The effects of periodic eutrophication on parasitism and stock identification of Trematomus bernacchii (Pisces: Nototheniidae) in McMurdo Sound, Antarctica

McMurdo Sound, Antarctica, is suited for the study of the effects of eutrophication on diet-related parasites of Trematomus bernacchii (Boulenger, 1902) (Pisces). It has been relatively well studied, the environment is predictable, and there are significant differences in primary productivity over small distances. The differences in parasitism for 5 helminth species can be attributed to the enrichment processes in the sound. The prevalences of the acanthocephalan Echinorhynchus sp., the nematode Ascarophis nototheniae, the digenean Dinosoma sp., and the cestode Phyllobothrium sp. are significantly higher on the east side of the sound than on the west. The prevalence of the digenean Lepidepedon garrardi was significantly higher on the west side. We believe that the differences in the prevalences of infection between the east and west sides suggest that at least the benthic, adult fish do not readily move from one side of the sound to the other. We report the following (new host records are marked by asterisks): Echinorhynchus sp., Corynosoma hamanni (Acanthocephala); Phyllobothrium sp., Diphyllobothrium sp., Trypanorhyncha (Cestoidea); Clavellopsis (Copepoda); Dinosoma sp., Lepidepedon garrardi (Digenea); Pseudobenedenoides brachicola, Polyopisthocotylea (Monogenea); Contracaecum sp., Ascarophis nototheniae (Nematoda); Myxoproteus sp., Henneguya sp., Ceratomyxa sp., and Zschokkella sp. (Protozoa).     

Cloning and expression of recombinant Aspergillus fumigatus allergen I/a (rAsp f I/a) with IgE binding and type I skin test activity.

Aspergillus fumigatus secretes an 18-kDa nonglycosylated IgE-binding protein. This protein was previously shown to be a ribotoxin, like alpha-sarcin and mitogillin. A 686-bp long A. fumigatus cDNA encoding an 18-kDa ribotoxin was cloned and expressed in Escherichia coli as a fusion protein with six adjacent histidines (rAsp f I/a). rAsp f I/a was purified to homogeneity by Ni(2+)-chelate affinity chromatography and refolded. The recombinant protein was enzymatically active resulting in the cleavage of 28S rRNA within a universally conserved region. rAsp f I/a was cytotoxic for EBV immortalized or PHA stimulated human PBMC. Furthermore, rAsp f I/a was recognized by murine mAb made against an 18-kDa ribotoxin. IgE of individuals allergic to A. fumigatus bound to rAsp f I/a as shown by ELISA, dot blots, and Western blots. rAsp f I/a elicited positive immediate type I skin reactions in individuals allergic to A. fumigatus but not in healthy control individuals. The results show that rAsp f I/a has similar functional characteristics when compared to the native 18-kDa ribotoxin. rAsp f I/a expressed in E. coli can therefore be used as a standardized Ag/allergen for serologic and clinical diagnosis of A. fumigatus-associated diseases.     

Endotoxin rapidly induces changes in lipid metabolism that produce hypertriglyceridemia: low doses stimulate hepatic triglyceride production while high doses inhibit clearance.

Hyperlipidemia frequently accompanies infectious diseases and may be due to increases in lipoprotein production or decreases in lipoprotein clearance. The administration of endotoxin (LPS) has been used to mimic infection and prior studies demonstrate that LPS produces hypertriglyceridemia. In the present study in rodents, the dose of LPS necessary to induce hyperlipidemia was orders of magnitude less than that necessary to induce shock and death. As little as 10 ng/100 g body weight induced hypertriglyceridemia and this increase in serum triglyceride levels occurred rapidly (78% increase at 2 h). At high doses of LPS (50 micrograms/100 g body weight), the clearance of triglyceride-rich lipoproteins was decreased. At low doses of LPS (100 ng/100 g body weight), triglyceride clearance was not altered but the hepatic secretion of triglyceride was increased. Low dose LPS stimulated hepatic de novo fatty acid synthesis and lipolysis, both of which provided a source of fatty acids for the increase in hepatic triglyceride production. High dose LPS did not increase hepatic fatty acid synthesis or peripheral lipolysis, and hepatic triglyceride secretion was not stimulated. Thus, low dose LPS produces hypertriglyceridemia by increasing hepatic lipoprotein production, while high dose LPS produces hypertriglyceridemia by decreasing lipoprotein catabolism. Administration of anti-tumor necrosis factor (TNF) antibodies or interleukin 1 (IL-1) receptor antagonist did not prevent the increase in serum triglyceride levels induced by LPS. However, anti-TNF antibodies and interleukin 1 receptor antagonist (IL-1ra) blocked the increase in serum triglycerides induced by TNF or IL-1, respectively. These data suggest that neither of these cytokines is absolutely required for the increase in serum triglycerides induced by LPS, raising the possibility that other cytokines, small molecular mediators, or LPS itself may play a crucial role.     

Anti-CD3 antibodies induce T cells from unprimed animals to secrete IL-4 both in vitro and in vivo

Recently, functional heterogeneity among Th cells has been recognized. Based on pattern of lymphokine secretion, two mutually exclusive subsets of CD4+ cells have been defined and designated Th1 (secreting IL-2 and IFN-gamma) and Th2 (secreting IL-4 and IL-5). Identification of these subsets was mostly based on the study of long term cultured T cell lines and clones, and little is known about the Th heterogeneity in vivo. In particular, it has been suggested that IL-4 producing cells cannot be detected in vivo or in primary stimulations in vitro unless responder cells had been previously primed. Our data however, indicate that anti-CD3 mediated stimulation can induce T cells isolated from unprimed animals to IL-4 production. An assay system based on the ability of IL-4 to increase Ia expression of B cells present in the environment of activated T cells was found to be more sensitive than detection of secreted IL-4 in the supernatant by conventional bioassays and was used to study IL-4 production by unprimed lymphocytes polyclonally stimulated in vivo and in vitro by anti-CD3 mAb. The results obtained indicate that CD4+ CD8- T cells able to produce IL-4 upon receptor-specific stimulation exist in the preimmune pool of adult animals. Remarkably, these cells can also be stimulated in vivo by treating animals with anti-CD3 mAb, as indicated by the in vivo induction of IL-4 specific mRNA and hyper-Ia expression on B cells. These results indicate that the inability to detect IL-4 in primary cultures is not due to different activation requirements of Th2 cells but may simply result from their lower frequency in unprimed animals.     

Uniparental origin of sex chromosome polysomies.

The parental origin of the additional sex chromosomes in 8 cases with high-order sex chromosome polysomies was determined using DNA polymorphisms. The additional sex chromosomes were paternally derived in 3 48,XXYY cases, and maternal in origin in 1 48,XXXY case and 4 49,XXXXY cases. Thus, all extra chromosomes, within a particular patient, were always derived from only one parent. Their most likely origin was successive nondisjunction at the first and second meiotic division in one germ cell. The mechanism involved remains unclear, but appears to be independent of parental ages.     

Lake restoration with and without dredging of phosphorus-enriched upper sediment layers

Human activity has been the cause of continuing decline of water quality in most Dutch lakes. Development of lake restoration programmes must take into account the lake functions. Major reduction of the nutrient and pollutant loading is the primary step in lake restoration. Still, the recovery of eutrophic lakes is retarded frequently because of internal phosphorus loading by the lakes' sediments. Sediment dredging, as an additional tool for water quality management to accelerate accomplishing the desired water quality, is studied. In this paper we evaluate the preliminary results of eight lake restoration projects in the Netherlands. The lakes are compared in order to estimate the magnitude of the internal phosphorus loading. Dredging as an additional measure was carried out twice in the peatlake Geerplas. In the Nieuwkoop Lakes only the external phosphorus loading was substantially reduced from 0.9 to 0.2 g P m^–2 y^–1. Provisional results of these two shallow peatlake restoration projects focussed on eutrophication abatement with and without dredging, are presented. Both show a decrease in phosphorus concentration in the lakes. The necessity to dredge the lakes is discussed.     

The Min (multiple intestinal neoplasia) mutation: its effect on gut epithelial cell differentiation and interaction with a modifier system

Min is a fully penetrant dominant mutation that leads to the development of multiple intestinal adenomas throughout the duodenal-to-colonic axis. Min/+ C57BL6/J mice have an average life-span of 120 d. Multi-label immunocytochemical studies of these lesions demonstrate patches of differentiated enterocytes, and scattered enteroendocrine, goblet and Paneth cells. Expression of endogenous marker genes within these differentiated cells can be directly correlated with the position occupied by the adenoma along the duodenal-to-colonic axis and mirrors the regional differentiation of the normal gut epithelium. The presence of multiple lineages in adenomas together with their retention of spatial information suggests that tumorigenesis in Min/+ mice may be initiated in a multipotent stem cell normally located at the base of intestinal crypts. To study the time-dependent properties of these tumors, genetic conditions were sought in which Min/+ animals could survive for up to 300 d. Min is fully penetrant in hybrids with either AKR/J or MA/MyJ. However, the hybrids demonstrate a reduction in the number of intestinal adenomas. Preliminary backcross analysis is consistent with a single major modifier locus unlinked to Min in both the AKR/J and MA/MyJ strains. The increased lifespan of the hybrid animals is also associated with the development of invasive tumors. New tumors do not arise continuously over the lifespan of these animals; instead all adenomas appear to be established by 100 d of age or sooner. These studies indicate that the Min/+ mouse is a powerful model system for analyzing the mechanisms that establish and maintain a balance between proliferation and differentiation in the continuously renewing gut epithelium and for an assessment of the multi-step hypothesis of intestinal neoplasia.     

Peroxisome assembly mutations in humans: structural heterogeneity in Zellweger syndrome.

Empty membrane ghosts of peroxisomes were found in fibroblasts from a patient with Zellweger's syndrome, a genetic disease of humans (Santos et al: Science 239:1536-1538, 1988). Import of soluble matrix proteins into the organelle was defective. We have now studied fibroblasts from seven patients representing five complementation groups of the syndrome (defined by complementation for peroxisome enzyme function). We find that empty peroxisome ghosts are present in all seven cell samples. Three patients, representing three complementation groups, give the same membrane pattern by immunofluorescence: few large ghosts. Three other patients, representing two complementation groups, give a second pattern: many large ghosts. The seventh patient's pattern is distinct. Thus, all seven of these patients exhibit Peroxisome IMport (PIM) mutations. Since membrane assembly occurs in these cells, the results indicate that biogenesis of organelle content and membrane proteins proceed by different mechanisms. Growth and division of the empty peroxisomal membrane must occur, but are modified by the mutations (ghost size and abundance vary). Cell fusion and immunofluorescence analyses of peroxisome size and catalase packaging formally demonstrate genetic complementation groups for peroxisome assembly in Zellweger syndrome.     

Explaining the doubling of N2O emissions under elevated CO2 in the Giessen FACE via in‐field 15N tracing

Rising atmospheric CO₂ concentrations are expected to increase nitrous oxide (N₂O) emissions from soils via changes in microbial nitrogen (N) transformations. Several studies have shown that N₂O emission increases under elevated atmospheric CO₂ (eCO₂), but the underlying processes are not yet fully understood. Here, we present results showing changes in soil N transformation dynamics from the Giessen Free Air CO₂ Enrichment (GiFACE): a permanent grassland that has been exposed to eCO₂, +20% relative to ambient concentrations (aCO₂), for 15 years. We applied in the field an ammonium‐nitrate fertilizer solution, in which either ammonium () or nitrate () was labelled with ¹⁵N. The simultaneous gross N transformation rates were analysed with a ¹⁵N tracing model and a solver method. The results confirmed that after 15 years of eCO₂ the N₂O emissions under eCO₂ were still more than twofold higher than under aCO₂. The tracing model results indicated that plant uptake of did not differ between treatments, but uptake of was significantly reduced under eCO₂. However, the and availability increased slightly under eCO₂. The N₂O isotopic signature indicated that under eCO₂ the sources of the additional emissions, 8,407 μg N₂O–N/m² during the first 58 days after labelling, were associated with reduction (+2.0%), oxidation (+11.1%) and organic N oxidation (+86.9%). We presume that increased plant growth and root exudation under eCO₂ provided an additional source of bioavailable supply of energy that triggered as a priming effect the stimulation of microbial soil organic matter (SOM) mineralization and fostered the activity of the bacterial nitrite reductase. The resulting increase in incomplete denitrification and therefore an increased N₂O:N₂ emission ratio, explains the doubling of N₂O emissions. If this occurs over a wide area of grasslands in the future, this positive feedback reaction may significantly accelerate climate change.