Mutational optimization of the coelenterazine-dependent luciferase from Renilla

Renilla luciferase (RLUC) is a popular reporter enzyme for gene expression and biosensor applications, but it is an unstable enzyme whose catalytic mechanism remains to be elucidated. We titrated that one RLUC molecule can turn over about one hundred molecules of coelenterazine substrate. Mutagenesis of active site residue Pro220 extended the half-life of photon emission, yielding brighter luminescence in E. coli. Random mutagenesis uncovered two new mutations that stabilized and increased photon emission in vivo and in vitro, while ameliorating substrate inhibition. Further amended with a previously identified mutation, a new triple mutant showed a threefold improved k_cat, as well as elevated luminescence in Arabidopsis. This advances the utility of RLUC as a reporter protein, biosensor, or resonance energy donor.     

Prevalence and risk factors for medical events following exercise at Australian Greyhound race meetings

Aim

A prospective, observational study to determine the prevalence of post‐exercise conditions at Australian Greyhound race meetings and to assess association with race performance and other environmental, race‐ and dog‐related factors was undertaken.

Methods

A total of 4020 starters were observed (2813 Greyhounds, 1009 trainers, 536 races, 52 race meets, 48 race dates and 11 race tracks) following a race. The presence of diaphragmatic flutter (DF), ataxia, seizure, collapse or sudden death was recorded. Risk factors were screened by univariable logistic regression prior to multivariable backward stepwise model building.

Results

In this study, 962 starters (n = 768 dogs) had DF (23.9%), 16 starters were ataxic (0.4%) and there were no observed cases of collapse, seizure or sudden death. Race track location, increasing race distance, race grade based on increasing 1st place prize value, lower (earlier) race number at the meeting, age, a previous observation of DF at the last start, females, colour (white) and better finishing position were all associated with an increased risk of a Greyhound being observed with DF. However, when logistic regression assessing the random effect of dog was performed, the presence of previous DF was not significant. In this cohort, DF was common following strenuous exercise in Greyhounds and on its own does not appear to result in reduced performance or distress to the animal.

Conclusion

The incidence of ataxia was low and collapse, seizure and sudden death were not observed. However, even though uncommon, ataxia has welfare concerns for racing Greyhounds that warrants further investigation.     

Bewertung der nährstoffgehalte im statischen düngungsversuch bad lauchstädt

Der Statische Düngungsversuch Bad Lauchstädt wurde 1902 auf Löß‐Schwarzerde (21% Ton, 8,7°C Jahresdurchschnittstemperatur und 484 mm Jahresniederschlag) angelegt. Nach 96 Versuchsjahren wurde im Jahre 1998 auf Schlag I von allen Varianten der Gehalt an Gesamt‐P, doppellaktatlöslichem P und K, Mg, Cu, Zn, Mn und pH‐Wert bestimmt. Die P‐Entzüge liegen zwischen 9 kg/ha.a (ungedüngt) und 30 kg/ha.a. Die Gehalte an Gesamt‐P liegen zwischen 47 und 100 mg/100g Boden. Mit 10t/ha.a Stalldung wird bereits eine optimale Zufuhr an den Nährstoffen P, K und Mg erreicht bzw. überschritten. Mg und Zn reagieren deutlich auf die Stalldunggabe, weniger Cu. Die pH‐Werte liegen mit Kalkdüngung im Mittel aller Prüfglieder bei 7,3 (Spannweite 7,2 bis 7,5), ohne Kalk bei 6,6 (6,3–7,3). Stalldung wirkt nicht auf die pH‐Werte. Es besteht eine deutliche Wechselwirkung zwischen pH‐Wert und Mn. Mit Kalk wird Mn festgelegt und liegt zwischen 27 und 40 ppm, ohne Kalk zwischen 32 und 100 ppm.     

Assessing natural hazards in forestry for risk management: a review

We address the problem of how to integrate risk assessment into forest management and therefore provide a comprehensive review of recent and past literature on risk analysis and modeling and, moreover, an evaluation and summary on these papers. We provide a general scheme on how to integrate concepts of risk into forest management decisions. After an overview of the risk management process and the main hazards in forests (storm, snow, insects, fire), the paper focuses on the principal methods used to assess risks from these hazards for commercial forestry. We review mechanistic models, empirical models, and expert systems and consider the needs for different spatial scales of risk assessment, from the regional to the single-tree level. In addition to natural hazards and their secondary effects, we deal with economic aspects of risk analysis. Monte Carlo simulations to deal with volatile timber prices and ways to include risk in classical Faustmann approaches are briefly discussed along with the integration of portfolio theory into forest management decision making and attitude toward risk. Special attention is paid to the implications for risk modeling under climate change.     

Crop damage by nematodes in improved-fallow fields in western Kenya

The rotation of leguminous shrubs and crops is being tested on farms and recommended as a means of improving soil fertility and increasing crop yield in eastern and southern Africa, including western Kenya. However, this improved fallow practice may also increase the nematode population in the soil. An experiment was conducted to monitor the effects of plant-parasitic nematodes on crops after improved fallow. Soil was collected from a maize (Zea mays L.)/bean (Phaseolus vulgaris L.) field, a natural fallow, a Crotalaria (Crotalaria grahamiana Wight & Arn.) fallow, a Tephrosia (Tephrosia vogelii Hook. f.) fallow and a Crotalaria — Tephrosia mixed fallow and used to fill plastic pots placed in a shade. Three successive crop cycles of 2 months were tested in these pots using maize and beans, the most important staple foods in western Kenya. In the first cycle, beans grew poorly on the Tephrosia and Crotalaria — Tephrosia soil due to the high incidence of root knot nematodes, Meloidogyne spp., while maize did not suffer any loss. Although the populations of root knot nematodes reduced drastically in the second and third cycles, both maize and beans experienced heavy losses on the soil under improved fallow probably due to the spiral nematodes, Scutellonema spp., which became dominant in the nematode communities. Despite the use of fertilisers (N, P, K), both crops became highly sensitive to spiral nematodes in the third cycle because of the degradation of the soil physical properties. The study showed that the benefits of improved fallows in terms of crop production may be limited by the high number of plant-parasitic nematodes they help develop in the process.This revised version was published online in November 2005 with corrections to the Cover Date.     

Biomarker evidence for a major preservation pathway of sedimentary organic carbon

Hydrogenation processes leading from biomolecules to fossil biomarkers in anoxic sediments are crucial for the preservation of organic matter. However, these processes are still poorly understood. The present identification of several reduced carotenoids in recent sediments attests that these processes operate at the earliest stages of diagenesis without structural or stereochemical specificity, implying a nonbiological reduction pathway. Sulfur species (e.g., H2S) are the hydrogen donors involved in such reduction, as demonstrated with laboratory experiments. These reactions allow the preservation of abundant organic carbon in the rock record.     

Doubled haploids in tropical maize: <Emphasis Type="SmallCaps">II</Emphasis>. Quantitative genetic parameters for testcross performance

Single crosses (SC) of elite inbreds and open-pollinated populations (OP) are suitable source germplasm for doubled haploid (DH) line development in hybrid maize breeding, given that they combine a high population mean ([`(x)] \overline{x} ) for testcross performance with adequate response to selection ( \Updelta G \Updelta G ). This is the first study reporting testcross grain yield (TCGY) and dry matter content (TCDMC) evaluations of 131 DH lines developed from ten tropical source germplasm comprising five OP (OP1–OP5) and five SC (SC1–SC5). Gene diversity (d) and the average number of alleles (a <sub>r</sub> ) per locus was estimated for DH lines based on 24 simple sequence repeat markers. Analysis across three environments revealed no significant differences between [`(x)] \overline{x} of OP- and SC-derived DH lines for TCGY and TCDMC. Significant genetic variance for both traits was only detected among OP-derived DH lines which may be explained by a larger number of segregating quantitative trait loci (QTL) as suggested by higher d and a _r values than in SC-derived DH lines. The usefulness criterion ( U = [`(x)] + \Updelta G U = \overline{x} + \Updelta G ) was higher for OP-derived DH lines for TCDMC, but higher for SC-derived DH lines for TCGY. DH lines from OP1, OP2, and OP3 showed high TCGY, suggesting that they may be useful in tropical hybrid breeding. We conclude that tropical OP represent a valuable source of untapped genetic variation that can efficiently be exploited with DH technology for hybrid maize breeding.