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The separation of germ cell populations from the soma is part of the evolutionary transition to multicellularity. Only genetic information present in the germ cells will be inherited by future generations, and any molecular processes affecting the germline genome are therefore likely to be passed on. Despite its prevalence across taxonomic kingdoms, we are only starting to understand details of the underlying micro‐evolutionary processes occurring at the germline genome level. These include segregation, recombination, mutation and selection and can occur at any stage during germline differentiation and mitotic germline proliferation to meiosis and post‐meiotic gamete maturation. Selection acting on germ cells at any stage from the diploid germ cell to the haploid gametes may cause significant deviations from Mendelian inheritance and may be more widespread than previously assumed. The mechanisms that affect and potentially alter the genomic sequence and allele frequencies in the germline are pivotal to our understanding of heritability. With the rise of new sequencing technologies, we are now able to address some of these unanswered questions. In this review, we comment on the most recent developments in this field and identify current gaps in our knowledge.

The quality of porcine oocytes for use in IVF is commonly graded according to the number of layers of cumulus cells (CCs) surrounding the oocyte; together these form the cumulus oocyte complex (COC). At least three compact layers of CCs is regarded as important for efficient IVP. To test this, oocytes were scored according to cumulus investment, with grade A representing COCs with three or more cumulus layers including granulosa cell‐cumulus oocyte complexes, grade B those with an intact corona radiata surrounded by another layer of cumulus cells and grades C and D representing COCs with lower CC investment. These oocytes were then monitored for in vitro maturation (IVM), as assessed by tubulin immunostaining for meiotic progression, the development of a cortical granule ring, and by glutathione levels. Results indicate that grading correlates closely with nuclear maturation and cytoplasmic maturation, suggesting that grading oocytes by cumulus investment is a reliable method to predict IVM success. Importantly, Grade A and B oocytes showed no significant differences in any measure and hence using a cut‐off of two or more CC layers may be optimal. We also determined the effect of assisted aspiration for oocyte retrieval, comparing the effect of needle size and applied pressure on the retrieval rate. These data indicated that both variables affected oocyte recovery rates and the quality of recovered oocytes. In combination, these experiments indicate that grade A and B oocytes have a similar developmental potential and that the recovery of oocytes of these grades is maximised by use of an 18‐gauge needle and 50 mmHg aspiration pressure., The global population is expected to rise dramatically over the next few decades. This means that the demand for meat, including pig derived products will also increase, and that a more sustainable way to produce such products is necessary. The establishment of production scale pig embryo in vitro production (IVP) could be one answer to this problem, however some challenges still exist, one of which is that pig IVP requires the consistent isolation of large numbers of high‐quality oocytes. Here, ways of maximising both the yield and the developmental potential of oocytes from abattoir derived ovaries has been investigated.

SUMMARYBackground Patterns of healthcare encounters by patients in each stage of chronic kidney disease (CKD) have not been fully described. Objective This study describes patterns of healthcare resource use by patients with CKD. Design A retrospective descriptive design was used. Participants Patients with Stages 1–5 CKD were identified in five existing de-identified healthcare insurance claims databases in the United States using codes from the International Classification of Diseases (ICD-9-CM). Measurements The databases contained more than 23,660,000 claims records from over 11 million subscribers who were continuously enrolled in a single 2014 health plan. All CKD patients’ 2014 claims were extracted, yielding 1,987 unique people with 110,594 healthcare encounters. Results Healthcare resources are used to manage the causes of CKD and its multiple effects on health, and thus the number of healthcare encounters among people with more advanced disease was, as expected, relatively higher. There were more hospitalisations, emergency department visits and specialist encounters in this group. Surprisingly, however, even people in earlier stages of kidney disease experienced a median of 14–17 healthcare encounters during a single calendar year. Conclusions Understanding patterns of healthcare encounters provides important information about the transition experiences of patients with CKD. Exploring ways to reduce the risks associated with transitions in care may prevent problems with home medication management, frequent emergency department visits and potentially avoidable hospitalisations.

The identification of “critical drainage areas” to quantify “hotspot” flood and pollution risks associated with extreme event urban surface runoff is central to Stormwater Management Plans (SWMPs) and Water Framework Directive (WFD) catchment planning. An innovative GIS-based 1D-2D modeling analysis coupled with a drainage assessment tool is described which addresses this methodological requirement. The modeling approach further integrates a Sustainable Urban Drainage System (SUDS) tool called SUDSLOC to provide a stakeholder-friendly surface water management framework. The modeling approach is illustrated by reference to two small urban catchments within the cities of Birmingham and Coventry in the Midlands region of England and the benefits of utilising ground-based LiDAR survey to build the surface micro-topography are demonstrated. The performance effectiveness of the selected SUDS controls are explored and the utility of the graphical animated outputs are discussed. Whilst the implemented SUDS controls exhibit substantial reductions (>57%) in total discharge volumes for storms up to 1:30 year return periods, there are very limited volumetric reductions for storm events exceeding this return period.

Objective To devise a kinematic technique to objectively ascertain the location and orientation of the centre of rotation of the canine elbow and to compare this axis following arthroplasty with the first generation TATE™ prosthesis in an ex vivo model. Methods Five pairs of cadaveric forelimbs were obtained and proximal limb soft tissues removed. Pin-mounted reflective markers were applied to the humerus and ulna. Limbs were mounted on a frame and six trials of the elbow manually cycled through its sagittal range of motion captured using 4 Qualisys cameras at 120 Hz. Radiography was performed to identify marker position. TATE™ cartridges were implanted and kinematic analysis repeated. Kinematic data were imported into custom software and the three-dimensional joint centre of rotation defined using a closed-form solution for absolute orientation. Paired t tests were performed to determine if the centre of rotation of the elbow differed significantly (P

Adult degree programs have been seen as a win-win solution for private colleges and adult learners, but their innovative and often-entrepreneurial postures are not a natural fit with governance structures in more traditional institutions. Through narrative and illustrative vignettes, this chapter presents an overview of efforts employed by some institutions to strike a balance between their historic missions and these nontraditional programs.

Background: The aims of this study are to investigate the expression of the main structural components of the tarsal extracellular matrix (ECM) in floppy eyelid syndrome (FES) focusing on elastic fibres and collagen types I and III, and also to identify possible cell-mediated inflammatory mechanisms in the pathogenesis of this condition. Methods: A histopathological case control study was conducted using 30 upper lid specimens from patients with FES and 15 undiseased upper lid control specimens. Structural ECM components were assessed using a combination of immunctorial ataining ohistochemical and techniques including antibodies to collagens I and III, Verhoeff's iron haematoxylin, Gomori's aldehyde fuchsin and Lillie's oxidised aldehyde fuchsin. The contribution of different cellular components of the inflammatory response was investigated by immunohistochemical techniques using antibodies to CD3, CD20, CD68. Slide scoring was performed using a semiquantitative technique on an ordinal scale. Statistical analysis was performed using matched ordinal regression analysis. Results: FES tarsal plate tissue demonstrated a decreased abundance of mature elastic fibres (P ≤ 0.001) and an increased abundance of oxytalan fibres (P = 0.006). Intensity of staining for collagens I (P = 0.012) and III (P

This study identified genes that distinguish Australian Fusarium oxysporum f.sp. vasinfectum (Fov) isolates from related co-localized non-pathogenic F. oxysporum isolates and from non-Australian Fov isolates. One gene is a homologue of the F. oxysporum f.sp. lycopersici (Fol) effector gene SIX6, encoding a 215-residue cysteine-rich secreted protein. The Six6 proteins from Fol and Fov contained eight conserved cysteine residues, five of which occurred in the highly diverged 48-amino-acid region where FovSix6 differs from FolSix6 at 32 residues. Two other potential effector genes, PEP1 and PEP2, were identified in a cDNA library of Fov genes expressed during infection of cotton. The presence of FovSIX6 and other differences in DNA fingerprints clearly distinguished Australian Fov isolates from non-Australian Fov isolates and these differences further support the hypothesis based on earlier phylogenetic analysis that Australian Fov is different from Fov in other cotton-growing areas. A specific diagnostic for Fov based on FovSIX6 is described.

The horse evolved to survive on rations high in forage. Many performance horses are fed rations containing reduced levels of forage, with a corresponding increase in concentrate supply. Such reductions in forage intake are widely established to be associated with a corresponding number of physiological and psychological adaptations. Therefore, the influence of forage intake on bodyweight (bwt) and performance was investigated. Four Thoroughbred-type geldings in light to moderate work received 4 diets (100% forage [100H]; 80% forage:20% concentrate [80H]; 60% forage:40% concentrate [60H] and 50% forage:50% concentrate [50H]) in a 4 x 4 Latin-square design. A submaximal standardised exercise test (SET) was performed for each diet. Rate of passage, bwt and water intakes were measured throughout the trial and maximum, recovery heart rates and postexercise rectal temperatures recorded for each SET. Mean +/- s.e. bwt was significantly (P

The importance of evaluating greenhouse gas (GHG) emissions from dairy cows within the whole farm setting is being realized as more important than evaluating these emissions in isolation. Current whole farm models aimed at evaluating GHG emissions make use of simple regression equations to predict enteric methane (CH4) production. The objective of the current paper is to evaluate the performance of nine CH4 prediction equations that are currently being used in whole farm GHG models. Data used to evaluate the prediction equations came from a collection of individual (IND) and treatment averaged (TRT) data. Equations were compared based on mean square prediction error (MSPE) and concordance correlation coefficient (CCC) analysis. In general, predictions were poor, with root MSPE (as a percentage of observed mean) values ranging from 20.2 to 52.5 for the IND database and from 24.0 to 38.2 for the TRT database and CCC values ranging from 0.009 to 0.493 for the IND database and from 0.000 to 0.271 for the TRT database. Overall, the equations of Moe & Tyrrell and IPCC Tier II performed best on the IND dataset, and the equations of Moe & Tyrrell and Kirchgesner et al., performed best on the TRT dataset. Results show that the simple more generalized equations performed worse than those that attempted to represent important aspects of diet composition, but in general significant amounts of bias and deviation of the regression slope from unity existed for all equations. The low prediction accuracy of CH4 equations in whole farm models may introduce substantial error into inventories of GHG emissions and lead to incorrect mitigation recommendations.

Brevetoxin A is a decacyclic ladder toxin that possesses five-,six-, seven-, eight-, and nine-membered oxacycles, as well as 22 tetrahedral stereocenters. Herein, we describe a unified approach to the B, E, G, and J rings predicated upon a ring-closing metathesis strategy from the corresponding dienes. The enolate technologies developed in our laboratory allowed access to the precursor acyclic dienes for the B, E, and G medium ring ethers. The strategies developed for the syntheses of these four monocycles ultimately provided multigram quantities of each of the rings, supporting our efforts toward the convergent completion of brevetoxin A.

It is felt, among zoo bird curators, that the European Union (EU) Regulation banning bird imports may have a hugely detrimental effect on European Association of Zoos and Aquaria (EAZA) bird populations. This paper discusses the possible effects of the legislation despite a derogation that allows some birds to be brought into the EU for zoos. The ban would only affect EAZA populations if reliant on large-scale commercial importers either directly or indirectly through affecting private collections or non-EAZA zoos. Data from the International Species Information System (ISIS) and EAZA Taxon Advisory Group (TAG) surveys were used to determine the sustainability and the main source of recruitment for these populations. The feeling that the ban will cause the demise of EAZA bird populations is not upheld by the data used, with the exception of white-eyes (Zosteropidae). However, this conclusion must be taken in context of the data available. The reality of commercial importers in underpinning EAZA populations through intermediates is possibly being masked by the way data have been entered by ISIS-member zoos. A positive outcome of discussions within EAZA Bird TAGs was to identify several other issues that need to be addressed which may be hindering population survival in EAZA collections.

Controlling the radiative lifetime of quantum dot excitons by using microcavities provides a means of improving the performance of single photon sources. By manipulating the density of optical states in the environment of the quantum dot, the rate of spontaneous recombination can be altered by the Purcell effect. The emitted light can also be concentrated into a smaller extraction cone, improving efficiency. We investigate the use of temperature to spectrally tune an InAs self-assembled quantum dot exciton and the mode of a photonic crystal defect cavity. A Purcell enhancement of the spontaneous emission rate of up to a factor of 11.4 is seen on-resonance, while suppression by up to a factor of 4.4 is seen off-resonance. Also, a two orders of magnitude increase in the intensity of light detected is measured when compared to quantum dots in bulk GaAs. (© 2006 WILEY-VCH Verlag GmbH & Co. KGaA, Weinheim)

The Water Framework Directive (WFD) provides a coherent structure within which the problems of diffuse urban pollution can be effectively tackled and managed. The context of impermeable surface water quality within the terms of the Directive are outlined, and the pollutant sources and loadings associated with various urban land uses are identified. Stormwater flows and quality are sufficient to prejudice any ‘good’ ecological potential, particularly in terms of chronic receiving water sediment toxicity. A simple volume–concentration method for the prediction of pollutant loadings and impact assessment on a subcatchment scale is described and discussed.

Aims To ascertain the frequency and identify predictors of self-reported hypoglycaemia in Type 1 and insulin-treated Type 2 diabetes. Methods A random sample of 267 people with insulin-treated diabetes were recruited from a population-based diabetes register in Tayside, Scotland. Each subject prospectively recorded the number of mild and severe hypoglycaemic episodes experienced over a 1-month period. Ordinal logistic regression was performed to identify potential predictors of hypoglycaemia. Results Five hundred and seventy-two hypoglycaemic events were reported by 155 patients. The participants with Type 1 diabetes had a total of 336 hypoglycaemic events with a rate of 42.89 events per patient per year. Of these, nine were severe hypoglycaemic events, with a rate of 1.15 events per patient per year. Participants with insulin-treated Type 2 diabetes experienced a total of 236 hypoglycaemic events with a rate of 16.37 events per patient per year. Of these, five were severe hypoglycaemic events, which would be equivalent to 0.35 events per patient per year. Predictors of hypoglycaemia in Type 1 diabetes were a history of previous hypoglycaemia (P = 0.006) and co-prescribing of any oral drug (P = 0.048). In patients with insulin-treated Type 2 diabetes, a history of previous hypoglycaemia (P

Our understanding of Lake Vostok, the huge subglacial lake beneath the East Antarctic Ice Sheet, has improved recently through the identification of key physical and chemical interactions between the ice sheet and the lake. The north of the lake, where the overlying ice sheet is thickest, is characterized by subglacial melting, whereas freezing of lake water occurs in the south, resulting in ∼210 m of ice accretion to the underside of the ice sheet. The accreted ice contains lower concentrations of the impurities normally found in glacier ice, suggesting a net transfer of material from meltwater into the lake. The small numbers of microbes found so far within the accreted ice have DNA profiles similar to those of contemporary surface microbes. Microbiologists expect, however, that Lake Vostok, and other subglacial lakes, will harbour unique species, particularly within the deeper waters and associated sediments. The extreme environments of subglacial lakes are characterized by high pressures, low temperatures, permanent darkness, limited nutrient availability, and oxygen concentrations that are derived from the ice that provides the meltwater. Copyright © 2002 John Wiley & Sons, Ltd.

Direct automated DNA sequencing was used to analyze exons 2 and 3 of HLA-B alleles present in forty-four unrelated individuals residing in the village of Adiopodoume, Cote d'Ivoire (Ivory Coast). Of the 23 HLA-B alleles observed, the most frequently detected allele was HLA-B*5301 (22.7%), which is believed to confer resistance to severe Plasmodium falciparum malaria. B*4501 (9.1%), B*1503 (8.0%), B*0705 (5.7%), B*1510 (5.7%) and B*3501 (5.7%) occurred frequently in the population. A second allele of B53 was identified; B*5302 contains a single amino acid variation at residue 171 (Y-->H). Two additional novel alleles, B* 1405 (a single amino acid variant of B*1402) and B*4410 (a five amino acid variant of B*4403) were characterized.

To examine the genetic diversity in west Africa, class I HLA-A and HLA-B alleles of 92 unrelated individuals from two areas in the Cameroon, the capital Yaounde and the village of Etoa, were identified by direct automated DNA sequencing of exons 2 and 3 of the HLA-B locus alleles and sequence-specific oligonucleotide probe (SSOP) and/or sequencing of the HLA-A locus alleles. HLA-A*2301 (18.7%), A*2902 (10.4%), B*5301 (10.9%), and B*5802 (10.9%) were the most frequently detected alleles, present in at least 10% of the population. A total of 30 HLA-A locus and 33 HLA-B locus alleles, including six novel alleles, were detected. The novel alleles were HLA-A*03012, A*2612, A*3006 and HLA-B*1403, B*4016, and B*4703. HLA-B*4703 contains a novel amino acid sequence that is a combination of the first 5 amino acids of the Bw6 epitope and the last 2 residues of the Bw4 epitope. The addition of 6 alleles to the ever-expanding number of known class I HLA alleles supports our hypothesis that extensive genetic diversity, including previously undescribed alleles, would be observed in this African population. In the Yaounde population, the allele frequency distribution at the HLA-A locus is consistent with distributions indicative of balancing selection. Extensive HLA-A-B haplotypes were observed in this population suggesting that only a fraction of the Cameroon HLA-A-B haplotype diversity has been observed.

This paper describes seventeen novel HLA-A locus alleles: A*0106, *0235, *0236, *0237, *1105, *2302, *2303, *24032, *2422, *2424, *2503, *2613, *3007, *3203, *3204, *6809 and *6810. All alleles were identified due to unexpected probe reaction patterns during routine SSOP typing. Exons 2 and 3 of these alleles were subsequently characterized by DNA sequencing. The alleles represent a shuffling of sequence motifs, likely by interallelic conversion, expanding the diversity of the HLA system.

Recombinant human monoclonal antibodies against CD152 have been generated by selecting a synthetic phage scFv library with purified CD152-Ig fusion protein. Sixteen scFv fragments were isolated which specifically react with CD152 by enzyme-linked immunoabsorbent assay (ELISA) and Western blot resulting in their clustering into two groups recognizing different antigenic determinants. One group of scFvs (#3, #13, #40, #44, #47, #51, #57, #80 #83) recognized an epitope on CD152 dimer whereas another group (#15, #18, #31, #35, #54, #72, #81) recognized an epitope on both dimeric and monomeric CD152 molecule suggesting their possible use in understanding the subunit structure of CD152 which is still controversial. Sequencing of the VH genes revealed that all the scFvs belonged to the VH3 gene family but they were different in CDR3 length and composition. It was possible to correlate specific CDR3 sequences with reactivity of the two groups of scFvs. Four scFvs, #3, #40, #81 and #83, each representative of one specific CDR3, were selected for further analysis. Competition ELISA experiments showed that they recognize CD152 in its native configuration and bound to different epitopes from the CD80/CD86 interaction site. The scFvs were able to stain human T lymphocytes stimulated either with anti-CD3 and CD28 antibodies or PHA, PMA and ionomycin by cytofluorimetry suggesting that they can be useful reagents for monitoring the kinetics of surface-bound and intracellular CD152.

1 The population density, diversity and productivity of the microbial plankton in an oligotrophic maritime Antarctic lake were studied for a 15-month period between December 1994 and February 1996. 2 In the lake, concentrations of nutrients and dissolved organic carbon were uniformly low, temperature varied over a small annual range of 0.1–3 °C, and the surface was ice-covered except during a period of approximately 6 weeks in summer. 3 The total of 57 morphotypes of protozoa observed during the study is a higher taxonomic diversity than previously reported from continental Antarctic lakes, but lower than that found in more eutrophic maritime Antarctic lakes. Likewise, planktonic abundance and productivity were lower than has been reported in other lakes on Signy Island, but generally higher than those of lakes on the Antarctic continent. 4 There were marked seasonal and interannual variations in planktonic population density. 5 Chlorophyll a concentrations ranged from undetectable to 4.2 µg L-1 and the greatest rate of primary productivity measured was 4.5 mg C m-3 h-1. The phytoplankton was dominated by small chlorophytes and chrysophytes, with phototrophic nanoflagellate abundance ranging from 1.1 × 103 to 1.2 × 107 L-1. 6 Bacterial densities of 3.6 × 108 to 1.9 × 1010 L-1 were recorded and bacterial productivity reached a peak of 0.36 µg C L-1 h-1. Numbers of heterotrophic nanoflagellates between 5.0 × 104 and 1.8 × 107 L-1, and of ciliates from undetectable to 1.1 × 104 L-1 were observed. Naked amoebae were usually rare, but occasionally reached peaks of up to 1.5 × 103 L-1.

The frequency of each A*10 allele was determined in 5 major United States ethnic populations randomly selected from a pool containing 82,979 unrelated individuals. The phenotype frequency of A10 was 10.5% in Caucasians, 14.0% in African-Americans, 21.1% in Asians/Pacific Islanders, 10.6% in Hispanics, and 9.8% in Native Americans. Fifty-nine individuals who had at least one A10 antigen were randomly chosen from each ethnic group for polymerase chain reaction using sequence-specific oligonucleotide probes (PCR-SSOP) typing. Thirteen of sixteen known A10 alleles were identified in this pool. The most common alleles observed were: A*2601 in Caucasians (55%), Hispanics (58%), and Native Americans (45%); A*3402 in African-Americans (34%); and A*3401 in Asians/Pacific Islanders (61%). The African-American and Asian/Pacific Islander populations differ from all other populations in the distribution of A*10 alleles, particularly, A*2601, A*3401, and A*3402.

The L6 rust resistance gene from flax generates at least four transcript classes by alternative splicing of the third intron. The most abundant transcript class encodes a resistance protein containing domains that include nucleotide binding site motifs and a leucine-rich repeat region (NBS-LRR). The remaining three transcript classes encode truncated products which lack most of the C-terminal part of the protein containing the leucine-rich region (LRR). The four transcript classes occur in all plant organs examined and no induction of L6 expression was observed following infection of resistant plants with an avirulent rust strain expressing the corresponding A-L6 avirulence gene. Flax plants transgenic for an intronless L6 gene, incapable of encoding truncated resistance proteins by alternative splicing, expressed L6 resistance indistinguishable from that of the wild-type gene. Therefore, a definitive role for alternative transcripts and their predicted truncated products could not be assigned in the flax/flax rust system.

Two spontaneous mutant alleles of the L6 flax rust resistance gene, 16-X3A and 16-X117, contain the same transposable element designated dLute (defective Linum usitatissimum transposable element). The element is 314 bp long, 70% AT-rich and, because it contains no extended open reading frame, is probably non-autonomous. It has 14 bp imperfect terminal inverted repeats related to those in the Ac family of plant transposons and, like Ac, causes 8 bp target site duplications upon insertion. Multiple copies of dLute-related sequences exist in the flax genome. Rust resistant revertants were recovered amongst the progeny of both mutants and reversion was associated with excision of dLute. Excision either restored the wild-type L6 sequence or was imprecise, leaving sequence alterations ('footprints') resulting in one to three amino acid alterations in the L6 protein. No phenotypic differences were discerned between plants containing the standard and revertant L6 alleles.

In the past few years large areas of the Midwest and Great Plains have been inundated with water and plant P deficiency is often a problem after such events. Soil and plant samples were collected in 1994 in Missouri and Iowa from sites flooded and fallowed in 1993. Plants were P deficient during early vegetative growth in spite of soil tests indicating adequate P availability and typical levels of starter fertilizer. The flood and fallow problem was associated with the loss of root colonizing vesicular-arbuscular mycorrhizal (VAM) fungi that benefit the plant by increasing uptake of P. The VAM fungi are dependent upon the plant for growth and reproduction and loss of host significantly reduces VAM fungal populations in soil. The VAM fungal populations increased with the presence of a crop in the year following the flood. A greenhouse study on the effect of flooding on VAM fungi indicates the number of VAM hyphae or spores producing colonization were not affected by an extended flooding period and therefore are related to the lack of host for an extended period of time in the same manner as long fallow disorder in Australia. The research conducted indicates that the Post Flood Syndrome was associated with the loss of VAM fungi. Use of starter fertilizer was beneficial in correcting Post Flood Syndrome but broadcast fertilizer application appeared to have an insignificant effect on reducing the P deficiency. The application of 80 lb P/acre as starter fertilizer produced plants that were not P deficient.

The eighteenth report of the Royal Commission on Environmental Pollution made no reference to any potential environmental impacts resulting from runoff pollution associated with transport activities. This paper identifies the magnitude and extent of pollution associated with discharges from various transport sources and identifies solids, metals, hydrocarbons, herbicides and dc-icing agents as the principal contaminants of environmental concern. The impacts of these pollutants on the receiving water are reviewed. They are primarily associated with highways which have a high traffic density, and herbicides are considered to be the only potential and widespread hazard to groundwaters. Highway and airport runoff are shown to have both acute and chronic effects on biotic diversity and organism mortality rates, although these effects appear to be largely confined to reaches immediately downstream from the discharge outfalls.

Three previously unreported DRB1*03 alleles are described, adding to the diversity of the DRB 1 family of alleles. DRB1*03022 contains a silent substitution at codon 77. DRB1*0307 differs from DRB1*03011 by a substitution at codon 26 resulting in a predicted change from tyrosine to phenylalanine. DRB1*0308 is almost identical to DRB1*03011 differing at codon 58 which specifies the glutamic acid residue commonly found in DRB1*11 alleles. The new alleles (DRB1*03022, *0307, *0308) may have arisen by gene conversion-like events and add to the increasing complexity of the HLA system.

intranasally with 2 x 107 TCID50. The calves were monitored daily for signs of disease, and blood samples and nasal and rectal swabs were collected on the day of inoculation and two, four, seven, nine, 11, 14, 17, 21, 24 and 28 days later. All procedures complied with the guidelines of the Canadian Council on Animal Care. Virus isolation was performed as described by Tessaro and others (1999). Two of the three calves developed fever (40·5 and 42·0°C), but none showed any other signs of disease. Virus was only isolated from serum and/or leucocytes four and seven days after inoculation. All three animals showed a decrease in total leucocyte counts of 50 per cent or more by day 7, and had seroconverted to BVDV at day 11, as determined by a serum neutralisation assay (Deregt and others 1992). Thus, although the isolate could infect cattle, it appeared to be innocuous, at least for calves of this age group. Both of the bison isolates were typed as BVDV-1 using a multiplex PCR that distinguishes between BVDV-1 and BVDV-2 strains (Gilbert and others 1999). This PCR uses primers to amplify sequences of the NS5B polymerase gene. Since this gene has not been used for phylogenetic analysis, as have the Npro and E2 genes, the entire E2 gene of each isolate was sequenced by the methods described by Deregt and others (1998) (Fig 1). The phylogenetic relationships between the E2 genes of pestiviruses representing all five recognised species were determined using the distance matrix programs PRODIST and FITCH (PHYLIP). Bootstrap resampling of the phylogenetic trees was carried out to test the robustness of the observed clades. The phylogenetic relationships of the two bison isolates to other pestiviruses are shown in Fig 2. Both bison isolates clustered in the species BVDV-1, with isolate V1127 in the BVDV Isolation of bovine viral diarrhoea viruses from bison

Male Scyliorhinus canicula possess a longer and narrower mouth than females resulting in pronouced sexual dimorphism with respect to the mouth length/mouth width ratio (0.49 and 0.43, respectively). Significant sexual differences in the girth of the head and pre-oral, pre-branchial and head lengths were also recorded. Males were found to have longer teeth than females. Reasons for these differences are discussed.

Soil fumigation is used to eliminate plant pathogens and weeds in commercial practice and to eliminate organisms in controlled experiments, e.g., vesicular arbuscular mycorrhizal fungi (VAMF). This study was conducted to study the effect of methyl bromide fumigation on plant accumulation of 14 elements and assess plant colonization by VAMF after fumigation. Field experiments were conducted near Mead, NE, on a Sharpsburg silty clay loam (fine, montmorillonitic, mesic Typic Argiudoll). Corn (Zea mays L.) sorghum [Sorghum bicolor (L.) Moench], and soybean [Glycine max (L.) Merr.] were used in the experiments. Fumigation increased Al, Br, Ca, Fe, Mg, Mn, and Si and decreased Cu, K, P, and Zn shoot concentrations in all three plants. Three weeks after germination, shoot Al was sevenfold greater in sorghum and soybean and 16-fold greater in corn from fumigated plots than in shoots from nonfumigated plots. Shoot bromide concentrations from fumigated plots were sixfold greater in sorghum and 17- to 18-fold greater in corn and soybean than shoots from nonfumigated plots. Lower P and Zn shoot concentrations from fumigated plots were attributed to fumigation-induced suppression of VAMF root colonization or an ion balance problem resulting from the enhanced uptake of Al, Br, and other ions after soil fumigation. At harvest, Al was still three- to 10-fold and Br eight- to 20-fold greater in fumigated plot stover compared with nonfumigated plots. This study shows that methyl bromide fumigation alteration of plant uptake of Al, Br, Ca, Fe, Mg, Mn, and Si should be considered when interpreting experiments using methyl bromide soil fumigation. Contribution from USDA-ARS in cooperation with the Agric. Res. Div., Univ. of Nebraska-Lincoln. Published as Paper no. 10973. Journal Series, Agric. Res. Div.

Two thousand horses of different ages, heights and breeds were divided into two height groups of up to 14.2 hands high (hh) and more than 14.2 hh, and weighed on a weighbridge; each horse then had its weight estimated by three weigh tapes, one height specific (tape 1 or 2, depending on the animal's height) and two for general use (tapes 3 and 4). For horses up to 14.2 hh, weigh tape 1 provided the most accurate estimate of mean (sd) bodyweight (100.5 [6.2] per cent), and weigh tapes 3 and 4 were 112 (6.8) and 97-0 (6.1) per cent accurate, respectively. For horses more than 14.2 hh, weigh tape 2 provided the most accurate estimate of bodyweight (98.6 [18.4] per cent), with weigh tapes 3 and 4 being 102.6 (17.4) and 90.8 (15.2) per cent accurate, respectively.

Ocs-elements, a family of 20 bp DNA sequences, are components of a number of promoters active in plants. In the maize BMS cell line the dominant ocs-element binding activity is the ocs-element transcription factor complex called OTF. The isolation of cDNA clones from a BMS cDNA expression library for two bZIP (basic region-leucine zipper) proteins that bind the ocs-element sequence and are good candidates for forming at least part of OTF is described. The two ocs-element binding proteins, called OBF3.1 and OBF3.2, are closely related, with the OBF3.1 protein sharing 95.8% amino acid homology with part of the OBF3.2 protein although there were significant differences in the 3' untranslated regions. Genomic Southern blot analysis revealed a small gene family with a minimum of two OBF3 loci mapping to chromosomes 3L105 and 8L075. The OBF3.1 protein shared considerable homology with the wheat HBP1b protein (80% amino acid identity) and to a lesser extent with the tobacco TGA1aa protein. OBF3.1 like HBP1b was able to bind well to the Hex sequence but poorly to G-box/ABRE sequences. Interestingly, OBF3.1 bound eightfold more efficiently to an ocs-element sequence than TGA1a, raising the possibility that OBF3.1 and TGA1a may be distinct members of an OBF3/TGA subfamily.

The energy intake of a group of 30 patients with cystic fibrosis (CF) was compared with 30 age/gender matched controls. Dietary intakes were determined from 7-day weighed food records using computerized food composition tables. Absolute energy intakes of the CF group were comparable to those of the controls: when expressed as energy per kg body-weight, the CF group consumed approximately 34% more energy (P

Brevetoxin A is a decacyclic ladder toxin that possesses 5-, 6-, 7-, 8-, and 9-membered oxacycles, as well as 22 tetrahedral stereocenters. Herein, we describe a unified approach to the B, E, G, and J rings based upon a ring-closing metathesis strategy from the corresponding dienes. The enolate technologies developed in our laboratory allowed access to the precursor acyclic dienes for the B, E, and G medium-ring ethers. The strategies developed for the syntheses of these four monocycles ultimately provided multigram quantities of each of the rings, supporting our efforts toward the completion of a convergent synthesis of brevetoxin A.

A highly convergent, enantioselective total synthesis of brevetoxin A is reported. The development of a [X + 2 + X] Horner–Wadsworth–Emmons/cyclodehydration/reductive etherification convergent coupling strategy allowed for a unified approach to the synthesis of two advanced tetracyclic fragments from four cyclic ether subunits. The Horner–Wittig coupling of the two tetracyclic fragments provided substrates that were explored for reductive etherification, the success of which delivered a late-stage tetraol intermediate. The tetraol was converted to the natural product through an expeditious selective oxidative process, followed by methylenation.

The C2-C11 cyclized cembranoids, which include the eunicellins (also known as the cladiellins), briarellins, asbestinins, and sarcodictyins, are secondary metabolites isolated from gorgonian octocorals and soft corals. An unusual oxatricyclic ring system containing a hydroisobenzofuran and an oxonene unit with stereogenic centers residing at C1-3, 9, 10, and 14 is common to the eunicellins, briarellins, and asbestinins. However, the location of the cyclohexyl methyl groups (C11 versus C12) and the oxidation level of the sixand nine-membered rings differ among the three classes. Faulkner has proposed that the cyclization of the cembranoid diterpene skeleton initiates a biosynthetic pathway that leads to all four subclasses of these unusual molecules (Figure 1). Beginning with the cembrane skeleton, C2-C11 cyclization provides the cladiellin framework. An intramolecular etherification of the cladiellin tricycle affords the tetracyclic framework of the briarellin subclass, and a subsequent 1,2-suprafacial methyl shift of the briarellin structure is postulated to deliver the asbestinins as the class that is furthest evolved from the cembrane skeleton. The presence of multiple structural types in a common organism provides circumstantial evidence for Faulkner’s proposed biosynthetic pathway. The isolation of a cembrane metabolite with cladiellin metabolites in Alcyonium molle and with asbestinin metabolites in Briareum steckii are specific examples. The sarcodictyins are also proposed to arise from a C2-C11 cyclization of the cembrane skeleton; however, in these systems, the cyclization results in a fused cyclohexane and oxonane in place of the hydroisobenzofuran of the cladiellins, briarellins, and asbestinins. As a result of this significant structural variation of the sarcodictyins, the synthetic approaches to these molecules are quite different than those for the other three related subclasses. This review will cover efforts toward the eunicellins, briarellins, and asbestinins but will not cover efforts toward the total synthesis of the sarcodictyins. Eunicellin was the first reported member of the C2-C11 cyclized cembranoid natural products, isolated in 1968 by Djerassi and co-workers from the soft coral Eunicella stricta found off the coast of Banyuls-sur-Mer in France. Since this discovery, over 100 unique secondary metabolites of gorgonian octocorals have been characterized, including the first asbestinin in 1980 and the first briarellin in 1995. A wide range of structural diversity is displayed by this group of marine natural products. The natural role of these cembranoids is proposed, based upon mollusk and fish lethality assays, to involve predation deterrence. Upon further investigation, several of the members of these subclasses have demonstrated significant pharmacological potential. Particularly, these diterpenes have been shown to exhibit in vitro cytotoxicity against various cancer cell lines, anti-inflammatory properties, antimicrobial activities, and histamine and acetylcholine antagonism. The fascinating molecular architecture of these cembranoids, as well as their potential as therapeutic agents, has sparked much interest in the synthetic community over the past decade. A variety of approaches toward these challenging structural motifs have been investigated and several total syntheses have been accomplished. Efforts toward the total synthesis of the cladiellins, briarellins, and asbestinins are the subject of this review.

A new synthesis of 3,3′-bioxindoles is reported that is well suited for the preparation of unsymmetrical structures. In the key step, 3-hydroxy-3,3′-bioxindoles are constructed by Mukaiyama aldol reaction of 2-siloxyindoles with isatins These tertiary carb inols are formed in high diastereoselectivities, with substitution at various positions of the isatin and the 2-siloxyindole being tolerated.

To effectively utilize resources in the measurement of canopy photosynthesis, acceptable statistical precision is necessary. Our objective was to determine the effect of sample number per lot on effective precision of canopy apparent photosynthesis (CAP) and leaf area index (LAI) measurements

Numerous fruits and vegetables are priced according to the level of one or more quality characteristics. From a grower's perspective there is a trade-off between quantity (yield) and the quality characteristics for which priced premiums are explicitly paid. This article develops a procedure to analyze such trade-offs and applies it to the situation of Concord grapes. The quality attributes included were soluble solids (sugar), pH, acid, and color. Relationships among these attributes and yield were estimated using regression procedures. The estimated coefficients were used in a separable programming model in order to impute values for the attributes.

Reasons for performing study A quantifiable measure of muscle activity related to the cervical spine may provide further understanding and evidence based support for chiropractic techniques. Surface electromyography (sEMG) is a noninvasive method of measuring muscle activity of the splenius muscle when the horse is at rest. Objectives To determine if there is a relationship between objective measurable muscle parameters and misalignments and muscle tension in the equine cervical spine. Study design Controlled paired randomised study. Methods Privately owned horses (n = 14), of mixed sex, age and mean height 157.8 cm were selected and assigned a group by matching work, management regime, age, sex and breed. The treatment group (n = 7) underwent manual chiropractic treatment following palpation. The control group underwent palpation only. A Delsys 4 sensor system was used for data collection. Probes were positioned on the muscle halfway between C1/C2 joint and the crest on the left and right sides, between the tendon insertion and the motor point to maximise signals. sEMG readings were taken at immediately before (0) and after palpation (PP) and 30 min later (30). Data were tested for normality and variance by one-way ANOVA and paired t test. Results Post treatment, there was a significant decrease (P

In a sample of 71 Lamna nasus from the Bristol Channel, most were juveniles. Males, which outnumbered females, matured at about 165 cm. Their diet was composed of fish and cephalopods.