Volatile constituents of male and female boll weevils and their frass

The volatile constituents of male and female boll weevils, Anthonomus grandis, and their frass were analysed by GLC-MS. The 4 previously identified components of the male pheromone were present only in the male volatile oil (3·9%) and in the male frass volatile oil (38·9%). The compounds found in one or both sexes and their frass include 26 carbonyls, 23 hydrocarbons, 12 alcohols, 6 phenols, 4 esters, 3 furans, 1 ether, and 1 lactone. Also found were 2 compounds containing nitrogen, 1 halogen, and 1 sulphur. There were 33 terpenes and 24 aromatic compounds. 3,7-Dimethyl-1-octanol comprised 15·6 per cent of the male frass oil. Carvone was found only in females (5·4%) and in female frass (6·8%). A series of monoterpene aldehydes (M⁺ 152) were found only in the female frass oil. A pheromone rôle for these components was suggested.     

Pecan weevil sex attractant: Bioassay and chemical studies

When the distillable oil from adult pecan weevils, Curculio caryae, of both sexes was investigated separately with an integrated gas chromatographymass spectrometry system, evidence was obtained for a number of di- and trisubstituted aromatic hydrocarbons, seven phenolic and carbonyl aromatic compounds, and at least three alcohols. The major compounds were a series of alkanes and alkenes, four esters and four heterocyclic compounds. Among these, eleven compounds were unique to males and four were unique to females. The volatiles were also bioassayed in the field and each attracted the opposite sex. This investigation was part of a study made to identify the possible rôle of these compounds as a pheromone produced by the pecan weevil.     

Hormonal regulation of the synthesis and mRNA content of the regulatory subunit of cyclic AMP-dependent protein kinase type II in cultured rat ovarian granulosa cells

These studies were undertaken to determine the molecular events by which estradiol and follicle-stimulating hormone (FSH) stimulate in ovarian granulosa cells the increase in the content of one of the regulatory subunits of cAMP-dependent protein kinase type II, RII51 (Mr = 51,000), and its electrophoretic variants RII51.5 (Mr = 51,500) and RII52 (Mr = 52,000). To analyze the de novo synthesis of RII51/52, granulosa cells were cultured (10(6) cells/ml) for 0, 12, 24, or 48 h with estradiol (10 nM) +/- FSH (12.5, 25, and 50 ng/ml), 8-bromo-cAMP (0.25-3 mM), or forskolin (0.5-100 microM) and then pulse-labeled with [35S]methionine (300 microCi/ml; 4 h). Labeled RII51, present either in urea extracts of total cellular protein or after partial purification from a soluble cell extract by cAMP-Sepharose chromatography, was quantitated by autoradiography of two-dimensional sodium dodecyl sulfate-polyacrylamide gel electrophoresis gels and by excision of the silver-stained spots of the RII51 variants from the gels and counting. Synthesis of RII51 and its electrophoretic variants was low in cells cultured with estradiol alone for 48 h, whereas it was increased 4-5-fold in cells cultured with estradiol and FSH. Changes in the synthesis of actin were minor throughout the culture period regardless of hormone treatment. Pulse-chase experiments using [35S]methionine provided evidence that the isoelectric variants RII51.5 and RII52 may be derived from RII51 by post-translation modification, such as phosphorylation. Labelling with [32P]orthophosphate showed that RII52 contained more radioactivity than RII51.5 and RII51. Northern and filter hybridization assays demonstrated a 6-10-fold dose- and time-dependent increase in the amount of RII51 mRNA in granulosa cells exposed to estradiol and FSH or estradiol and forskolin compared to those cultured with estradiol alone. In vitro translation of poly(A)+ mRNA of granulosa cells from estradiol- and FSH-treated hypophysectomized rats also demonstrated an increase in the content of translatable RII51 mRNA. These studies indicate that in cultured rat granulosa cells the synthesis of RII51 and the content of its mRNA are selectively increased by estradiol and cAMP in a time- and dose-dependent manner. Based on these observations, RII51 appears to be a useful marker to determine the molecular (genomic?) sites of estradiol and FSH action in differentiating rat granulosa cells.     

Molecular cloning, cDNA structure, and regulation of the regulatory subunit of type II cAMP-dependent protein kinase from rat ovarian granulosa cells

One isoform of the regulatory subunit of type II cAMP-dependent protein kinase (R-II51; Mr = 51,000) and its electrophoretic variants (R-II51.5 and R-II52; Mr = 51,500 and 52,000, respectively) are selectively induced by estradiol and follicle-stimulating hormone (cAMP) in rat ovarian granulosa cells. To ascertain the amino acid sequence of R-II51 and to gain insight into the molecular events regulating the intracellular content of ovarian follicular R-II51, we constructed a lambda gt11 cDNA expression library from poly(A)+ RNA of hormone-primed rat granulosa cells. A 1.5-kilobase (kb) cDNA insert, isolated from a plaque-purified R-II antibody positive bacteriophage clone, selectively bound R-II51 mRNA as demonstrated by analysis of the hybrid-selected translation product. Restriction maps and sequence analyses of the 1.5-kb cDNA insert and of the 1.8- and 2.2-kb cDNA inserts from two additional clones showed overlapping sequences which span a region of 3065 nucleotides in size. The 1.5- and 1.8-kb cDNA inserts each contained poly(A) addition signals (1508 and 1761 nucleotides, respectively), terminal poly(A) sequences, and the entire coding region for R-II51 (1204 nucleotides) except for a small number of nucleotides at the 5' end. The 2.2-kb cDNA insert contained 394 nucleotides of the coding region a long 3' untranslated region and two more poly(A) addition signals (3041 and 3059 nucleotides). An amino acid microsequence surrounding the autophosphorylation site of pure rat ovarian R-II51 agreed with the amino acid sequence deduced from the nucleotide sequence of the cDNA. Northern blot analyses demonstrated two major mRNA species (1.8 and 3.2 kb in size) in hormone-primed rat ovaries which were approximately 10- and 50-fold greater than the R-II mRNA content in rat brain and rat heart, respectively. Southern blot analysis of rat liver DNA indicated that a single gene codes for R-II51 mRNA. Structural differences among rat ovarian R-II51, rat heart R-II54, and the known amino acid sequences of bovine R-II and R-I subunits also indicate that the rat ovarian R-II51 subunit is the product of a distinct gene.     

A new monster from southwest Oregon forests: Cryptomaster behemoth sp. n. (Opiliones,Laniatores, Travunioidea)

The monotypic genus Cryptomaster Briggs, 1969 was described based on individuals from a single locality in southwestern Oregon. The described species Cryptomaster leviathan Briggs, 1969 was named for its large body size compared to most travunioid Laniatores. However, as the generic name suggests, Cryptomaster are notoriously difficult to find, and few subsequent collections have been recorded for this genus. Here, we increase sampling of Cryptomaster to 15 localities, extending their known range from the Coast Range northeast to the western Cascade Mountains of southern Oregon. Phylogenetic analyses of mitochondrial and nuclear DNA sequence data reveal deep phylogenetic breaks consistent with independently evolving lineages. We use discovery and validation species delimitation approaches to generate and test species hypotheses, including a coalescent species delimitation method to test multi-species hypotheses. For delimited species, we use light microscopy and SEM to discover diagnostic morphological characters. Although Cryptomaster has a small geographic distribution, this taxon is consistent with other short-range endemics in having deep phylogenetic breaks indicative of species level divergences. Herein we describe Cryptomaster behemoth sp. n., and provide morphological diagnostic characters for identifying Cryptomaster leviathan and Cryptomaster behemoth.     

Genetic diversification without obvious genitalic morphological divergence in harvestmen (Opiliones, Laniatores, Sclerobunus robustus) from montane sky islands of western North America

The southern Rocky Mountains and adjacent Intermontane Plateau Highlands region of western North America is a geographically diverse area with an active geologic history. Given the topological complexity and extensive geologic activity, organisms inhabiting this region are expected to show some degree of morphological and genetic divergence, especially populations found on the southern montane 'sky islands' of this region. Here we examine the phylogeographic history and diversification of a montane forest inhabiting harvestmen, Sclerobunus robustus, using a combination of genetic and morphological data. Divergence time estimates indicate that much of the diversification within and between major groups S. robustus predate the Pleistocene glacial cycles. The most widespread subspecies, Sclerobunus robustus robustus, is recovered as six genetically distinct, geographically cohesive mitochondrial phylogroups. Gene tree data for a single nuclear gene reveals congruent, albeit slightly more conservative, patterns of genetic divergence. Despite high levels of genetic divergence throughout their distribution, phylogroups show extreme conservation in somatic and reproductive morphology. This uncoupling of morphological and genetic differentiation may be due to morphological conservatism associated with a conserved microhabitat preference. Based on these data, it is obvious that S. robustus has undergone some level of cryptic diversification.     

MHC class II epitope nesting modulates dendritic cell function and improves generation of antigen-specific CD4 helper T cells

CD4 Th cells are critical to the development of coordinated immune responses to infections and tumors. Th cells are activated through interactions of the TCR with MHC class II complexed with peptide. T cell activation is dependent on the density of MHC peptide complexes as well as the duration of interaction of the TCR with APCs. In this study, we sought to determine whether MHC class II peptides could be modified with amino acid sequences that facilitated uptake and presentation with the goal of improving Th cell activation in vitro and in vivo. A model epitope derived from the murine folate receptor α, a self- and tumor Ag, was modified at its carboxyl terminus with the invariant chain-derived Ii-Key peptide and at its N terminus with a peptide that enhances uptake of Ag by APC. Modification of a peptide resulted in enhanced generation of high-avidity murine folate receptor α T cells that persisted in vivo and homed to sites of Ag deposition. The nesting approach was epitope and species independent and specifically excluded expansion of CD4 regulatory T cells. The resulting Th cells were therapeutic, enhanced in vivo helper activity and had an increased ability to resist tolerizing immune microenvironments. In addition to improved immunoadjuvants, this epitope modification strategy may be useful for enhancing ex vivo and in vivo generation of Th cells for preventing and treating diseases.     

Seasonal shifts in habitat use and diet by eland confined in a small fenced reserve

Large‐scale environmental changes create challenges for conservation of wildlife, particularly in fenced, insular protected areas where many wildlife populations persist. Moreover, large mammalian herbivores inhabiting spatially and temporally heterogeneous environments face the challenge of securing highly variable forage resources. Mixed feeders like the eland (Taurotragus oryx) can switch between browse and grass, but the cues that elicit that switch are not well understood. We investigated the seasonal diet shift of eland confined to a small fenced reserve and the role of greenness to elicit that shift. Eland changed from a diet in the wet season, consisting of grasses and browse found in woodland and grassland vegetation types, to a diet in the dry season dominated by the greenest browse species still available in woodland vegetation types, as greenness of dry season forage decreased. Our results suggest that eland switch from browsing to grazing in response to phenophase of the grass sward, which could explain the varying selection of grasses versus browse observed across the species range.     

Habitat preferences and conservation of the marbled jewel beetle Poecilonota variolosa (Buprestidae)

Detailed knowledge on habitat requirements is a key to successful conservation actions. The marbled jewel beetle Poecilonota variolosa (Buprestidae) has a wide global distribution but populations are often scarce and typically fragmented. In Sweden it is monophagous on aspen Populus tremula and is classified as near threatened on the Swedish Red List due to its rapid population decline. This study aimed to investigate habitat preferences and regional-scale distribution patterns of P. variolosa in southern Sweden in order to suggest conservation measures. Aspen trees in four study areas in the province of Småland were surveyed for exit holes during late summer 2011. The occurrence and number of exit holes (both new and old ones) per tree were compared between study areas and habitat types, and were related to the sun exposure and bark thickness of individual trees. Further, the occurrence of new and older exit holes was related to tree sun exposure and bark thickness. The most preferred habitat types were aspens on clear-cuts, followed by roadside aspens, aspens in pastures, and aspens in closed forest. Thick bark and high sun exposure were consistently significant as predictors for both occurrence and number of new exit holes per tree. The majority of exit holes were located towards south. Our results indicate several useful management measures: to retain aspen on clear-cuts, to cut alongside roads and around some selected coarse aspens in closed forests and in pastures.