Compensatory spleen growth and protective function in rats

To quantify compensatory spleen growth after reduction in splenic mass and to determine the effect of age and vascular supply on remnant growth, we examined adult and weanling rats for remnant regeneration and adult rats for host protection against bloodstream pneumococcal challenge at intervals up to 180 days after partial resection or autograft implantation. In adults, subtotally resected spleens with remnant blood flow and segmental anatomy otherwise intact (eutopic remnants) displayed prompt but relatively limited gains in remnant weight and nucleated cell number, and growth after 60 days was minimal; survival after pneumococcal challenge was directly related to initial remnant weight and unrelated to length of postoperative interval. Splenic autografts (ectopic remnants), by comparison, underwent necrosis early after devascularization, recovered initial remnant weight slowly, fell far short of restoring original cellularity, and even 6 months after implantation barely attained initial size; moreover, host protection against pneumococcaemia was no different in rats with autografts than in those with no residual spleen. Remnants in situ (both partial resection and whole spleens) grew more vigorously in weanling than adult rats. The results suggest that splenic regeneration after either partial resection or implantation is much more limited than previously suggested, at least in disease-free rats, and is regulated by size and integrity of the initial remnant spleen, adequacy of vascular inflow, and age of the host. Whereas eutopic remnants provide protection against bloodborne pneumococcal challenge in proportion to initial residual size, ectopic remnants even with prolonged maturation fail to display immunoprotective function.     

Molecular polymorphism as a tool for differentiating ground beetles (Carabus species): application of ubiquitin PCR/SSCP analysis

Differentiation between Carabus species (ground beetle) and subspecies is difficult, although there have been extensive studies. To address this problem we have applied PCR in combination with SSCP analysis focussing on the evolutionally conservative ubiquitin gene to elaborate a new approach to molecular differentiation between species. We report that Carabidae possess an ubiquitin gene and that its gene has a multimeric structure. Differential SSCP analysis was performed with the monomeric form of the gene to generate a clear SSCP pattern. Such PCR/SSCP resulted in reproducible patterns throughout our experiments. Comparing different Carabus species (Carabus granulatus, C. irregularis, C. violaceus and C. auronitens) we could observe clear interspecies differences but no differences between genders. Some species showed some remarkable differences between the individuals. We suggest that the ubiquitin PCR-SSCP technique might be an additional tool for the differentiation of ground beetles.     

Costs and benefits of a leaf beetle Integrated Pest Management (IPM) program. I. Modelling changes in wood volume yields from pest management

The Tasmanian leaf beetle Paropsisterna bimaculata is a species native to Tasmania that can cause severe defoliation of eucalypt plantations. High populations of P. bimaculata, capable of causing severe defoliation if unmanaged, can periodically occur through a substantial proportion of a plantation rotation. Some exotic insect pests not yet established in Australia, most notably Asian Gypsy Moth (Lymantria dispar), share the similar characteristic of imposing a threat for a substantial proportion of a plantation rotation. Managing such a threat is operationally complex and requires committing adequate resources to sustain a robust management system. However, the costs and benefits to the plantation owner of sustaining such a management system have not been quantified. The first step of such an analysis is to quantify the gains in additional wood yields from management that protects against severe defoliation of plantations.

Forestry Tasmania has conducted considerable research to develop an Integrated Pest Management (IPM) program to protect its plantations from damaging populations of P. bimaculata, and has been using this leaf beetle IPM program since the early 2000s. The program has generated many years of operational records from leaf beetle population monitoring. These historical population monitoring data coupled with knowledge of the impact of defoliation by leaf beetles on tree growth were used to estimate how much wood volume has been saved by protecting the Forestry Tasmania plantation estate from damaging large leaf beetle populations over an entire rotation. The probability of above-threshold leaf beetle populations (defined as populations that cause severe defoliation, >50%, if uncontrolled) could be predicted from (1) plantation age (the likelihood of above-threshold populations peaks at age 4–5 years and declines to a low value by age 12 years), and (2) site-level leaf beetle risk (sites of high leaf beetle risk were at higher elevation and closer to native grassland than sites of low leaf beetle risk). Based on these relationships, the occurrence of above-threshold leaf beetle populations was predicted in Forestry Tasmania’s 50 000 ha plantation estate between 2003 and 2034. The leaf beetle IPM program, through controlling these above-threshold populations, was then predicted to be able to avert losses of 2.18 million m³ (9.4% of merchantable wood volume).     

Phylogeny of the Coleoptera based on mitochondrial cytochrome oxidase I sequence data

A 400 base pair region of the mitochondrial cytochrome oxidase I gene (COI) was sequenced for thirty-seven species of beetle, representing fifteen families. The sequence was found to be highly variable, with 12–18% divergence within families, and up to 27% divergence between families. Phylogenetic analysis using the neighbour-joining method shows the Carabidae (Adephaga) as a distinct clade, but also shows that all other beetles (Polyphaga) diverged soon after the Adephaga/Polyphaga split. Whilst some species are grouped within their respective families, others are too diverged for easy resolution. The main disagreement with trees constructed using comparative morphology and the fossil record is the position of the Coccinelli-dae. Whilst COI may be an informative gene for molecular systematics at lower taxonomic levels, or in other insects, improved resolution of this particular phylogeny will require a more highly conserved sequence.     

Elongation Factor 1 alpha resolves the monophyly of the haplodiploid ambrosia beetles Xyleborini (Coleoptera: Curculionidae)

Elongation Factor 1-alpha was used to test the monophyly of the wood boring beetle tribe Xyleborini, where all species are haplodiploid and perform regular inbreeding by brother-sister mating. Due to their feeding requirements, being highly dependent on ophiostomatoid fungi which they cultivate in wood tunnels, monophyly may be expected due to nutritional constraints. During the course of analyses, two copies of EF-1alpha were amplified in these beetles, differing in intron structure. The high similarity between paralogous amino acid sequences (93-94%) indicates a rather recent duplication in beetles, but phylogenetic analyses of different copies in insects rejected this hypothesis. Subsequent phylogenetic analyses of eighty orthologous sequences from Xyleborini and allied taxa, using the single-intron bearing copy, were greatly improved in resolution and node support by including the intron sequences (c. 60 bp). Most analyses resulted in a monophyletic Xyleborini, implying one origin of fungus feeding in this tribe. However, clear evidence for a polyphyletic Xyleborus and three more xyleborine genera calls for further revision of xyleborine classification.     

Chymase in exocytosed rat mast cell granules effectively proteolyzes apolipoprotein AI-containing lipoproteins, so reducing the cholesterol efflux-inducing ability of serum and aortic intimal fluid.

Degranulated mast cells are present in human fatty streaks. Chymase in granules released from degranulated rat serosal mast cells, i.e., in granule remnants, proteolyzes human high density lipoprotein3 (HDL3), and so reduces its ability to induce cholesterol efflux from macrophage foam cells in vitro. In this study we found that remnant chymase, by proteolyzing human serum and human aortic intimal fluid, prevents these two physiologic fluids from effectively inducing cholesterol efflux from cultured macrophage foam cells. Inhibition was strongest when remnants were added to apolipoprotein AI (apoAI)-containing lipoproteins; the remnants had no effect on the weaker efflux produced by apoAI-deficient serum. Western blot analysis showed that granule remnants degrade apoAI in serum and in internal fluid. When released from remnants, chymase lost its ability to proteolyze HDL3 in the presence of serum. Thus, remnant chymase (but not isolated chymase) was able to resist the natural protease inhibitors present in serum and in intimal fluid. The results imply participation of exocytosed mast cell granules in foam cell formation in atherogenesis.     

LDL receptor-related protein mediates cell-surface clustering and hepatic sequestration of chylomicron remnants in LDLR-deficient mice

It has been proposed that in the liver, chylomicron remnants (lipoproteins carrying dietary lipid) may be sequestered before being internalized by hepatocytes. To study this, chylomicron remnants labeled with a fluorescent dye were perfused into isolated livers of LDL receptor-deficient (LDLR-deficient) mice (Ldlr(-/-)) and examined by confocal microscopy. In contrast to livers from normal mice, there was clustering of the chylomicron remnants on the cell surface in the space of DISSE: These remnant clusters colocalized with clusters of LDLR-related protein (LRP) and could be eliminated by low concentrations of receptor-associated protein, an inhibitor of LRP. When competed with ligands of heparan sulfate proteoglycans (HSPGs), the remnant clusters still appeared but were fewer in number, although syndecans (membrane HSPGs) colocalized with the remnant clusters. This suggests that the clustering of remnants is not dependent on syndecans but that the syndecans may modify the binding of remnants. These results establish that sequestration is a novel process, the clustering of remnants in the space of DISSE: The clustering involves remnants binding to the LRP, and this may be stabilized by binding with syndecans, eventually followed by endocytosis.     

Development of a homogeneous assay to measure remnant lipoprotein cholesterol

BACKGROUND: Quantification of triglyceride-rich lipoprotein (TRL) remnants is useful for risk assessment of coronary artery disease and the diagnosis of type III hyperlipoproteinemia. Although an immunoseparation procedure for remnant-like particle cholesterol has been evaluated extensively in recent years, available methods for measuring TRL remnants have not achieved wide use in routine laboratory practice, suggesting a need for a homogeneous assay that can measure TRL remnant cholesterol in serum or plasma without pretreatment. METHODS: We screened for suitable surfactants that exhibited favorable selectivity toward the VLDL remnant (VLDLR) fraction, including intermediate-density lipoproteins (IDLs). We investigated the principal characteristics of this assay by gel filtration of lipoproteins and their particle size distribution. We developed a simple assay and evaluated its performance with the Hitachi-7170 analyzer. RESULTS: Polyoxyethylene-polyoxybutylene block copolymer (POE-POB) exhibited favorable selectivity toward VLDLR and IDL fractions. POE-POB removed apolipoprotein (apo) E and apo C-III from IDL particles in the presence of cholesterol esterase (CHER), and the particle size distribution of IDLs became smaller after the reaction. These results revealed that IDL particles are specifically modified in the presence of CHER and POE-POB, making their component cholesterol available for enzymatic assay. Addition of phospholipase D improved the reactivity toward chylomicron remnants (CMRs). We found a high correlation [y = 1.018x- 0.01 mmol/L, r = 0.962 (n = 160)] between the proposed assay and the immunoseparation assay in serum from healthy individuals. CONCLUSION: The homogeneous assay described in this report can measure TRL remnant cholesterol, including CMRs, VLDLRs, and IDLs, with high sensitivity and specificity.     

Landscape Level Biodiversity Databases in India: Status and the Scope

Conservation of biodiversity requires management higher than the species level of organization, particularly at the landscape scale. It is, however, difficult to manage each threatened species individually. Alternatively, management can focus on the ecosystems that contain these species, and on the landscapes containing the ecosystems. The relatively new discipline of landscape ecology provides insight into both landscape diversity and the species diversity, and suggests a theoretical and practical basis for conservation planning. The information on the biodiversity characteristics such as species richness and their spatial distribution, economic, and the ethno-botanical importance is of great significance to any nation. The Department of Biotechnology and Department of Space, Government of India funded a nationwide project between 1998 and 2010 to characterize and map the flowering plants richness in the natural (forests, grasslands, scrub, etc.) and man-made (forest plantations) vegetation formations at landscape level. The spatial database on vegetation types generated using wet and dry season satellite imagery and ancillary data such as topographic maps and the species richness through field inventory were used to generate the spatially-explicit species distribution maps and statistics. A customized software package, SPLAM (Spatial Landscape Modeling) was developed for landscape analysis and spatial data integration. This, first of its kind unique study spanning over 12 years, has resulted in a large baseline spatial database on vegetation types, porosity, patchiness, interspersion, juxtaposition, fragmentation, disturbance regimes, ecosystem uniqueness, terrain complexity and the species richness. The field inventory involved 16,581 geo-referenced 0.04 ha plots across India and 7,761 plant species. The geospatially-tagged species database, created in the project, provides information on the endemic, rare, endangered, threatened, and medicinally/economically important species. The database, disseminated to large number of organizations including State Forest Departments, has found extensive applications in policy planning, operational management, biodiversity conservation, bio-prospecting, and the climate change studies.     

Binding and uptake of chylomicron remnants by primary and THP-1 human monocyte-derived macrophages: determination of binding proteins

The binding and uptake of chylomicron remnants by human macrophages was studied in order to resolve paradoxical observations that have described the putative mechanisms by which postprandial lipoproteins induce foam cell formation. Chylomicron remnants bound to human monocyte-derived macrophages (HMMs) and to the transformed monocytic cell line THP-1 with high affinity (Kd of approx. 5.5 microg of chylomicron remnant protein/ml). Binding was found to be saturable for both cell types, and was strongly inhibited in the presence of unlabelled chylomicron remnants. Ligand blot studies with colloidal-gold-labelled chylomicron remnants identified two cell surface binding sites on both HMMs and THP-1 cells, with molecular masses of approx. 128 kDa and 43 kDa. The high-molecular-mass binding site was found to be the low-density lipoprotein (LDL) receptor, based on the strong inhibition of chylomicron remnant binding in the presence of unlabelled LDL, Fab2 antibody fragments to the LDL receptor or calcium chelators. Competition studies suggested that, in HMMs, the LDL receptor appeared to facilitate approximately half of the total chylomicron remnant uptake. In contrast, the LDL receptor was not significantly involved in macrophage uptake of chylomicron remnants by THP-1 cells. The identity of the 43 kDa binding site is presently unknown, but, importantly, expression was not inhibited as a consequence of sterol loading, which was induced by incubating HMMs and THP-1 cells with 25-hydroxycholesterol. In contrast, the expression of the LDL receptor was substantially attenuated following lipid loading. Collectively, our data suggest that, while the macrophage LDL receptor can bind chylomicron remnants and facilitate uptake in non-lipid-loaded HMMs, other sterol-insensitive sites are responsible for the unabated uptake of chylomicron remnants by macrophages. We propose that the 43 kDa macrophage chylomicron remnant binding protein may be a candidate for the sterol loading of macrophages.     

Characteristics of chylomicron remnant uptake into rat liver

We have investigated uptake of 125I-labeled chylomicron remnants into livers of rats in the presence of lactoferrin. This glycoprotein possesses a cluster of four arginines at the N-terminus similar to the arginine rich binding sequence of apoprotein E (apoE) to the LDL-receptor. We found that this protein inhibits uptake of 125I-chylomicron remnant radioactivity by 50% when measured as accumulation of radioactivity into the intact organ, and even more pronounced, over 75%, when measured as uptake into an endosomal fraction prepared therefrom. Provided that the arginine rich sequence is responsible for the inhibition, a similarity in the characteristics of binding of apoE to the LDL (low density lipoprotein)- and chylomicron remnant-receptor is likely. Second, transferrin having sequence homologies with lactoferrin, but lacking the arginine cluster does not interfere with chylomicron remnant uptake. Third, lactoferrin does not inhibit the uptake of chylomicron remnants by the spleen, which is most likely mediated through scavenger cells by a mechanism different from the chylomicron remnant uptake system of the liver. We hypothesize from this that lactoferrin specifically interferes with the physiologically relevant chylomicron remnant uptake system of the liver. Investigation of the mechanism of this inhibition will provide information about the physical characteristics of the remnant receptor system.     

A secondary structural model of the 28S rRNA expansion segments D2 and D3 from rootworms and related leaf beetles (Coleoptera: Chrysomelidae; Galerucinae)

We analysed the secondary structure of two expansion segments (D2, D3) of the 28S rRNA gene from 229 leaf beetles (Coleoptera: Chrysomelidae), the majority of which are in the subfamily Galerucinae. The sequences were compared in a multiple sequence alignment, with secondary structure inferred primarily from the compensatory base changes in the conserved helices of the rRNA molecules. This comparative approach yielded thirty helices comprised of base pairs with positional covariation. Based on these leaf beetle sequences, we report an annotated secondary structural model for the D2 and D3 expansion segments that will prove useful in assigning positional nucleotide homology for phylogeny reconstruction in these and closely related beetle taxa. This predicted structure, consisting of seven major compound helices, is mostly consistent with previously proposed models for the D2 and D3 expansion segments in insects. Despite a lack of conservation in the primary structure of these regions of insect 28S rRNA, the evolution of the secondary structure of these seven major motifs may be informative above the nucleotide level for higher-order phylogeny reconstruction of major insect lineages.     

Chylomicron remnant concentrations in patients with coronary artery disease.

Persisting chylomicron remnants have been linked to premature atherosclerosis. The analysis of chylomicron remnant concentrations by an oral triglyceride tolerance test, however, is time-consuming for the study subjects and requires large resources in the laboratory. Therefore, only small numbers of subjects have been studied in the past. The aim of this study was to elucidate the prevalence of elevated chylomicron remnants, to identify effectors of chylomicron remnant clearance and to compare chylomicron remnants in the prediction of coronary artery disease with other risk factors. We applied a novel oral triglyceride tolerance test to 423 patients (368 males, 55 females) with a confirmed diagnosis of coronary artery disease (CAD) and to 390 control subjects (295 males, 95 females) in a case-control setting. This study revealed that elevated chylomicron remnant concentrations (retinyl esters > 1.5 micromol/l) are present in 20% of all subjects. Male gender, the apolipoprotein E2 isoform, and higher body mass index were associated with increased chylomicron remnant concentrations. However, chylomicron remnants were lower and plasma triglycerides higher in patients with CAD. We conclude that screening for a delayed clearance of chylomicron remnants is of little clinical value in CAD.     

Sonographic features of ovarian remnants.

Ovarian remnants occur after a portion of ovarian tissue is left behind unintentionally after oophorectomy. The ovarian remnant may be functional and cystic, producing pelvic pain and, in some patients, extrinsic compression of the distal ureter. Ovarian remnants frequently are associated with adhesions from previous pelvic surgery for endometriosis or pelvic inflammatory disease. Ovarian remnants also may be included within pelvic peritoneal inclusion cysts. In this retrospective study, the sonographic features of ovarian remnants in 10 patients with surgical proof or clinical follow-up data are described. Most ovarian remnants were simple cysts (seven of 10), three had multiple septations, and six had a rim of presumably ovarian tissue with arterial and venous flow. Three patients with ovarian remnant masses that were aspirated had symptomatic relief without recurrence. In one patient, guided aspiration was unsuccessful, probably owing to the presence of organized hemorrhage within the mass. Extrinsic compression of the distal ureter was observed in one patient, who was treated with gonadotropin releasing hormone agonist (Lupron). The sonographic findings of a completely cystic or multiseptated pelvic mass with a rim of vascularized solid tissue in a postoophorectomy patient, although such cases are rare, suggest the diagnosis of an ovarian remnant. If the diagnosis can be established with a high degree of certainty, sonographically guided aspiration may be attempted in an effort to provide symptomatic relief. Otherwise, sonography is useful in serial assessment of these masses in patients receiving medical treatment.     

Characterization of remnants produced during the metabolism of triglyceride-rich lipoproteins of blood plasma and intestinal lymph in the rat.

The metabolism of intravenously injected large and small chylomicrons from intestinal lymph and of very low density lipoproteins from blood plasma was studied in functionally eviscerated "supradiaphragmetic" rats. For studies with lymph lipoproteins, recipient animals were injected with 4-amino-pyrazolopyrimidine 18 h before injection of lipoprotein to prevent secretion of very low density lipoproteins into their blood plasma. In all cases, most of the triglycerides (labeled with 14C) were rapidly metabolized, whereas cholesteryl esters (labeled with 3H) persisted in the blood. Most of the cholesteryl esters remained in smaller "remnant" lipoproteins, less dense that 1.006, which retained an apparently spherical shape, as determined by electron microscopy of negatively stained preparations. Whereas the diameters and chemical compositions of large chylomicrons were substantially different from those of small chylomicrons and very low density lipoproteins, all remnants were similar in these respects. Average remnant diameters were 400-600 A and remnants were enriched in cholesteryl esters and in protein insoluble in tetramethylurea. In addition to triglycerides, remnants were depleted of phospholiarticle size, the composition of remnants, like that of their precursors, was consistent with the "pseudomicellar" model of lipoproteins, in which a core of nonpolar lipids is covered by a monolayer of polar lipids and protein. These results domonstrate the fundamental similarity of the initial step in the metabolism of triglyceride-rich lipoproteins from intestinal mucosa and liver and show that loss of triglycerides from the core of the particles is accompanied by removal of polar components from the surface.     

Increase in atrial size in long-term survivors of heart transplant.

Atrial size in recipients of orthotopic heart transplant (OHTX) who have long-term survival is not well described in the literature. We reviewed 2-dimensional echocardiograms of 14 recipients of OHTX who survived at least 5 years. Apical 4-chamber images were used for measurements starting at 1-year post-OHTX. The recipients of OHTX were 8 (57%) men and 6 (43%) women with a mean age of 43.6 +/- 12.1 (20-60) years and mean survival of 9.5 +/- 2.6 (5.8-14.4) years. All chambers increased with time post-OHTX (r > 0.83, P <.002). The changes in left atrium (LA) and right atrium (RA), remnants of donor LA and RA, and remnants of recipients' LA areas correlated inversely with patient survival post-OHTX (r > 0.83, P <.002). The change in RA recipient remnant area did not correlate with survival (r = 0.58, P =.06). In recipients of OHTX with long-term survival, there is a significant correlation between the echocardiographic size of LA and RA, and donor and recipient remnants with time and survival.     

Effects of hypothyroidism and cholesterol feeding on the clearance of chylomicron remnants in vivo and by rat hepatocyte monolayers

Abstract. The hepatic catabolism of chylomicron remnants in normal rats and in hypothyroid rats which were either normocholesterolaemic or made hypercholesterolaemic by feeding cholesterol and cholic acid was studied in vivo and in hepatocyte monolayers.
In vivo , the clearance of injected chylomicron remnants labelled with either cholesteryl- or retinyl ester was delayed in the hypercholesterolaemic hypothyroid rats, but not in normocholesterolaemic hypothyroid rats.
Cholesteryl-ester-rich hepatocytes from hypercholesterolaemic hypothyroid rats took up remnants less efficiently than did normal hepatocytes. Hepatocytes from normocholesterolaemic hypothyroid rats had a lower cholesteryl ester content and took up remant particles to a greater degree than did normal hepatocytes. When normal hepatocytes were incubated with hypercholesterolaemic serum or with mevalonolactone, which increased cell cholesteryl ester content, there was a slight suppression of remnant uptake. Also, addition of triiodothyronine to hepatocyte monolayers suppressed rather than increased uptake of chylomicron remants in hepatocytes.
Thus, the study suggests that chylomicron remnant uptake by the liver is not inhibited by hypothyroidism per se but by the cholesterol accumulation in hepatocytes that is the consequence of cholesterol and bile acid feeding of the rats. Although the cholesteryl ester content in hepatocytes seems to determine remnant uptake, the regulation of uptake does not seem to be as effective as that of the LDL receptor in extrahepatic cells.     

Atherogenic remnant lipoproteins: role for proteoglycans in trapping, transferring, and internalizing

Unraveling the mechanisms controlling remnant lipoprotein clearance is important, as these lipoproteins are highly atherogenic. The most critical molecule in this process is apoE, which mediates high-affinity binding of remnant lipoproteins to members of the LDL receptor (LDLR) family and cell-surface heparan sulfate proteoglycans (HSPGs), which have been shown to play major independent as well as cooperative roles in remnant lipoprotein clearance. While all the players may have been identified, our understanding of how they interact and function together continues to evolve. In this issue of the JCI, MacArthur et al. (see the related article beginning on page 153) demonstrated that HSPGs under normal physiological conditions are critically important in the clearance of remnant lipoproteins, independent of LDLR family members. The complexity of VLDL and chylomicron remnant clearance was exemplified by the studies of Jones et al., also in this issue (see the related article beginning on page 165). Despite defective clearance of LDL in mice with a deficiency in the adaptor protein controlling internalization of the LDLR, called autosomal recessive hypercholesterolemia (ARH), remnant lipoprotein clearance was not grossly abnormal. A likely explanation is that the abnormal LDLRs bind the remnants and then transfer them to another acceptor for internalization. While the studies clearly demonstrate that the LDLR-related protein 1 is not involved and suggest a role for an additional unidentified receptor, it remains a possibility that HSPGs are responsible for remnant uptake by hepatocytes in the presence of defective LDLR internalization.     

Book Review

The effects of three prescribed low intensity fires varying from means of 80 to 209 kWm^?1 within six years on invertebrates in litter/upper soil were assessed. The 8.6 year study was based on 88 934 arthropod specimens, representing 35 ordinal or lower level taxa contained in 3520 pitfall trap samples from a site burnt three times in spring and an unburnt control site within a 30.7 ha area.

The three fires had no discernible effect on total taxa, total non-insects, Araneae (spiders), Acarina (mites), Collembola (springtails), total insects and Formicidae (ants), although activity of the Coleoptera (beetles) decreased significantly after the second and third fires while that of the Diptera (flies) increased over the same period. However, as Coleoptera and Diptera activity on the unburnt control site decreased and increased respectively over the same period, it is uncertain whether the changes in activity were due to the fires or other non fire-related environmental factors. Futhermore, these changes in activity resulted from one or two of the spring fires only, and not all three, further questioning whether the fires were the contributing causal factors.

Three prescribed low-intensity fires in spring appear therefore, not to impact adversely on the activity of major forest litter arthropod groups, although the situation of the Coleoptera and Diptera is less clear due to similar activity on the unburnt control site in both cases. This therefore suggests that any fuel reduction burning in spring be conducted at intervals of not less than three years in order to minimize any impact on litter arthropods. Where a period of drought has occurred, burning should be delayed until the drought period has ended.     

An improved method for the rapid assessment of persisting chylomicron remnant concentrations

Persisting chylomicron remnant concentrations have been linked to premature atherosclerosis. The analysis of persisting chylomicron remnant concentrations by an oral triglyceride tolerance test, however, is time-consuming for the study subjects and requires large resources in the laboratory. Therefore, only small numbers of subjects have been studied in the past.We describe major improvements of the testing procedure in regard of composition of the fatty meal, of patient testing, and measurement of postprandial remnants. Shifting the time of the (ready-to-use) fatty drink from the morning hours to bedtime was well accepted by the study subjects and allowed the analysis of blood samples drawn at the morning with minimal impact on the participants' time and with minimal interferences by confounding factors (e.g. smoking, additional food intake, physical activity). Chylomicron remnants were measured by fluorometry of the supernatant after ultracentrifugation. This procedure was sensitive, was specific for chylomicron remnants, and was easy to perform. The biological validity of the improved procedure was evaluated by studying type III hyperlipoproteinemia patients and normolipemic apolipoprotein (Apo) E2 homozygotes.In conclusion, this improved test permits the rapid testing for persisting chylomicron remnants in the clinical routine and in large epidemiological studies.