Pulmonary O2 toxicity in lambs: physiological and biochemical effects of endotoxin infusion

Hyperoxic adult rats have prolonged survival and reduced morphological evidence of lung injury when treated with a single dose of bacterial endotoxin; this effect is mediated by an augmentation of antioxidant enzyme activity in lung homogenate. To determine whether endotoxin would prolong survival and influence antioxidant enzyme levels in lambs whose physiological response to O2 breathing can be serially measured, we administered a single intravenous dose of endotoxin (0.75 microgram/kg body wt) to 13 lambs before exposing them to greater than 95% O2 (n = 11) or air (n = 2). Seven additional lambs were placed in O2 after receiving only saline vehicle. All lambs had been instrumented to measure pulmonary vascular pressures and cardiac output, and 10 lambs had lung lymph fistulas. O2-exposed control lambs developed noncardiogenic pulmonary edema and respiratory failure within 85 +/- 10 h (range 76-110 h); antioxidant enzymes were not increased, but reduced glutathione (GSH) levels fell and oxidized glutathione (GSSG) increased, reflecting the oxidant stress of O2 exposure. By contrast, endotoxin-treated O2-exposed lambs had a delayed increase in microvascular permeability to protein, a reduced rate of lung edema formation, normal gas exchange after 72 h in O2, and prolonged survival (136 +/- 15 h; range 90-160 h; all variables P less than 0.05). Despite prolonged survival, postmortem lung water content was no greater in the lambs that received endotoxin. Treatment with endotoxin did not increase antioxidant enzyme levels in lung homogenate, but levels of GSH relative to GSSG were significantly elevated.(ABSTRACT TRUNCATED AT 250 WORDS)     

Scope for use of stable carbon isotopes in discerning the incorporation of forest detritus into aquatic foodwebs

Stable isotope analysis of carbon has been proposed as a means for discerning the incorporation of terrestrial forest detritus into aquatic foodwebs, and as such, has the potential to be used as a biomonitor of the aquatic effects of riparian deforestation. A synthesis of ¹³C/¹²C data from the literature indicates, however, that the scope for successful use of carbon isotope analysis in separating allochthonous and autochthonous food provenance is much more limited than was once thought. This occurs due the overlap in carbon isotope ratios between terrestrial forest detritus and those of both lotic attached algae and lentic filamentous attached algae. Only within rockyshored, oligotrophic lakes without macrophytes, and forest-fringed estuaries and lagoons, where the carbon isotope ratios for attached algae and forest detritus are significantly different, is there any likelihood of discerning the incorporation of allochthonous carbon into aquatic foodwebs using ¹³C/¹²C values alone.     

Structural analysis of an outer surface protein from the Lyme disease spirochete, Borrelia burgdorferi, using circular dichroism and fluorescence spectroscopy.

The etiological agent of Lyme disease is the tick-borne spirochete, Borrelia burgdorferi. A major antigen of B. burgdorferi is a 31 kDa lipoprotein called outer surface protein A (OspA). Recently, a truncated form of OspA (lacking 17 amino acids at the N-terminus) was cloned, expressed and purified in large quantities (Dunn, J.J., Lade, B.A. and Barbour, A.G. (1990) Protein Expression and Purification 1, 159-168). The truncated protein (OspA-257) is water-soluble, retains the ability to bind antibodies from the sera of Lyme disease patients and may prove useful in development of a vaccine against Lyme disease. We have used far UV circular dichroism (CD) and fluorescence spectroscopy to characterize the secondary structure of and to study conformational changes in OspA-257. CD spectra from 260 to 178 nm predict five classes of secondary structure: alpha-helix (11%), anti-parallel beta-sheet (32%), parallel beta-sheet (10%), beta-turns (18%) and aperiodic structures (including 'random coil') (30%). Analysis of the primary sequence of OspA yielded the most likely sites for alpha-helical regions (residues 100-107, 121-134, 253-273) and for antigenic determinants (Lys-46, Asp-82, Lys-231). CD spectra of the native protein show little change from pH 3 to 11. Thermal denaturation curves, indicate that 'salt bridges' play a role in stabilizing the native protein. Both thermal and chemical denaturations that eliminate all secondary structure as judged by CD or fluorescence are reversible. Denaturation by guanidine-HCl (gdn-HCl) appears to be a cooperative, two-state transition, as indicated by a sudden change in the CD spectrum at approximately 0.75 M gdn-HCl, and an isodichroic point at 208 nm in all CD spectra measured from 0.0-1.75 M gdn-HCl.     

Nonimmunochemical quantitation of mammalian apolipoprotein A-I in whole serum or plasma by nonreducing gel electrophoresis

A rapid and convenient method for the quantitation of mammalian apoA-I has been developed. The method involves nonreducing sodium dodecyl sulfate gel electrophoresis and Coomassie blue staining, and takes advantage of the relative abundance of apoA-I in whole serum or plasma. ApoA-I was sufficiently resolved to allow quantitation by laser densitometry or spectrophotometry. The assay was linear from 0.25 to 4.0 micrograms of apoA-I. Analytic recovery was 98%. Within-assay variability was 3.1% and between-assay variability was 7.5%. A high degree of positive correlative (r = 0.98) was observed with a human apoA-I radioimmunoassay. For several species investigated, the apoA-I values obtained correlated strongly and positively with high density lipoprotein cholesterol values. When applied to a study of nutritional perturbation in the Mongolian gerbil, the method detected sensible and significant changes in serum apoA-I that paralleled changes in HDL cholesterol.     

Contamination and health risk assessment of trace elements in soil at play centers of urban low-income settings

Abstract

Young children are considered critical receptors of potentially toxic trace elements (PTEs) by non-dietary ingestion of contaminated soil. The study assessed the potential enrichment of soil and the health risk of PTEs to 471 children less than seven years via non-dietary soil ingestion at six Early Childhood Development Centers (ECDCs) in urban low-income settings. The total concentrations of PTEs were determined by ICP-AES after wet acid digestion. The extent of soil contamination with PTEs and their source apportionment were assessed by the enrichment factor (EF). The US-EPA risk assessment model was used to determine the risk of PTE exposure by children. Multivariate statistical analyses and the EF suggested anthropogenic origin of PTEs in playgrounds and indoors, especially Cd and Pb from atmospheric deposition. Indoor floor dust at ECDCs was enriched (significant to extreme) with PTEs of anthropogenic origin imported from the outside environment. Children at the six ECDCs were not at significant non-carcinogenic risk of PTEs in soil and dust through non-dietary ingestion. The study setting is typical of urban child play centers in low-income countries which needs regular risk assessment and the enforcement of legislation in order to reduce the exposure of children to PTEs.     

Longitudinal variation in zooplankton δ13C through the Northwest Passage: inference for incorporation of sea-ice POM into pelagic foodwebs

Previous research has demonstrated a west-to-east decrease in zooplankton δ¹³C along the Alaskan coast. We extended the transect into the Canadian Arctic archipelago through the Northwest Passage in order to investigate whether this longitudinal gradient persists eastward. Contrary to the previous observations, zooplankton δ¹³C was found to be essentially invariate, despite a sampling survey length double that of the earlier studies. This resulted from zooplankton at the coldest and ice-bound stations being found to be enriched in ¹³C relative to what would be expected from the ambient water temperatures and calculated [CO₂aq]. We suggest that this was due to the assimilation of ice POM which, as has been demonstrated for other forms of benthic algae, is enriched in ¹³C due to restraints on boundary layer diffusion. The present results therefore support other traditional, non-isotope research in suggesting that ice-bound polar foodwebs may be examples of aquatic systems where there is a functional energy connectivity between the attached and openwater environments. Received: 21 January 1998 / Accepted: 27 April 1998     

Heligmosomoides polygrus (equals nematospiroides dubius): humoral and intestinal immunologic responses to infection in mice

Measurements of serum IgG₁, IgG_2a, IgM, and IgA levels and antibody titers in these immunoglobulin classes were made at intervals after initial infection and challenge infection of mice immunized by two or three previous infections. Identical measurements were made on the content of the small intestine in mice which had been exposed to the same infection schedule. Sections of small intestine taken after initial infection and challenge infection were examined by the fluorescent antibody technique for changes in populations of immunoglobulin-containing cells and by routine histologic procedures for histopathologic changes.In serum, only IgG₁ was consistently increased after initial infection, and antibody in IgG₁ was detected within the first 2 wk of infection. In immunized animals, only IgG₁ and antibody of this class always responded to challenge infection, although antibody in other immunoglobulin classes was detected.IgA concentration of the intestinal content did not differ significantly after initial infection or challenge infection of immunized mice. Immunized mice had about twice the IgG₁ concentration in intestinal content as singly infected animals. No intestinal antibody was detected after initial infection; only IgG₁ antibody was detected in the intestinal content of immunized and challenged mice.Cell infiltrates in the intestinal mucosa and submucosa of immunized animals contained numerous IgG₁-containing cells. Mast cells and globular leukocytes were observed in the intestine of immunized animals.     

Cortisol and cortisone levels in umbilical cord plasma and maternal plasma of normal pregnancies

A method for assaying cortisol and cortisone using chromatography on either paper or Sephadex LH-20 columns for isolation, followed by competitive protein binding, has been applied to umbilical cord and maternal plasma samples. In mixed cord plasma the mean cortisol concentration was 6.0 ± 0.8 μg/100 ml (n = 9) and the mean cortisone concentration was 13.5 ± 2.9 μg/100 ml (n = 9). In cord arterial plasma the mean cortisol concentration was 6.3 ± 2.9 μg/100 ml (n = 6) and the mean cortisone level was 10.1 ± 2.5 μg/100 ml (n = 6). For cord venous plasma, the mean level of cortisol was 5.6 ± 1.5 μg/100 ml (n = 6) and of cortisone was 13.5 ± 2.4 μg/100 ml (n = 6). Maternal plasma gave a mean value of cortisol of 42.3 ± 4.5 μg/100 ml (n = 6) and of cortisone of 6.2 ± 0.9 μg/100 ml. The results of this study suggest that the fetus at term-gestation produces cortisol. The significance of this production compared with placental transfer of maternal cortisol into the fetal circulation however is uncertain.