The 28-day repeated arsenic exposure increases tau phosphorylation in the rat brain.

Herein, we examined whether prolonged arsenic exposure altered tau phosphorylation in the brain of Sprague Dawley rats expressing endogenous wild-type tau. The results showed that daily intraperitoneal injections of 2.5 mg/kg BW sodium arsenite over 28 days caused arsenic accumulation in the rat brain. Interestingly, we found an increase in tau phosphorylation at the Tau 1 region (189-207) and S202 in the hippocampus, S404 in the cerebral cortex, and S396 and S404 in the cerebellum of arsenic-treated rats. Additionally, arsenic increased active ERK1/2 phosphorylation at T202/Y204 in the hippocampus, cerebral cortex, and cerebellum. Meanwhile, we detected increasing active JNK phosphorylation at T183/Y185 in the hippocampus and cerebellum. Moreover, p35, a neuron-specific activator of CDK5, was also elevated in the cerebellum of arsenic-treated rats, suggesting that CDK5 activity may be increased by arsenic. These results suggested that arsenic may induce tau phosphorylation through the activation of tau kinases, ERK1/2, JNK, and CDK5. Together, the findings from this study demonstrated that prolonged arsenic exposure is implicated in neurodegeneration by promoting tau phosphorylation in the rat brain and points toward a possible prevention strategy against neurodegeneration induced by environmental arsenic exposure.

Sulfur amendments to soil decrease inorganic arsenic accumulation in rice grain under flooded and nonflooded conditions: Insights from temporal dynamics of porewater chemistry and solid-phase arsenic solubility.

Rice cultivation under flooded conditions enhances arsenic (As) solubility and favors As accumulation in rice grain that poses an indisputable threat to human health worldwide. The reduction of sulfur may induce processes that decrease As solubility, but its impact on rice grain As species remains unresolved. Herein, we investigated the influence of sulfur (S)-containing materials, including chicken manure and elemental sulfur powder on As accumulation and speciation in rice grain as well as the dynamics of the porewater chemistry and solid-phase As solubility throughout the entire growth stage under continuous flooding and intermittent flooding conditions in pot experiments. The S amendments (200 mg S kg-1) to the soil significantly decreased inorganic As in rice grain under continuous flooding (~65% decrease) as well as under intermittent flooding (~70% decrease). The chicken manure amendment promoted sulfur reduction and enhanced dissolvable Mn, Fe, and As at an earlier growth stage. The sequential extraction results corroborated a decrease in the soluble and exchangeable As (F1) and an increase in residual As (F5) fractions in the S-amended treatments. Solubility data suggested that As adsorption onto Fe oxides was the primary mechanism controlling As solubility rather than the formation of AsFe sulfides. Porewater As, considered to represent the most bioavailable As fraction, failed to explain the grain As accumulation. The time-averaged concentration of oxalate-extractable As explained grain arsenite best, suggesting that poorly crystalline Fe oxides may be the primary dissolvable reactive phases that control As bioavailability in the soil-rice system. Our results suggest that the application of S-containing soil amendments can effectively decrease inorganic As accumulation in rice grains grown under the flooded conditions, which are most widely applied in paddy rice production.

Biochar and ash derived from silicon-rich rice husk decrease inorganic arsenic species in rice grain.

Exposure to arsenic (As) through rice consumption potentially threatens millions of people worldwide. Understanding is still lacking the recycling impacts of rice residues on As phytoavailability in paddy soils and is of indisputable importance in providing a sustainable and effective measure to decrease As accumulation in rice grain. Herein, we examined the effects of rice husk biochar (RHB) and rice husk ash (RHA) on As grain speciation, and As dynamics in the soil porewater and solid-phase fractions. The results corroborated that both the RHB and RHA (0.64% w/w) treatments significantly (p < 0.05) decreased inorganic As accumulation in rice grain to 0.27-0.29 mg kg-1, which was below the maximum inorganic As level in husked rice (0.35 mg kg-1) established by the Codex. The residual phase (F6 = 90% of total soil As) as quantified by the sequential extraction was the dominant As pool; the fractions were subsequently transformed into several As pools associated with soluble and exchangeable (F1), organically bound (F2), Mn oxides (F3), poorly crystalline (F4) and crystalline (F5) Fe oxides during the rice growing periods. The Si-rich amendments enhanced the residual phase formation upon soil flooding, which decreased the As availability to rice plant. The inorganic grain-As concentrations were well explained by the soil-extractable As concentrations in the F2, F3, F5, and F6 fractions. The pore-water analysis indicated that Mn oxides were important sources and sinks for As released to the soil solution. Our findings shed light on the beneficial role of RHB and RHA in alleviating inorganic As uptake in paddy rice.

Sodium arsenite exposure impairs B cell proliferation and enhances vascular inflammation in Plasmodium berghei mouse model.

Arsenic exposure has been linked to an impaired immune response and inflammation. Our study investigated the effects of sodium arsenite on host immune response and vascular inflammation during malarial infection. Mice were divided into three groups: control (C), Plasmodium berghei infection (I) and sodium arsenite exposure with Plasmodium berghei infection (As-I). The results showed that splenocyte proliferation stimulated by lipopolysaccharide (LPS) and pokeweed mitogen (PWM) was suppressed in the I group, and the suppression was more pronounced in the As-I group, suggesting that acquired immunity in infected mice was worsening following arsenic exposure. ICAM-1, an adhesion protein involved in parasite-infected red blood cell (iRBC) binding to endothelium, and HIF-1α, a hypoxia marker protein in the descending aorta, were increased in the As-I group compared to the I group. Collectively, our results suggest that arsenic may increase host susceptibility to malaria through suppression of B cell proliferation and enhancement of adhesion between iRBC and endothelium by increasing ICAM-1.

Arsenic Speciation and Accumulation in Selected Organs after Oral Administration of Rice Extracts in Wistar Rats.

Despite its nutritional values, rice also contains arsenic. There has been increasing concern about health implications associated with exposure to arsenic through rice consumption. The present study evaluated arsenic accumulation and its speciation in selected organs of Wistar rats after 28 day repeated oral administrations of polished or unpolished rice and their control arsenic compounds (sodium arsenite or dimethylarsinic acid; DMA). Only the treatment of sodium arsenite (2 µg/kg body weight), significantly increased total arsenic concentrations in blood when compared to the distilled water control group. In all groups, total arsenic concentrations were highest in kidney (1.54-1.90 mg/kg) followed by liver (0.85-1.52 mg/kg), and the predominant arsenic form in these organs was DMA. However, there was no significant difference in arsenic accumulation in the measured organs among the control and rice-treated groups. Therefore, the repeated 28 day administration of arsenic-contaminated rice did not cause significant arsenic accumulation in the animal organs.

Hijiki and sodium arsenite stimulate growth of human colorectal adenocarcinoma cells through ERK1/2 activation.

Hijiki seaweed (Hijikia fusiformes) contains high levels of inorganic arsenic, a known carcinogen. However, scientific reports on carcinogenic risks associated with the consumption of this seaweed are limited. This study investigated the effects of seaweed extracts contaminated with arsenic on two colorectal cancer cell lines. Two seaweed extracts, including Hijiki and red seaweed, induced H508 but not HT29 cell proliferation. Growth induction of H508 cells after treatments with Hijiki and sodium arsenite at concentrations equivalent to arsenic found in Hijiki was observed by both MTT and BrdU assays. Hijiki and sodium arsenite induced epidermal growth factor receptor (EGFR) and ERK1/2 activations. AG1478, an EGFR inhibitor, decreased the activation of EGFR and ERK1/2 induced by Hijiki and sodium arsenite. U0126, an ERK1/2 upstream inhibitor, and atropine, a muscarinic acetylcholine receptor (mAChR) antagonist, but not AG1478 completely inhibited the proliferative effect of Hijiki. Altogether, the results suggest that the presence of arsenic in seaweed may partly contribute to the proliferation of colorectal cancer cells. EGFR-dependent, and -independent ERK1/2 signaling pathways, and mAChR may be involved in the growth stimulation by Hijiki. These results raise concern regarding the potential colorectal cancer risks from regular consumption of Hijiki containing high contents of inorganic arsenic.

Regulation of the Cobalt/Nickel Efflux Operon dmeRF in Agrobacterium tumefaciens and a Link between the Iron-Sensing Regulator RirA and Cobalt/Nickel Resistance.

UNLABELLED: The Agrobacterium tumefaciens C58 genome harbors an operon containing the dmeR (Atu0890) and dmeF (Atu0891) genes, which encode a transcriptional regulatory protein belonging to the RcnR/CsoR family and a metal efflux protein belonging to the cation diffusion facilitator (CDF) family, respectively. The dmeRF operon is specifically induced by cobalt and nickel, with cobalt being the more potent inducer. Promoter-lacZ transcriptional fusion, an electrophoretic mobility shift assay, and DNase I footprinting assays revealed that DmeR represses dmeRF transcription through direct binding to the promoter region upstream of dmeR A strain lacking dmeF showed increased accumulation of intracellular cobalt and nickel and exhibited hypersensitivity to these metals; however, this strain displayed full virulence, comparable to that of the wild-type strain, when infecting a Nicotiana benthamiana plant model under the tested conditions. Cobalt, but not nickel, increased the expression of many iron-responsive genes and reduced the induction of the SoxR-regulated gene sodBII Furthermore, control of iron homeostasis via RirA is important for the ability of A. tumefaciens to cope with cobalt and nickel toxicity. IMPORTANCE: The molecular mechanism of the regulation of dmeRF transcription by DmeR was demonstrated. This work provides evidence of a direct interaction of apo-DmeR with the corresponding DNA operator site in vitro The recognition site for apo-DmeR consists of 10-bp AT-rich inverted repeats separated by six C bases (5'-ATATAGTATACCCCCCTATAGTATAT-3'). Cobalt and nickel cause DmeR to dissociate from the dmeRF promoter, which leads to expression of the metal efflux gene dmeF This work also revealed a connection between iron homeostasis and cobalt/nickel resistance in A. tumefaciens.

Concentrations of Trace Elements in Organic Fertilizers and Animal Manures and Feeds and Cadmium Contamination in Herbal Tea (Gynostemma pentaphyllum Makino).

Thailand is predominantly an agriculture-based country. Organic farming is enlisted as an important national agenda to promote food safety and international export. The present study aimed to determine the concentrations of trace elements in commercial organic fertilizers (fermented and nonfermented) composed of pig and cattle manures available in Thailand. Pig and cattle manures as well as animal feeds were also collected from either animal farms or markets. The results were compared to the literature data from other countries. Fermented fertilizer composed of pig manure contained higher concentrations of nitrogen (N) and phosphorus (P) than fertilizer composed of cattle manure. High concentrations of copper (Cu) and zinc (Zn) were also found in fertilizers and manures. Some organic fertilizers had high concentrations of arsenic (As), cadmium (Cd), and lead (Pb). The range of As concentration in these fertilizers was 0.50-24.4 mg/kg, whereas the ranges of Cd and Pb were 0.10-11.4 and 1.13-126 mg/kg, respectively. Moreover, pig manure contained As and Cd (15.7 and 4.59 mg/kg, respectively), higher than their levels in cattle manure (1.95 and 0.16 mg/kg, respectively). The use of pig manure as soil supplement also resulted in high Cd contamination in herbal tea (Gynostemma pentaphyllum Makino; GP). The Cd concentration in GP plants positively correlated with the Cd concentration in the soil. Therefore, the application of some organic fertilizers or animal manures to agricultural soil could increase some potentially toxic elements in soil, which may be absorbed by plants and, thus, increase the risk of contamination in agricultural products.

Agrobacterium tumefaciens Zur Regulates the High-Affinity Zinc Uptake System TroCBA and the Putative Metal Chaperone YciC, along with ZinT and ZnuABC, for Survival under Zinc-Limiting Conditions.

UNLABELLED: Agrobacterium tumefaciens has a cluster of genes (Atu3178, Atu3179, and Atu3180) encoding an ABC-type transporter, here named troA, troB, and troC, respectively, which is shown here to be a zinc-specific uptake system. Reverse transcription (RT)-PCR analysis confirmed that troA, troB, and troC are cotranscribed, with troC as the first gene of the operon. The yciC (Atu3181) gene is transcribed in the opposite orientation to that of the troCBA operon and belongs to a metal-binding GTPase family. Expression of troCBA and yciC was inducible under zinc-limiting conditions and was controlled by the zinc uptake regulator, Zur. Compared to the wild type, the mutant strain lacking troC was hypersensitive to a metal chelator, EDTA, and the phenotype could be rescued by the addition of zinc, while the strain with a single yciC mutation showed no phenotype. However, yciC was important for survival under zinc limitation when either troC or zinT was inactivated. The periplasmic zinc-binding protein, ZinT, could not function when TroC was inactivated, suggesting that ZinT may interact with TroCBA in zinc uptake. Unlike many other bacteria, the ABC-type transporter ZnuABC was not the major zinc uptake system in A. tumefaciens However, the important role of A. tumefaciens ZnuABC was revealed when TroCBA was impaired. The strain containing double mutations in the znuA and troC genes exhibited a growth defect in minimal medium. A. tumefaciens requires cooperation of zinc uptake systems and zinc chaperones, including TroCBA, ZnuABC, ZinT, and YciC, for survival under a wide range of zinc-limiting conditions. IMPORTANCE: Both host and pathogen battle over access to essential metals, including zinc. In low-zinc environments, physiological responses that make it possible to acquire enough zinc are important for bacterial survival and could determine the outcome of host-pathogen interactions. A. tumefaciens was found to operate a novel pathway for zinc uptake in which ZinT functions in concert with the high-affinity zinc importer TroCBA.

Roles of Agrobacterium tumefaciens C58 ZntA and ZntB and the transcriptional regulator ZntR in controlling Cd2+/Zn2+/Co2+ resistance and the peroxide stress response.

The putative zinc exporters ZntA (a P1B-type ATPase) and ZntB (2-TM-GxN family) in Agrobacterium tumefaciens were characterized. The expression of the zntA gene is inducible by CdCl2, ZnCl2 and CoCl2, of which CdCl2 is the most potent inducer, whereas zntB is constitutively expressed. The metal-induced expression of zntA is controlled by the MerR-like regulator ZntR. The zntA and zntR mutants were highly sensitive to CdCl2 and ZnCl2, and CoCl2 sensitivity was demonstrated to a lesser extent. By contrast, the zntB mutant showed similar levels of metal resistance to the WT strain. Even in the zntA mutant background, zntB did not play an apparent role in metal resistance under the conditions tested. The inactivation of zntA increased the accumulation of intracellular cadmium and zinc, and conferred hyper-resistance to H2O2. Thus, the metal transporter ZntA and its regulator ZntR are important for controlling zinc homeostasis and cadmium and cobalt detoxification. The loss of either the zntA or zntR gene did not affect the virulence of A. tumefaciens in Nicotiana benthamiana.

Arsenic, cadmium, and manganese levels in shellfish from Map Ta Phut, an industrial area in Thailand, and the potential toxic effects on human cells.

Map Ta Phut Industrial Estate is a major industrial area in Thailand for both petrochemical and heavy industries. The release of hazardous wastes and other pollutants from these industries increases the potential for contamination in foods in the surrounding area, especially farmed shellfish. This study determined the arsenic (As), cadmium (Cd), and manganese (Mn) concentrations in the edible flesh of farmed shellfish, including Perna viridis, Meretrix meretrix, and Scapharca inaequivalvis, around the Map Ta Phut area using inductively coupled plasma mass spectrometry. The results showed that shellfish samples contained high levels of total As [1.84-6.42 mg kg(-1) wet weight (ww)]. High Mn concentrations were found in P. viridis and M. meretrix, whereas S. inaequivalis contained the highest Cd. Arsenobetaine (AsB) was found to be the major As species in shellfish (>45% of total As). The in vitro cytotoxicity of these elements was evaluated using human cancer cells (T47D, A549, and Jurkat cells). An observed decrease in cell viability in T47D and Jurkat cells was mainly caused by exposure to inorganic As (iAs) or Mn but not to AsB or Cd. The combined elements (AsB+Mn+Cd) at concentrations predicted to result from the estimated daily intake of shellfish flesh by the local people showed significant cytotoxicity in T47D and Jurkat cells.

Control of zinc homeostasis in Agrobacterium tumefaciens via zur and the zinc uptake genes znuABC and zinT.

The Agrobacterium tumefaciens zinc uptake regulator (Zur) was shown to negatively regulate the zinc uptake genes znuABC, encoding a zinc transport system belonging to the ATP-binding cassette (ABC) transporter family, and zinT, which encodes a periplasmic zinc-binding protein. The expression of znuABC and zinT was inducible when cells were grown in medium containing a metal chelator (EDTA), and this induction was shown to be specific for zinc depletion. The expression of znuABC was reduced in response to increased zinc in a dose-dependent manner, and zinT had a less pronounced but similar pattern of zinc-regulated expression. The inactivation of zur led to constitutively high expression of znuABC and zinT. In addition, a zur mutant had an increased total zinc content compared to the WT NTL4 strain, whereas the inactivation of zinT caused a reduction in the total zinc content. The zinT gene is shown to play a dominant role and to be more important than znuA and znuB for A. tumefaciens survival under zinc deprivation. ZinT can function even when ZnuABC is inactivated. However, mutations in zur, znuA, znuB or zinT did not affect the virulence of A. tumefaciens.

Determination of arsenic species in rice from Thailand and other Asian countries using simple extraction and HPLC-ICP-MS analysis.

Human exposure to arsenic (As) via rice consumption is of increasing concern. In the present study, the extraction and HPLC-ICP-MS analysis for As speciation in rice were investigated. A simple extraction with water and digestion with α-amylase followed by the analysis using ion-paring mode HPLC separation was developed. The method showed good extraction efficiencies (generally >80%) and column efficiencies (>90%) for rice samples. The optimization of mobile phase showed well separated peaks of As species. The limits of quantification (LOQ) of As(III), As(V), MMA, and DMA that were calculated based on sample mass were 1.6, 2.0, 2.0, and 1.6 µg kg(-1), respectively. A total of 185 rice samples (various types of rice) collected from different four regions in Thailand and some other Asian countries were analyzed. The total As and inorganic As in rice samples were in the ranges of 22.51-375.39 and 13.89-232.62 µg kg(-1), respectively. The estimated weekly intake of inorganic As from rice by Thai people accounted for 13.91-29.22% of the provisional tolerable weekly intake (PTWI).

Immunomodulatory effects of cadmium and Gynostemma pentaphyllum herbal tea on rat splenocyte proliferation.

Gynostemma pentaphyllum Makino (GP) is a herbal tea widely grown in Southeast Asia. However, this herbal tea can be contaminated with some heavy metals, especially cadmium (Cd), from agricultural areas, which may affect human health. The objective of this study is to evaluate the immunomodulatory effects of Cd contaminated in GP herbal tea and inorganic Cd on rat splenocytes. Rats were divided into groups and treated with drinking water (control), high CdCl 2 in drinking water (HCd; 0.05 mg/L), GP herbal tea containing 0.05 mg/L Cd (GP-HCd) for 4 months, low CdCl 2 in drinking water (LCd; 0.006 mg/L), and GP herbal tea containing 0.006 mg/L Cd (GP-LCd) for 6 months. After the treatments, Cd accumulation in organs and blood was detected by using a graphite furnace atomic absorption spectrophotometer. In spleen, HCd-treated rats had 4-fold higher Cd accumulations than GP-HCd-treated rats. Cd accumulation in liver and kidney in the HCd group also increased significantly. There were no significant changes in total leucocyte and lymphocyte counts; however, these parameters tended to decrease slightly in LCd, GP-LCd, and GP-HCd groups. The HCd group (ex vivo) significantly produced suppressive effects on T cell mitogen-induced splenocyte proliferation, with 1 mug/mL Con A and PHA-P. In addition, 0.5 mug/mL PWM-induced B cell proliferation, through T cell functions, was also significantly inhibited by HCd as compared to the control group, while GP-HCd had no effects. However, both GP-LCd- and LCd-treated rats had a slight increase in Con A-stimulated splenocyte proliferation. This study indicated that high Cd contamination in drinking water alone had suppressive effects on T cell functions, but these effects could not be found with the same Cd level contamination in GP herbal tea.

Activation of inflammation/NF-kappaB signaling in infants born to arsenic-exposed mothers.

The long-term health outcome of prenatal exposure to arsenic has been associated with increased mortality in human populations. In this study, the extent to which maternal arsenic exposure impacts gene expression in the newborn was addressed. We monitored gene expression profiles in a population of newborns whose mothers experienced varying levels of arsenic exposure during pregnancy. Through the application of machine learning-based two-class prediction algorithms, we identified expression signatures from babies born to arsenic-unexposed and -exposed mothers that were highly predictive of prenatal arsenic exposure in a subsequent test population. Furthermore, 11 transcripts were identified that captured the maximal predictive capacity to classify prenatal arsenic exposure. Network analysis of the arsenic-modulated transcripts identified the activation of extensive molecular networks that are indicative of stress, inflammation, metal exposure, and apoptosis in the newborn. Exposure to arsenic is an important health hazard both in the United States and around the world, and is associated with increased risk for several types of cancer and other chronic diseases. These studies clearly demonstrate the robust impact of a mother's arsenic consumption on fetal gene expression as evidenced by transcript levels in newborn cord blood.

Determination of trace elements in herbal tea products and their infusions consumed in Thailand.

Nineteen elements, Mg, Al, Ca, V, Cr, Mn, Fe, Co, Ni, Cu, Zn, Se, Sr, Sb, Ba, As, Cd, Hg, and Pb, were determined in three types of popular herbal tea products, Gynostemma pentaphyllum, Camellia sinensis, and Morus alba. These herbal tea products, both imported and locally made products, are widely consumed in Thailand and worldwide. Microwave-assisted acid digestion was used for all of the samples, and the element contents were determined by ICP-MS. The concentrations of all elements varied among these herbal teas. Ca and Mg were the most abundant elements in all herbal samples (1384-34070 and 783-7739 mg/kg, respectively). Most elements in these herbal tea powders were also released into the infusions at different percentages depending on types of herbs. G. pentaphyllum infusion contained essential elements (Mg, Ca, V, and Fe) at higher levels than C. sinensis and M.alba infusions. Al and Ni were present at high levels in C. sinensis infusion, and Cd level was high in M. alba infusion. The daily intake of all elements from these herbal tea infusions (three cups/day) is still within the average daily intake. Therefore, it may not produce any health risks for human consumption, if other sources of toxic metal contaminated food are not taken at the same time.