Proteomics in Drosophila melanogaster: first 2D database of larval hemolymph proteins

A proteomic approach was used for the identification of larval hemolymph proteins of Drosophila melanogaster. We report the initial establishment of a two-dimensional gel electrophoresis reference map for hemolymph proteins of third instar larvae of D. melanogaster. We used immobilized pH gradients of pH 4-7 (linear) and a 12-14% linear gradient polyacrylamide gel. The protein spots were silver-stained and analyzed by nanoLC-Q-Tof MS/MS (on-line nanoscale liquid chromatography quadrupole time of flight tandem mass spectrometry) or by Matrix assisted laser desorption time of flight MS (MALDI-TOF MS). Querying the SWISSPROT database with the mass spectrometric data yielded the identity of the proteins in the spots. The presented proteome map lists those protein spots identified to date. This map will be updated continuously and will serve as a reference database for investigators, studying changes at the protein level in different physiological conditions.     

Teen at work: the burden of a double shift on daily activities

The purpose of this study was to the evaluate time spent by working and nonworking adolescents on daily activities (work, home duties, school, transportation, other activities, leisure, sleep, and naps). Twenty-seven students, 8 male workers, 8 female workers, 5 male nonworkers, and 6 female nonworkers, ages 14-18 yrs participated in the study. They attended evening classes Monday-Friday (19:00-22:30h) in a public school in the city of S?o Paulo, Brazil. The students answered a comprehensive questionnaire on the characterization of their life, work, and health conditions. Simultaneously, they wore actigraphs (Ambulatory Monitoring, Inc.) and completed a diary of their daily activities (time spent at work, on home duties, commuting, leisure, other activities) for a minimum of 10 to a maximum of 17 consecutive days. The means of the variables were tested for differences by a two-factor (work and sex) ANOVA and Student-t test applied to pair-wise samples (weekdays and weekends). The average duration during weekdays of working time was 7 h 09 min and home duties 0 h 48 min. As for commuting time, there was a work effect [F(1,23) = 4.9; p = 0.04]; mean commuting time was 2 h 22 min for workers (males and females) and 1 h 25 min for nonworkers. There was a significant difference between workers and nonworkers [F(1,23) = 4.6; p = 0.04] regarding extra-cirricular class activities; workers spent a mean of 3 min/day on them as opposed to 1 h 14 min by nonworkers. The average daily time spent on leisure activities by workers was 6h 31 min; whereas, for nonworkers it was 7h 38min. Time spent in school amounted to 2h 47min for workers in comparison to 3h 22min by nonworkers. There was a significant work effect upon sleep [F(1,23)= 10.0; p <0.01]. The work effect upon nighttime sleep duration was significant [F(1,23)= 16.7; p <0.01]. Male workers showed a mean night sleep of 6 h 57 min and female workers 07h 15min. The average nighttime sleep duration for nonworkers was 9 h 06 min. There was a significant interactive effect between work and sex [F(1,23)= 5.6; p=0.03] for naps. Female workers showed took shortest nap on average (36 min; SD = 26 min), and female nonworkers the longest naps (1 h 45min; SD= 35min). Study and employment exert significant impact on the life and activities of high school students. Work affects sleep and nap duration plus the amount of time spent in school and other extra-curricular activities.     

The interaction of a glycosaminoglycan heparin,with HIV-1 major envelope glycoprotein

We demonstrate in vitro the occurence of a specific but low-affinity interaction between soluble tetrameric rgp160 or soluble monomeric or tetrameric rgp120 and heparin-agarose (HA). This interaction is saturable, pH and temperature-dependent, and can be inhibited by soluble heparin, but not by soluble dextran. In buffer supplemented with 10 mM CaCl₂, the C₅₀ of soluble heparin, i.e., the concentration of soluble heparin which leads to 50% inhibition of the binding of [¹²⁵I]rgp160 or [¹²⁵I]rgp120 to HA, is 1.1. · 10^?4 disaccharidic molar concentration for rgp160 and 3.2 · 10^?4 disaccharidic molar concentration for rgp120, which indicates low-affinity interactions. Upon chromatography on HA, [¹²⁵I]rgp160 is repeatedly eluted as a retarded fraction when compared to the elutions volume of [¹²⁵I]rgp160-soluble heparin complex. Under the same experimental conditions, [¹²⁵I]rgp120 is also eluted, but as a less retarded fraction than [¹²⁵I]rgp160. Taken together, these results suggest that, at least part of the described anti HIV-1 activity of heparin might be mediated by interaction with HIV-1 major envelope glycoprotein.     

Metal X-ray microanalysis in the olfactory system of rainbow trout exposed to low level of copper

Summary— It has recently been shown that a chronic copper exposure induces specific degeneration of olfactory receptor cells in rainbow trout; however, the exact mechanism of action of the metal is not yet known. Using X-ray microanalysis in transmission electron microscopy, we have studied the distribution of metal in the olfactory system of fish exposed for 15, 30 and 60 days to 20 μg/l of copper. This was done in order to determine if it was accumulated in receptor cells and transported into the central nervous system via the olfactory nerve. No copper accumulation was detected either in the olfactory epithelium, in the olfactory nerve or in the olfactory bulb. The heavy metal was exclusively found within melanosomes of melanophores located in the lamina propria. After 60 days of exposure, the copper content in melanosomes was about two-fold higher than that in controls. As far as some morphological recovery took place in the olfactory organ during the metal exposure, which lets us suppose that some detoxication mechanism occurs, it could be suggested that melanophores might be somehow involved in such a mechanism.     

On the occurrence of twisted fibrillar structures in the cytoplasm of the red algaAlsidium helminthochorton (La Tourette) Kütz., ultrastructural and cytochemical observations

Summary The cytoplasm ofAlsidium cells contains unusual structures showing various fibrillar arrangements, either stacked rows of arcs or parallel fibrils in longitudinal view alternating with fibrils in cross-section. These regions, located exclusively in the cytoplasmic matrix, are highly proteinaceous and do not seem to be complexed with polysaccharides. Tilting observations under the electron microscope show that the appearance of arcs is an illusion and that the pattern fits the explanation given byBouligand and collaborators for various twisted biological structures.     

FLUCTUATING ASYMMETRY DOES NOT CONSISTENTLY REFLECT SEVERE DEVELOPMENTAL DISORDERS IN HUMAN FETUSES

Developmental instability (DI), as measured by fluctuating asymmetry (FA), may reflect fitness and facilitate the expression of morphological variation. Insights in the underlying mechanisms and magnitude of DI during early development would increase our understanding of its role in evolutionary biology. We studied associations between FA and congenital abnormalities of different origins and functional systems in deceased human fetuses. Major congenital abnormalities corresponded to severe, often-lethal developmental disorders disrupting normal development from early organogenesis onward, but only moderately increased FA. Lower FA with age also supported the hypothesis that more severe abnormalities, leading to an earlier death, increased DI. Although FA related significantly to measures of fitness or health, we anticipated stronger associations because fetal health problems were detrimental. Furthermore, elevated FA occurred in only 4 of 17 disorders (left–right patterning, limb defects, and problems of bronchopulmonary and urogenital system). Fetuses experiencing major abnormalities other than these four types did not show increased FA. This suggests that the functional importance of symmetry in limbs has resulted in strong selection for symmetry and reduced its sensitivity to stress. Finally, the observed patterns suggest that specific developmental pathways have a stronger effect on DI than others do.     

The alkaline hydrolysis of nucleic acid for removal of RNA associated fluorescence in phenantridium related flow through cytofluorometry

Summary Flow through microfluorometry of phenantridium and fluoresceinisothiocyanate stained HeLa cells showed the absence of extranuclear RNA related fluorescence after prolonged hydrolysis of cells in ethanolic barium hydroxide. DNA distribution were as good as in RNAse treated samples. Distribution pattern of cellular protein/DNA ratios yielded greater resolution power than those after RNAse digestion. The cellular protein content has not been affected. The ethanolic hydrolysis has the advantage to be of less expense and to allow cell conservations for further measurements with other well preserving fixatives. Thus, application of glutaraldehyde has been proved to be valuable.     

Insect lysozymes from three species of lepidoptera: Their structural relatedness to the c (chicken) type lysozyme

Summary Sequence studies of the N-terminal halves of the lysozymes isolated fromBombyx mori, Galleria mellonella andSpodoptera littoralis (Lepidoptera) allow us to classify these enzymes among the c (chicken) type lysozymes.114th communication on lysozymes.     

Étude comparée des protéines des polyèdres et des virions de trois isolats deBaculovirus deSpodoptera littoralis

Résumé Trois isolats de polyèdres nucléaires deSpodoptera littoralis Boisduval (isolats Egypte, Maroc et Lyon) sont comparés entre eux et avec des préparations de polyèdres nucléaires des lépidoptèresBombyx mori L.,Galleria mellonella L.,Lymantria dispar L. etAutographa californica Speyer. L'analyse porte à la fois sur la composition peptidique des virions, sur le poids moléclaire des protéines des polyèdres et sur les parentés antigéniques entre ces protéines. Nous montrons que chaque espèce de lépidoptères possède un type différent de virus et que les trois isolats de polyèdres nucléaires obtenus chezS. littoralis sont très semblables. Cependant l'isolat Lyon diffère des deux autres et se compose d'un mélange de deux souches. La première souche commune aux trois isolats se propage à la fois sur larves deS. littoralis etSpodoptera exigua Hübner, tandis que la seconde souche propre à l'isolat Lyon ne se multiplie que sur larves deS. littoralis.

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Summary Three NPV isolates ofSpodoptera littoralis Boisduval (Egypt, Maroc and Lyon isolates) are studied and compared to NPV ofBombyx mori L.,Galleria mellonella L.,Porthetria dispar L. andAutographa californica Speyer. Polyhedron protein and peptide analyses as well as immunological comparisons were performed. Each of lepidopterous species possesses a different virus type. The 3 isolates coming fromS. littoralis are similar. Nevertheless, Lyon isolate is a mixture of 2 closely related strains. The 1st strain multiplies inS. littoralis andSpodoptera exigua Hbn. larvae and is found in the 3 isolates. The 2nd strain belongs only to Lyon isolate and does not develop inS. exigua larvae. The polyhedron protein molecular weight of the strain 1 is 30 590 and that of the strain 2 is 28 533. Strain 2 has 1 extrapeptide of MW 80 000 in addition to 11 virion peptides common to the both strains. Attention is drawn to the importance of NPV isolates differences for the bioinsecticide characterization.