Application of the Red List Index as an indicator of habitat change

For the first time ever, the International Union for Conservation of Nature Red List Index for habitat types was calculated for an entire country, Finland. The RLIs were based on species threat assessments from 2000 and 2010 and included habitat definitions for all 10,131 species of 12 organism groups. The RLIs were bootstrapped to track statistically significant changes. The RLI changes of species grouped by habitats were negative for all habitat types except for forests and rural biotopes which showed a stable trend. Trends of beetles and true bugs were positive in rural and forest habitats. Other 16 observed trends of species group and habitat combinations were negative. Several trends observed were in accordance with studies focusing on particular taxa and habitats, and drivers for their change. This study demonstrates the usefulness of the RLI as a tool for observing habitat change based on species threat assessment data.     

Isotype concentrations of human antibodies to Haemophilus influenzae type b polysaccharide (Hib) in young adults immunized with the polysaccharide as such or conjugated to a protein (diphtheria toxoid)

Antibody responses of young adults to Haemophilus influenzae type b polysaccharide (Hib) or its protein conjugate were studied with special attention to the isotype composition of the antibodies. Three conclusions of interest can be made: 1) Immunoglobulin G (IgG) antibodies in polysaccharide-immunized volunteers displayed the subclass pattern previously found in antibodies to meningococcal type A polysaccharide. IgG1 was the predominant subclass in IgG antibodies of some individuals, IgG2 in others. Still others had the two subclasses in varying but more even proportions. 2) The conjugate vaccine induced a geometric mean response 2 to 3 times higher and an IgG response 4 times higher to Hib than the polysaccharide vaccine. 3) Anti-Hib antibodies induced by the conjugate vaccine still had essentially the same IgG subclass composition as anti-Hib antibodies induced by the polysaccharide. This composition was strikingly different from the composition of the anti-diphtheria toxoid response induced by the same conjugate vaccine.     

Structural organization of the beta-globin locus of B-haplotype sheep

Goats and some sheep synthesize a juvenile hemoglobin, Hb C (alpha 2 beta C2), at birth and produce this hemoglobin exclusively during severe anemia. Sheep that synthesize this juvenile hemoglobin are of the A haplotype. Other sheep, belonging to a separate group, the B haplotype, do not synthesize hemoglobin C and during anemia continue to produce their adult hemoglobin. To understand the basis for this difference we have determined the structural organization of the beta- globin locus of B-type sheep by constructing and isolating overlapping genomic clones. These clones have allowed us to establish the linkage map 5' epsilon I-epsilon II-psi beta I-beta B-epsilon III-epsilon IV- psi beta II-beta F3' in this haplotype. Thus, B sheep lack four genes, including the BC gene, and have only eight genes, compared with the 12 found in the goat globin locus. The goat beta-globin locus is as follows: 5' epsilon I-epsilon II-psi beta X-beta C-epsilon III-epsilon IV-psi beta Z-beta A-epsilon V-epsilon VI-psi beta Y-beta F3'. Southern blot analysis of A-type sheep reveals that these animals have a beta- globin locus similar to that of goat, i.e., 12 globin genes. Thus, the beta-globin locus of B-haplotype sheep resembles that of cows and may have retained the duplicated locus of the ancestor of cows and sheep. Alternatively, the B-sheep locus arrangement may be the result of a deletion of a four-gene set from the triplicated locus.      

The growth of potential food poisoning organisms on chicken and pork muscle surfaces

Muscle surfaces of pork were inoculated with a mixture of Yersinia enterocolitica and Staphylococcus aureus , and chicken muscle with Campylobacter jejuni or a mixture of Salmonella typhimurium and Staph. aureus . The surface growth at 20°C was followed microscopically. Organisms grew as discrete colonies bound together by a glycocalyx which differed between bacterial species. On prolonged incubation colonies spread peripherally and tended to coalesce, while still retaining their colony structure. Staphlycoccus aureus colonies were very small and remained so. The glycocalyx was considered critical in maintaining the dense populations of bacteria on the meat surfaces.     

Fidelity of DNA synthesis by the Thermococcus litoralis DNA polymerase--an extremely heat stable enzyme with proofreading activity

We demonstrate that the DNA polymerase isolated from Thermococcus litoralis (VentTM DNA polymerase) is the first thermostable DNA polymerase reported having a 3'----5' proofreading exonuclease activity. This facilitates a highly accurate DNA synthesis in vitro by the polymerase. Mutational frequencies observed in the base substitution fidelity assays were in the range of 30 x 10(-6). These values were 5-10 times lower compared to other thermostable DNA polymerases lacking the proofreading activity. All classes of DNA polymerase errors (transitions, transversions, frameshift mutations) were assayed using the forward mutational assay (1). The mutation frequencies of Thermococcus litoralis DNA polymerase varied between 15-35 x 10(-4) being 2-4 times lower than the respective values obtained using enzymes without proofreading activity. We also noticed that the fidelity of the DNA polymerase from Thermococcus litoralis responds to changes in dNTP concentration, units of enzyme used per one reaction and the concentration of MgSO4 relative to the total concentration of dNTPs present in the reaction. The high fidelity DNA synthesis in vitro by Thermococcus litoralis DNA polymerase provides good possibilities for maintaining the genetic information of original target DNA sequences intact in the DNA amplification applications.     

Recombinant E-selectin-protein mediates tumor cell adhesion via sialyl-Le(a) and sialyl-Le(x).

E-selectin (previously known as ELAM-1) is one of the adhesion molecules expressed on activated endothelium. Here we show that HL-60 cells express sialyl-Le(x), but not Sialyl-Le(a) on their surface, a colon carcinoma cell line COLO 205 express both these epitopes and another colon carcinoma COLO 320 does not express either one of them. HL-60 and COLO 205 cell adhere strongly to E-selectin coated microwells, whereas COLO 320 does not adhere at all to E-selectin. Finally we provide evidence that monoclonal anti-sialyl-Le(x) can abolish part of the adherence of HL-60 cells to recombinant E-selectin. The adherence of COLO 205 cells can be decreased by either monoclonal anti-sialyl-Le(a) or anti-sialyl-Le(x) antibodies. These results indicate that cell-associated sialylated carbohydrate moieties can act as ligands for recombinant E-selectin.     

Conformational analysis of a toxic peptide from Trimeresurus wagleri which blocks the nicotinic acetylcholine receptor.

The 22-residue toxic peptide (WTX1) from the venom of the Southeast Asian snake Trimeresurus wagleri has multiple sites of action, but its lethal effect has been attributed to blocking the postsynaptic acetylcholine receptor at the neuromuscular junction. The 3-dimensional structure of WTX1 was studied using 2-dimensional nuclear magnetic resonance spectroscopy, circular dichroism, and computer simulations. In aqueous solution, WTX1 was shown to have extended and flexible "tails" defined by a short, rigid disulfide-bonded loop. The flexible regions can undergo structural rearrangement when moved from an aqueous to a less polar environment and may contribute to its effectiveness at different receptor sites. By substituting Gly or Phe for His at position 10, significant effects on the disulfide bond formation and, thereby, the activity of the peptide were observed. These results suggest that even subtle differences in single residues can have profound effects on the dynamics of folding, disulfide bond formation, and activity of this toxic peptide.     

Proton-induced X-ray emission analysis of atherosclerotic plaques of the carotid bifurcation

The trace elements of both calcified atherosclerotic plaques and plaque-free vessel walls of the carotid bifurcation from 31 autopsies were investigated using the proton-induced X-ray emission (PIXE) method. The trace elements studied were phosphorus (P), calcium (Ca), chrome (Cr), iron (Fe), copper (Cu), zinc (Zn), lead (Pb), selenium (Se), bromine (Br), strontium (Sr), and rubidium (Rb). All samples contained Fe and Zn. Mercury (Hg) was not detected in any of the samples studied. All plaque-free samples contained Cu and almost all Br and Ca, none Sr. All calcified atherosclerotic plaques contained Ca and almost all Br and Sr. The relative levels of Ca were higher in the calcified plaques than in the plaque-free vessel walls. The relative value of Ca in calcified and uncalcified samples was greatest in the group who had died because of cardiovascular disorders and smallest in the group who had died from other causes. There was a strong positive correlation between the Ca and Sr of the plaque samples and between the P and Br of the plaque-free samples.