Nature and extent of genetic diversity of dengue viruses determined by 454 pyrosequencing

Dengue virus (DENV) populations are characteristically highly diverse. Regular lineage extinction and replacement is an important dynamic DENV feature, and most DENV lineage turnover events are associated with increased incidence of disease. The role of genetic diversity in DENV lineage extinctions is not understood. We investigated the nature and extent of genetic diversity in the envelope (E) gene of DENV serotype 1 representing different lineages histories. A region of the DENV genome spanning the E gene was amplified and sequenced by Roche/454 pyrosequencing. The pyrosequencing results identified distinct sub-populations (haplotypes) for each DENV-1 E gene. A phylogenetic tree was constructed with the consensus DENV-1 E gene nucleotide sequences, and the sequences of each constructed haplotype showed that the haplotypes segregated with the Sanger consensus sequence of the population from which they were drawn. Haplotypes determined through pyrosequencing identified a recombinant DENV genome that could not be identified through Sanger sequencing. Nucleotide level sequence diversities of DENV-1 populations determined from SNP analysis were very low, estimated from 0.009-0.01. There were also no stop codon, frameshift or non-frameshift mutations observed in the E genes of any lineage. No significant correlations between the accumulation of deleterious mutations or increasing genetic diversity and lineage extinction were observed (p>0.5). Although our hypothesis that accumulation of deleterious mutations over time led to the extinction and replacement of DENV lineages was ultimately not supported by the data, our data does highlight the significant technical issues that must be resolved in the way in which population diversity is measured for DENV and other viruses. The results provide an insight into the within-population genetic structure and diversity of DENV-1 populations.

The carbon dioxide hydration activity of the sulfonamide-resistant carbonic anhydrase from the liver of male rat: pH independence of the steady-state kinetics

The steady-state kinetic constants for the catalysis of CO2 hydration by the sulfonamide-resistant and testosterone-induced carbonic anhydrase from the liver of the male rat has been determined by stopped-flow spectrophotometry. The turnover number was 2.6 ± 0.6 × 103 s− at 25 °C, and was invariant with pH ranging from 6.2 to 8.2 within experimental error. The Km at 25 °C was 5 ± 1 mImage , and was also pH independent. These data are in quantitative agreement with earlier findings of pH-independent CO2 hydration activity for the mammalian skeletal muscle carbonic anhydrase isozyme III. The turnover numbers for higher-activity isozymes I and II are strongly pH dependent in this pH range. Thus, the kinetic status of the male rat liver enzyme is that of carbonic anhydrase III. This finding is consistent with preliminary structural and immunologic data from other laboratories.

Dynamics of plant populations in Heteropogon contortus(black speargrass) pastures on a granite landscape in southern Queensland. 1. Dynamics of H. contortus populations

The dynamics of Heteropogon contortus (black speargrass) populations were measured in a subset of treatments contained within an extensive grazing study conducted between 1990 and 1996 in H. contortus pasture in southern Queensland. This subset included 2 landscape positions and 3 stocking rates in both native pasture and legume-oversown native pasture. Severe drought conditions throughout much of the study necessitated ongoing adjustments to the original stocking rates and, as a result, drought was the major influence on the dynamics of H. contortus populations. Plant density and basal area in the silver-leaved ironbark landscape were consistently higher than those in the narrow-leaved ironbark landscape. There was limited evidence of any impact by either light or moderate stocking rate but there was evidence of an impact at the heaviest stocking rate. There was minimal impact of legume oversowing. Relatively large fluctuations in plant density occurred during this study resulting from the death of existing plants, due mainly to drought, and seedling recruitment. Similarly, there were relatively large fluctuations in basal area caused mainly by changes in plant size. Rates for turnover of plant numbers were relatively high whereas plant turnover rates of basal areas were relatively low. Regular seedling recruitment appeared necessary to ensure the persistence of this species. Despite the high turnover, populations were maintained at reasonable levels indicating the overall resilience of H. contortus.

Development of a stable isotope index to assess decadal-scale vegetation change and application to woodlands of the Burdekin catchment, Australia

Forty-four study sites were established in remnant woodland in the Burdekin River catchment in tropical north-east Queensland, Australia, to assess recent (decadal) vegetation change. The aim of this study was further to evaluate whether wide-scale vegetation 'thickening' (proliferation of woody plants in formerly more open woodlands) had occurred during the last century, coinciding with significant changes in land management. Soil samples from several depth intervals were size separated into different soil organic carbon (SOC) fractions, which differed from one another by chemical composition and turnover times. Tropical (C4) grasses dominate in the Burdekin catchment, and thus δ13C analyses of SOC fractions with different turnover times can be used to assess whether the relative proportion of trees (C3) and grasses (C4) had changed over time. However, a method was required to permit standardized assessment of the δ13C data for the individual sites within the 13 Mha catchment, which varied in soil and vegetation characteristics. Thus, an index was developed using data from three detailed study sites and global literature to standardize individual isotopic data from different soil depths and SOC fractions to reflect only the changed proportion of trees (C3) to grasses (C3) over decadal timescales. When applied to the 44 individual sites distributed throughout the Burdekin catchment, 64% of the sites were shown to have experienced decadal vegetation thickening, while 29% had remained stable and the remaining 7% had thinned. Thus, the development of this index enabled regional scale assessment and comparison of decadal vegetation patterns without having to rely on prior knowledge of vegetation changes or aerial photography.

Forest health surveillance methodology in hardwood plantations in Queensland, Australia

Forest health surveillance (FHS) of hardwood plantations commenced in Queensland in 1997 as plantations expanded following a state government planting initiative arising from the national 2020 forest policy vision. The estate was initially characterised by a large number of small plantations (10-50 ha), although this has changed more recently with the concentration of larger plantations in the central coast and South Burnett regions. Due to the disparate nature of the resource, drive- and walkthrough surveys of subsets of plantations have been undertaken in preference to aerial surveys. FHS has been effective in detecting a number of new hardwood pests in Queensland including erinose mites (Rhombacus and Acalox spp.), western white gum plate galler (Ophelimus sp.), Creiis psyllid and bronzing bug (Thaumastocoris sp.), in evaluating their potential impact and assisting in focussing future research efforts. Since 2003 there has been an increased emphasis on training operational staff to take a greater role in identifying and reporting on forest health issues. This has increased their awareness of forest health issues, but their limited time to specifically survey and report on pests and diseases, and high rates of staff turnover, necessitate frequent ongoing training. Consequently, common and widespread problems such as quambalaria shoot blight (Quambalaria pitereka), chrysomelid leaf beetles (mainly Paropsis atomaria) and erinose mites may be under-reported or not reported, and absence data may often not be recorded at all. Comment is made on the future directions that FHS may take in hardwood plantations in Queensland.

Models of reserve manpower

Absenteeism is one of the major problems of Indian industries. It necessitates the employment of more manpower than the jobs require, resulting in the increase of manpower costs, and lowers the efficiency of plant operation through lowered performance and higher rejects. It also causes machine idleness, if extra manpower is not hired, resulting in disrupted work schedules and assignments. Several studies have investigated the causes of absenteeism (Vaid 1967) for example and their remedy and relationship between absenteeism and turnover with a suggested model for diagnosis and treatment (Hawk 1976) However, the production foremen and supervisor will face the operating task of determining how many extra operatives are to be hired in order to stave off the adverse effects of absenteeism on the man-machine system. This paper deals with a class of reserve manpower models based on the reject allowance model familiar in quality control literature. The present study considers, in addition to absenteeism, machine failures and the graded nature of manpower met within production systems and seeks to find optimal reserve manpower through computer simulation.

A Theoretical Investigation of the Ni(II)-Catalyzed Hydrovinylation of Styrene

We report a detailed and full computational investigation on the hydrovinylation reaction of styrene with the Ni(II)-phospholane catalytic system, which was originally presumed to proceed through a cationic mechanism involving a nickel hydride intermediate. The following general features emerge from this study on a specific catalyst complex that was found to give quantitative yield and moderate selectivity: (a) the activation barrier for the initiation (18.8 kcal/mol) is higher than that for the reaction due to a low-lying square-planar pentenyl chelate intermediate originating from a Ni(II)-allyl catalyst precursor. Consequently there is an induction period for the catalysis; (b) the exit of product from the catalyst is via a β-H-transfer step instead of the usual β-H elimination pathway, which has a very high activation energy due to a trans effect of the phospholane ligand; (c) the turnover-limiting and enantio- determining transition state is also the β-H-transfer; (d) because of the absence of a hydride intermediate, the unwanted isomerization of the product is prevented; (e) since the enantio-discrimination is decided at the H-transfer stage itself, the configuration of the product in a catalytic cycle influences the enantioselectivity in the subsequent cycle; (f) the trans effect of the sole strong ligand in the d8 square-planar Ni(II), the stability of the η3-benzyl intermediate, and the availability of three coordination sites enable regioselective hydrovinylation over the possible oligomerization/polymerization of the olefin substrates and linear hydrovinylation. This work has also confirmed the previously recognized role of the hemilabile group at various stages in the mechanism.

Senior House Operations in Finland

This article analyzes the operations of a sample of rental independent living senior apartments in Tampere, Finland. We compare 10 properties containing 421 units owned by one nonprofit and two for-profit housing providers. We examine costs and revenues across properties using data collected through interviews and a survey of company representatives and property managers. The results indicate that until the recent economic downturn, these senior houses generally experienced fast initial lease-up and low turnover but relatively long vacancies for some units when re-leasing. Performance varies among properties.

The relationship between the determinants of rental housing satisfaction and considering moving in Finland

Purpose The purpose of this paper is to discuss the relation between dissatisfaction with housing conditions and considering moving among residents of Finnish rental multifamily buildings. The paper examines physical attributes, socioeconomic factors, and subjective opinions related to housing conditions and satisfaction with housing. Design/methodology/approach Logistic regression analysis is used to examine survey data to analyse which factors contribute to dissatisfaction with the housing unit and the apartment building and whether dissatisfaction is related to consideration of moving. Findings The findings indicate that dissatisfaction with the building and individual housing unit are associated with greater probability of considering moving. Satisfaction with kitchen, living room, storage, and building age are the most important indicators of satisfaction with the housing unit, and satisfaction with living room, bathroom, storage, and building age are associated with satisfaction with the apartment building. These are the areas in which landlords could invest in renovations to increase satisfaction in an attempt to reduce turnover. Research limitations/implications The study is conducted with Finnish data only. The sample is not a representative sample of the Finnish population. A longitudinal study would be needed to determine whether dissatisfied residents indending to move actually change residence. Originality/value This study is the first of its kind in the Finnish housing market. It tests a general model that has been suggested to be customized to local conditions. In addition, much of the research on this topic is more than 20 years old. Examination of the model under current housing and socioeconomic conditions is necessary to determine if relationships have changed over time.

Direct electrochemical formation of nanostructured amorphous Co(OH)2 on gold electrodes with enhanced activity for the oxygen evolution reaction.

The oxides of cobalt have recently been shown to be highly effective electrocatalysts for the oxygen evolution reaction (OER) under alkaline conditions. In general species such as Co3O4 and CoOOH have been investigated that often require an elevated temperature step during their synthesis to create crystalline materials. In this work we investigate the rapid and direct electrochemical formation of amorphous nanostructured Co(OH)2 on gold electrodes under room temperture conditions which is a highly active precursor for the OER. During the OER some conversion to crystalline Co3O4 occurs at the surface, but the bulk of the material remains amorphous. It is found that the underlying gold electrode is crucial to the materials enhanced performance and provides higher current density than can be achieved using carbon, palladium or copper support electrodes. This catalyst exhibits excellent activity with a current density of 10 mA cm-2 at an overpotential of 360 mV with a high turnover frequency of 2.1 s-1 in 1 M NaOH. A Tafel slope of 56 mV dec-1 at low overpotentials and a slope of 122 mV dec-1 at high overpotentials is consistent with the dual barrier model for the electrocatalytic evolution of oxygen. Significantly, the catalyst maintains excellent activity for up to 24 hr of continuous operation and this approach offers a facile way to create a highly effective and stable material.

Estimation of red blood cell lifespan from alveolar carbon monoxide measurements

Measurement of alveolar carbon monoxide (CO) presents a facile technique to estimate the lifespan, L, of red blood cells (RBCs) in vivo. Several recent studies employ this technique and calculate L (in days) using the expression, L = 13.8 (Hb)/P-CO(end), where (Hb) is the concentration (in g/dL) of hemoglobin in blood, and P-CO(end) is the endogenous production of CO (in ppm). Implicit in this calculation is the assumption that the fraction, f, of endogenous CO production due to RBC turnover is a constant equal to 0.7, which yields the expected RBC lifespan, L approximate to 120 days, in normal controls. In anemic patients, however, enhanced RBC turnover may increase f substantially above 0.7. The above expression then overestimates L. Here, we deriv an alternative tive expression, L = 3390[Hb]/322P(CO (end)-110, that accounts explicitly for the dependence of f on the rate of RBC turnover and thereby provides more accurate estimates of L without requiring additional measurements. Using the latter expression, we recalculate L from recent measurements on hepatitis C virus infected patients undergoing treatment with ribavirin. We find that our estimates of L in these patients (39 +/- 13 days) are significantly lower than current estimates (46 +/- 14 days), indicating that ribavirin affects RBC survival more severely than expected from current studies. Our expression for L is simple to employ in a clinical setting and would render the broadly applicable technique of alveolar CO measurement for the estimation of RBC lifespan more accurate.

Losing the rural nursing workforce: Lessons learnt from resigning nurses

Background Australian workforce planning predicts a shortfall of nurses by 2025 with rural areas being most at risk. Rural areas have lower retention rates of nurses than metropolitan areas, with remote communities experiencing an even higher turnover of nursing staff. There have been few studies that examine the impact of nurse resignations on rural nursing workforces. Objective This paper is abstracted from a larger study into the reasons why nurses resign from rural hospitals and explores the resignation period. Design A qualitative study using grounded theory methods. Following in-depth interviewing and transcription, data analysis occurred with the assistance of NVivo software. Setting Rural NSW. Participants Twelve registered nurses who had resigned from rural NSW hospitals and not for reasons of retirement, maternity leave or relocation; two participants were re-interviewed. Results While the overall study identified a grounded theory which explained rural nurses resign from hospitals due to a conflict of values, three additional themes emerged about the resignation practices at rural hospitals. The first theme identified a ‘window period’ which was an opportunity for the nurse to be retained. The second theme identified that nurses who had resigned were not involved in formal exit processes such as exit interviews. The third theme captured the flow-on effect from rural nurse resignations resulting in nurses leaving the profession of nursing. Conclusion To facilitate nurse retention, it is important that rural hospitals manage nurse resignations more effectively. This includes re-examining resignation procedures, how nurses are treated and collecting meaningful data to inform retention strategies.

Periodontitis and peri-implantitis biomarkers in human oral fluids and the null-allele mouse model

Tissue destruction associated with the periodontal disease progression is caused by a cascade of host and microbial factors and proteolytic enzymes. Aberrant laminin-332 (Ln-332), human beta defensin (hBD), and matrix metalloproteinase (MMP) functions have been found in oral inflammatory diseases. The null-allele mouse model appears as the next step in oral disease research. The MMP-8 knock-out mouse model allowed us to clarify the involvement of MMP-8 in vivo in oral and related inflammatory diseases where MMP-8 is suggested to play a key role in tissue destruction. The cleaved Ln-332 γ2-chain species has been implicated in the apical migration of sulcular epithelial cells during the formation of periodontal pockets. We demonstrated that increased Ln-332 fragment levels in gingival crevicular fluid (GCF) are strongly associated with the severity of inflammation in periodontitis. Porphyromonas gingivalis trypsin-like proteinase can cleave an intact Ln-332 γ2-chain into smaller fragments and eventually promote the formation of periodontal pockets. hBDs are components of an innate mucosal defense against pathogenic microbes. Our results suggest that P. gingivalis trypsin-like proteinase can degrade hBD and thus reduce the innate immune response. Elevated levels and the increased activity of MMPs have been detected in several pathological tissue-destructive conditions where MMPs are shown to cleave extracellular matrix (ECM) and basement membrane (BM) molecules and to facilitate tissue destruction. Elevated levels of MMP-8 have been reported in many inflammatory diseases. In periodontitis, MMP-8 levels in gingival crevicular fluid (GCF) and in peri-implant sulcular fluid (PISF) are elevated at sites of active inflammation, and the increased levels of MMP-8 are mainly responsible for collagenase activity, which leads to tissue destruction. MMP-25, expressed by neutrophils, is involved in inflammatory diseases and in ECM turnover. MMP-26 can degrade ECM components and serve as an activator of other MMP enzymes. We further confirmed that increased levels and activation of MMP-8, -25, and -26 in GCF, PISF, and inflamed gingival tissue are associated with the severity of periodontal/peri-implant inflammation. We evaluated the role of MMP-8 in P. gingivalis-induced periodontitis by comparing MMP-8 knock-out (MMP8-/-) and wild-type mice. Surprisingly, MMP-8 significantly attenuated P. gingivalis-induced site-specific alveolar bone loss. We also evaluated systemic changes in serum immunoglobulin and lipoprotein profiles among these mouse groups. P. gingivalis infection increased HDL/VLDL particle size in the MMP-8-/- mice, which is an indicator of lipoprotein responses during systemic inflammation. Serum total LPS and IgG antibody levels were enhanced in both mice groups. P. gingivalis-induced periodontitis, especially in MMP-8-/- mice, is associated with severe alveolar bone loss and with systemic inflammatory and lipoprotein changes that are likely to be involved in early atherosclerosis.

Chemical probes into the active centre of a heme thiolate monoxygenase

Linalool-8-monoxygenase, a typical bacterial P-450 heme thiolase, shows a high degree of substrate specificity towards linalool. The active site of the pure enzyme has been probed with a large number of substrate analogues with systematic alterations or conformational variations in the linalool molecule. The comparison of three parameters, the mo→mos conversion of the enzyme as a result of substrate binding monitored at 392 nm, theK D of the analogues giving information about energies of association and the relative turnover as substrate have given information about the space-filling characteristics of the substrates in the enzyme cleft, the number of contacts the molecules make with the respective domains of the enzyme and the distance of the site undergoing hydroxylation from the oxygen site, respectively. The data permit the conclusion that linalool makes contact with the enzyme by hydrogen bonding with the hydroxyl group as well through hydrophobic association with all the eight carbons carrying hydrogen in the molecules.

Role of conformational dynamics in kinetics of an enzymatic cycle in a nonequilibrium steady state

Enzyme is a dynamic entity with diverse time scales, ranging from picoseconds to seconds or even longer. Here we develop a rate theory for enzyme catalysis that includes conformational dynamics as cycling on a two-dimensional (2D) reaction free energy surface involving an intrinsic reaction coordinate (X) and an enzyme conformational coordinate (Q). The validity of Michaelis-Menten (MM) equation, i.e., substrate concentration dependence of enzymatic velocity, is examined under a nonequilibrium steady state. Under certain conditions, the classic MM equation holds but with generalized microscopic interpretations of kinetic parameters. However, under other conditions, our rate theory predicts either positive (sigmoidal-like) or negative (biphasic-like) kinetic cooperativity due to the modified effective 2D reaction pathway on X-Q surface, which can explain non-MM dependence previously observed on many monomeric enzymes that involve slow or hysteretic conformational transitions. Furthermore, we find that a slow conformational relaxation during product release could retain the enzyme in a favorable configuration, such that enzymatic turnover is dynamically accelerated at high substrate concentrations. The effect of such conformation retainment in a nonequilibrium steady state is evaluated.