A taste of space: Remote animal observations and discrete-choice models provide new insights into foraging and density dynamics for a large subarctic herbivore.

Competition for resources and space can drive forage selection of large herbivores from the bite through the landscape scale. Animal behaviour and foraging patterns are also influenced by abiotic and biotic factors. Fine-scale mechanisms of density-dependent foraging at the bite scale are likely consistent with density-dependent behavioural patterns observed at broader scales, but few studies have directly tested this assertion. Here, we tested if space use intensity, a proxy of spatiotemporal density, affects foraging mechanisms at fine spatial scales similarly to density-dependent effects observed at broader scales in caribou. We specifically assessed how behavioural choices are affected by space use intensity and environmental processes using behavioural state and forage selection data from caribou (Rangifer tarandus granti) observed from GPS video-camera collars using a multivariate discrete-choice modelling framework. We found that the probability of eating shrubs increased with increasing caribou space use intensity and cover of Salix spp. shrubs, whereas the probability of eating lichen decreased. Insects also affected fine-scale foraging behaviour by reducing the overall probability of eating. Strong eastward winds mitigated negative effects of insects and resulted in higher probabilities of eating lichen. At last, caribou exhibited foraging functional responses wherein their probability of selecting each food type increased as the availability (% cover) of that food increased. Space use intensity signals of fine-scale foraging were consistent with density-dependent responses observed at larger scales and with recent evidence suggesting declining reproductive rates in the same caribou population. Our results highlight potential risks of overgrazing on sensitive forage species such as lichen. Remote investigation of the functional responses of foraging behaviours provides exciting future applications where spatial models can identify high-quality habitats for conservation.

Evaluation of a 49 InDel Marker HID panel in two specific populations of South America and one population of Northern Africa.

The majority of STR loci are not ideal for the analysis of forensic samples with degraded and/or low template DNA. One alternative to overcome these limitations is the use of bi-allelic markers, which have low mutation rates and shorter amplicons. Human identification (HID) InDel marker panels have been described in several countries, including Brazil. The commercial kit available is, however, mostly suitable for Europeans, with lower discrimination power for other population groups. Recently, a combination of 49 InDel markers used in four different ethnic groups in the USA has been shown to be more informative than another panel from Portugal, already tested in a Rio de Janeiro sample. However, these 49 InDels have yet to be applied to other admixed or isolated populations. We assessed the efficiency of this panel in two urban admixed populations (Rio de Janeiro, Brazil; Tripoli, Libya) and one isolated Native Brazilian community. All markers are in Hardy-Weinberg equilibrium (HWE) after the Bonferroni correction, and no Linkage disequilibrium was detected. Assuming loci independence and no substructure effect, cumulative RMP was 2.7×10(-18), 1.5×10(-20), and 4.5×10(-20) for Native Brazilian, Rio de Janeiro, and Tripoli populations, respectively. The overall Fst value was 0.05512. Rio de Janeiro and Tripoli showed similar admixture levels, however for Native Brazilians one parental cluster represented over 60 % of the total parental population. We conclude that this panel is suitable for HID on these urban populations, but is less efficient for the isolated group.

Molecular comparison of Quebec and Newfoundland populations of the blackfly, Simulium vittatum, species complex.

Specific haplotypes at five positions in the COI and COII mitochondrial genes allowed a partial differentiation of Simulium vittatum Zetterstedt (Diptera: Simuliidae) populations from Quebec-Ontario and Newfoundland, respectively. This geographical signature was superimposed on about 40 other polymorphic sites such that sequence divergence alone did not enable a clear-cut distinction between the two populations. Together with the sporadic occurrence of haplotypes intermediate to the Newfoundland and Quebec-Ontario consensus, this suggested that one peculiar sequence among many found in populations from the North American landmass predominates in Newfoundland as a result of a founder effect. The internal transcribed spacer (ITS1) sequence from the nuclear rDNA transcription unit was no more able to resolve populations along geographical lines than the COI/COII criteria.

Development and characterization of a continuous cell line, AFKM-On-H, from hemocytes of the European corn borer Ostrinia nubilalis (Hübner) (Lepidoptera, Pyralidae).

The corn borer, Ostrinia nubilalis, is a very important pest in different countries, and the in vitro system of the insect could be a useful tool for isolation and characterization of the pathogens and physiological responses of the insect. In this context, a cell line was derived from the hemocytes of the European corn borer and was named AFKM-On-H for, respectively, O. nubilalis, Armand Frappier, King Mongkut Institutes, and Hemocytes. This cell line was initiated and maintained in Ex-Cell 400 medium supplemented with 10% heat-inactivated fetal bovine serum. The cells, mostly spherical in shape, not firmly attached to the plastic culture flasks, were passaged up to 200 times by repeated gentle pipetting of the cells. The doubling times at the 80th and 125th passages at 28 degrees C and at the 122th and 169th passages at 25 degrees C were 40, 29, 35, and 34 h, respectively. The AFKM-On-H cell line was further characterized by the morphology, karyotype, random amplified polymorphic DNA analysis, and isozyme profiles. Susceptibility of the cell line to cytoplasmic polyhedrosis viruses (CPV) Euxoa scandens (EsCPV), Dendrolimus punctatus (DpCPV), and Choristoneura fumiferana (CfCPV); nuclear polyhedrosis viruses [Autographa californica (AcMNPV) wild type and recombinant, Antherea yammamai (AnyaNPV)]; and Chilo iridescent virus was demonstrated. Relative sensitivities of the cell line to Bacillus thuringiensis and Metarhizium anisopliae toxins and effects of the molting hormone 20-hydroxyecdysone on this new hemocyte cell line were characterized.

Health care delivery reorganization innovative outcome: universal computerized patient identification.

Twenty-three large employers in the Minneapolis/St. Paul area formed the business health care action coalition group (BHCAC) to obtain medical care for their employees and families. Requirements of BHCAC included: 1) detailed reporting of outcomes of both preventive and standard health care, 2) development and successful implementation of practice guidelines, and 3) development of a functioning automated medical record. The coalition was contracted to meet these requirements. A nonprofit foundation--the Institute for Clinical Systems Integration (ICSI)--was formed and the data standards committee created. To ensure registration information transmitted by network would be attached to the correct patient's computer file; a standardized registration data set was agreed upon to be recorded in a uniform manner. This innovative change has allowed, encouraged, and required that clinical data be transferable electronically among organizations. The following are the ICSI data standard requirements for ICSI member organizations: the Health Level 7 (HL7) is to be used for data transmission [1]. Fields to be used include: 1) Social Security Number (SSN): The SSN was chosen as the patient identifier for the reasons listed in [2]. Record the official SSN assigned by the Social Security Administration; record a pseudo-SSN if a social security number is not available following the Veterans Administration (VA) Hospital algorithm. 2) Name: Use the American National Standards Institute (ANSI) Healthcare Informatics Standards Planning Panel (HISPP) and Message Standards Developers Subcommittee (MSDS) Standard for representation of a person's name [3]; completely record all of the person's legal name (including any punctuation, hyphenated and double last names); nicknames and appellations are to be recorded separately; salutations and suffixes are optional, but must be placed in separate computer fields from other name parts. 3) Gender: Allowable values include FEMALE, MALE, OTHER, UNKNOWN. 4) Date of Birth: Include the day, month, year (including century); permitted is approximate or estimated birth dates provided they are indicated as such. Use the ANSI HISPP MSDS standard for date representation [4]. 5) Address: Record at least one address; multiple addresses desirable identified by type or usage (i.E., home, business, office). Allow two or more lines for street, city, U.S. state or Canadian Province; identify by standard two-letter postal abbreviation--zip code (at least 5-digits, preferably 9) or postal code and country. 6) Home ID or Medical Record Number (MRN): Record if different than Social Security Number.

Primordial reading of genetic information.

From the consideration of general features of the anticodon loop and stem in tRNA and the properties of present-day translation, we put forward a plausible scenario to explain the evolution of the genetic code from a highly ambiguous triplet code to the present refined decoding system. Our model based on the reading of the code suggests that the anticodon of primordial tRNA could adopt either the 3' or the 5' stacked conformation permitting the formation of the "best two out of three" base pairs, either the first and second codon position or the second and third. Progressive acquisition of precise structural constraint and the modification of bases in the anticodon loop would give way eventually to the less ambiguous "two out of three" reading mechanism having only the 3' stacked conformation. Further adjustments of base composition and modification leads inevitably to the present generalized code. In this way the primordial code encoding 4-8 amino acids or related derivates evolves smoothly to the present code having 20 amino acids.

Photoacoustic spectroscopy of Anacystis nidulans. III. Detection of photosynthetic activities.

Photosynthetic activities of Anacystis nidulans can be detected by photoacoustic spectroscopy. Algae treated by a photosynthetic inhibitor are used to provide the signal from the photochemically inactive sample. The results of these measurements correspond well with the activities which can be monitored by conventional biochemical assays. Acoustic data from A. nidulans are used to obtain the action spectrum for photochemical energy storage. It is concluded that phycocyanin harvests light for both photoreactions but that chlorophyll alpha molecules convey most of their excitation energy to photoreaction I. As judged from the relationship between the modulation frequency and the acoustic signal intensity, at least 60% of the photons absorbed at 630 nm perform photochemical work and about half of the useful energy is stored at stable products. Although it cannot be separated from the purely thermal effect, the contribution of modulated oxygen evolution to the acoustic signal of algae is estimated to be relatively small. Due to structural peculiarities, the opposite situation predominates in low frequency measurements performed with leaves from Impatiens petersiana.

Photoacoustic spectroscopy of Anacystis nidulans. I. Effect of sample thickness on the photoacoustic signal.

A photoacoustic spectroscopy study of the cyanobacteria Anacystis nidulans has been undertaken. It is demonstrated, by using a filter deposition technique, that the photoacoustic signal intensity becomes progressively saturated as the thickness of the algal layer is increased. This saturation effect originates mostly from the limited optical penetration of the sample and distorts the photoacoustic spectrum from its true shape. A theoretical model is proposed to explain these results, and practical means to obviate the limitations of this spectroscopic technique are suggested.

Photoacoustic spectroscopy of Anacystis nidulans. II. Characterization of pigment holochroms and thermal deactivation spectrum.

The photoacoustic spectrum of Anacystis nidulans recorded at room temperature is qualitatively similar to low-temperature absorption or fluorescence excitation spectra. The bands of pigment holochroms are well resolved compared to room-temperature absorption spectra. The thermal deactivation spectrum obtained by extrapolating acoustic data for an infinitely thin sample indicates that the photosynthetic efficiency decreases from phycocyanin to chlorophyll a and carotenoids.

The evolution of multi-isoacceptor tRNA families. Sequence of tRNA Leu CAA and tRNA Leu CAG from Anacystis nidulans.

Two leucine tRNAs from the cyanophyte Anacystis nidulans have been isolated, and their complete nucleotide sequences have been determined by combining data from oligonucleotide fingerprints and sequencing gels. The two sequences are 87 nucleotides long, have the anticodons CAA and CAG, and differ from each other at a total of 28 positions. They have been compared to other known tRNA Leu sequences and incorporated into a phylogenetic tree comprising prokaryotic and chloroplastic tRNA Leu sequences. Mutations inferred from the tree show that some parts of the tRNA molecule are highly variable (the extra arm and the acceptor stem) while others are much more conserved (the D and T arms). The topology of the tree supports the idea that blue-green algae and chloroplasts share a common prokaryotic ancestor and show a basic divergence between XAA and XAG anticodon-containing tRNAs, suggesting that these two subfamilies result from an ancient gene duplication. Finally, comparison of this phylogenetic tree with those of other multi-isoacceptor tRNA families shows no common scheme, which may be due to independent refinement of codon-reading patterns in different tRNA families.

The evolving tRNA molecule.

The study of tRNA molecular evolution is crucial to understanding the origin and establishment of the genetic code as well as the differentiation and refinement of the machinery of protein synthesis in prokaryotes, eukaryotes, organelles, and phage systems. The small size of the molecule and its critical involvement in a multiplicity of roles distinguish its study from classical protein molecular evolution with respect to goals and methods. Here, the authors assess available and missing data, existing and needed methodology, and the impact of tRNA studies on current theories both of genetic code evolution and of the evolution of species. They analyze mutational "hot spots", the role of base modification, synthetase recognition, codon-anticodon interactions and the status of organelle tRNA.

Convergence and minimal mutation criteria for evaluating early events in tRNA evolution.

The convergence of ancestral sequences independently constructed from different branches of a phylogenetic tree can be used as a test of homology of data sequences. This criterion has shown that all phenylalanine tRNAs are related to a common ancestor, whereas eukaryotic and prokaryotic tyrosine tRNAs may have independent origins. All glycine tRNAs share a common ancestor. The glycine tRNA family splits according to the purine or pyrimidine nature of the first anticodon base prior to the divergence of eukaryotes and prokaryotes. The structural similarity between some prokaryotic glycine and and valine tRNAs is the result of their derivation from a common ancestor that existed previous to the divergence of the different glycine tRNAs. These results support models of genetic code evolution involving the incremental elaboration of earlier, simpler codes.

Evolution of methionine initiator and phenylalanine transfer RNAs.

Sequence data from methionine initiator and phenylalanine transfer RNAs were used to construct phylogenetic trees by the maximum parsimony method. Although eukaryotes, prokaryotes and chloroplasts appear related to a common ancestor, no firm conclusion can be drawn at this time about mitochondrial-coded transfer RNAs. tRNA evolution is not appropriately described by random hit models, since the various regions of the molecule differ sharply in their mutational fixation rates. "Hot" mutational spots are identified in the Tpsic, the amino acceptor and the upper anticodon stems; the D arm and the loop areas on the other hand are highly conserved. Crucial tertiary interactions are thus essentially preserved while most of the double helical domain undergoes base pair interchange. Transitions are about half as costly as transversions, suggesting that base pair interchanges proceed mostly through G-U and A-C intermediates. There is a preponderance of replacements starting from G and C but this bias appears to follow the high G + C content of the easily mutated base paired regions.

In vitro synthesis of transfer RNA. II. Identification of required enzymatic activities.

We have shown that the synthesis of active su+III tRNATyr from a phi80psu+III DNA template requires the action of four distinct enzymatic activities. The first of these, DNA-dependent RNA polymerase, catalyzes the formation of a large molecular weight transcript, initiating synthesis at a specific site 41 nucleotides proximal to the 5' end of the su+III tRNATyr structural gene and continuing at least 100 nucleotides beyond the 3' terminus of the su+III tRNATyr sequence. The second required component, designated Fraction V, allows purified DNA-DEPENDENT RNA polymerase to function in tRNA synthesis. We have shown that this fraction contains an endonuclease that together with DNA-dependent RNA polymerase is responsible for the synthesis of su+III tRNATyr "precursor". Thus, su+III tRNATyr precursor is not itself the primary transcription product of the su+III tRNATyr gene, but rather, it arises as a result of post-transcriptional cleavage of a much larger transcript by the action of the nuclease present in Fraction V. The third enzymatic activity required for synthesis of active su+III tRNATyr is a ribonuclease (RNase P III) that specifically catalyzes the removal of the 3' extra nucleotides from the su+III tRNATyr precursor. The fourth activity required for synthesis of tRNA is a previously identified endonuclease, RNase P, that specifically catalyzes the removal of the 5' extra nucleotides from tRNA precursors. The properties of RNase P purified according to the procedure developed in this laboratory have been compared with those of the enzyme purified from ribosomes according to the procedure described by Robertson et al. (Robertson, H.D., Altman, S., and Smith, F.D. (1972) J.Biol. Chem. 247, 5243-5251.).