Inhibitory Effect Of Humic Acid Research Articles

Coating TiO2 with MIL-101(Fe) to decarboxylate D-Glu and D-MeAsp of microcystin-LR from HA-rich water: Diminishing the inhibitory effect of humic acids

Humic acids (HA) released during algal bloom poses a key challenge for efficient photocatalytic degradation of Microcystin-LR (MC-LR) due to their scavenging function on active species. Herein, MIL-101(Fe)/TiO2-P25 (M/P) was constructed to diminish the inhibitory effect of HA on the photocatalytic degradation of MC-LR. Results showed that MC-LR could be efficiently removed assigning to the photogenerated holes (h+) and superoxide radical (O2•−) oxidation under visible light irradiation, where the carboxy groups of D-glutamic acid (Glu) and methyl-D-aspartic acid (MeAsp), and Adda side chain related to MC-LR biotoxicity were selectively destroyed to achieve MC-LR detoxication. More importantly, HA exerted non-inhibitory effect on MC-LR removal by M/P. The counteracting mechanisms were studied. Firstly, the location of TiO2 in the pore structure of MIL-101(Fe) could occupy the adsorption sites of HA, and decrease the HA adsorption on M/P. Furthermore, XPS analyses suggested the coordination of unsaturated Fe metal sites on the surface of M/P hybrids complexed with –OH and –COOH of HA, which could accelerate the electron transfer and reduce hole consumption. Thus, the inhibitory effect of HA was alleviated during photocatalytic MC-LR treatment. This work offers a possible way to remove MC-LR in an abundant HA matrix water system efficiently.

Citric acid treatment directly on anaerobic digestor sludge alleviates the inhibitory effect of in-situ generated humic acids by their deconstruction and redistribution

Humic acids were known to inhibit sewage sludge anaerobic digestion (AD) and many studies have been investigated to deal with the inhibitory effect of humic acids (HA) in raw sludge feeding the digestor. However, HA could also be in-situ produced during the sludge AD process with a significant amount, but the relieving of the inhibitory effect posed by the in-situ produced HA has been historically ignored. Thus, this study attempted to use citric acid (CA), an easily biodegradable polyatomic acid, to directly treat sludge in the anaerobic digestor instead of the feeding sludge, for alleviating inhibition caused by the in-situ produced HA. Results showed that methane production of AD batches with CA pretreatment at 0.06∼0.10 g/g TS on digestor sludge were effectively enhanced, with the highest production being 2.89 times of that observed in control AD without CA treatment and 2.08 times of that achieved by contrast AD with CA pretreatment on raw sludge. However, the AD batches with CA treatment at low dosages of 0.02∼0.04 g/g TS had lower methane production, during which sludge solubilization and hydrolysis were enhanced initially associated with released HA surfactant and hydrolytic enzymes; then later inhibited since higher structurally sound HA in SP easily rebind proteinaceous substrates and enzymes again. Whilst CA treatment at 0.06∼0.10 g/g TS could promote sludge solubilization and hydrolysis throughout the sludge AD process. This dosage-dependent effect of CA treatment can be caused by its discriminating impact on HA structure and distribution. At low dosages, complexes of HA-proteins (including hydrolases) can be decrosslinked and transferred to the soluble phase (SP), thus enhancing solubilization and hydrolysis initially. However, the released HA in SP could rebind more organics and hydrolases during AD, resulting in augmented inhibition. While the dosage increased, HA were drastically deconstructed with aromatic cores in SP migrating to extracellular polymeric substances, preventing binding effects on organics and hydrolases. Accordingly, a smart mutex-switch operational strategy of CA treatment was proposed. The results suggested, to alleviate the inhibitory effect of HA on sludge AD, more attention should be given to HA produced and accumulated in AD rather than raw sludge so that the CA treatment directly on sludge in the digestor can be a promising method.

Percarbonate mediated advanced oxidation completely degrades recalcitrant pesticide imidacloprid: Role of reactive oxygen species and transformation products

Pesticides are vital and indispensable especially in agricultural production. However, pesticide contamination is also a serious threat to human health and the environment. Imidacloprid (IMD), an archetypal neonicotinoid insecticide, is commonly used to protect a variety of crops worldwide. This study examined the destruction of IMD contaminant in wastewater via catalytic oxidation with sodium percarbonate (SPC). The process exemplified 100% removal at optimum condition. Rates of reaction were expressed in terms of the apparent rate constants (kapp) and were observed to satisfactorily follow pseudo-first-order kinetics. Inhibitory effects of humic acid and various coexisting ions on IMD degradation were examined and the trend was observed as follows: NO2− > PO43− > NH4+ > Cl− > NO3−. In addition, we elucidated the participation of OH (hydroxyl), O2− (superoxyl), and HO2 (perhydroxyl) radicals in the process by performing scavenging experiments for these reactive oxygen species. A likely IMD degradation pathway was proposed based on the intermediate species identified via gas chromatography/mass spectroscopy electron ionization procedure. Overall, the present study established SPC as a cleaner alternative to other oxidants for pesticide degradation as it is non-toxic, safe to handle, and produces by-products that inherently exist in the natural water matrix.

Chlorination of bisphenol S: Kinetics, products, and effect of humic acid.

Bisphenol S (BPS), as a main alternative of bisphenol A for the production of industrial and consumer products, is now frequently detected in aquatic environments. In this work, it was found that free chlorine could effectively degrade BPS over a wide pH range from 5 to 10 with apparent second-order rate constants of 7.6-435.3 M-1s-1. A total of eleven products including chlorinated BPS (i.e., mono/di/tri/tetrachloro-BPS), 4-hydroxybenzenesulfonic acid (BSA), chlorinated BSA (mono/dichloro-BSA), 4-chlorophenol (4CP), and two polymeric products were detected by high performance liquid chromatography and electrospray ionization-tandem quadrupole time-of-flight mass spectrometry. Two parallel transformation pathways were tentatively proposed: (i) BPS was attacked by stepwise chlorine electrophilic substitution with the formation of chlorinated BPS. (ii) BPS was oxidized by chlorine via electron transfer leading to the formation of BSA, 4CP and polymeric products. Humic acid (HA) significantly suppressed the degradation rates of BPS even taking chlorine consumption into account, while negligibly affected the products species. The inhibitory effect of HA was reasonably explained by a two-channel kinetic model. It was proposed that HA negligibly influenced pathway i while appreciably inhibited the degradation of BPS through pathway ii, where HA reversed BPS phenoxyl radical (formed via pathway ii) back to parent BPS.

Phosphate Changes Effect of Humic Acids on TiO2 Photocatalysis: From Inhibition to Mitigation of Electron-Hole Recombination.

A major challenge for photocatalytic water purification with TiO2 is the strong inhibitory effect of natural organic matter (NOM), which can scavenge photogenerated holes and radicals and occlude ROS generation sites upon adsorption. This study shows that phosphate counteracts the inhibitory effect of humic acids (HA) by decreasing HA adsorption and mitigating electron-hole recombination. As a measure of the inhibitory effect of HA, the ratios of first-order reaction rate constants between photocatalytic phenol degradation in the absence versus presence of HA were calculated. This ratio was very high, up to 5.72 at 30 mg/L HA and pH 4.8 without phosphate, but was decreased to 0.76 (5 mg/L HA, pH 8.4) with 2 mM phosphate. The latter ratio indicates a surprising favorable effect of HA on TiO2 photocatalysis. FTIR analyses suggest that this favorable effect is likely due to a change in the conformation of adsorbed HA, from a multiligand exchange arrangement to a complexation predominantly between COOH groups in HA and the TiO2 surface in the presence of phosphate. This configuration can reduce hole consumption and facilitate electron transfer to O2 by the adsorbed HA (indicated by linear sweep voltammetry), which mitigates electron-hole recombination and enhances contaminant degradation. A decrease in HA surface adsorption and hole scavenging (the predominant inhibitory mechanisms of HA) by phosphate (2 mM) was indicated by a 50% decrease in the photocatalytic degradation rate of HA and 80% decrease in the decay rate coefficient of interfacial-related photooxidation in photocurrent transients. These results, which were validated with other compounds (FFA and cimetidine), indicate that anchoring phosphate - or anions that exert similar effects on the TiO2 surface - might be a feasible strategy to counteract the inhibitory effect of NOM during photocatalytic water treatment.

Transformation of Flame Retardant Tetrabromobisphenol A by Aqueous Chlorine and the Effect of Humic Acid.

In this work, it was found that the most widely used brominated flame retardant tetrabromobisphenol A (TBrBPA) could be transformed by free chlorine over a wide pH range from 5 to 10 with apparent second-order rate constants from 138 to 3210 M(-1)·s(-1). A total of eight products, including one quinone-like compound (i.e., 2,6-dibromoquinone), two dimers, and several simple halogenated phenols (e.g., 4-(2-hydroxyisopropyl)-2,6-dibromophenol, 2,6-dibromohydroquinone, and 2,4,6-tribromophenol), were detected by high-performance liquid chromatography tandem mass spectrometry (HPLC-MS/MS) using a novel precursor ion scan (PIS) approach. A tentative reaction pathway was proposed: chlorine initially oxidized TBrBPA leading to the formation of a phenoxy radical, and then this primary radical and its secondary intermediates (e.g., 2,6-dibromo-4-isopropylphenol carbocation) formed via beta-scission subsequently underwent substitution, dimerization, and oxidation reactions. Humic acid (HA) considerably inhibited the degradation rates of TBrBPA by chlorine even accounting for oxidant consumption. A similar inhibitory effect of HA was also observed in permanganate and ferrate oxidation. This inhibitory effect was possibly attributed to the fact that HA competitively reacted with the phenoxy radical of TBrBPA and reversed it back to parent TBrBPA. This study confirms that chlorine can transform phenolic compounds (e.g., TBrBPA) via electron transfer rather than the well-documented electrophilic substitution, which also have implications on the formation pathway of halo-benzoquinones during chlorine disinfection. These findings can improve the understanding of chlorine chemistry in water and wastewater treatment.

Photoelectrocatalytic degradation of the antibiotic sulfamethoxazole using TiO2/Ti photoanode

Photoelectrocatalysis (PEC) of sulfamethoxazole (SMZ) was investigated using TiO2/Ti (UV-A) photoanode. Complete photoelectrocatalytic degradation of SMZ was achieved after 70min at 0.5V, while SMZ resisted to photolysis and electrolysis. Langmuir-Hinshelwood analysis yielded an adsorption constant (K) of 0.519L/mg for the studied PEC-TiO2 system, with K being 3–4 times of those from photocatalysis. PEC degradation of SMZ accelerated with increasing anodic potential (0–0.5V), declining pH (from 10.1 to 2.7), and increasing NaCl concentration (1–100mM). These trends were consistent with the understanding that higher potential gradient reduces recombination of holes and electrons, lower pH favors photoadsorption through modulation of SMZ speciation and TiO2 surface charge, and higher Cl− level encourages formation of chlorine radicals. Molar ratios of HO and chlorine radicals showed that the effects of pH and Cl− levels were coupled, with the level of PEC activity linked to the relative dominance of [Cl2−] over [HO]. In the presence of radical scavengers, SMZ photoelectrocatalysis was lowered by 30% at merely 10mg/L of humic acid, and was reduced to <5% of the original rate at 10% methanol. The inhibitory effect of humic acid and methanol on PEC was consistent with the view that HO and Cl2−/active chlorine species were the main oxidants in a Cl−-present environment. A 3-times above-average photocurrent was observed in 10% methanol, whose unusual photocurrent increase was attributed to sequential current doubling – additional electron flow due to reactive adsorbate-hole interaction – through a series of proposed reactions: conversion of hVB+-bound methanol through formaldehyde and formic acid to CO2. Implications of the findings with respect to practical application of PEC are presented.

Effects of humic acid on adhesion of Bacillus subtilis to phyllosilicates and goethite

Bacterial attachment is a physicochemical interfacial process probably affected by the presence of humic acid (HA) in natural environments. However, the effect of HA coating on bacterial adhesion to soil minerals remains unknown, despite many studies focusing on cell attachment to pure or iron oxide-coated quartz in the presence of HA. The influence of HA on Bacillus subtilis adhesion to kaolinite, montmorillonite, and goethite under neutral pH and 1mM ionic strength was examined using batch experiments coupled with Fourier-transform infrared spectroscopy (FTIR), isothermal titration calorimetry, and Derjaguin, Landau, Verwey, and Overbeek (DLVO) theory. Results showed a reduction in cell adhesion on goethite and montmorillonite with increasing HA content from 0% to 2%, indicating an inhibitory effect of HA coating on the interaction. However, a promotive effect of HA was observed for kaolinite, which disagreed with previous findings based on cell–quartz systems. In addition, the presence of HA reduced the exothermic adhesion enthalpy for kaolinite and goethite, but increased that for montmorillonite. For goethite, the HA coating depressed its positive charge and increased its aggregation, thereby leading to a weaker electrostatic attraction and lesser available surface area, which probably contributed to the observed decreasing adhesion. For montmorillonite, the reduced attachment was likely caused by the increase in electrostatic repulsion that overcompensated for the appeared chemical interactions as suggested by FTIR analysis. The enhanced cell adhesion on kaolinite, which was unexpected from DLVO theory based on its surface charge change, is likely ascribed to the HA-induced decrease in its aggregation. Additionally, the effect of HA vanishes at HA contents higher than 2%, likely due to a saturation effect. Overall, the influence of HA on bacterial attachment is likely a combined result of multiple factors, including not only electrostatic forces and chemical interactions, but also mineral aggregation.

Mitigation of Humic Acid Inhibition in Anaerobic Digestion of Cellulose by Addition of Various Salts.

Humic compounds are inhibitory to the anaerobic hydrolysis of cellulosic biomass. In this study, the impact of salt addition to mitigate the inhibitory effects of humic compounds was investigated. The experiment was conducted using batch tests to monitor the anaerobic hydrolysis of cellulose in the presence of humic acid. Sodium, potassium, calcium, magnesium and iron salts were tested separately for their efficiency to mitigate humic acid inhibition. All experiments were done under mesophilic conditions (30 °C) and at pH 7. Methane production was monitored online, using the Automatic Methane Potential Test System. Methane production, soluble chemical oxygen demand and volatile fatty acid content of the samples were measured to calculate the hydrolysis efficiencies. Addition of magnesium, calcium and iron salts clearly mitigated the inhibitory effects of humic acid and hydrolysis efficiencies reached up to 75%, 65% and 72%, respectively, which were similar to control experiments. Conversely, potassium and sodium salts addition did not mitigate the inhibition and hydrolysis efficiencies were found to be less than 40%. Mitigation of humic acid inhibition via salt addition was also validated by inductively coupled plasma atomic emission spectroscopy analyses, which showed the binding capacity of different cations to humic acid.

Effects of Natural Organic Matter on TCE Removal in Groundwater by Zero-Valent Iron: Batch Study

An investigation on the effects of humic acid (representing NOM) on TCE (a typical organic contaminant) removal by Fe0in batch settings was carried out. Inhibitory effects of humic acid on Fe0towards TCE removal were observed. At early stage of the experiments, humic acid might partition with TCE, and the adsorption or deposition of humic acid onto Fe0surface would further facilitated TCE immobilization. Once the reduction reactive sites on Fe0surfaces were covered by accumulated humic acid and the partition of TCE to humic acid became saturated, TCE removal in Fe0was observed retarded.

Remediation of Trichloroethylene by FeS-Coated Iron Nanoparticles in Simulated and Real Groundwater: Effects of Water Chemistry

The reactivity of FeS-coated iron nanoparticles (nFe/FeS) toward trichloroethylene (TCE) reduction was examined in both synthetic and real groundwater matrices to evaluate the potential performance of nFe/FeS in field treatment. The rate of TCE reduction increased with increasing pH, which is consistent with the pH effect reported previously for iron sulfide systems, but opposite that has been observed for (nonsulfidic) Fe0 systems. The rates of TCE reduction were unaffected by ionic strength over the range of 0.1–10 mM NaCl, increased with Ca2+ or Mg2+ concentrations, and inhibited by the presence of humic acid. The inhibitory effect of humic acid on the reactivity of nFe/FeS was largely alleviated when humic acid was combined with Ca2+/Mg2+, presumably due to decreased adsorption of humic acid onto nFe/FeS surface by the formation of humic acid–Ca2+/Mg2+ complexes.

Abiotic reduction of nitroaromatic compounds by Fe(II) associated with iron oxides and humic acid

Experiments were conducted to examine the reduction of nitroaromatic compounds (NACs) by Fe(II) associated with iron oxides (goethite, hematite and magnetite) and humic acid. The reduction rate of nitrobenzene decreased in the order of Fe(II) associated with magnetite>Fe(II) associated with goethite>Fe(II) associated with hematite. We proposed a four-step model (adsorption, electron transfer to conduction band, electron transfer to nitrobenzene and electron transfer to crystal lattice) for nitrobenzene reduction by Fe(II) associated with iron oxides. Fe(II)–humic acid complexes did not present reduction capability of nitrobenzene. Furthermore, Humic acid significantly inhibited nitrobenzene reduction by Fe(II) associated with iron oxides. The inhibitory effect of humic acid toward the reduction of nitrobenzene decreased in the order of magnetite>goethite>hematite.

Techniques for minimizing the effects of PCR inhibitors in the chytridiomycosis assay

Chytridiomycosis is an amphibian disease of global conservation concern that is caused by the fungal pathogen Batrachochytrium dendrobatidis (Bd). Since the discovery of Bd in 1998, several methods have been used for detection of Bd; among these polymerase chain reaction (PCR) from skin swabs is accepted as the best method due to its noninvasiveness, high sensitivity and ease of use. However, PCR is not without problems - to be successful, this technique is dependent upon the presence of nondegraded DNA template and reaction contents that are free from inhibitors. Here, we report on an investigation of several techniques aimed at improving the reliability of the Bd PCR assay by minimizing the effects of humic acid (HA), a potent PCR inhibitor. We compared the effectiveness of four DNA extraction kits (DNeasy, QIAamp DNA Stool, PowerLyzer Power Soil and PrepMan Ultra) and four PCR methods (Amplitaq Gold, bovine serum albumin, PowerClean DNA Clean-up and inhibitor resistant Taq Polymerase). The results of this and previous studies indicate that chytridiomycosis studies that use PCR methods for disease detection may be significantly underestimating the occurrence of Bd. Our results suggest that to minimize the inhibitory effects of HA, DNeasy should be used for sample DNA extraction and Amplitaq Gold with bovine serum albumin should be used for the Bd PCR assay. We also outline protocols tested, show the results of our methods comparisons and discuss the pros and cons of each method.

Quantification of Escherichia coli O157:H7 in soils using an inhibitor-resistant NanoGene assay

Humic acids are ubiquitous and abundant in terrestrial environments; therefore, they are often co-extracted with nucleic acids and interfere with quantitative PCR (qPCR) assays. In this study a recently developed NanoGene assay that is resistant to interference by humic acids was evaluated for gene detection in soil samples. The NanoGene assay utilizes a combination of magnetic beads, dual quantum dots labels, and DNA hybridization in solution. Seven soil samples containing different amounts of organic matter were tested to compare NanoGene and qPCR assays for their respective ability to detect a bacterial pathogen. We spiked the soils with Escherichia coli O157:H7, extracted genomic DNA, and conducted NanoGene and qPCR assays targeting the E. coli O157:H7-specific eaeA gene. To prevent the inhibition of PCR that is common when using DNA extracted from soils, we used a range of template DNA concentrations and BSA addition in the qPCR assay. Compared to the qPCR assay the NanoGene assay was significantly more resistant to the inhibitory effect of humic acids, successfully quantifying the eaeA gene within a linear (R2 = 0.99) range of 105 through 108 CFU/g soil for all seven soil samples tested. In contrast, the qPCR assay was significantly inhibited using the same template DNA isolated from soils containing a range of organic content (2.0%–12%). Interestingly, the qPCR assay was still inhibited despite additional purification steps, suggesting that humic acids were still associated with DNA at a level that was inhibitory to qPCR. This study demonstrated that the NanoGene assay is suitable for quantitative gene detection in diverse soil types and is not susceptible to inhibition by humic acids and other organic compounds that commonly lead to false negative results in qPCR assays.

Effects and mechanism of humic acid on chromium(VI) removal by zero-valent iron (Fe 0) nanoparticles

The present study dealt with the performance evaluation of the nanoscale Fe 0 systems for the remediation of chromium contaminated groundwater in the ambient environment. The role of humic acid (HA) in the Cr(VI) removal and the reduction mechanism were investigated. HA was found to exert an obvious inhibitory effect on Cr(VI) removal by Fe 0 nanoparticles, and the Cr(VI) removal efficiencies decreased from 71.6%, 58.4%, 57.8% to 38.5% with the increasing HA concentrations (0, 5, 10, 20 to 40 mg L −1). A dual effect of humic acid on chromium(VI) reduction by Fe 0 nanoparticles was observed. HA adsorbed on the surface of Fe 0 nanoparticles and occupied the active surface sites, leading to the decrease in Cr(VI) reduction rates. Greater was the adsorbed HA, the more obvious was the inhibitory effect. However, the HA adsorption on iron surface areas was one of the factors leading to the decreased reduction rate. The appropriate starch dosage (0.5 g per 0.3 g nanoscale Fe 0 particles) could definitely eliminate the inhibitory effect of humic acid.

Bimodal effect of humic acids on the LPS-induced TNF-α release from differentiated U937 cells

Humic substances (HS) have been reported to possess anti-inflammatory as well as pro-inflammatory properties. The anti-inflammatory activity was demonstrated in the rat paw edema model and we found a preliminary explanation in the 5-lipoxygenase inhibitory effect of humic acids (HA). The pro-inflammatory activity is reflected by the production and release of pro-inflammatory cytokines in HA-treated neutrophilic granulocytes. With regard to the potential use of HA as antiviral and UV-protective agents it appears advisable to investigate the role of HS in the inflammation process in more detail. Hence we tested four different HS preparations - two naturally occurring HA from the Altteich peatland in Germany, one fulvic acid (FA) preparation from a Finnish spruce forest and a synthetic HA-like polymer (caffeic acid oxidation product, KOP) for their influence on the lipopolysaccharide (LPS)-induced TNF-alpha release in human U937 cells. In addition, the cytotoxicity of HS was determined. The results demonstrate a concentration-dependent bimodal effect of HA on the TNF-alpha release of differentiated LPS-stimulated U937 cells for both the natural black peat HA from the Altteich peatland and the HA-like polymer KOP. Low HA concentrations (10-80 microg/ml) enhanced the TNF-alpha release by up to threefold (pro-inflammatory activity), while HA concentrations >100 microg/ml reduced it about 10-fold (anti-inflammatory activity). FA failed to enhance TNF-alpha release, but reduced it at higher concentrations (>200 microg/ml) by the half. Brown water HA did not exert any significant effect on TNF-alpha release. No HS-stimulated TNF-alpha release was also observed in the absence of exogenously supplied LPS. This means that HS, unlike endotoxin, are no inflammation-causing agents for LPS-untreated cells. Differences in the effect of individual HS on TNF-alpha release are discussed in connection with the polyanionic character of HS, their molecular mass distribution and the hitherto imperfectly known chemical structure.

Effects of Natural Organic Matter, Anthropogenic Surfactants, and Model Quinones on the Reduction of Contaminants by Zero-Valent Iron

Recent studies of contaminant reduction by zero-valent iron metal (Fe 0) have highlighted the role of iron oxides at the metal–water interface and the effect that sorption has at the oxide–water interface on contaminant reduction kinetics. The results suggest that a variety of organic surface-active substances might enhance or inhibit contaminant degradation, depending on the degree to which they promote solubilization, sorption, and/or reaction. Of particular interest is the effect of natural organic matter (NOM), because of its ubiquitous presence in natural waters and amphiphilic properties; anthropogenic surfactants, because of their use in groundwater remediation; and certain quinones, which represent the redox-active functional groups associated with NOM. In this study, no well-defined effects of these substances were found on the reduction of nitro benzene by Fe 0, but the reduction of carbon tetrachloride and trichloroethylene (TCE) was inhibited by NOM. Results with carbon tetrachloride showed that the inhibitory effect of humic acids was greater than fulvic acids, but that several quinonoid NOM model compounds (juglone, lawsone, and anthraquinone disulfonate) increased the rate of reduction by Fe 0. Isotherms for adsorption of TCE and NOM onto Fe 0 showed evidence of competition for surface sites.

Inhibition of human plasmin activity using humic acids with arsenic

In this study, we examined the effects of natural humic acid (HA) purified from drinking well-water in Blackfoot disease (BFD) endemic areas, using synthetic humic acid (SHA), such as protocatechuic acid, ferulic acid, vanillic acid or catechol, and trivalent arsenic on human plasmin activity. Data in this report indicated that both HA and SHA inhibited human plasmin activity by 20–80 and 5–95%, respectively, at concentrations of 20–480 μg/ml. Organometallic complexes composed of HA and arsenic show enhanced inhibition of plasmin activity as compared with either HA or arsenic alone. Monomers of HA or arsenic alone do not inhibit plasma activity. Oxidative stress may play a role in the inhibition of plasma activity, as various free-radical scavengers, such as ascorbic acid, α-tocopherol, catalase and superoxide dismutase (SOD), abrogate the inhibitory effects of HA and HA–arsenic complexes. The notion that HA/organometallic complexes (HA/OR) impaired plasmin activity was significant, due to the fact that both of these agents (HA and arsenic) are etiological factors in the development of peripheral vascular diseases, such as BFD. This report substantiates the inhibitory effects of HA/OR on plasmin activity, and thus provides evidence for the partial mechanism of action of HA/OR in BFD.

Oxidation of Substituted Anilines by Aqueous MnO2: Effect of Co-Solutes on Initial and Quasi-Steady-State Kinetics

The effect of pH, Mn(II), and humic acid (HA) concentration as well as aromatic ring substituents on initial and quasi-steady-state rates of oxidation of monosubstituted anilines by MnO2 has been investigated in batch and completely mixed flow-through reactors. The reaction rates were strongly pH-dependent and increased with decreasing pH. Mn(II), Ca(II), HA, and other organic solutes inhibited the oxidation of substituted anilines. Inhibition by adsorbed Mn(II) was due to blocking of reactive Mn(IV) surface sites. The inhibitory effect of HA and some other organic solutes was in part due to reductive dissolution of MnO2 and, thus, due to formation of adsorbed Mn(II). Initial rates of oxidation of a series of substituted anilines were linearly correlated to polarographic half-wave oxidation potentials, E1/2, indicating that electron-transfer kinetics were relevant for the overall rate of oxidation. However, results from flow-through experiments demonstrate that such correlations may change or disappear, s...

Microscale detection of specific bacterial DNA in soil with a magnetic capture-hybridization and PCR amplification assay

A magnetic capture-hybridization PCR technique (MCH-PCR) was developed to eliminate the inhibitory effect of humic acids and other contaminants in PCRs targeting specific soil DNA. A single-stranded DNA probe, which was complementary to an internal part of the target gene, was used to coat magnetic beads. After hybridization in a suspension of soil DNA, magnetic extraction of the beads separated the hybrid DNA from all other soil DNA, humic acids, and other interfering soil components. The MCH was followed by PCR amplification of the specific target DNA. In barley rhizosphere soil, detection of a lux gene inserted in a Pseudomonas fluorescens strain could be demonstrated in nonsterile soil samples (0.5 mg). This corresponded to a detection of fewer than 40 bacterial cells per cm of barley root. The MCH-PCR technique greatly improves the current protocols for PCR detection of specific microorganisms or genes in soil because specific target DNA sequences from very small soil samples can be extracted and determined.