Characterization of chicken gizzard calcyclin and examination of its interaction with caldesmon

Using a procedure developed to purify calcyclin from mouse Ehrlich ascites tumor cells calcyclin was purified from smooth muscle of chicken gizzard. Chicken gizzard calcyclin bound to phenyl-Sepharose in a calcium dependent manner as did mouse EAT cells and rabbit lung calcyclin but appeared to be more acidic than its mammalian counterparts as revealed by ion exchange chromatography on Mono Q. Chicken gizzard calcyclin bound ⁴⁵Ca²⁺ on nitrocellulose filters and exhibited a shift in electrophoretic mobility on urea-PAGE depending on Ca²⁺ concentration. Crosslinking experiments with BS³ showed that chicken gizzard calcyclin was able to form noncovalent dimers. As indicated by a decrease in maximum tryptophan fluorescence emission of caldesmon (about 14% at 1:1 molar ratio) and displacement of calmodulin from its complex with caldesmon, chicken gizzard calcyclin binds caldesmon. This binding was, however, much weaker than that of calmodulin and could not influence the interaction of caldesmon with actin. In consequence, calcyclin was unable to reverse the inhibitory effect of caldesmon on actin-activated Mg²⁺-ATPase activity of myosin in the presence of Ca²⁺.     

CHRONOCRISIS: When Cell Cycle Asynchrony Generates DNA Damage in Polyploid Cells

Polyploid cells contain multiple copies of all chromosomes. Polyploidization can be developmentally programmed to sustain tissue barrier function or to increase metabolic potential and cell size. Programmed polyploidy is normally associated with terminal differentiation and poor proliferation capacity. Conversely, non-programmed polyploidy can give rise to cells that retain the ability to proliferate. This can fuel rapid genome rearrangements and lead to diseases like cancer. Here, the mechanisms that generate polyploidy are reviewed and the possible challenges upon polyploid cell division are discussed. The discussion is framed around a recent study showing that asynchronous cell cycle progression (an event that is named “chronocrisis”) of different nuclei from a polyploid cell can generate DNA damage at mitotic entry. The potential mechanisms explaining how mitosis in non-programmed polyploid cells can generate abnormal karyotypes and genetic instability are highlighted.     

Studies of insect peptides alloferon,Any‐GS and their analogues. Synthesis and antiherpes activity

The subject of these studies was synthesis and determination of biological properties of a series of insect peptides, such as alloferon, Any‐GS and their analogues. The synthesis of 14 peptides was performed by the solid‐phase method. Biological effect of these peptides was evaluated by the antiviral test against Human Herpes Virus type 1 (HHV‐1) in vitro using a Vero cell line. It was found that the investigated peptides inhibit the replication of HHV‐1 in Vero cells. Copyright © 2010 European Peptide Society and John Wiley & Sons, Ltd.     

Beta-carotene storage, conversion to retinoic acid, and induction of the lipocyte phenotype in hepatic stellate cells

Hepatic stellate cells (HSCs) are the major site of retinol (ROH) metabolism and storage. GRX is a permanent murine myofibroblastic cell line, derived from HSCs, which can be induced to display the fat-storing phenotype by treatment with retinoids. Little is known about hepatic or serum homeostasis of beta-carotene and retinoic acid (RA), although the direct biogenesis of RA from beta-carotene has been described in enterocytes. The aim of this study was to identify the uptake, metabolism, storage, and release of beta-carotene in HSCs. GRX cells were plated in 25 cm(2) tissue culture flasks, treated during 10 days with 3 micromol/L beta-carotene and subsequently transferred into the standard culture medium. beta-Carotene induced a full cell conversion into the fat-storing phenotype after 10 days. The total cell extracts, cell fractions, and culture medium were analyzed by reverse phase high-performance liquid chromatography for beta-carotene and retinoids. Cells accumulated 27.48 +/- 6.5 pmol/L beta-carotene/10(6) cells, but could not convert it to ROH nor produced retinyl esters (RE). beta-Carotene was directly converted to RA, which was found in total cell extracts and in the nuclear fraction (10.15 +/- 1.23 pmol/L/10(6) cells), promoting the phenotype conversion. After 24-h chase, cells contained 20.15 +/- 1.12 pmol/L beta-carotene/10(6) cells and steadily released beta-carotene into the medium (6.69 +/- 1.75 pmol/ml). We conclude that HSC are the site of the liver beta-carotene storage and release, which can be used for RA production as well as for maintenance of the homeostasis of circulating carotenoids in periods of low dietary uptake.     

Effects of forest fragmentation on small mammals in an Atlantic Forest landscape

Recent studies on the effects of tropical forest fragmentation indicate that fragmented landscapes are complex and heterogeneous systems influenced by factors other than the size or degree of isolation of forest remnants: of particular importance are the quality of the matrix and the edge-induced habitat changes. In order to investigate the influence of these factors, small mammals were surveyed in 36 sites in the landscape of Una, a region that encompasses some of the last and largest Atlantic Forest remnants in northeastern Brazil. Six sites were distributed on each of six landscape components – the interiors and edges of small remnants, the interiors and edges of large remnants, and the most common forested habitats found in the matrix. The survey comprised 46,656 trap-nights and yielded 1725 individuals of 20 species of rodents and marsupials. Results revealed: an increase in beta-diversity caused by fragmentation; the contrasting effects of the altered forested habitats of the matrix, which harbor both forest and disturbance-adapted species; a greater importance of edge effect than of patch size to the observed changes in small mammal community in remnants; an association among terrestrial forest species and among arboreal forest species in terms of the distribution and abundance in the Una mosaic; and a distinctive vulnerability of these two groups of species to fragmentation. Results emphasize the biological importance and conservation value of both fragmented landscapes and small remnants in the Atlantic Forest, as well as the importance of management techniques to control and attenuate edge effects.     

On the way from morphology to phonology: German linking elements and the role of the phonological word

German linking elements are sometimes classified as inflectional affixes, sometimes as derivational affixes, and in any case as morphological units with at least seven realisations (e.g. -s-, -es-, -(e)n-, -e-). This article seeks to show that linking elements are hybrid elements situated between morphology and phonology. On the one hand, they have a clear morphological status since they occur only within compounds (and before a very small set of suffixes) and support the listener in decoding them. On the other hand, they also have to be analysed on the phonological level, as will be shown in this article. Thus, they are marginal morphological units on the pathway to phonology (including prosodics). Although some alloforms can sometimes be considered former inflectional endings and in some cases even continue to demonstrate some inflectional behaviour (such as relatedness to gender and inflection class), they are on their way to becoming markers of ill-formed phonological words. In fact, linking elements, above all the linking -s-, which is extremely productive, help the listener decode compounds containing a bad phonological word as their first constituent, such as Geburt+s+tag ‘birthday’ or Religion+s+unterricht ‘religious education’. By marking the end of a first constituent that differs from an unmarked monopedal phonological word, the linking element aids the listener in correctly decoding and analysing the compound. German compounds are known for their length and complexity, both of which have increased over time—along with the occurrence of linking elements, especially -s-. Thus, a profound instance of language change can be observed in contemporary German, one indicating its typological shift from syllable language to word language.     

A comparative study of different glycosylation methods for the synthesis of D-mannopyranosides of Nalpha-fluorenylmethoxycarbonyl-trans-4-hydroxy-L-proline allyl ester

The synthesis of Nalpha-fluorenylmethoxycarbonyl-trans-4-hydroxy-4-O-[(2,3,4,6-tetra-O-acetyl)-alpha-d-mannopyranosyl]-l-proline allyl ester and Nalpha-fluorenylmethoxycarbonyl-trans-4-hydroxy-4-O-[(2,3,4,6-tetra-O-benzoyl)-alpha-d-mannopyranosyl]-l-proline allyl ester is described. Glycosylation using K?nigs-Knorr conditions with a benzoyl protected glycosyl donor provided the optimum method. Removal of the allyl ester gave two mannosylated building blocks suitable for solid phase glycopeptide synthesis.     

Identification of calmodulin-binding proteins in brain of worker honeybees

Calmodulin is a Ca(+2)-binding protein important in a variety of cell functions. The Ca(+2)/calmodulin complex interacts with and regulates various enzymes and target proteins, known as calmodulin-binding proteins (CaMBPs). In this study, we revealed a comparative identification of the CaMBPs composition in the worker honeybee (Apis mellifera) brain, considering two different honeybee behaviors in the colony. To this end, the CaMBPs of forager and nurse workers were purified by affinity chromatography, separated in 1D gel, digested and submitted to peptide mass fingerprinting (PMF). In the PMF analysis, 15 different proteins, considered behavior-specific proteins, were identified, one of them exclusively in forager workers and 10 in nurses. All the proteins were classified in terms of their function and cell localization, revealing a greater expression of metabolism-related CaMBPs in both worker subcastes. Protein sequences were then analyzed for the presence of the calmodulin-binding sites. Therefore, the honeybee brain CaMBPs profiles presented differences between worker subcastes. This is the first identification of calmodulin-binding proteins in the brain of A. mellifera upon nursing and foraging behaviors in the colony and this diversity of target proteins for Ca(+2)/CaM may be involved in terms of the function of these proteins in the nervous system.