Partitioning of Lead in Urban Street Dust Based on the Particle Size Distribution and Chemical Environments

The objective of the present study was to investigate the distribution of lead among the physical fractions and between the various chemical forms of urban street dust. In order to achieve this aim, street dust samples were collected from three major roads with high traffic volume and one minor road with a low traffic density in urban areas of Az Zarqa City, Jordan. The dust samples (N = 6 for each site) were split into two portions. One part was employed for physical size fractionation and the other portion for chemical and physical analyses. A sequential extraction procedure was used to determine Pb associated with various chemical and physical fractions. It was found that about half of the lead was associated with the carbonate fraction and Fe-Mn oxides were ranked second, followed by the exchangeable and organic fractions. A general trend of increasing lead levels with decreasing particle size in street dust was observed. The current study showed that leaded gasoline was the major source for the elevated lead levels in street dust. Therefore, for those countries still employing leaded gasoline, such as Jordan, substantial reductions in lead contamination of street dust and roadside soils could be achieved by prohibiting the use of Pb additives. Consequently, many health benefits could be expected for the entire population and especially for children. The accuracy of lead results was checked by periodic analysis of SRM Soil 7. Observed concentrations were within 5% of certified value in analyzed SRM for lead.     

Trace Elements in Street and Household Dusts in Amman,Jordan

This research aims at quantifying the concentrations of heavy metals within the home environment in Amman, the capital city of Jordan, and to compare the total concentrations of indoor dusts to that of exterior dusts and soils. Housedust samples were collected from different zones of Amman. Street dust samples and garden soil samples were collected in the immediate vicinity within 10–50 m of each residence. The geometric mean concentrations of metals in the household dust were Pb, 169 mg/kg; Cd, 2.92 mg/kg; Zn, 1985 mg/kg; Cu, 133 mg/kg; Cr, 66 mg/kg; Co, 21 mg/kg; Ni, 31 mg/kg; Mn, 284 mg/kg; Be, 3.0 mg/kg; Ba, 43 mg/kg; B, 697 mg/kg and Al, 1441 mg/kg. Comparisons of household dust, garden soil and street dust were based on the same particle size fraction. Results showed housedust samples to contain higher concentrations for Pb, Zn, Cr, Ni, Cd, Cu and B, than either street dust or garden soil samples. However, the differences between Pb and Cr levels in the three different sample categories were insignificant. Enrichment factor calculations and the enrichment factor ratios indicated that patterns of enrichment of indoor dust differ from that of exterior dusts.     

堆肥处理对污泥腐殖物质形态及其重金属分配的影响

采用透析、凝胶色谱 (SephadexG 75 )研究了污泥堆肥前后腐殖质分子大小的变化及重金属Cu和Zn在各级组分中的分配。透析结果表明 ,污泥经过堆腐以后 ,腐殖质中小分子物质 (<10 0 0Da)组分的含量下降 6 4 % ,而相对高分子组分 (>2 5 0 0 0Da)却增加了 6 8%。凝胶色谱进一步证实 ,污泥经过 4 9d堆腐后 ,腐殖质中大于 2 0 0 0KDa的大分子组分是堆肥起始时的2 3倍。而小分子组分明显减少 ,表现在小分子组分的凝胶洗脱体积明显减少。堆肥腐熟以后 ,腐殖质吸附的Cu、Zn元素含量增加 ,其中Cu主要被吸附在大分子物质上 ,而Zn主要与小分子物质结合     

The influence of humic substances on the molecular weight distributions of phosphate and iron in epilimnetic lake waters

1. The influence of humic substances on the association of free inorganic iron and phosphate with material of larger molecular weight was investigated in epilimnetic samples from two humus‐rich lakes of contrasting ionic strength. After modification of the molecular weight distribution of the humic substances in the samples using dialysis and ultrafiltration, the molecular weight distribution of added radioisotopes ( Fe3+ and PO43?) was assessed using gel filtration chromatography.
2. The association of Fe3+ and PO43? with larger molecular weight fractions (>50 000 and 10 000–50 000 Da) was not in general related to the quantity of humic substances of the same molecular weight in the samples. However, the proportions of Fe3+ and PO43? observed in higher molecular weight peaks were strongly correlated to the quantity of humic substances of the same molecular weight in (a) the 10 000–50 000 Da peak in the sample of low ionic strength at pH 5.5 and pH 7.0, and (b) the> 50 000 Da peak in the sample of higher ionic strength at pH 4.0.
3. It was concluded that humic substances promote the association of Fe3+ and PO43? with higher molecular weight fractions primarily by acting as peptizing agents for inorganic colloids containing Fe and P. Association of Fe3+ and PO43? with material of higher molecular weight via the formation of humic substance‐Fe3+–PO43? complexes was identified but only at specific pH and within specific molecular weight ranges for each of the epilimnetic lake water samples studied.     

Overexpression of hyaluronan synthases alters vascular smooth muscle cell phenotype and promotes monocyte adhesion

Hyaluronan (HA) accumulates in vascular disease but its functional role is not fully understood. To investigate the impact of HA enriched extracellular matrices (ECM) on cell phenotype, arterial smooth muscle cells (ASMCs) were transduced with retroviral constructs (LXSN) encoding murine has-1, has-2, and has-3. HA synthesis was significantly elevated in has transduced ASMCs. Metabolically labeled HA from has-1 and has-2 transduced cells was present mostly in high molecular weight (HWA) fractions (2-10x10(6) Da), whereas HA produced by has-3 and control cells was present in lower molecular weight fractions (approximately 2x10(6) Da). Both has-1 and has-3 transduced ASMCs accumulated more pericellular HA than has-2 transduced ASMCs. All has transduced ASMCs had attenuated growth and migration rates, and a decreased detachment response. Affinity histochemistry revealed that has-1 transduced ASMCs accumulated the greatest amount of HA containing ECM than the other transduced ASMCs. This ECM was hyaluronidase sensitive and bound a significantly greater number of monocytes than the ECM generated by has-2 or has-3 transduced ASMCs. Confocal microscopy showed CD44 positive monocytes bound to hyaluronidase sensitive ECM in has-1 transduced ASMCs. These data implicate specific has isoforms in the formation of HA enriched pro-inflammatory ECMs.     

Subcellular distribution of tissue radiocopper following intravenous administration of 67Cu-labeled Cu-PTSM

The subcellular distribution of radiocopper in the brain and liver of rats has been determined following i.v. administration of Cu-PTSM, pyruvaldehyde bis(N⁴-methylthiosemicarbazonato)copper(II), labeled with copper-67. Homogenized tissue samples were separated by differential centrifugation into four subcellular fractions: (I) cell membrane + nuclei; (II) mitochondria; (III) microsomes; and (IV) cell cytosol. Upon sacrifice at 10 min post-Cu-PTSM injection, brain fractions, I, II, III and IV contain 35 ± 12, 11 ± 3, 2.8 ± 1.3 and 51 ± 7% of brain activity, respectively (n = 4). In animals sacrificed 24 h post-injection the subcellular fractions of brain tissue show little change from the radiocopper distribution seen at 10 min post-injection, although the mitochondrial fraction may contain slightly more tracer and the cytosolic fraction slightly less (I, 40 ± 10%; II, 18 ± 5%; III, 3.4 ± 1.5%; and IV, 38 ± 5%; n = 5). Subcellular fractions I, II, III and IV of liver contain 25 ± 5, 12 ± 3, 17 ± 4 and 46 ± 6% of ⁶⁷Cu tracer in animals sacrificed 10 min post-Cu-PTSM injection. An identical subcellular distribution of ⁶⁷Cu, was found in the liver following i.v. administration of ionic radiocopper (as Cu-citrate). The liver and brain cytosolic fractions at 10 min post-injection were further separated by Sephadex column chromatography. In liver cytosol, three different radiocopper components with molecular weights of about 140,000, 41,000–46,000 and 10,000–16,000 Da were found. In the brain supernatant fraction, most of the radiocopper was bound to a single low molecular weight cytosolic component (14,000–16,000 Da). These results suggest that the intracellular decomposition of tracer Cu-PTSM may result in the radiocopper entering the normal cellular pools for copper ions.     

Physico-chemical characteristics of humic substances of major soil associations of Bihar (India)

Humic acid (HA) and fulvic acid (FA) extracted from fourteen surface soil samples (0–20 cms) belonging to nine major Soil Association Groups scattered over different agro-climatic situations, were characterized by elemental and functional group analysis, E₄/E₆ ratios, coagulation behaviour and distribution of carbon in different soil humus fractions. The E₄/E₆ ratio of FA extracted from different soils was wider than that of HA. The coagulation behaviour of HA and FA fractions and also of cultivated and forest soils showed marked differences. The variations in the ratios of HAC:FAC (0.31 to 1.0) among different soils were indicative of the degree of humification under the influence of vegetation and agroclimatic conditions. The elemental composition of HA and FA, in general, indicated a higher carbon and nitrogen content and C/N ratio in the former than in the latter fraction. On the contrary, the oxygen content of FA was higher compared to that of HA. The carbon contents of HA extracted from the cultivated and forest soils of Hazaribagh were almost equal, as were also the carbon contents of HA from the cultivated and forest soils from Ranchi. Total acidity of FA of the soils selected in the present study was higher than that of HA. The functional groups such as carboxyl, phenolic hydroxyl and carbonyl were present in the two fractions in varying proportions.     

Bioavailability and speciation of arsenic in urban street dusts from Baoding city,China

Twenty-one street dust samples were collected from the urban sites in Baoding, China, to investigate the species and bioavailabilty of arsenic in them. The ecological risk and bioavailability of arsenic were evaluated using three models including Bioavailability Factor (BF), Contamination Factors (C_f)and Geoaccumulation Index model (I_geo). The species of arsenic in the dust samples were analyzed using an optimized BCR sequential extraction method. The total concentrations of As in the street dust samples ranged from 13.16 mg kg^?1 to 67.26 mg kg^?1. Geoaccumulation index (I_geo) of As ranged from 0.28 to 1.99. The speciation analysis indicated that As in the street dust samples were mainly in the residual fraction (F4), and the proportion ranged from 84.35% to 87.07%. Moreover, the ranges of the BF and C_f were 0.650–0.129 and 0.119–0.186, respectively. The results indicated that arsenic contained in the street dust samples was with low bioavailability.     

Fractionation of phosphorus and trace elements species in soybean flour and common white bean seeds by size exclusion chromatography-inductively coupled plasma mass spectrometry

Soluble species of phosphorus, sulfur, selenium and eight metals (Mn, Fe, Co, Ni, Cu, Zn, Mo and Cd) in soybean flour and common white bean seeds were investigated by size exclusion chromatography (SEC) and inductively coupled plasma mass spectrometry (ICP-MS). Samples were extracted by 0.02 mol l(-1) Tris-HCI buffer solution (pH 7.5). Fractionation of sample extracts by preparative scale SEC was accomplished using a Fractogel EMD BioSEC column (600 x 16 mm) and 0.02 mol l(-1) Tris-HCl buffer solution (pH 7.5) as mobile phase (flow rate: 2 ml min(-1)). A 2-ml sample was injected. Contents of elements in chromatographic fractions were determined by AAS, ICP-AES and ICP-MS. The elution profiles of P, Fe, Co, Ni, Cu, Zn and Mo in both samples were similar. Main species of Co, Ni, Cu, Zn and Mo were found in the low molecular weight region (2-5 kDa), whereas Fe is predominantly bound to high molecular weight compounds (180 kDa). The dominant phosphorus fraction was detected in the medium molecular weight region (10-30 kDa) and the other fraction in the low molecular weight region. Isotachophoretic analysis of chromatographic fractions revealed that the main phosphorus compound in the medium molecular weight region is phytic acid. SEC on Superdex 75 and Superdex Peptide columns (300 x 10 mm) was performed in on-line hyphenation with ICP-MS. The same mobile phase was used with a flow rate of 0.5 ml min(-1); volume of injected sample was 200 microl. Element specific chromatograms were obtained by continuous nebulization of effluent into ICP-mass spectrometer measuring intensities of 47(PO)+ and 48(SO)+ oxide ions and 55Mn, 57Fe, 59Co, 62Ni, 65Cu, 66Zn, 82Se, 95Mo and 114Cd nuclides. Chromatographic profiles of elements are generally analogous to those obtained with a Fractogel column, but better chromatographic resolution of separated species was achieved so that slight differences between samples were revealed. Estimated molecular weights of major phosphorus species in soybean flour and common white bean seed extracts are 6 and 3.6 kDa, respectively, whereas those of minor phosphorus species in both samples are 0.7 kDa. Traces of phosphorus were also detected in the high molecular weight region (130 kDa). Chromatograms of P, Ni, Cu, Zn and Mo compounds in both extracts are similar but not identical. Molecular weights of major Cu and Zn species are approximately 1 and 0.4 kDa for soybean flour and white bean seeds, respectively. In cases of Mn, Fe, Co and Se, the element profiles of soybean flour and white bean seed extracts are significantly different.     

Electron and X-ray microscopic analyses of reaggregated materials obtained after fractionation of dissolved sporopollenin

Summary Exines fromTypha angustifolia L. pollen were dissolved in hot 2-aminoethanol. The solubilisate was successively fractionated and reaggregated via a dialysis cascade with dialysis tubings of different exclusion volumina. Four fractions of reaggregated material with different molecular mass were obtained. Fraction 1 with a molecular mass above 25,000 Da, fraction 2 with a molecular mass between 10,000–25,000 Da, fraction 3 with a molecular mass between 5,000–10,000 Da, and fraction 4 of a molecular mass lower than 5,000 Da. The fractions were comparatively analysed by scanning and transmission electron microscopy and X-ray microscopy. The material of the fractions with a molecular mass above 10,000 Da exhibit high congruence to the initial material. Analysis of the reaggregated material with the lowest molecular mass revealed special distinct substructures which in form and size showed high similarities to substructures of exines described in literature. In detail, spherical substructures consisting of an electron-dense core surrounded by an electron-transparent corona and in addition elongated substructures with a distinctive surface sculpture were detected.Abbreviations SEM scanning electron microscopy - TEM transmission electron microscopy     

Interaction of platinum with metallothionein-like ligands in the rat kidney after administration of cis-dichlorodiammine platinum II

After the administration of the anticancer drug cis-dichlorodiammine platinum II (cisplatin) to male rats, the Pt in the soluble fraction of the kidney is isolated, by gel filtration, in association with a high molecular weight component and a low molecular weight fraction. At 24 h, Pt is also recovered in a metallothionein-like fraction which elutes from Sephadex G-50 with a lower apparent molecular weight than endogenous (Cu, Zn)-thionein or Cd-thionein isolated from the kidneys of Cd2+-treated rats. None of these low molecular weight metal-binding fractions binds to Octyl Sepharose CL-4B. On DE-52 ion exchange chromatography, Cd-thionein is resolved into two isometallothioneins whereas the low molecular weight Pt-binding fraction is only partially purified and contains at least six components which elute at higher gradient concentrations than metallothionein. Pretreatment with Cd2+ which stimulates the synthesis of renal and hepatic metallothionein has no effect on the uptake and subcellular distribution of Pt in the liver and kidneys. Cisplatin treatment reduces the concentration of Cu and Zn in the renal metallothionein and other soluble protein fractions in the kidney. When administered to Cd2+-pretreated rats, cisplatin promotes the loss of Zn from the soluble protein fractions but causes the redistribution of Cd from the metallothionein to the high molecular weight fraction and fails to inhibit the Cd2+-induced accumulation of Cu in the kidneys and the binding of Cu to the soluble protein fractions. It is suggested that metallothionein probably does not have a significant role in the renal metabolism of Pt following the administration of cisplatin to rats.     

Land use intensity controls actinobacterial community structure

Actinobacteria are major producers of secondary metabolites; however, it is unclear how they are distributed in the environment. DNA was extracted from forest, pasture and cultivated soils, street sediments (dust and material in place), and sediments affected by animal activity (e.g. guano, vermicompost) and characterised with two actinobacterial and a bacterial-specific 16S rDNA primer set. Amplicons (140/156) generated with the two actinobacterial-specific and amplicons (471) generated with bacterial-specific primers were analysed. Amplicons from actinobacterial-specific primer were disproportionately actinomycetal from animal-affected (soil) samples and street sediments and either verrucomicrobial (i.e. non-actinobacterial) and from a novel non-actinomycetal actinobacterial group for soils. Actinobacterial amplified ribosomal DNA restriction analysis and terminal restriction fragment length polymorphism fingerprints clustered by land use, with cultivated soils clustering apart from uncultivated soils. Actinobacterial amplicons generated with eubacterial primers were overwhelmingly from (116/126) street sediments; acidobacterial amplicons from soils (74/75). In two street samples, >90% of clones were actinomycetal. Actinomycetes are selected in terrestrial soils and sediments by cultivation, urbanisation and animal activity.     

Fate and Mobility of Copper in Soil of Cocoa Plantations in Two Southwestern States of Nigeria Treated with Copper-Based Fungicides

Soils from cocoa plantations treated with Boudreaux mixture in two southwestern states of Nigeria were collected at different depths, 0–15 cm and 15–30 cm, and subjected to five-stage sequential extraction to obtain the speciation forms of copper: exchangeable, carbonate, manganese and iron oxides, organic and residual fractions. The Cu content in the extracts from the sequential extraction was read with an Atomic Absorption Spectrophotometer (AAS). The total Cu content of the soil and the physicochemical parameters of the soils were also determined. The results from the study showed that the soils had high organic matter and copper is mostly bounded to the more mobile exogenic phase much more than the stable lithogenic phase, indicating higher mobility. Within the exogenic species, carbonate fraction was the highest followed by the organic bound and the exchangeable fraction in decreasing order. Cu was not detected in the Fe/Mn bound fraction. The implication is that the fate of the administered Cu-based pesticide is more in the relatively stable carbonate bound species than the other, more mobile phase. The results showed variation in the distribution of the copper species from one depth to another. The most transported metal from the surface to the lower layer is the exchangeable fraction. The carbonate bound species is less mobile and is not readily transported into the bottom soil layer. The organic bound Cu has nearly equal distribution between the top and bottom soils and there was little or no transport of the residual metal specie from the top to the bottom. The existence of copper in the soil largely in the anthropogenic (exogenous) phase is not the most desirable for the ecosystem. This may increase the availability of Cu in the cocoa plant and bean and may lead to potential exposure risk.     

Risk Assessment of Heavy Metals in Street Dust Particles to a Stream Network

Runoff with contaminated urban soil has an environmental risk to the aquatic environment. An assessment of heavy metals in street dust particles from a small town and their risks to the township stream network were conducted at Yangtze River delta. This assessment is based on measurement of heavy metal contents in dust particles with different particle sizes, river sediments, and suspended solids of urban runoff. The ranges of heavy metal content were 0.8–4.3, 16–380, 69–240, 9.3–350, 9.6–863 and 67–1170 mg/kg dry street dust, for Cr, Cu, Ni, Pb and Zn, respectively. Approximately 63%-71% of heavy metals were associated with particles less than 250 μ m; this particle size accounted for 40% of the total mass of street dusts. Of the five land use areas, the industrial areas had the highest heavy metal level. The smaller particle size fraction has a higher heavy metal content, low density, high mobility in runoff, and thus is a higher risk to the stream network. The topographical and hydrological features of the landscape also influence the transport of the contaminated street dusts to the aquatic environment.     

High and low molecular weight fractions of humic and fulvic acids

Summary The present investigation was initiated to study molecular weight fractionation of humic compounds isolated from surface samples of an Ultisol (Red Yellow Pc lzolic soil) and a Spodosol (Podzol), using fine (mol. wt. cutoff 3500) and coarse (mol. wt. cutoff 12000) dialysis membranes and sephadex gel filtration. Characterization of the humic fractions was conducted by elemental analysis and infrared spectroscopy. The results confirmed that fulvic acid (FA) was higher in ash and elemental content than humic acid (HA). With careful purification the amount of ash was found to be reduced to a minimum, but not to zero. Sephadex gel filtration revealed that fine HA (obtained with fine membranes) was composed of smaller amounts of HA-I (high mol. wt.) and large amounts of HA-II (low mol. wt. fraction). Coarse HA (obtained with coarse membranes) had almost equal amounts of HA-I and HA-II. Fine or coarse FA yielded only low molecular weight components after elution through sephadex. Infrared spectra of the humic fractions were indicative for the presence of phosphoglyceric acid as a possible constituent of the low molecular weight fraction of humic compounds.Contribution of the Univ. of Georgia, Agric. Expt. Sta., College Sta., Athens, Ga. Permission for the publication herein of Sadtler Standard spectrum has been granted, and all rights are reserved by Sadtler Res., Inc.Contribution of the Univ. of Georgia, Agric. Expt. Sta., College Sta., Athens, Ga. Permission for the publication herein of Sadtler Standard spectrum has been granted, and all rights are reserved by Sadtler Res., Inc.     

Hyaluronan accumulation is elevated in cultures of low density lipoprotein receptor-deficient cells and is altered by manipulation of cell cholesterol content

The extracellular matrix molecule hyaluronan (HA) accumulates in human atherosclerotic lesions. Yet the reasons for this accumulation have not been adequately addressed. Because abnormalities in lipid metabolism promote atherosclerosis, we have asked whether disrupted cholesterol homeostasis alters HA accumulation in low density lipoprotein receptor-deficient cell cultures. Cultured aortic smooth muscle cells (ASMC) from Watanabe heritable hyperlipidemic (WHHL) rabbits and skin fibroblasts from homozygous patients with familial hypercholesterolemia accumulated 2-4-fold more HA than corresponding cells from age- and sex-matched normolipidemic rabbits and individuals. This occurred in both cell-associated and secreted HA fractions and was independent of cell density or medium serum concentration. WHHL ASMC cultures synthesized twice the proportion of high molecular mass HA (>2x10(6) Da) as normal rabbit ASMC but showed a lower capacity to degrade exogenous [3H]HA. Most importantly, cholesterol depletion or blocking cholesterol synthesis markedly reduced HA accumulation in WHHL ASMC cultures, whereas cholesterol replenishment or stimulation of cholesterol synthesis restored elevated HA levels. We conclude the following: 1) maintaining normal HA levels in cell cultures requires normal cell cholesterol homeostasis; 2) HA degradation may contribute to but is not the predominant mechanism to increase high molecular mass HA accumulation in low density lipoprotein receptor-deficient WHHL ASMC cultures; and 3) elevated accumulation of HA depends on cellular or membrane cholesterol content and, potentially, intact cholesterol-rich microdomains.     

Thermal/Optical Methods for Elemental Carbon Quantification in Soils and Urban Dusts: Equivalence of Different Analysis Protocols

Quantifying elemental carbon (EC) content in geological samples is challenging due to interferences of crustal, salt, and organic material. Thermal/optical analysis, combined with acid pretreatment, represents a feasible approach. However, the consistency of various thermal/optical analysis protocols for this type of samples has never been examined. In this study, urban street dust and soil samples from Baoji, China were pretreated with acids and analyzed with four thermal/optical protocols to investigate how analytical conditions and optical correction affect EC measurement. The EC values measured with reflectance correction (ECR) were found always higher and less sensitive to temperature program than the EC values measured with transmittance correction (ECT). A high-temperature method with extended heating times (STN120) showed the highest ECT/ECR ratio (0.86) while a low-temperature protocol (IMPROVE-550), with heating time adjusted for sample loading, showed the lowest (0.53). STN ECT was higher than IMPROVE ECT, in contrast to results from aerosol samples. A higher peak inert-mode temperature and extended heating times can elevate ECT/ECR ratios for pretreated geological samples by promoting pyrolyzed organic carbon (PyOC) removal over EC under trace levels of oxygen. Considering that PyOC within filter increases ECR while decreases ECT from the actual EC levels, simultaneous ECR and ECT measurements would constrain the range of EC loading and provide information on method performance. Further testing with standard reference materials of common environmental matrices supports the findings. Char and soot fractions of EC can be further separated using the IMPROVE protocol. The char/soot ratio was lower in street dusts (2.2 on average) than in soils (5.2 on average), most likely reflecting motor vehicle emissions. The soot concentrations agreed with EC from CTO-375, a pure thermal method.     

Lead Distribution in Plant Residues Amended Calcareous Soils as a Function of Incubation Time

We investigated the effect on lead (Pb) fractionation of the addition of plant residues and incubation time in three metal-amended calcareous soils of Iran. Four amended treatments with plant residues (helianthus, potato, rape, and wheat) were established at the rate of 2% and incubated for four weeks at 25°C and constant moisture. Lead was added to amended-soils at the rate of 200 mg kg^?1 as chloride. A control without amendments but with Pb (control) was also set up. The samples were incubated for 3, 72, 168, 336, 672, 1008, and 1344 h at 25°C and constant moisture. After incubation, the soils were sequentially fractionated into water-soluble plus exchangeable (EXCH), organically bound (OM), inorganically bound (CARB), and residual (RES) forms. There were changes in the proportional distribution of Pb in all three studied soils during 1344 h of incubation with spiked Pb. In general, the proportions of Pb associated with the most weakly bound fraction (EXCH) tended to decrease, with corresponding increases in the other three more strongly binding fractions during the incubation. Application of plant residues increased Pb significantly in EXCH fractions in studied soils. Among the plant residues, application of potato residue gave the highest increase in EXCH fractions in two soils, which may be related to its lower C/N ratio. The higher proportions of EXCH fraction of spiked and amended soils in these calcareous soils indicate the higher potential of downward leaching and runoff transport, especially at the early stage of pollution.     

Molecular heterogeneity of the SHAP-hyaluronan complex. Isolation and characterization of the complex in synovial fluid from patients with rheumatoid arthritis

We previously found that a covalent complex of SHAPs (serum-derived hyaluronan-associated proteins), the heavy chains of inter-alpha-trypsin inhibitor family molecules, with hyaluronan (HA) is accumulated in synovial fluid of patients with rheumatoid arthritis, and the complex is circulated in patient plasma at high concentrations. How the SHAP-HA complex participates in this disease is unknown. To address this question, it is essential to clarify the structural features of this macromolecule. The SHAP-HA complex purified from synovial fluid of the patients by three sequential CsCl isopycnic centrifugations was heterogeneous in density, and the fractions with different densities had distinct SHAP-to-HA ratios. Agarose gel electrophoresis and column chromatography revealed that there was no apparent difference in the size distribution of HA to which SHAPs were bound between the fractions with different densities. The SHAP-HA complex in the higher density fraction had fewer SHAP molecules per HA chain. Therefore, the difference between the fractions with different densities was due to a heterogeneous population of the SHAP-HA complex, namely the different number of SHAP molecules bound to an HA chain. Based on the SHAP and HA contents of the purified preparations, we estimated that an HA chain with a molecular weight of 2 x 106 has as many as five covalently bound SHAPs, which could give a proteinaceous multivalency to HA. Furthermore, we also found that the SHAP-HA complex tends to form aggregates, judging from the migration and elution profiles in agarose gel electrophoresis and gel filtration, respectively. The multivalent feature of the SHAP-HA complex was also confirmed by the negative staining electron micrographic images of the purified fractions. Taken together, those structural characteristics may underlie the aggregate-forming and extracellular matrix-stabilizing ability of the SHAP-HA complex.     

Homotypic Versican G1 Domain Interactions Enhance Hyaluronan Incorporation into Fibrillin Microfibrils

Versican G1 domain-containing fragments (VG1Fs) have been identified in extracts from the dermis in which hyaluronan (HA)-versican-fibrillin complexes are found. However, the molecular assembly of VG1Fs in the HA-versican-microfibril macrocomplex has not yet been elucidated. Here, we clarify the role of VG1Fs in the extracellular macrocomplex, specifically in mediating the recruitment of HA to microfibrils. Sequential extraction studies suggested that the VG1Fs were not associated with dermal elements through HA binding properties alone. Overlay analyses of dermal tissue sections using the recombinant versican G1 domain, rVN, showed that rVN deposited onto the elastic fiber network. In solid-phase binding assays, rVN bound to isolated nondegraded microfibrils. rVN specifically bound to authentic versican core protein produced by dermal fibroblasts. Furthermore, rVN bound to VG1Fs extracted from the dermis and to nondenatured versican but not to fibrillin-1. Homotypic binding of rVN was also seen. Consistent with these binding properties, macroaggregates containing VG1Fs were detected in high molecular weight fractions of sieved dermal extracts and visualized by electron microscopy, which revealed localization to microfibrils at the microscopic level. Importantly, exogenous rVN enhanced HA recruitment both to isolated microfibrils and to microfibrils in tissue sections in a dose-dependent manner. From these data, we propose that cleaved VG1Fs can be recaptured by microfibrils through VG1F homotypical interactions to enhance HA recruitment to microfibrils.