framing and agenda setting, help to justify racial prejudice, discrimination against minorities and their marginalized status, while maintaining the status quo. Taking the social constructionist position, this case study examines the opinions expressed by a sample of undecided voters selected from different geographic locations at various stages of the 1992 US presidential campaign under the themes 'Candidates' racial prejudice' and 'Race is used as political strategy by candidates.'

Non-market strategy and the 'race to the bottom': lessons from the Baywatch experience

This article examines how international firms operate strategically in the non-market environment to secure advantages which improve their cost and/or revenue structures and, hence, their economic performance. It centres on a detailed case study of the globally popular Baywatch television show's efforts during 1999 to secure attractive locational subsidies by placing the states of New South Wales, Queensland (both in Australia) and Hawaii into competition with each other in a 'race to the bottom'. The article is organized into three main sections: the first briefly reviews the concept of non-market strategy; the second examines the Baywatch case in detail; and the conclusion presents some lessons for various stakeholders involved in this and similar non-market contexts.

Low-frequency electrical stimulation combined with a cooling vest improves recovery of elite kayakers following a simulated 1000-m race in a hot environment

This study compared the effects of a low-frequency electrical stimulation (LFES; Veinoplus® Sport, Ad Rem Technology, Paris, France), a low-frequency electrical stimulation combined with a cooling vest (LFESCR) and an active recovery combined with a cooling vest (ACTCR) as recovery strategies on performance (racing time and pacing strategies), physiologic and perceptual responses between two sprint kayak simulated races, in a hot environment (∼32 wet-bulb-globe temperature). Eight elite male kayakers performed two successive 1000-m kayak time trials (TT1 and TT2), separated by a short-term recovery period, including a 30-min of the respective recovery intervention protocol, in a randomized crossover design. Racing time, power output, and stroke rate were recorded for each time trial. Blood lactate concentration, pH, core, skin and body temperatures were measured before and after both TT1 and TT2 and at mid- and post-recovery intervention. Perceptual ratings of thermal sensation were also collected. LFESCR was associated with a very likely effect in performance restoration compared with ACTCR (99/0/1%) and LFES conditions (98/0/2%). LFESCR induced a significant decrease in body temperature and thermal sensation at post-recovery intervention, which is not observed in ACTCR condition. In conclusion, the combination of LFES and wearing a cooling vest (LFESCR) improves performance restoration between two 1000-m kayak time trials achieved by elite athletes, in the heat.

Down, down but not different: Australia’s supermarkets in a race to the bottom

For 25 years, Woolworths told shoppers they were “the fresh food people”. It was a very clear point of difference and delivered the group a sustainable competitive advantage. Any attempt by a competitor to replicate it would have been dismissed as lacking credence; a simple market-follower strategy.

Woolies private label strategy will play directly into the hands of Aldi

Woolworths’ plan to rebrand its private label ranges in an attempt to meet changing shopper demands and combat the growth of Aldi will simply play into the hands of this German discounter. This new strategy, a replication of what Coles did in November 2015, will see their existing “Homebrand” and “Woolworths Essential” product ranges combined under the “Essentials” brand name. However, in focusing on price and private label ranges, supermarkets are in a race to the bottom where there are no points of difference in products except for price.

Promiscuous restriction is a cellular defense strategy that confers fitness advantage to bacteria

Most bacterial genomes harbor restriction-modification systems, encoding a REase and its cognate MTase. On attack by a foreign DNA, the REase recognizes it as nonself and subjects it to restriction. Should REases be highly specific for targeting the invading foreign DNA? It is often considered to be the case. However, when bacteria harboring a promiscuous or high-fidelity variant of the REase were challenged with bacteriophages, fitness was maximal under conditions of catalytic promiscuity. We also delineate possible mechanisms by which the REase recognizes the chromosome as self at the noncanonical sites, thereby preventing lethal dsDNA breaks. This study provides a fundamental understanding of how bacteria exploit an existing defense system to gain fitness advantage during a host-parasite coevolutionary ``arms race.''

The Influence of Performance Level, Age and Gender on Pacing Strategy During a 100-km Ultramarathon

The aim of this study is to analyse the influence of performance level, age and gender on pacing during a 100-km ultramarathon. Results of a 100-km race incorporating the World Masters Championships were used to identify differences in relative speeds in each 10-km segment between participants finishing in the first, second, third and fourth quartiles of overall positions (Groups 1, 2, 3 and 4, respectively). Similar analyses were performed between the top and bottom 50% of finishers in each age category, as well as within male and female categories. Pacing varied between athletes achieving different absolute performance levels. Group 1 ran at significantly lower relative speeds than all other groups in the first three 10-km segments (all P < 0.01), and significantly higher relative speeds than Group 4 in the 6th and 10th (both P < 0.01), and Group 2 in the 8th (P = 0.04). Group 4 displayed significantly higher relative speeds than Group 2 and 3 in the first three segments (all P < 0.01). Overall strategies remained consistent across age categories, although a similar phenomenon was observed within each category whereby ‘top’ competitors displayed lower relative speeds than ‘bottom’ competitors in the early stages, but higher relative speeds in the later stages. Females showed lower relative starting speeds and higher finishing speeds than males. ‘Top’ and ‘bottom’ finishing males displayed differing strategies, but this was not the case within females. Although pacing remained consistent across age categories, it differed with level of performance within each, possibly suggesting strategies are anchored on direct competitors. Strategy differs between genders and differs depending on performance level achieved in males but not females.