Rosaceae conserved orthologous sequences marker polymorphism in sweet cherry germplasm and construction of a SNP-based map

The Rosaceae Conserved Orthologous Set (RosCOS) provides a gene-based genome-wide set of markers that have been used in comparative analyses of peach (Prunus persica), apple (Malus × domestica), and strawberry (Fragaria spp.). In order to extend the use of these RosCOS to sweet cherry (Prunus avium L.), we identified markers that are polymorphic in breeding germplasm. Ninety-five percent (595/627) of previously designed RosCOS primer pairs amplified a product in six sweet cherry cultivars predicted to represent the range of genetic diversity in breeding germplasm. A total of 45% (282/627) RosCOS were polymorphic among the six cultivars, and allele number ranged from 2 to 6, with a genome-wide mean of 2.35. A subset of 92 genome-wide single nucleotide polymorphisms (SNPs) corresponding to 76 RosCOS was analyzed in 36 founder accessions and progeny. The expected and observed heterozygosity suggested that 83% of the RosCOS were in Hardy–Weinberg equilibrium, implying that most RosCOS behave as neutral markers. Principal coordinate analysis (PCO) identified one wild accession and two Spanish landraces that clustered differently from the other accessions. The relatively high number of unique alleles found in the three differentially clustered selections suggested that their use as parents has potential to increase the genetic diversity in future US-bred cultivars. Of the 92 RosCOS SNPs, 81 SNPs that represented 68 genome-wide RosCOS segregated in four mapping populations. These RosCOS were mapped in four F₁ populations, thereby greatly improving the genetic linkage map of sweet cherry.     

A tale of two cyclists: a cross-cultural comparison between Taiwanese and Filipino perceptions on cycling infrastructure landscapes

How is cycling culture defined? Because the word “culture” brings with it deep complexities, there is a need to understand varying contexts in looking for suitable strategies toward the advancement of cycling culture. The stage for cycling culture is the landscape where cycling infrastructure can be properly provided. With development, an influential element would be economic capacity. The paper explores the influence of the economic development divide by comparing cyclist perception between developed and developing countries, namely Taiwan and the Philippines. An online survey between 122 Taiwanese and 111 Filipino cyclists was conducted to find out the landscape needs of people to consider cycling transport based on affordances in the landscape. The variables selected were based on landscape elements for cycling as a commuting activity. The data were processed through factor analysis to reveal latent landscape needs to profile cycling needs. Two factors were identified as ‘environmental’ and ‘civil facilities.’ The factor loadings were then compared based on the nationalities which revealed that the bike riding motivation were different with relating to the context of their respective environment and similar with the basic infrastructural demands.

     

Attenuation of induced bronchoconstriction in healthy subjects: effects of breathing depth.

The effects of breathing depth in attenuating induced bronchoconstriction were studied in 12 healthy subjects. On four separate, randomized occasions, the depth of a series of five breaths taken soon (approximately 1 min) after methacholine (MCh) inhalation was varied from spontaneous tidal volume to lung volumes terminating at approximately 80, approximately 90, and 100% of total lung capacity (TLC). Partial forced expiratory flow at 40% of control forced vital capacity (V(part)) and residual volume (RV) were measured at control and again at 2, 7, and 11 min after MCh. The decrease in V(part) and the increase in RV were significantly less when the depth of the five-breath series was progressively increased (P < 0.001), with a linear relationship. The attenuating effects of deep breaths of any amplitude were significantly greater on RV than V(part) (P < 0.01) and lasted as long as 11 min, despite a slight decrease with time when the end-inspiratory lung volume was 100% of TLC. In conclusion, in healthy subjects exposed to MCh, a series of breaths of different depth up to TLC caused a progressive and sustained attenuation of bronchoconstriction. The effects of the depth of the five-breath series were more evident on the RV than on V(part), likely due to the different mechanisms that regulate airway closure and expiratory flow limitation.     

Effects of chemical modifications of crotoxin B, the phospholipase A(2) subunit of crotoxin from Crotalus durissus terrificus snake venom, on its enzymatic and pharmacological activities.

Crotoxin B, the basic Asp49-PLA(2) subunit from crotoxin, the main component of Crotalus durissus terrificus venom, displays myotoxic, edema-inducing, bactericidal (upon Escherichia coli), liposomal-disrupting and anticoagulant activities. Chemical modifications of His (with 4-bromophenacyl bromide, BPB), Tyr (with 2-nitrobenzenesulphonyl fluoride, NBSF), Trp (with o-nitrophenylsulphenyl chloride, NPSC) and Lys (with acetic anhydride) residues of this protein, in addition to cleavage with cyanogen bromide (CNBr) and inhibition with ethylenediaminetetraacetic acid (EDTA), were carried out in order to study their effects on enzymatic and pharmacological activities. Lethality was reduced after modification of His or Lys residues, as well as after cleavage with CNBr, while enzymatic activity was completely abolished after modification of His or incubation with EDTA. Modification of Lys or Tyr, or cleavage with CNBr, partially reduced enzymatic activity. Anticoagulant activity was modified similarly to enzymatic activity, evidencing the dependency of this pharmacological effect on catalytic activity. Myotoxicity was reduced after modification of His or Lys, as well as after cleavage with CNBr, whereas EDTA reduced this effect to a lesser extent. Bactericidal effect was significantly reduced only after modification of Lys and after cleavage with CNBr. Edema-inducing activity was partially inhibited after treatment with EDTA and strongly reduced after acetylation of Lys residues and cleavage with CNBr, being only partially reduced after His alkylation. On the other hand, liposome disrupting activity was only partially reduced after modification of His and Tyr or after cleavage with CNBr. Modification of Trp residue partially reduced lethality and myotoxicity but did not affect enzymatic or anticoagulant activities. These data indicate that enzymatic activity is relevant for some pharmacological effects induced by crotoxin B (mainly lethal, myotoxic and anticoagulant activities), and also evidence that this subunit of crotoxin displays regions different from the active catalytic site which are involved in some of the toxic and pharmacological effects induced by this phospholipase A(2).     

Islands within islands: a molecular phylogenetic study of the Leucocroton alliance (Euphorbiaceae) across the Caribbean Islands and within the serpentinite archipelago of Cuba

Aim Our aim was to investigate the historical biogeography of the three genera of the Leucocroton alliance (i.e. Garciadelia Jestrow & Jiménez Rodr., Lasiocroton Griseb., and Leucocroton Griseb., Euphorbiaceae). Location  The alliance is restricted to the Bahamas, Cuba, Hispaniola and Jamaica. Methods Members of the Leucocroton alliance, along with representatives from tribe Adelieae (Adelia L. and Philyra Klotzsch.), were included in a molecular phylogenetic analysis based upon nucleotide sequences of the internal transcribed spacer region of the nuclear ribosomal DNA and the non‐coding chloroplast regions psbM–trnD and ycf6–pcbM. The program s‐diva was used to calculate ancestral areas based on the phylogenetic trees and present species distributions. Results Phylogenetic analyses support the monophyly of the three genera. The ancestral area of the Leucocroton alliance is eastern Cuba and Hispaniola. Ancestral forms of Leucocroton arose on eastern Cuba and underwent two migrations across the island. The ancestor of Lasiocroton also originated on eastern Cuba followed by later dispersal to and speciation events on the other islands. Our study also suggests that ancestral forms of the Leucocroton alliance probably occurred on limestone soils. Main conclusions Our study concurs with previous hypotheses suggesting that the flora of serpentinite regions of the Caribbean derives from other types of soils. The serpentine endemics of the Leucocroton alliance have a single origin and represent one of the most extraordinary examples of speciation in this unique environment of the New World. The high colonization success achieved by the members of Leucocroton on serpentine soils was not attained by the other genera of the alliance, which occur on limestone areas.     

Patient and impatient punishers of free-riders

Costly punishment of cheaters who contribute little or nothing to a cooperating group has been extensively studied, as an effective means to enforce cooperation. The prevailing view is that individuals use punishment to retaliate against transgressions of moral standards such as fairness or equity. However, there is much debate regarding the psychological underpinnings of costly punishment. Some authors suggest that costly punishment must be a product of humans'' capacity for reasoning, self-control and long-term planning, whereas others argue that it is the result of an impulsive, present-oriented emotional drive. Here, we explore the inter-temporal preferences of punishers in a multilateral cooperation game and show that both interpretations might be right, as we can identify two different types of punishment: punishment of free-riders by cooperators, which is predicted by patience (future orientation); and free-riders'' punishment of other free-riders, which is predicted by impatience (present orientation). Therefore, the picture is more complex as punishment by free-riders probably comes not from a reaction against a moral transgression, but instead from a competitive, spiteful drive. Thus, punishment grounded on morals may be related to lasting or delayed psychological incentives, whereas punishment triggered by competitive desires may be linked to short-run aspirations. These results indicate that the individual''s time horizon is relevant for the type of social behaviour she opts for. Integrating such differences in inter-temporal preferences and the social behaviour of agents might help to achieve a better understanding of how human cooperation and punishment behaviour has evolved.     

NAADP+ is an agonist of the human P2Y11 purinergic receptor

Nicotinic acid adenine dinucleotide phosphate (NAADP+) is an intracellular second messenger releasing Ca2+ from intracellular stores in different cell types. In addition, it is also active in triggering [Ca2+](i) increase when applied extracellularly and various underlying mechanisms have been proposed. Here, we used hP2Y(11)-transfected 1321N1 astrocytoma cells to unequivocally establish whether extracellular NAADP+ is an agonist of the P2Y(11) receptor, as previously reported for beta-NAD+ [I. Moreschi, S. Bruzzone, R.A. Nicholas, et al., Extracellular NAD+ is an agonist of the human P2Y11 purinergic receptor in human granulocytes, J. Biol. Chem. 281 (2006) 31419-31429]. Extracellular NAADP+ triggered a concentration-dependent two-step elevation of [Ca2+](i) in 1321N1-hP2Y(11) cells, but not in wild-type 1321N1 cells, secondary to the intracellular production of IP(3), cAMP and cyclic ADP-ribose (cADPR). Specifically, the transient [Ca2+](i) rise proved to be related to IP(3) overproduction and to consequent Ca2+ mobilization, while the sustained [Ca2+](i) elevation was caused by the cAMP/ADP-ribosyl cyclase (ADPRC)/cADPR signalling cascade and by influx of extracellular Ca2+. In human granulocytes, endogenous P2Y(11) proved to be responsible for the NAADP+-induced cell activation (as demonstrated by the use of NF157, a selective and potent inhibitor of P2Y(11)), unveiling a role of NAADP+ as a pro-inflammatory cytokine. In conclusion, we provide unequivocal evidence for the activation of a member of the P2Y receptor subfamily by NAADP+.