The intracellular storage and turnover of apolipoprotein B of oxidized LDL in macrophages.

We have studied the effect of several chemical modifications to low-density lipoprotein (LDL) on its intracellular fate in macrophages. Native, acetylated and oxidized 125I-LDL were supplied to cultured peritoneal macrophages and the accumulation and distribution of labelled protein was measured both during uptake and a subsequent chase period. The intracellular accumulation of macromolecular oxidized LDL protein greatly exceeded that of acetylated LDL, despite similar rates of uptake and common endocytic receptors. The accumulation of intracellular apoprotein was proportional to the extent to which the LDL was first oxidized. ApoB of oxidized LDL was more resistant to proteolysis by lysosomal enzymes than native apoB. Interestingly, acetylated apoB is more rapidly hydrolysed than the native protein. 125I-LDL modified with 4-hydroxynonenal (HNE) and myricetin, but not with malondialdehyde (MDA), was also accumulated within macrophages in a high-molecular weight fraction, and was resistant to cell-free lysosomal proteolysis. These forms of LDL also contained crosslinked apoB molecules. It is suggested that the accumulation of oxidized LDL within macrophages may he due, at least in part, to the formation of inter- or intra-molecular crosslinks in apoB which render it less accessible to proteolysis.     

X-ray structure of the gamma-subunit of a dissimilatory sulfite reductase: fixed and flexible C-terminal arms

The X-ray structure of the gamma-subunit of the dissimilatory sulfite reductase (DsrC) from Archaeoglobus fulgidus was determined at 1.12 and 2.1A resolution, in the two crystal forms named DsrC(nat) and DsrC(ox) the latter being cocrystallized with the oxidizing agent tert-butyl hydroperoxide. The fold corresponds to that of the homologous protein from Pyrobaculum aerophilum but is significantly more compact. The most interesting, highly conserved C-terminal arm adopts a well-defined conformation in A. fulgidus DsrC in contrast to the completely disordered conformation in P. aerophilum DsrC. The functional relevance of both conformations and of a potentially redox-active disulfide bond between the strictly invariant Cys103 and Cys114 are discussed.     

Emissions of methane from northern peatlands: a review of management impacts and implications for future management options

Northern peatlands constitute a significant source of atmospheric methane (CH₄). However, management of undisturbed peatlands, as well as the restoration of disturbed peatlands, will alter the exchange of CH₄ with the atmosphere. The aim of this systematic review and meta‐analysis was to collate and analyze published studies to improve our understanding of the factors that control CH₄ emissions and the impacts of management on the gas flux from northern (latitude 40° to 70°N) peatlands. The analysis includes a total of 87 studies reporting measurements of CH₄ emissions taken at 186 sites covering different countries, peatland types, and management systems. Results show that CH₄ emissions from natural northern peatlands are highly variable with a 95% CI of 7.6–15.7 g C m^?2 year^?1 for the mean and 3.3–6.3 g C m^?2 year^?1 for the median. The overall annual average (mean ± SD) is 12 ± 21 g C m^?2 year^?1 with the highest emissions from fen ecosystems. Methane emissions from natural peatlands are mainly controlled by water table (WT) depth, plant community composition, and soil pH. Although mean annual air temperature is not a good predictor of CH₄ emissions by itself, the interaction between temperature, plant community cover, WT depth, and soil pH is important. According to short‐term forecasts of climate change, these complex interactions will be the main determinant of CH₄ emissions from northern peatlands. Drainage significantly (p < .05) reduces CH₄ emissions to the atmosphere, on average by 84%. Restoration of drained peatlands by rewetting or vegetation/rewetting increases CH₄ emissions on average by 46% compared to the original premanagement CH₄ fluxes. However, to fully evaluate the net effect of management practice on the greenhouse gas balance from high latitude peatlands, both net ecosystem exchange (NEE) and carbon exports need to be considered.     

Blinded and unblinded sample size reestimation in crossover trials balanced for period

The determination of the sample size required by a crossover trial typically depends on the specification of one or more variance components. Uncertainty about the value of these parameters at the design stage means that there is often a risk a trial may be under‐ or overpowered. For many study designs, this problem has been addressed by considering adaptive design methodology that allows for the re‐estimation of the required sample size during a trial. Here, we propose and compare several approaches for this in multitreatment crossover trials. Specifically, regulators favor reestimation procedures to maintain the blinding of the treatment allocations. We therefore develop blinded estimators for the within and between person variances, following simple or block randomization. We demonstrate that, provided an equal number of patients are allocated to sequences that are balanced for period, the proposed estimators following block randomization are unbiased. We further provide a formula for the bias of the estimators following simple randomization. The performance of these procedures, along with that of an unblinded approach, is then examined utilizing three motivating examples, including one based on a recently completed four‐treatment four‐period crossover trial. Simulation results show that the performance of the proposed blinded procedures is in many cases similar to that of the unblinded approach, and thus they are an attractive alternative.     

Full carbon and greenhouse gas balances of fertilized and nonfertilized reed canary grass cultivations on an abandoned peat extraction area in a dry year

Bioenergy crop cultivation on former peat extraction areas is a potential after‐use option that provides a source of renewable energy while mitigating climate change through enhanced carbon (C) sequestration. This study investigated the full C and greenhouse gas (GHG) balances of fertilized (RCG‐F) and nonfertilized (RCG‐C) reed canary grass (RCG; Phalaris arundinacea) cultivation compared to bare peat (BP) soil within an abandoned peat extraction area in western Estonia during a dry year. Vegetation sampling, static chamber and lysimeter measurements were carried out to estimate above‐ and belowground biomass production and allocation, fluxes of carbon dioxide (CO₂), methane (CH₄) and nitrous oxide (N₂O) in cultivated strips and drainage ditches as well as the dissolved organic carbon (DOC) export, respectively. Heterotrophic respiration was determined from vegetation‐free trenched plots. Fertilization increased the above‐ to belowground biomass production ratio and the autotrophic to heterotrophic respiration ratio. The full C balance (incl. CO₂, CH₄ and DOC fluxes from strips and ditches) was 96, 215 and 180 g C m^?2 yr^?1 in RCG‐F, RCG‐C and BP, respectively, suggesting that all treatments acted as C sources during the dry year. The C balance was driven by variations in the net CO₂ exchange, whereas the combined contribution of CH₄ and DOC fluxes was <5%. The GHG balances were 3.6, 7.9 and 6.6 t CO₂ eq ha^?1 yr^?1 in RCG‐F, RCG‐C and BP, respectively. The CO₂ exchange was also the dominant component of the GHG balance, while the contributions of CH₄ and N₂O were <1% and 1–6%, respectively. Overall, this study suggests that maximizing plant growth and the associated CO₂ uptake through adequate water and nutrient supply is a key prerequisite for ensuring sustainable high yields and climate benefits in RCG cultivations established on organic soils following drainage and peat extraction.     

CASSIOPE: An expert system for conserved regions searches

Background  

Understanding genome evolution provides insight into biological mechanisms. For many years comparative genomics and analysis of conserved chromosomal regions have helped to unravel the mechanisms involved in genome evolution and their implications for the study of biological systems. Detection of conserved regions (descending from a common ancestor) not only helps clarify genome evolution but also makes it possible to identify quantitative trait loci (QTLs) and investigate gene function.     

Conceptual framework and rationale

The sterile insect technique (SIT) has been shown to be an effective and sustainable genetic approach to control populations of selected major pest insects, when part of area-wide integrated pest management (AW-IPM) programmes. The technique introduces genetic sterility in females of the target population in the field following their mating with released sterile males. This process results in population reduction or elimination via embryo lethality caused by dominant lethal mutations induced in sperm of the released males. In the past, several field trials have been carried out for mosquitoes with varying degrees of success. New technology and experience gained with other species of insect pests has encouraged a reassessment of the use of the sterility principle as part of integrated control of malaria vectors. Significant technical and logistic hurdles will need to be overcome to develop the technology and make it effective to suppress selected vector populations, and its application will probably be limited to specific ecological situations. Using sterile males to control mosquito vector populations can only be effective as part of an AW-IPM programme. The area-wide concept entails the targeting of the total mosquito population within a defined area. It requires, therefore, a thorough understanding of the target pest population biology especially as regards mating behaviour, population dynamics, dispersal and level of reproductive isolation. The key challenges for success are: 1) devising methods to monitor vector populations and measuring competitiveness of sterile males in the field, 2) designing mass rearing, sterilization and release strategies that maintain competitiveness of the sterile male mosquitoes, 3) developing methods to separate sexes in order to release only male mosquitoes and 4) adapting suppression measures and release rates to take into account the high reproductive rate of mosquitoes. Finally, success in area-wide implementation in the field can only be achieved if close attention is paid to political, socio-economic and environmental sensitivities and an efficient management organization is established taking into account the interests of all potential stakeholders of an AW-IPM programme.     

Historical applications of induced sterilisation in field populations of mosquitoes

Research on sterile mosquito technology from 1955 to the 1980s provided a substantial body of knowledge on propagation and release of sterile mosquitoes. Radiation sterilisation and chemosterilisation have been used effectively to induce dominant lethality and thereby sterilise important mosquito vectors in the laboratory. Experimental releases of chemosterilised males provided complete control of Anopheles albimanus in a small breeding population (14-15 sq km) in El Salvador. Releases of radiation sterilised males failed to control either Aedes aegypti or Anopheles quadrimaculatus in the USA. Releases of radiation-sterilised and chemosterilised male Culex quinquefasciatus in the USA and India were successful in some instances. Development of genetic sexing systems for Anopheles and improved physical separation methods for Culex have made it possible to rear and release males almost exclusively (> 99%) minimizing the release of potential vectors, the females. Factors that affected efficacy in some field programmes included reduction of competitiveness by radiation, immigration of fertilized females from outside the release zones, and inability of laboratory-bred males to perform in the wild. Despite significant progress, institutional commitments to carry the process further were generally lacking in the late 1970s and until recently. Now, with renewed interest and support for further assessment of this technology, this paper summarizes the current knowledge base, prioritizes some areas of investigation, and challenges scientists and administrators to maintain an awareness of progress, remain realistic about the interpretation of new findings, and make decisions about the sterile insect technique on the basis of informed scientific documentation. Areas recommended for priority research status include the establishment of genetic sexing mechanisms that can be transferred to other mosquito species, re-examination of radiation sterilisation, aerial release technology and mass rearing.