Insolubility of Cr2O3 in Bioaccessibility Tests Points to Requirement for a New Human Oral Reference Dose for Trivalent Chromium

Bioaccessibility measurements have the potential to improve the accuracy of risk assessments and reduce the potential costs of remediation when they reveal that the solubility of chemicals in a matrix (e.g., soil) differs markedly from that in the critical toxicity study (i.e., the key study from which a toxicological or toxicity reference value is derived). We aimed to apply this approach to a brownfield site contaminated with chromium, and found that the speciation was Cr^III, using a combination of alkaline digestion/diphenylcarbazide complexation and X-ray absorption near edge structure analysis. The bioaccessibility of Cr₂O₃, the compound on which a reference dose for Cr^III is based, was substantially lower (<0.1%) than that of the Cr^III in the soils, which was a maximum of 9%, giving relative bioaccessibility values of 13,000% in soil. This shows that the reference dose is based on essentially an insoluble compound, and thus we suggest that other compounds be considered for toxicity testing and derivation of reference dose. Two possibilities are CrCl₃·6H₂O and KCr(SO₄)₂·12H₂O, which have been used for derivation of ecological toxicity reference values and are soluble at a range of dosing levels in our bioaccessibility tests.     

A Non-Synonymous HMGA2 Variant Decreases Height in Shetland Ponies and Other Small Horses

The identification of quantitative trait loci (QTL) such as height and their underlying causative variants is still challenging and often requires large sample sizes. In humans hundreds of loci with small effects control the heritable portion of height variability. In domestic animals, typically only a few loci with comparatively large effects explain a major fraction of the heritability. We investigated height at withers in Shetland ponies and mapped a QTL to ECA 6 by genome-wide association (GWAS) using a small cohort of only 48 animals and the Illumina equine SNP70 BeadChip. Fine-mapping revealed a shared haplotype block of 793 kb in small Shetland ponies. The HMGA2 gene, known to be associated with height in horses and many other species, was located in the associated haplotype. After closing a gap in the equine reference genome we identified a non-synonymous variant in the first exon of HMGA2 in small Shetland ponies. The variant was predicted to affect the functionally important first AT-hook DNA binding domain of the HMGA2 protein (c.83G>A; p.G28E). We assessed the functional impact and found impaired DNA binding of a peptide with the mutant sequence in an electrophoretic mobility shift assay. This suggests that the HMGA2 variant also affects DNA binding in vivo and thus leads to reduced growth and a smaller stature in Shetland ponies. The identified HMGA2 variant also segregates in several other pony breeds but was not found in regular-sized horse breeds. We therefore conclude that we identified a quantitative trait nucleotide for height in horses.     

d-Aspartate in Human Brain

The presence of the biologically uncommon D-aspartic acid (D-aspartate) in human brain white matter has been previously reported. The earlier study has now been expanded to include D/L-aspartate ratios from 67 normal brains. The data show that the D-aspartate content increases rapidly from 1 year to approximately 35 years of age, levels off in middle age, and then appears to decrease somewhat. The D-aspartate content in gray matter remains at a consistently low level (half of that found in white matter) throughout the human life span. Within the limitations of current analytical methods, there was no detectable difference in D/L-aspartate ratios in white and gray matter of brains with Alzheimer's disease and several other pathologies when compared with brains of normal subjects. However, the presence of a significant D-aspartate level in white matter during the adult life span may lead to changes in protein configuration related to dysfunctions associated with the aging brain.     

Determination of the enantiomeric composition of (R*,S*)-1-ethylpropyl 2-methyl-3-hydroxypentanoate, the male-produced aggregation pheromone of Sitophilus granarius

The enantiomeric composition of sitophilate, the granary weevil [Sitophilus granarius (L.)] male-produced aggregation pheromone [(R^*,S^*)-1-ethylpropyl 2-methyl-3-hydroxypentanoate)], was determined by three methods: (1) bioassaying the synthetic (2S,3R) and (2R,3S) enantiomers of the active (R^*,S^*) diastereomer; (2) ¹H NMR spectroscopy of Mosher ester derivatives of the natural pheromone and synthetic (2S,3R)-and (2R,3S)-sitophilate; and (3) capillary GLC comparisons of the retention times of derivatized natural pheromone and the two synthetic enantiomers. The combined methods confirmed the (2S,3R) enantiomer as the active form of sitophilate. Male granary weevils were shown to produce >96% (2S,3R)-sitophilate. No significant attraction of S. granarius by the (2R,3S) enantiomer was observed. Rice and maize weevils [S. oryzae (L.) and S. zeamais Motschulsky] were not attracted by (2S,3R)-sitophilate. S. granarius L. est un déprédateur important des grains stockés. Le (R^*,S^*)-1-éthylpropyl 2-méthyl-3-hydroxypentanoate a été identifié en 1987 comme le principal composé du sitophilate, la phéromone mâle d'agrégation de S. granarius. La composition énantiométrique du sitophilate a été déterminée par 3 méthodes:

Calcium-dependent protease of the cyanobacterium Anabaena: molecular cloning and expression of the gene in Escherichia coli, sequencing and site-directed mutagenesis

Summary It has been suggested that a calcium-dependent intracellular protease of the cyanobacterium, Anabaena sp., participates in the differentiation of heterocysts, cells that are specialized for fixation of N₂. Clones of the structural gene (designated prcA) for this protease from Anabaena variabilis strain ATCC 29413 and Anabaena sp. strain PCC 7120 were identified via their expression in Escherichia coli. The prcA gene from A. variabilis was sequenced. The genes of both strains, mutated by insertion of a drug resistance cassette, were returned to these same strains of Anabaena on suicide plasmids. The method of sacB-mediated positive selection for double recombinants was used to achieve replacement of the wild-type prcA genes by the mutated forms. The resulting mutants, which lacked Ca²⁺-dependent protease activity, were not impaired in heterocyst formation and grew on N₂ as sole nitrogen source.     

EFFECT OF TREE CANOPY REMOVAL BY GYPSY MOTH LARVAE ON THE MACROALGAE OF A RHODE ISLAND HEADWATER STREAM1

A spring-fed, headwater stream in central Rhode Island was examined during the period from June to October, 1979 to 1982. In the first two summers, a dense riparian canopy reduced the light penetration at the stream surface to a range of 5 to 18% of incident radiation. The lotic macroalgal community during this period was limited to 1 to 4 species covering < 1 to 35% of the stream bottom. However, in June and July, 1981, the surrounding leaf canopy was removed by a massive gypsy moth larval outbreak. Light penetration to the stream during this summer increased to 73% by early July, thereby resulting in a rise in water temperatures by 3.7°C. Even though there was a partial regrowth of leaves in late July and August of 1981, macroalgal cover values continued to rise to an early August peak of 80%. During the third summer, 88% of the macroalgal abundance could be attributed to illumination and water temperature. The filamentous diatom Funotia pectinalis ( O.F. Müll.) Rabh. was the predominant species in the midsummer of all four years, accounting for at least 60% of the total cover. In 1981. an important taxon was the desmid Hyalotheca dissiliens (S. Smith) Bréb., a species which was not seen in other years. A less severe gypsy moth defoliation occurred in 1982 but did not produce significant differences in light, temperature or macroalgal cover from 1979 and 1980. The results indicate that light and temperature can be limiting during the summer in spring-fed, headwater streams and that seed populations of some species are present in undetect-able levels during these periods of suboptimal growth conditions. In addition, it appears that stream macroalgal communities can be quite resilient, recovering rapidly following a major perturbation .     

Characteristics of softwater streams in Rhode Island II. Composition and seasonal dynamics of macroalgal communities

Forty stream segments in Rhode Island, U.S.A., were examined seasonally from June 1979 to March 1982. Thirty-nine species of macroalgae were collected, respresenting 25 genera. The composition of the lotic flora was 54% green algae, 31% red algae, 5% blue-green algae, 5% xanthophytes, 3% chrysophytes and 3% diatoms. The majority of these taxa (85%) were filamentous. From a biweekly examination of five stream segments, macroalgal communities could be grouped according to light regime. Species in unshaded streams exhibited little seasonality, whereas in streams shaded by one or more layers of riparian canopy, maxima in species numbers and abundance occurred during colder seasons. The most widespread and abundant species were the blue-green alga Phormidium retzii, the green alga Draparnaldia acuta, and the diatom Eunotia pectinalis. P. retzii and E. pectinalis were aseasonal annuals, while D. acuta was primarily a winter-spring form. It appears that pH is a major factor affecting broad geographic distribution patterns of stream macroalgae, whereas the light regime established by overhanging canopy is an important factor which influences localized abundance and seasonality of lotic macroalgal communities. Niche pre-emption appears to be a common mode of resource space division among stream macroalgae in Rhode Island. E. pectinalis is the strongly developed dominant in this drainage system.     

Aerobic spore-forming bacteria as detrimental infectants in plant tissue cultures

Summary Aerobic spore-forming bacteria were isolated from plant tissue cultures from a commercial plant cultivation station. Bacillus circulans was found to be a detrimental infectant as a serious consequence of the heat-resistance of the endospores of these bacteria. They were extremely motile, utilized several growth-promoting factors, and could be eliminated by early microscopical identification, killing by heat treatment, or by using antibiotics or disinfecants.Dedicated to Professor Dr. Gustav Kortüm on the occasion of his 80th birthday     

The effects of sodium and magnesium-ion interactions on chromatin structure and solubility

The effects of sodium and magnesium-ion interactions on chromatin structure and solubility were examined in isolated mouse liver nuclei. To facilitate this study, a simple assay of chromatin structure was developed, based on the absorbances at 260 nm (A260) and 320 nm (A320) of nuclei in test solutions. By subtracting the A320 from the A260, a single "spectral index" was obtained which served as a useful, but not absolute, indicator of chromatin structure. Electron microscopy verified the validity of this approach. The results indicate that either 200 mM NaCl or 0.5 mM MgCl2 were capable of preserving the native 20 to 30 nm chromatin fiber structure. Below 200 mM NaCl, the native fiber progressively uncoiled to the 10 nm unit fiber. The presence of 0.5 mM MgCl2 inhibited this uncoiling. Only divalent cations stabilized condensed chromatin (heterochromatin) within the nucleus. Monovalent and divalent cations interacted with one another at critical concentrations and modified their individual effects on chromatin structure; e.g., 10 to 25 mM NaCl interfered with the action of 0.5 to 1.5 mM MgCl2, causing a complete loss of condensed chromatin. Maximum solubility of micrococcal nuclease-digested chromatin occurred at 10 mM NaCl, which treatment allowed the chromatin to unfold to the 10 nm fiber. However, ionic conditions that disrupted condensed chromatin but maintained the native chromatin fiber morphology still resulted in relatively high yields of soluble chromatin. Minimum solubility occurred under conditions which preserved the structure of condensed chromatin.