Purification of intact chloroplasts from spruce (Picea abies) by free-flow electrophoresis

Highly purified chloroplasts were isolated from spruce ( Picea abies [L.] Karst.) needles by free-flow electrophoresis. The chloroplast crude suspension was separated into two structurally different populations. The two populations could not be distinguished according to their protein/chlorophyll ratios and protein patterns from two-dimensional gel electrophoresis. The O² evolution, however, showed differences: the chloroplast population deflected towards the cathode contained a major part of structurally intact chloroplasts in contrast to the population deflected towards the anode. The two populations were not contaminated by endoplasmatic membranes or mitochondria.     

The hatching mechanism of 130‐million‐year‐old insects: an association of neonates,egg shells and egg bursters in Lebanese amber

Hatching is a pivotal moment in the life of most animals. Diverse chemical, behavioural and mechanical methods have evolved in metazoans to break the egg membranes. Among them, many arthropod and vertebrate embryos hatch using ephemeral, frequently convergent structures known as egg bursters. However, the evolutionary processes by which hatching mechanisms and related embryonic structures became established in deep time are poorly understood due to a nearly complete absence from the fossil record. Herein we describe an exceptional c. 130‐million‐year‐old association in Lebanese amber composed of multiple neonate green lacewing larvae, Tragichrysa ovoruptora gen. et sp. nov. (Neuroptera, Chrysopoidea), and conspecific egg remains. Egg bursters with a serrated blade bearing a short process are attached to three longitudinally split egg shells. Embryos of extant green lacewing relatives (Chrysopidae) utilize this egg burster morphotype to open a vertical slit on the egg, after which the burster is moulted and left joined to the empty egg shell. Additionally, the new larval species has extremely elongate dorsal tubercles, an adaptation to carry exogenous debris for protection and camouflage also known from other Cretaceous chrysopoids but absent in modern relatives. The present discovery demonstrates that the hatching mechanism of modern green lacewings was established in the chrysopoid lineage by the Early Cretaceous and proves through direct fossil evidence how some morphological traits related to hatching and linked behaviours, at least in insect embryos, have been subject to a high degree of evolutionary conservatism.     

Molecular analysis of the CYP2F1 gene: identification of a frequent non-functional allelic variant

The CYP2F1 is a human cytochrome P450 that is selectively expressed in lung tissue and involved in the metabolism of various pneumotoxicants with potential carcinogenic effects. In the present study, we report the first systematic investigation of the genetic polymorphism of this enzyme. We analyzed the nucleotidic sequence of the CYP2F1 gene in DNA samples from 90 French Caucasians consisting in 44 patients with lung cancer and 46 control individuals, using single-strand conformation polymorphism analysis of PCR products (PCR-SSCP). We identified 24 novel mutations distributed in the promoter region of the gene, as well as in the coding regions and their flanking intronic sequences. In addition to the wild-type CYP2F1*1 allele, seven allelic variant, CYP2F1*2A, *2B, *3, *4, *5A, *5B and *6, were characterized. The most frequent allelic variant, CYP2F1*2A (25.6%), harbors a combination of 9 mutations, including 2 missense mutations (Asp218Asn and Gln266His) and a 1-bp insertion (c.14_15insC) that creates a premature stop codon in exon 2, probably leading to the synthesis of a severely truncated protein with no catalytic activity. The identification of around 7% of homozygotes for the frameshift mutation in our Caucasian population suggests the existence of an interindividual variation of the CYP2F1 activity and, consequently, the possibility of interindividual differences in the toxic response to some pneumotoxicants and in the susceptibility to certain chemically induced diseases. However, our preliminary results did not show any evidence that the CYP2F1 genetic polymorphism has implications in the pathogenesis of lung cancer.     

Angiotensin II Type I and Prostaglandin F2α Receptors Cooperatively Modulate Signaling in Vascular Smooth Muscle Cells

The angiotensin II type I (AT1R) and the prostaglandin F2α (PGF2α) F prostanoid (FP) receptors are both potent regulators of blood pressure. Physiological interplay between AT1R and FP has been described. Abdominal aortic ring contraction experiments revealed that PGF2α-dependent activation of FP potentiated angiotensin II-induced contraction, whereas FP antagonists had the opposite effect. Similarly, PGF2α-mediated vasoconstriction was symmetrically regulated by co-treatment with AT1R agonist and antagonist. The underlying canonical Gα_q signaling via production of inositol phosphates mediated by each receptor was also regulated by antagonists for the other receptor. However, binding to their respective agonists, regulation of receptor-mediated MAPK activation and vascular smooth muscle cell growth were differentially or asymmetrically regulated depending on how each of the two receptors were occupied by either agonist or antagonist. Physical interactions between these receptors have never been reported, and here we show that AT1R and FP form heterodimeric complexes in both HEK 293 and vascular smooth muscle cells. These findings imply that formation of the AT1R/FP dimer creates a novel allosteric signaling unit that shows symmetrical and asymmetrical signaling behavior, depending on the outcome measured. AT1R/FP dimers may thus be important in the regulation of blood pressure.     

Essential oils from wild populations of Algerian Lavandula stoechas L.: composition, chemical variability, and in vitro biological properties

In an effort to develop local productions of aromatic and medicinal plants, a comprehensive assessment of the composition and biological activities of the essential oils (EOs) extracted from the aerial flowering parts of wild growing Lavandula stoechas L. collected from eleven different locations in northern Algeria was performed. The oils were characterized by GC‐FID and GC/MS analyses, and 121 compounds were identified, accounting for 69.88–91.2% of the total oil compositions. The eleven oils greatly differed in their compositions, since only 66 compounds were common to all oils. Major EO components were fenchone ( 2 ; 11.27–37.48%), camphor ( 3 , 1.94–21.8%), 1,8‐cineole ( 1 ; 0.16–8.71%), and viridiflorol ( 10 ; 2.89–7.38%). The assessed in vitro biological properties demonstrated that the DPPH‐based radical‐scavenging activities and the inhibition of the β‐carotene/linoleic acid‐based lipid oxidation differed by an eight‐fold factor between the most and the least active oils and were linked to different sets of molecules in the different EOs. The eleven EOs exhibited good antimicrobial activities against most of the 16 tested strains of bacteria, filamentous fungi, and yeasts, with minimum inhibitory concentrations (MICs) ranging from 0.16 to 11.90 mg/ml.     

Do changes in gene expression contribute to sexual isolation and reinforcement in the house mouse?

Expression divergence, rather than sequence divergence, has been shown to be important in speciation, particularly in the early stages of divergence of traits involved in reproductive isolation. In the two European subspecies of house mice, Mus musculus musculus and Mus musculus domesticus, earlier studies have demonstrated olfactory‐based assortative mate preference in populations close to their hybrid zone. It has been suggested that this behaviour evolved following the recent secondary contact between the two taxa (~3,000 years ago) in response to selection against hybridization. To test for a role of changes in gene expression in the observed behavioural shift, we conducted a RNA sequencing experiment on mouse vomeronasal organs. Key candidate genes for pheromone‐based subspecies recognition, the vomeronasal receptors, are expressed in these organs. Overall patterns of gene expression varied significantly between samples from the two subspecies, with a large number of differentially expressed genes between the two taxa. In contrast, only ~200 genes were found repeatedly differentially expressed between populations within M. m. musculus that did or did not display assortative mate preferences (close to or more distant from the hybrid zone, respectively), with an overrepresentation of genes belonging to vomeronasal receptor family 2. These receptors are known to play a key role in recognition of chemical cues that handle information about genetic identity. Interestingly, four of five of these differentially expressed receptors belong to the same phylogenetic cluster, suggesting specialization of a group of closely related receptors in the recognition of odorant signals that may allow subspecies recognition and assortative mating.     

Biosynthesis and regulation of juvenile hormone III,juvenile hormone III bisepoxide,and methyl farnesoate during the reproductive cycle of the grey fleshfly,Neobellieria (Sarcophaga) bullata

To study the effect of brain signals on the biosynthesis of juvenile hormone by the corpora allata of the grey fleshfly Neobellieria bullata, exposed corpora allata connected to the brain were surgically removed from sugar-fed flies and incubated in vitro with L -[³H-methyl]methionine. After incubation, the media together with the tissues were analyzed by HPLC. [³H]Juvenile hormone III (JH III), [³H]JH III bisepoxide (BE), [³H]methyl farnesoate (MF) and an unknown [³H]labeled metabolite (Un) were identified as the primary products. The rate of synthesis of [³H]JH III bisepoxide was higher than that of [³H]JH III, [³H]MF and [³H]Un. Two days after a liver meal, female flies synthesized more JH III, MF, BE, and the Un than did males. Synthesis of JH III, BE, and MF in females was lower during the previtellogenic, sugar-feeding period than during the vitellogenic liver-feeding period. Isolated corpus cardiacum–corpus allatum (CC-CA) complexes that were incubated in vitro synthesized less JH III, MF, and BE, as compared to complexes that were attached to the brain, indicating that the brain probably modulates the biosynthesis of JH III, MF, and BE in the corpora allata. Upon incubation of brain–CC–CA complexes with Neb-TMOF (10^–8 M), Neb-colloostatin (10^–8 M), ovarian, or brain extracts resulted in significant inhibition of JH III and BE biosynthesis in the presence of ovarian extracts. These results indicate that allatostatin-like factors are present in the ovary of the flesh fly. Arch. Insect Biochem. Physiol. 37:248–256, 1998. © 1998 Wiley–Liss, Inc.