Enrichment of human bone marrow mononuclear phagocytes and characterization of macrophage subpopulations by immunoenzymatic double staining

In order to isolate and enrich bone marrow mononuclear phagocytes, we performed magnetic-activated cell sorting using beads coupled to a monoclonal antibody directed against the monocyte/macrophage surface molecule CD14. Co-localization of antigens in single cells was achieved by combining an alkaline phosphatase--anti-alkaline phosphatase and an avidin--biotin complex immunoassay, avoiding the use of peroxidase. Bone marrow macrophages were first labelled by the monoclonal antibody PG-M1 (anti-CD68). Subsequently, cytoplasmic and/or surface double staining by the monoclonal antibodies against HLA-DR and Mac-2 antigen or the lectin GSA-I-B4 was carried out. Whereas HLA-DR was co-expressed by the great majority of PG-M1+ macrophages (84.9% +/- 6.9%), only a subpopulation exhibited Mac-2 (69.9% +/- 5.9%) antigen or galactoside structures detected by GSA-I-B4 (65.0% +/- 6.7%). The latter result differed only slightly from the percentage of GSA-I-B+4 macrophages determined in a previous comparative immunomorphometrical study. Therefore, using our method of isolation and enrichment by magnetic-activated cell sorting, only a negligible portion of macrophages is apparently stimulated, as shown by GSA-I-B4 staining. This methodology seems to be a valuable tool for further studies on the monocyte--macrophage system. © 1998 Chapman & Hall     

Preparation, Properties, and Composition of Nereis vitellin, the Yolk Protein of the Annelid, Nereis virens

In the literature, oogenesis in nereid annelids is considered to be a model system because, unlike other system studied, nereid oocytes are thought to synthesize the bulk of their yolk protein themselves. As the first step to test the validity of this hypothesis, nereid yolk protein was characterized biochemically. Vitellin, the main fraction of the soluble yolk proteins, was prepared from Nereis virens oocytes. Preparation, purification, and some physical characteristics of this green-colored protein Nereis vitellin are described. The molecular weight was determined by gel chromatography as 420,000 daltons. With regard to the amino acid composition, Nereis vitellin was found to resemble both insect vitellins and an average protein, as defined by other authors. Methionine and cysteine were found in traces only. By staining procedures, Nereis vitellin was characterized as lipoglycoprotein. Nereis vitellin was also prepared from the coelomic fluid of gravid females of Nereis virens .     

Wildlife and climate change: assessing the sensitivity of selected species to simulated doubling of atmospheric CO2

We explored, using computer simulations, the sensitivity of four mammal species (elk, Cervus canadensis ; white-tailed deer, Odocoileus virginianus ; Columbian ground squirrel, Spermophilus columbianus ; and chipmunk, Tamias striatus ) within the continental USA to the effect of anticipated levels of global climate change brought about by a doubling of atmospheric CO₂. Sensitivity to the direct effects of climate change were evaluated using a climate-space approach to delineate the range of thermal conditions tolerable by each species. Sensitivity to indirect effects were evaluated by quantifying the association of each species to the current vegetation distribution within the continental USA and using this association to assess whether wildlife species distributions might shift in response to vegetation shifts under climate change. Results indicate that altered thermal conditions alone should have little or no effect on the wildlife species' distributions as physiological tolerance to heat load would allow them to survive. Analyses of the effects of vegetation change indicate that deer and chipmunks should retain their current distributions and possibly expand westward in the USA. For Elk and ground squirrels, there is a possibility that their current distributions would shrink and there is little possibility that each species would spread to new regions. This work emphasizes that the distributions of the four mammalian species are likely to be influenced more by vegetation changes than by thermal conditions. Future efforts to understand the effects of global change on wildlife species should focus on animal–habitat and climate–vegetation linkages.     

Cambrian high-resolution biostratigraphy and carbon isotope chemostratigraphy in Scania, Sweden: first record of the SPICE and DICE excursions in Scandinavia

A core drilling (Andrarum‐3), from the classical locality at Andrarum, Scania, southernmost Sweden, penetrated a 28.90‐m‐thick Cambrian succession. The core comprises dark grey to black, finely laminated mudstones and shales with early concretionary carbonate lenses (stinkstones or orsten) and a few primary carbonate beds. The middle Cambrian (provisional Series 3) part of the core comprises 17.35 m, whereas the Furongian Series (upper Cambrian) part covers the remaining 11.55 m. Nineteen trilobite and two phosphatocopine genera are present in the middle Cambrian, whereas the less diverse Furongian interval yielded four trilobite and three phosphatocopine genera. Other, less frequent, faunal elements include conodonts (s. l.), brachiopods, sponge spicules, bradoriids, and coprolites. Trilobites and phosphatocopines were used to subdivide the core into seven biozones ranging from the Ptychagnostus atavus Zone to the Parabolina spinulosa Zone (P. spinulosa Subzone). Carbon isotopic analyses (δ¹³C_org) through the core show two important excursions, the negative DrumIan Carbon isotope Excursion (DICE) in the Pt. atavus Zone, and the Steptoean Positive Carbon Isotope Excursion (SPICE) beginning near the first appearance of Glyptagnostus reticulatus and extending upward into the Olenus and Agnostus (Homagnostus) obesus Zone. The DICE displays a peak value, in the samples at hand, of –30.45‰δ¹³C_org in the lower part of the P. atavus Zone. The δ¹³C_org values increase through the overlying L. laevigata and A. pisiformis zones and display peak values of c. –28.00‰δ¹³C_org in the lowermost Furongian Olenus wahlenbergi and O. attenuatus subzones. Thereafter the values decrease significantly through the O. scanicus Subzone. Both isotopic excursions have been documented from several palaeocontinents, but never before from Baltica. Moreover, for the first time these excursions are recorded from organic matter in an alum shale setting. The recorded shift of +1.50–2.00‰δ¹³C_org is approximately half the magnitude of the SPICE documented from other regions. This discrepancy may be related to temporal variations in the type, origin, or diagenesis of the organic fraction analysed.     

The Hawaiian amphibious caterpillar guild: new species of Hyposmocoma (Lepidoptera: Cosmopterigidae) confirm distinct aquatic invasions and complex speciation patterns

Although aquatic caterpillars are a globally rare lifestyle, we have found them in multiple, independent lineages of the endemic moth genus Hyposmocoma across all of the Hawaiian Islands with flowing water. We formally describe 11 new species of Hyposmocoma that belong to four different larval case types: cone, bugle, medium burrito, and large burrito: Hyposmocoma kahamanoa sp. nov. from Oahu Island, Hyposmocoma kamakou sp. nov. from Molokai Island, Hyposmocoma kahaiao sp. nov. , Hyposmocoma waihohonu sp. nov. , and Hyposmocoma moopalikea sp. nov. from Maui Island, and Hyposmocoma aumakuawai sp. nov. , Hyposmocoma eepawai sp. nov. , Hyposmocoma ipowainui sp. nov. , Hyposmocoma kawaikoi sp. nov. , Hyposmocoma uhauiole sp. nov. , and Hyposmocoma wailua sp. nov. from Kauai Island. We also illustrate and describe in detail the aquatic case‐bearing larva of Hyposmocoma kahamanoa. Despite having similar ecologies as algae and lichen grazers at and below the water line of streams, prior research indicates that species with each case type constitute an independent lineage, with terrestrial sister taxa, and therefore the different groups of species bearing unique case types each represent an independent aquatic invasion. The case‐bearing larvae often occur sympatrically, and on Kauai even species with similar case‐types occur together, suggesting complex patterns of speciation and either past periods of isolation or sympatric speciation. Phylogenetic analysis of 2243 base pairs from two nuclear and one mitochondrial gene for 18 species confirm that each species is endemic to a single volcano, and that morphological divergence within case‐types has not been dramatic. Diversification has been complex, and superficially similar case type lineages are not all monophyletic. Kauai, the oldest but smallest of the major high islands, supports more species in the aquatic guild than any other island, thus island age, rather than size, may be important in generating diversity in this group. © 2011 The Linnean Society of London, Zoological Journal of the Linnean Society, 2011, 162 , 15–42.