Rice calcium‐dependent protein kinase OsCPK17 targets plasma membrane intrinsic protein and sucrose‐phosphate synthase and is required for a proper cold stress response

Calcium‐dependent protein kinases (CDPKs) are involved in plant tolerance mechanisms to abiotic stresses. Although CDPKs are recognized as key messengers in signal transduction, the specific role of most members of this family remains unknown. Here, we test the hypothesis that OsCPK17 plays a role in rice cold stress response by analysing OsCPK17 knockout, silencing and overexpressing rice lines under low temperature. Altered OsCPK17 gene expression compromises cold tolerance performance, without affecting the expression of key cold stress‐inducible genes. A comparative phosphoproteomic approach led to the identification of six potential in vivo OsCPK17 targets, which are associated with sugar and nitrogen metabolism, and with osmotic regulation. To test direct interaction, in vitro kinase assays were performed, showing that the sucrose‐phosphate synthase OsSPS4 and the aquaporin OsPIP2;1/OsPIP2;6 are phosphorylated by OsCPK17 in a calcium‐dependent manner. Altogether, our data indicates that OsCPK17 is required for a proper cold stress response in rice, likely affecting the activity of membrane channels and sugar metabolism.     

Genetic diversity and relationships among Nopalea sp. and Opuntia spp. accessions revealed by RAPD,ISSR and ITS molecular markers

Molecular Biology Reports - Dactylopius opuntiae (Cockerell) (carmine cochineal) is an insect pest highly noxious that has spread through cactus pear crops in the Brazilian semiarid region....     

The Biology and Thermal Requirements of the Fennel Aphid Hyadaphis foeniculi (Passerini) (Hemiptera: Aphididae)

The relationship between the insect development rate and temperature was established very early and represents an important ecological variable for modeling the population dynamics of insects. The accurate determination of thermal constant values and the lower and upper developmental thresholds of Hyadaphis foeniculi (Passerini) (Hemiptera: Aphididae) on fennel (Foeniculum vulgare Miller (Apiales: Apiaceae)) crops would obviously benefit the effective application of control measures. This paper is a study of the biology and thermal requirements of H. foeniculi. Winged insects were collected from fennel crops at the Embrapa Algodão in Campina Grande, Paraíba. Nymphs (age ≤24 h) produced by winged insects were subjected to constant temperatures of 15, 20, 25, 28, 30 or 33°C, a photophase of 12 h and a relative humidity of 70±10%. The results of the study showed that at temperatures between 15 and 30°C, H. foeniculi nymphs were able to develop normally. The four instars were found at all temperatures tested. However, temperatures of 3 and 33°C were lethal to the nymphs. The nymph stage development time varied from 5 (30°C) to 19 (15°C) days. The influence of temperature on the development time is dependent on the instar. The base temperature (Tb) and the thermal constant (K) for the nymph stage were estimated at 11.2°C and 107.5 degree-days, respectively. The shortest nymph development stage was observed at 30°C, and the highest nymph viability (85.0%) was observed at 28°C. This information can be used for developing phenological models based on the temperature and development rate relationships so that outbreaks of H. foeniculi in the fennel crop can be predicted, therefore improving the application of control programs targeting this fennel pest.     

Effects of lipid environment on the conformational changes of an ABC importer

In order to shuttle substrates across the lipid bilayer, membrane proteins undergo a series of conformation changes that are influenced by protein structure, ligands, and the lipid environment. To test the effect of lipid on conformation change of the ABC transporter MolBC, EPR studies were conducted in lipids and detergents of variable composition. In both a detergent and lipid environment, MolBC underwent the same general conformation changes as detected by site-directed EPR spectroscopy. However, differences in activity and the details of the EPR analysis indicate conformational rigidity that is dependent on the lipid environment. From these observations, we conclude that native-like lipid mixtures provide the transporter with greater activity and conformational flexibility as well as technical advantages such as reconstitution efficiency and protein stability.     

Two-year citations of JAPPL original articles: evidence of a relative age effect

Several indicators have been used to analyze scientific journals, with the impact factor and the number of citations in a 2-yr calendar time frame (2-YRC) being the most common factors. However, considering that the Journal of Applied Physiology (JAPPL) appears monthly and that calculations of these indicators are based on citations of papers published in previous years, we hypothesized that articles published at the beginning of the year would be cited more in the 2-YRC compared with those appearing in the last issues of the year, a phenomena known as a relative age effect. Our objective was to confirm the existence of a relative age effect in the 2-YRC for original articles published in JAPPL. From 2005 to 2008, a total of 1,726 original articles were published, according to the Web of Science, and 9,973 citations in 2-YRC, varying from 0 to 45, with a mean of 5.78 for individual papers. Although there were no differences in the number of original articles published in a given month (P = 0.99), the 2-YRC varied considerably throughout the year, being higher for those earlier issues of the year, as shown by the linear regression analysis (r(2) = 0.76; P < 0.001). The 2-YRC began at 6.62 during the first 3 mo of the year, dropping by 10% at each 3-mo period. In summary, the longer an article has been out there, the more citations it collects. The relative age effect is a potential confounding variable for the assessment and interpretation of 2-YRC (using calendar years) from JAPPL original articles.     

Past and future approaches to ischemia-reperfusion lesion associated with liver transplantation

Ischemia-reperfusion (I/R) injury associated with liver transplantation remains a serious complication in clinical practice, in spite of several attempts to solve the problem. The present review focuses on the complexity of I/R injury, summarizing conflicting results obtained from the literature about the mechanisms responsible for it. We also review the therapeutic strategies designed in past years to reduce I/R injury, attempting to explain why most of them have not been applied clinically. These strategies include improvements in pharmacological treatments, modifications of University of Wisconsin (UW) preservation solution based on a variety of additives, and gene therapy. Finally, we will consider new potential protective strategies using trimetazidine, 5-amino-4-imidazole carboxamide riboside (AICAR), melatonin, modulators of the renin-angiotensin system (RAS) and the phosphatidylinositol-3-OH kinase (PI3K)-Akt and the p42/p44 extracellular signal-regulated kinases (Erk 1/2) pathway. These strategies have shown promising results for I/R injury but have not been tested in experimental liver transplantation to date. Moreover, we will review ischemic preconditioning, taking into account the recent clinical studies that suggest that this surgical strategy could be appropriate for liver transplantation.     

Distribution of cell surface saccharides on pancreatic cells

We describe here a simple, general procedure for the purification of a variety of lectins, and for the preparation of lectin-ferritin conjugates of defined molar composition and binding properties to be used as probes for cell surface saccharides. The technique uses a “universal” affinity column for lectins and their conjugates, which consists of hog sulfated gastric mucin glycopeptides covalently coupled to agarose. The procedure involes: (a) purification of lectins by chromatography of aqueous extracts of seeds or other lectin-containing fluids over the affinity column, followed by desorption of the desired lectin with its hapten suge; (b) iodination of the lectin to serve as a marker during subsequent steps; (c) conjugation of lectin to ferritin with glutaraldehyde; (d) collection of active lectin-ferritin conjugates by affinity chromatography; and (e) separation of monomeric lectin-ferritin conjugates from larger aggregates and unconjugated lectin by gel chromatography. Based on radioactivity and absorbancy at 310 nm for lectin and ferritin, respectively, the conjugates consist of one to two molecules of lectin per ferrritin molecule. Binding studies of native lectins and their ferritin conjugates to dispersed pancreatic acinar cells showed that the conjugation procedure does not significantly alter either the affinity constant of the lectin for its receptor on the cell surface or the number of sites detected.     

Constitutive and blood meal-induced trypsin genes in Lutzomyia longipalpis

Trypsins constitute some of the most abundant midgut digestive proteases expressed by hematophagous insects upon blood feeding. In addition to their role in the digestion of the blood meal, these proteases also have been implicated in the ability of certain pathogens to infect their natural vector. In sand flies, digestive proteases including trypsins were associated with early killing of Leishmania and are believed to play a role in the species-specificity dictating sand fly vectorial capacity. Our group is involved in studies of midgut digestive proteases in the sand fly Lutzomyia longipalpis, the principal vector of visceral leishmaniasis in Brazil. Here we report on the identification of two cDNAs, Lltryp1 and Lltryp2, which code for putative midgut trypsins in L. longipalpis. Analyses of RNA abundance using semi-quantitative RT-PCR show a different pattern of expression between the two genes. Lltryp1 expression remains undetected until blood feeding and reaches a peak at 12 h post-blood meal (PBM), returning to pre-blood meal levels at 72 h PBM. Additionally, Lltryp1 expression is undetected during larval development. Lltryp2, on the other hand, is constitutively expressed as high levels in the non-blood fed female, but is reduced upon blood feeding. At the end of the digestive cycle, Lltryp2 regains its pre-blood meal levels. This cDNA also is present in all developmental stages and in adult males. This pattern of expression is reminiscent of what is seen in mosquitoes and Old World sand flies, but has characteristics that are unique to L. longipalpis.     

Taste perception in honeybees: just a taste of honey?

The advent of the genomic era has opened new doors to understand the fundamental organization of living organisms and has therefore promoted a fertile field of comparative research that intends to identify similarities and differences between related and unrelated species at the genomic level. One of the organisms whose genome has been recently decoded is that of the honeybee Apis mellifera, enabling a direct comparison with another well-studied insect, the fruit fly Drosophila melanogaster. It was reported that the honeybee has only ten gustatory receptors and thus a very poor taste perception compared to Drosophila, which presents 68 gustatory receptors, and the mosquito Anopheles gambiae, which presents 76 gustatory receptors. In this forum article, we discuss the implications of these findings taking into account previous and new discoveries on honeybee gustation based on behavioral and neurobiological studies by several authors and us. We conclude that the world of taste of a honeybee might not be as poor as proposed and that further studies should integrate molecular, neurobiological, behavioral and ecological approaches to better characterize taste perception in bees. Handling editor: Lars Chittka