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PubMed1996

Multiple enhancer elements mediate induction of c-fos in vascular smooth muscle cells.

Previous work from this and other laboratories has demonstrated that the vasoconstrictor peptide angiotensin II results in hypertrophy of rat aortic smooth muscle cells that is associated with an increase in transcription of the early growth response gene c-fos. To explore the molecular mechanism responsible for c-fos induction in rat aortic smooth muscle cells, we used a series of reporter constructs linked to the chloramphenicol acetyl transferase gene in transient transfection experiments in rat aortic smooth muscle cells. Constructs containing both the serum response element and cAMP response element exhibited a 20-fold increase in chloramphenicol acetyl transferase activity in response to either serum or angiotensin II, whereas no increase was seen in vehicle-treated cells. Mutations in either the serum response element or cAMP response element alone, which have been demonstrated to inactivate these elements in other cell types, had no effect on chloramphenicol acetyl transferase inducibility. In contrast, if both elements were mutated, inducibility was almost abolished. Electrophoretic mobility shift assays with oligonucleotides corresponding to either serum response element or cAMP response element demonstrated that these oligonucleotides are capable of forming specific complexes with proteins from rat aortic smooth muscle cell nuclear extracts. One of the proteins binding to the serum response element is the previously described serum response factor, since it was supershifted by a monospecific antibody. These studies demonstrate that c-fox induction in smooth muscle occurs by a dual mechanism that can activate transcription via the serum response element or cAMP response element. These elements appear to act equally and independently, involving a distinct set of transacting factors.

Angiotensin II↗

PubMed1993

Optical tearing in spatiotemporal boundary formation: when do local element motions produce boundaries, form, and global motion?

Perception of continuous boundaries, shape, and global motion can be produced by transformations in local elements separated in both space and time, a process here called spatiotemporal boundary formation (SBF). Prior research has shown that a broad class of element transformations gives rise to SBF. The present work used the transformation of local element displacement to explore the initiating conditions for SBF. Three experiments assessed SBF using a 10-alternative, forced-choice, shape identification task. Experiment 1a showed that large element displacements, but not small ones, produced high accuracy in shape identification. Experiment 1b tested the detectability of the small and large element displacements in an unrelated task, indicating that the results of Experiment 1a were not due to poor detectability for small displacements. Experiment 2 found no variation in SBF performance with changes in viewing distance. Experiment 3 provided evidence that initiating SBF depends on a ratio of element displacement to element separation. These results support an interpretation of SBF as a process geared to detection of object boundaries from spatiotemporal change. Initiating SBF requires transformations in local elements that are classified as spatiotemporal discontinuities (STDs). Small element displacements in a display of a given density do not register as STDs because they are classified as local deformations in an intact, implicit surface connecting visible elements. Complementarity is suggested between element changes which preserve continuity with their neighbors (optic flow) and those comprising spatiotemporal discontinuities (optic tearing). Classification of element transformations as optic flow or tearing may determine whether they provide information about surface form (e.g., through structure-from-motion) or about object boundaries, through SBF.

Form Perception↗

PubMedDate not supplied

Major and trace elements of selected pedons in the USA.

Few studies of soil geochemistry over large geographic areas exist, especially studies encompassing data from major pedogenic horizons that evaluate both native concentrations of elements and anthropogenically contaminated soils. In this study, pedons (n = 486) were analyzed for trace (Cd, Co, Cr, Cu, Hg, Mn, Ni, Pb, Zn) and major (Al, Ca, Fe, K, Mg, Na, P, Si, Ti, Zr) elements, as well as other soil properties. The objectives were to (i) determine the concentration range of selected elements in a variety of U.S. soils with and without known anthropogenic additions, (ii) illustrate the association of elemental source and content by assessing trace elemental content for several selected pedons, and (iii) evaluate relationships among and between elements and other soil properties. Trace element concentrations in the non-anthropogenic dataset (NAD) were in the order Mn > (Zn, Cr, Ni, Cu) > (Pb, Co) > (Cd, Hg), with greatest mean total concentrations for the Andisol order. Geometric means by horizon indicate that trace elements are concentrated in surface and/or B horizons over C horizons. Median values for trace elements are significantly higher in surface horizons of the anthropogenic dataset (AD) over the NAD. Total Al, Fe, cation exchange capacity (CEC), organic C, pH, and clay exhibit significant correlations (0.56, 0.74, 0.50, 0.31, 0.16, and 0.30, respectively) with total trace element concentrations of all horizons of the NAD. Manganese shows the best inter-element correlation (0.33) with these associated total concentrations. Total Fe has one of the strongest relationships, explaining 55 and 30% of the variation in total trace element concentrations for all horizons in the NAD and AD, respectively.

Geological Phenomena↗

PubMed1991

Synergism between steroid response and promoter elements during cell-free transcription.

We have analyzed quantitatively the influence of distal promoter elements on steroid-responsive gene expression in vitro. Functional synergism between enhancer and distal promoter elements was examined using two model promoters, one containing a natural promoter (mouse mammary tumor virus long terminal repeat) and one constructed artificially. Human glucocorticoid receptor (GR) expressed in baculovirus induces transcription from a mouse mammary tumor virus long terminal repeat-containing DNA template. Transcription is diminished by oligonucleotides containing a nuclear factor 1 (NF-1)-binding site or a glucocorticoid/progesterone response element. Quantitative analysis indicates that NF-1 and GR act synergistically during transcriptional activation. In contrast, efficient activation by GR or purified chick progesterone receptor of a glucocorticoid/progesterone response element-linked ovalbumin promoter does not require interaction with the chicken ovalbumin upstream promoter (COUP) element in the distal promoter. Lack of synergism is not related to enhancer strength, since the glucocorticoid/progesterone response elements can be moved further from the promoter or reduced to a single copy response element without increasing the dependence upon COUP. Strong synergism is restored following substitution of an NF-1 distal promoter element for the COUP element in this construct. Our results suggest that synergism between steroid response and distal promoter elements is dependent upon the identity of the promoter element rather than upon the inherent strength of the enhancer element.

Animals↗

PubMed1990

Islet cell-specific regulatory domain in the gastrin promoter contains adjacent positive and negative DNA elements.

The gastrin gene is expressed in fetal pancreatic islet cells, but after birth expression is selectively repressed as the islets terminally differentiate. DNA transfection studies identified a cis regulatory domain between -108 and -76 in the gastrin promoter which controls gastrin transcription in islet cells. This cis regulatory domain comprises adjacent positive and negative elements. The negative element (-108 to -82) contains the sequence ATTCCTCT, which is also found in the negative element of the beta-interferon promoter. Gel retardation assays and DNase footprinting studies demonstrated that specific islet nuclear protein(s) bind to the gastrin negative element. In vivo competition studies demonstrated that the trans-acting factors which bind to this element specifically repress gastrin promoter activity in islet cells. Immediately downstream of the negative element lies a positive element (-82 CATATGG -76), which activates gastrin transcription in islet cells. The sequence of the positive element resembles the islet-specific enhancer elements of the insulin gene (CATCTGG/C). Gel mobility shift assays and in vivo competition studies indicate that this positive element activates the gastrin promoter by binding to the same islet cell transcription factor which binds enhancer elements in the rat insulin gene. The tandem organization of the negative and positive elements suggests that this regulatory domain may act as a switch controlling the transient transcription of the gastrin gene during fetal islet development.

Animals↗

PubMed1989

[Application of the finite element method to craniofacial growth analysis. 4. Three-dimensional application of tensor analysis].

The purpose of this study is the 3-dimensional application of finite element method to analyze the craniofacial growth. In our previous studies, 2-dimensional finite element method was applied to analyze and to predict the craniofacial growth. In case of 2-dimensional application, the normal and shear strains were given by the displacements u, v that were linear functions of coordinates x, y. In the 3-dimensional case, six strain components are required to obtain the extension ratio and the directions of three-principal axes. In this study, for the 3-dimensional application, four-noded, pyramidal elements were used and twelve 3-dimensional elements were constructed. Materials were longitudinal frontal and lateral cephalometric X-rays of 4 males and 4 females from 7 to 10 years with normal occlusions. 3-dimensional coordinates of each nodal points were calculated by transforming 2-dimensional coordinates on the frontal and lateral cephalometric X-ray films. Coordinates of the nodal points at 7 years were standards against that results of the finite element method from 8 to 10 years were obtained. They were subjects of the analysis that elements equivalent to the cranial base, the maxillary portion, the maxillary alveolar portion and the posterior pharyngeal portion. Summarized results were as follows. 1. It was observed that the similar figured extension of the anterior cranial base element. 2. The posterior cranial base element extended to right, anterior and upwards direction. 3. On the maxillaly elements, especially, lateral extensions appeared. 4. The element of the posterior pharyngeal portion extended to left and downwards direction. 5. The elements of the maxillary alveolar portion extended to downwards directions. From the above, the directions of transformation of the elements that are selected with voluntary nodal points can be observed. Therefore, 3-dimensional tensor analysis is a method of great significance for obtaining new findings of the craniofacial growth.

Cephalometry↗

PubMed2004

Evolution and distribution of RNA polymerase II regulatory sites from RNA polymerase III dependant mobile Alu elements.

BACKGROUND: The primate-specific Alu elements, which originated 65 million years ago, exist in over a million copies in the human genome. These elements have been involved in genome shuffling and various diseases not only through retrotransposition but also through large scale Alu-Alu mediated recombination. Only a few subfamilies of Alus are currently retropositionally active and show insertion/deletion polymorphisms with associated phenotypes. Retroposition occurs by means of RNA intermediates synthesised by a RNA polymerase III promoter residing in the A-Box and B-Box in these elements. Alus have also been shown to harbour a number of transcription factor binding sites, as well as hormone responsive elements. The distribution of Alus has been shown to be non-random in the human genome and these elements are increasingly being implicated in diverse functions such as transcription, translation, response to stress, nucleosome positioning and imprinting. RESULTS: We conducted a retrospective analysis of putative functional sites, such as the RNA pol III promoter elements, pol II regulatory elements like hormone responsive elements and ligand-activated receptor binding sites, in Alus of various evolutionary ages. We observe a progressive loss of the RNA pol III transcriptional potential with concomitant accumulation of RNA pol II regulatory sites. We also observe a significant over-representation of Alus harboring these sites in promoter regions of signaling and metabolism genes of chromosome 22, when compared to genes of information pathway components, structural and transport proteins. This difference is not so significant between functional categories in the intronic regions of the same genes. CONCLUSIONS: Our study clearly suggests that Alu elements, through retrotransposition, could distribute functional and regulatable promoter elements, which in the course of subsequent selection might be stabilized in the genome. Exaptation of regulatory elements in the preexisting genes through Alus could thus have contributed to evolution of novel regulatory networks in the primate genomes. With such a wide spectrum of regulatory sites present in Alus, it also becomes imperative to screen for variations in these sites in candidate genes, which are otherwise repeat-masked in studies pertaining to identification of predisposition markers.

Alu Elements↗

PubMed2002

Mobile genetic element activation and genotoxic cancer therapy: potential clinical implications.

Approximately one-quarter of the human genome is composed of short and long interspersed elements (SINEs and LINEs, respectively). These elements have spread throughout the genome by a process termed retrotransposition, consisting of transcription of an element into RNA, reverse transcription into cDNA, and reinsertion of the copied element into a new genomic location. Recombination events involving these elements, including novel insertions into active genes, have been associated with a number of human diseases. Despite the fact that these elements have replicated to hundreds of thousands of copies in the genome, under most conditions they remain transcriptionally silent, and therefore are not actively replicating. The signals controlling retrotransposable element activation in the genome have not been defined. Our laboratory recently found that exposure of cells to a variety of DNA-damaging agents, including several common chemotherapeutic drugs and gamma-radiation, is associated with dramatic induction of SINE transcription, and of a concomitant endogenous reverse transcriptase activity. As SINEs do not encode for reverse transcriptase, the latter finding suggests a more global activation of retrotransposable elements in response to DNA damage. Together these observations suggest that genotoxic exposure may lead to genomic mutation not only through direct DNA damage, but also through indirect activation of potentially mutagenic mobile elements in the genome. The nonrandom distribution of retrotransposable elements in the human genome may contribute to the pattern of characteristic translocation events associated with secondary malignancies in patients exposed to genotoxic agents. Here we describe these and other mechanisms by which retrotransposable elements can contribute to disease, and present an overview of what is known about this large, and largely unexplored, segment of the genome. Understanding the cellular responses to genotoxic stress may permit the development of a means of predicting the risks and preventing the development of secondary malignancy following cancer therapy.

Antineoplastic Agents↗

PubMed1987

Discrete elements within the SV40 enhancer region display different cell-specific enhancer activities.

The SV40 enhancer contains three genetically defined elements, called A, B and C, that can functionally compensate for one another. By using short, synthetic DNA oligonucleotides, we show that each of these elements can act autonomously as an enhancer when present as multiple tandem copies. Analysis of a progressive series of B element oligomers shows a single element is ineffective as an enhancer and that the activity of two or more elements increases with copy number. Assay in five different cell lines of two separate enhancers containing six tandem copies of either the B or C element shows that these elements possess different cell-specific activities. Parallel oligomer enhancer constructs containing closely spaced double point mutations display no enhancer activity in any of the cell lines tested, indicating that these elements represent single units of enhancer function. These elements contain either a 'core' or 'octamer' consensus sequence but these consensus sequences alone are not sufficient for enhancer activity. The different cell-specific activities of the B and C elements are consistent with functional interactions with different trans-acting factors. We discuss how tandem duplication of such dissimilar elements, as in the wild-type SV40 72-bp repeats, can serve to expand the conditions under which an enhancer can function.

Animals↗

PubMed2003

Distribution and sequence analysis of a novel Ty3-like element in natural Saccharomyces paradoxus isolates.

Little is known about the transposable elements of species closely related to Saccharomyces cerevisiae. We present a novel transposable element in Saccharomyces paradoxus, a close congener of S. cerevisiae. Sequence analysis of this element, designated Ty3-1p, indicates that it is a homologue of the S. cerevisiae Ty3 element. Ty3-1p shares 82% nucleotide identity with an S. cerevisiae Ty3 element and appears to be structured identically to Ty3, containing two overlapping open reading frames, six retroviral-like domains, a J domain, and flanking sigma-like elements. A sigma element from Ty3-1p is 75% identical to a Ty3 sigma element. There is no evidence of horizontal transfer of Ty3 in Saccharomyces sensu stricto. We assess the distributions of Ty3p and Ty3 element insertions in natural population samples of S. paradoxus and S. cerevisiae. The S. paradoxus population sample exhibits Ty3p insertions present at a variety of sites at low frequency; this suggests that Ty3p elements are active in the sampled population. The S. cerevisiae population sample exhibits a uniform Ty3 hybridization profile in which all element insertions appear to be fixed. We comment on the possible causes of these contrasting observed distributions (GenBank Accession Nos AY198186 and AY198187).

Amino Acid Sequence↗

PubMed1998

Analysis of two transcription activation elements in the promoter of the developmentally regulated con-10 gene of Neurospora crassa.

The con-10 gene of Neurospora crassa is activated during conidiation. CRS-B (conidiation response sequence-B) and CGE (con-10 general enhancer) elements have been proposed to function as sites of transcriptional activation and as possible elements that confer developmental regulation to con-10. In a specific analysis of the roles of these elements we found that two CRS-B elements are necessary for full activation of con-10 during macroconidiation, whereas two CGE elements are functionally redundant, such that a single CGE is sufficient for maximal expression. However, CRS-B and CGE elements are not sufficient for developmental activation of a reporter gene. The CRS-B element was further dissected and one of the CRS-B elements appears to function in repression as well as activation. con-10 is also highly expressed during microconidiation, a different form of asexual sporulation, but we show here that CRS-B elements do not play a significant role in con-10 expression during microconidiation. Both CRS-B elements contribute to basal con-10 expression during mycelial growth. con-10 is also regulated by light, and CRS-B and CGE elements may play minor roles in controlling con-10 expression in response to light.

Base Sequence↗

PubMed1994

An activator element within the preprotachykinin-A promoter.

The rat Preprotachykinin-A promoter (PPT) directs high levels of expression in dorsal root ganglia (DRG) neurons in culture either endogenously or when linked to a receptor construct. It is not active in any of the established tissue culture cell lines which we have analyzed. To search for transcriptional regulators within this promoter we have started to dissect the promoter into individual elements to determine their function. A DNA element which had previously been suggested to regulate transcription from DNA sequence analysis of the rat PPT promoter occurs at position -200 relative to the major start of transcription within the PPT promoter. The equivalent element from the bovine PPT promoter had previously been proposed to be a cAMP responsive element (CRE). The sequence of this enhancer has similarities with both the AP1 and CRE DNA consensus sequences. We have demonstrated that one copy of this rat PPT element linked to a heterologous basal promoter will enhance transcription in HeLa and PC12 cell lines as well as adult rat DRG neurons grown in culture. It is also demonstrated that the rat PPT element will bind proteins in HeLa nuclear extract distinct from those binding to the well-characterized Gibbon Ape Leukemia Virus (GALV) AP1 or somatostatin CRE sites by gel retardation analysis. This PPT element, when cloned in a heterologous reporter construct, although showing properties of both AP1 and CRE elements, was functionally distinguished from both the somatostatin CRE element and the GALV AP1 enhancer when these elements were tested in the same reporter construct. This PPT element has a constitutive level of activity in adult rat DRG neurons, which is fivefold higher than that driven by the reporter construct promoter. It is also significantly different from the same reporter construct linked to the somatostatin CRE and analyzed in DRG neurons.

Animals↗

PubMed1993

Minimal enhancer elements of the leghemoglobin lba and lbc3 gene promoters from Glycine max L. have different properties.

The characteristics of the soybean leghemoglobin lba gene promoter were analyzed and important promoter elements from the lba and lbc3 promoters were compared using transgenic Lotus corniculatus plants. A 5' deletion analysis of the lba promoter delimited two cis-acting elements controlling expression: a distal positive element (-1254, -884) required for expression and a proximal element (-285, -60) essential for full-level activity. In contrast to the corresponding region of the lbc3 promoter, the lba proximal element is unable to control expression from the heterologous CaMV 35S enhancer. The upstream positive element of the lba gene contains a position- and orientation-independent enhancer between positions (-1091, -788). The sequence of this enhancer region is conserved in the lbc3 gene upstream (-1333, -1132) of the previously assigned strong positive element (SPE; -1090, -947). The present analysis revealed some of the properties of this extended lbc3 SPE element. The extended element (-1364, -947) functions in both orientations from 5' locations whereas the SPE2 subcomponent (-1364, -1154) containing the conserved sequence is only active in the correct orientation. Removal of the SPE2 by internal deletion demonstrates that the SPE2 subcomponent is indispensable for the activity of the lbc3 upstream positive element. These results indicate that the upstream positive elements of the lba and lbc3 genes possess different properties although their conserved minimal enhancer sequence has similar function. This may reflect the differential expression of the two lb genes of Glycine max L.

Base Sequence↗

PubMed1992

The pogo transposable element family of Drosophila melanogaster.

A 190 bp insertion is associated with the white-eosin mutation in Drosophila melanogaster. This insertion is a member of a family of transposable elements, pogo elements, which is of the same class as the P and hobo elements of D. melanogaster. Strains typically have many copies of a 190 bp element, 10-15 elements 1.1-1.5 kb in size and several copies of a 2.1 kb element. The smaller elements all appear to be derived from the largest by single internal deletions so that all elements share terminal sequences. They either always insert at the dinucleotide TA and have perfect 21 bp terminal inverse repeats, or have 22 bp inverse repeats and produce no duplication upon insertion. Analysis by DNA blotting of their distribution and occupancy of insertion sites in different strains suggests that they may be less mobile than P or hobo. The DNA sequence of the largest element has two long open reading frames on one strand which are joined by splicing as indicated by cDNA analysis. RNAs of this strand are made, whose sizes are similar to the major size classes of elements. A protein predicted by the DNA sequence has significant homology with a human centrosomal-associated protein, CENP-B. Homologous sequences were not detected in other Drosophila species, suggesting that this transposable element family may be restricted to D. melanogaster.

Amino Acid Sequence↗

PubMed1994

An unstable allele at the maize Opaque2 locus is caused by the insertion of a double Ac element.

An unstable allele designated o2-m55, was isolated as a derivative of the o2-m5 allele. Whereas the o2-m5 allele is caused by the insertion of one copy of the transposable element Activator (Ac) into the first exon of the maize Opaque2(O2) gene, the o2-m55 allele contains two Ac elements. The position of one copy is identical to the position of the Ac element in the o2-m5 allele. The second copy is present within the first copy, thereby interrupting its structure shortly before the first ATG of the major reading frame of Ac. Both Ac sequences have the same orientation. Excision of the internal Ac element as well as excision of the complete double Ac element was detectable. Truncated double Ac elements comprising the complete internal Ac element and either the proximal or distal fragment of the interrupted Ac element can also be excised. The Ac elements exhibit a strong negative dosage effect: kernels which display revertant sectors in a mutant background are rarely seen in plants homozygous for the o2-m55 allele. If only one dose of the o2-m55 allele is present in endosperm tissue, revertant sectors can be detected. The amount of the transcript expressed from the internal Ac element of the o2-m55 allele is less than that derived from the single Ac element present in the o2-m5 allele.

Alleles↗

PubMed2000

Ancient and recent horizontal invasions of drosophilids by P elements.

P elements of two different subfamilies designated as M- and O-type are thought to have invaded host species in the Drosophila obscura group via horizontal transmission from external sources. Sequence comparisons with P elements isolated from other species suggested that the horizontal invasion by the O-type must have been a rather recent event, whereas the M-type invasion should have occurred in the more distant past. To trace the phylogenetic history of O-type elements, additional taxa were screened for the presence of O- and M-type elements using type-specific PCR primers. The phylogeny deduced from the sequence data of a 927-bp section (14 taxa) indicate that O-type elements have undergone longer periods of regular vertical transmission in the lineages of the saltans and willistoni groups of Drosophila. However, starting from a species of the D. willistoni group they were transmitted horizontally into other lineages. First the lineage of the D. affinis subgroup was infected, and finally, in a more recent wave of horizontal spread, species of three different genera were invaded by O-type elements from the D. affinis lineage: Scaptomyza, Lordiphosa, and the sibling species D. bifasciata/D. imaii of the Drosophila obscura subgroup. The O-type elements isolated from these taxa are almost identical (sequence divergence <1%). In contrast, no such striking similarities are observed among M-type elements. Nevertheless, the sequence phylogeny of M-type elements is also not in accordance with the phylogeny of their host species, suggesting earlier horizontal transfer events. The results imply that P elements cross species barriers more frequently than previously thought but require a particular genomic environment and thus seem to be confined to a rather narrow spectrum of host species. Consequently, different P element types acquired by successive horizontal transmission events often coexist within the same genome.

Animals↗

PubMed2003

Rapid inactivation of the maize transposable element En/Spm in Medicago truncatula.

Transposable elements have been widely used as mutagens in many organisms. Among them, the maize transposable element En/Spm has been shown to transpose efficiently in several plant species including the model plant Arabidopsis, where it has been used for large-scale mutagenesis. To determine whether we could use this transposon as a mutagen in the model legume plant Medicago truncatula, we tested the activity of the autonomous element, as well as two defective elements, in this plant, and in Arabidopsis as a positive control. In agreement with previous reports, we observed efficient excision of the autonomous En/Spm element in A. thaliana. This element was also active in M. truncatula, but the transposition activity was low and was apparently restricted to the tissue culture step necessary for the production of transgenic plants. The activity of one of the defective transposable elements, dSpm, was very low in A. thaliana and even lower in M. truncatula. The use of different sources of transposases suggested that this defect in transposition was associated with the dSpm element itself. Transposition of the other defective element, I6078, was also detected in M. truncatula, but, as observed with the autonomous element, transposition events were very rare and occurred during tissue culture. These results suggest that the En/Spm element is rapidly inactivated in the regenerated plants and their progeny, and therefore is not suitable for routine insertion mutagenesis in M. truncatula.

Amino Acid Sequence↗

PubMed2004

Microbial acidification and pH effects on trace element release from sewage sludge.

Leaching of sludge-borne trace elements has been observed in experimental and field studies. The role of microbial processes in the mobilization of trace elements from wastewater sludge is poorly defined. Our objectives were to determine trace element mobilization from sludge subjected to treatments representing microbial acidification, direct chemical acidification and no acidification, and to determine the readsorption potential of mobilized elements using calcareous sand. Triplicate columns (10-cm diameter) for incubation and leaching of sludge had a top layer of digested dewatered sludge (either untreated, acidified with H2SO4, or limed with CaCO3; all mixed with glass beads to prevent ponding) and a lower glass bead support bed. Glass beads in the sludge layer, support layer or both were replaced by calcareous sand in four treatments used for testing the readsorption potential of mobilized elements. Eight sequential 8-day incubation and leaching cycles were operated, each consisting of 7.6 d of incubation at 28 degrees C followed by 8 h of leaching with synthetic acid rain applied at 0.25 cm/h. Leachates were analyzed for trace elements, nitrate and pH, and sludge layer microbial respiration was measured. The largest trace element, nitrate and S losses occurred in treatments with the greatest pH depression and greatest microbial respiration rates. Cumulative leaching losses from both microbial acidification and direct acidification treatments were > 90% of Zn and 64-80% of Cu and Ni. Preventing acidification with sludge layer lime or sand restricted leaching for all trace elements except Mo. Results suggested that the primary microbial role in the rapid leaching of trace elements was acidification, with results from direct acidification being nearly identical to microbial acidification. Microbial activity in the presence of materials that prevented acidification mobilized far lower concentrations of trace elements, with the exception of Mo. Trace elements mobilized by acidification were readsorbed by calcareous sand when present.

Calcium↗