Milk transfer of inorganic mercury to suckling rats

The transport of mercury into rat milk, and uptake in the suckling offspring was studied after peroral administration of inorganic mercury to lactating control rats, and to rats fed selenite in the diet. On day 8, 9, 10, or 11 of lactation, dams were administered a single oral dose of 0.1, 0.4, 0.7, 1.3, or 5.8 mg Hg/kg bw labeled with 203mercuric acetate. There was a linear relationship between mercury concentrations in dam's plasma and milk. The level of mercury in milk was approximately 25% of the level in plasma. After 3 d, milk levels were reduced to half the levels at 24 h. In the suckling offspring, exposed to mercury via milk during 3 d, the mercury level in blood was approximately 1% of the level in maternal blood. Mercury concentration in milk was linearly correlated to the levels in kidney, liver, and brain in the suckling offspring after 3 d exposure to mercury via milk. Selenite treatment of rats, 1.3 micrograms Se/g diet for 5 mo, resulted in increased transport of mercury to milk, probably because of increased plasma levels of mercury. However, selenite treatment of the dams did not cause any increased tissue levels of mercury in the suckling offspring.     

Growth inhibitors in oat grains. I. Leakage of inhibiting factors from oat grains during imbibition and the effect of these on germination and growth

Spikes of barley ( Hordeum vulgare L.) cultivar Bomi and high-lysine mutants Riso 1508 and Riso 56 were cultured on liquid media at varying N and sucrose levels. Bomi accumulated N in response to increasing N levels in the medium and a higher level was reached than in spikes of intact plants. The distribution of N in salt-soluble, hordein, and non-protein N fractions appeared to be normal. Endosperm dry weight and starch were lower than in intact plants and declined at higher N levels. A linear relationship was observed between starch content and the concentration of sucrose in the endosperm water. Uptake of culture medium by the spikes was affected by both N and sucrose concentration. The mutants had lower dry weights and starch contents, and higher sucrose contents than Bomi. At high N levels, the mutants accumulated less hordein, and more non-protein N than Bomi.     

Interaction of the nicotinic agonist (R,S)-3-pyridyl-1-methyl-2-(3-pyridyl)-azetidine (MPA) with nicotinic acetylcholine receptor subtypes expressed in cell lines and rat cortex

The interaction of the nicotinic agonist (R,S)-3-pyridyl-1-methyl-2-(3-pyridyl)-azetidine (MPA) with different nicotinic acetylcholine receptor (nAChR) subtypes was studied in cell lines and rat cortex. MPA showed an affinity (Ki = 1.21 nM) which was higher than anatoxin-a > (−)-nicotine > (+)-[R]nornicotine > (−)-[S]nornicotine > and (+)-nicotine, but lower than cytisine (Ki = 0.46 nM) in competing for (−)-[³H]nicotine binding in M10 cells, which stably express the recombinant ₄β₂ nAChR subtype. A one-binding site model was observed in all competing experiments between (−)-[³H]nicotine binding and each of the agonists studied in M10 cells. MPA showed a 13-fold higher affinity for (−)-[³H]nicotine binding sites compared to the [³H]epibatidine binding sites in rat cortical membranes. In human neuroblastoma SH-SY5Y cells, which predominantly express the ₃ nAChR subunit mRNA, MPA displaced [³H]epibatidine binding from a single population of the binding sites with an affinity in the same nM range as that observed MPA in displacing [³H]epibatidine binding in rat cortical membranes. Chronic treatment of M10 cells with MPA significantly up-regulated the number of (−)-[³H]nicotine binding sites in a concentration dependent manner. Thus MPA appears to have higher affinity to 4-subunit containing receptor subtype than ₃-subunit containing receptor subtype of nAChRs. Furthermore MPA binds to ₄β₂ receptor subtype with higher affinity than (−)-nicotine and behaves, opposite to cytisine, as a full agonist in up-regulating the number of nAChRs. © 1998 Elsevier Science Ltd. All rights reserved.     

Heterologous protein secretion in Lactococcus lactis is enhanced by the Bacillus subtilis chaperone-like protein PrsA

The Bacillus subtilis lipoprotein PrsA enhances the yield of several homologous and heterologous exported proteins in B. subtilis by being involved in the posttranslocational stage of the secretion process. In this work, we have studied the effect of B. subtilis PrsA on the secretion of Bacillus amyloliquefaciens α-amylase (AmyQ), a target protein for PrsA, and Bacillus licheniformis penicillinase (PenP) a nontarget protein for PrsA, in Lactococcus lactis. Two compatible plasmids were constructed and introduced into L. lactis strain NZ9000: one high copy plasmid, expressing the AmyQ gene (amyQ) or the PenP gene (penP), and one low copy plasmid, expressing the PrsA encoding gene (prsA). When amyQ and prsA were simultaneously expressed under the nisin-inducible promoter P _nisA , Western blotting experiments revealed a 15- to 20-fold increase in the total yield of AmyQ and a sixfold increase in secreted AmyQ activity, compared to a control strain lacking prsA. When expressed under the same induction conditions, PrsA had no effect on the secretion or total yield of PenP. These results show that the secretion yield of some heterologous proteins can be significantly increased in L. lactis when coproduced with the B. subtilis PrsA protein.     

The relative importance of tryptophan-dependent and tryptophan-independent biosynthesis of indole-3-acetic acid in tobacco during vegetative growth

A quantitative study of indole-3-acetic acid (IAA) turnover, and the contribution of tryptophan-dependent and tryptophan-independent IAA-biosynthesis pathways, was carried out using protoplast preparations and shoot apices obtained from wild-type and transgenic, IAA-overproducing tobacco (Nicotiana tabacum L.) plants, during a phase of growth when the level of endogenous IAA was stable. Based on the rate of disappearance of [¹³C₆]IAA, the half-life of the IAA pool was calculated to be 1.1 h in wild-type protoplasts and 0.8 h in protoplasts from the IAA-overproducing line, corresponding to metabolic rates of 59 and 160 pg IAA (μg Chl)⁻¹ h⁻¹, respectively. The rate of conversion of tryptophan to IAA was 15 pg IAA (μg Chl)⁻¹ h⁻¹ in wild-type protoplasts and 101 pg IAA (μg Chl)⁻¹ h⁻¹ in protoplasts from IAA-overproducing plants. In both instances, IAA was metabolised more rapidly than it was synthesised from tryptophan. As the endogenous IAA pools were in a steady state, these findings indicate that IAA biosynthesis via the tryptophan-independent pathway was 44 pg IAA (μg Chl)⁻¹ h⁻¹ and 59 pg IAA (μg Chl)⁻¹ h⁻¹, respectively, in the wild-type and transformed protoplast preparations. In a parallel study with apical shoot tissue, the presumed site of IAA biosynthesis, the rate of tryptophan-dependent IAA biosynthesis exceeded the rate of metabolism of [¹³C₆]IAA despite the steady state of the endogenous IAA pool. The most likely explanation for this anomaly is that, unlike the protoplast system, injection of substrates into the apical tissues did not result in uniform distribution of label, and that at least some of the [²H₅]tryptophan was metabolised in compartments not normally active in IAA biosynthesis. This demonstrates the importance of using experimental systems where labelling of the precursor pool can be strictly controlled. Received: 18 January 2000 / Accepted 24 February 2000     

The influence of ectomycorrhiza on nitrogen nutrition and growth of Pinus sylvestris seedlings

Ectomycorrhizal seedlings of Scots pine ( Pinus sylvestris L. cv.), inoculated with the fungus Suillus bovinus (L. ex Fr.) O. Kuntze, and non-mycorrhizal controls were grown in growth units with a circulating culture solution. Steady-state nutrition and constant relative growth rates were achieved by means of varied relative nutrient addition rates and free access of nutrients. Typical mycorrhizas always formed within a short period of time after inoculation. The nutrition/growth relationships were in principle similar to previous studies under steady-state conditions: there were close linear relationships between relative addition rate, relative growth rate and internal nitrogen concentration, i.e. an equilibrium established between nutrients added and taken up. This occurred when infected and uninfected seedlings were grown separately. When grown together in the same growth unit, there are indications that the fungus influenced the exudation pattern of the uninfected seedlings. More carbon was thus provided to the unspecified microflora in the cultivation system, and it was able to grow and withhold nitrogen from the seedlings. The mycorrhizal infection did not increase the specific uptake capacity of the roots, and the fungus constituted a sink for carbon. However, the nitrogen productivity (growth rate per unit of nitrogen per unit of time) was similar for mycorrhizal and non-mycorrhizal seedlings, so that there might be mechanisms which compensate for the carbon cost.     

Mercury and selenium in whole blood and serum in relation to fish consumption and amalgam fillings in adolescents

Mercury and selenium in whole blood and serum of 245 17-year old Swedish adolescents were analysed. The relationships between these elements' concentrations and the consumption of fish as well as the number of dental amalgam fillings were studied. The geometric means (GM) of the mercury concentrations were 1.1 microg/L in blood and 0.43 microg/L in serum. The mean selenium concentration in blood was 110 microg/L and the GM of the serum selenium concentration 110 microg/L. Fish species with dietary restrictions due to elevated mercury Levels (i.e. pike, perch, pikeperch, burbot, eel and halibut) were consumed on average 0.7 times/month and fish species without such restrictions 4.1 times/month. Despite this comparatively low fish consumption, the adolescents' blood mercury concentrations were positively correlated with fish consumption. Of the adolescents, 39% had amalgam fillings (mean 2 +/- 1.5). Serum mercury was influenced by the number of amalgam fillings, by fish consumption, blood and serum levels of selenium and the residential area. Blood and serum selenium concentrations were not influenced by fish consumption, but were positively associated with the serum mercury concentration.     

Receptor binding domain of Escherichia coli F18 fimbrial adhesin FedF can be both efficiently secreted and surface displayed in a functional form in Lactococcus lactis

Adherence of F18 fimbrial Escherichia coli to porcine intestinal epithelial cells is mediated by the adhesin (FedF) of F18 fimbriae. In a previous study, we demonstrated the specificity of the amino acid residues between 60 and 109 as the receptor binding domain of FedF. In this study, different expression, secretion, and anchoring systems for the receptor binding domain of the FedF adhesin in Lactococcus lactis were evaluated. Two partially overlapping receptor binding domains (42 and 62 amino acid residues) were expressed as fusions with L. lactis subsp. cremoris protein PrtP for evaluation of secretion efficiency. To evaluate the cell surface display of these FedF-PrtP fusions, they were further combined with different lengths of PrtP spacers fused with either the L. lactis AcmA anchor or the PrtP cell wall binding domain. An HtrA-defective L. lactis NZ9000 mutant was constructed to determine its effect on the level of secreted or anchored fusion proteins. Recombinant L. lactis clones secreting the receptor binding domain of F18 fimbriae as a fusion with the H domains of L. lactis protein PrtP were first constructed by using two different signal peptides. FedF-PrtP fusions, directed by the signal sequence of L. brevis SlpA, were throughout found to be secreted at significantly higher quantities than corresponding fusions with the signal peptide of L. lactis Usp45. In the surface display systems tested, the L. lactis AcmA anchor performed significantly better, particularly in the L. lactis NZ9000DeltahtrA strain, compared to the L. lactis PrtP anchor region. Of the cell surface display constructs with the AcmA anchor, only those with the longest PrtP spacer regions resulted in efficient binding of recombinant L. lactis cells to porcine intestinal epithelial cells. These results confirmed that it is possible to efficiently produce the receptor binding domain of the F18 adhesin in a functionally active form in L. lactis.