The effect of particle size on oral bioavailability and bioaccessibility of soil Ni from different sources.

The goal of the work was to contribute to a unified approach to assessing the risk to human health of soil ingestion, for contaminated sites with elevated [Ni]. Robust relationships between in vitro bioaccessibility and in vivo bioavailability of Ni in various soils, with mechanistic understanding, would enable site-specific assessments of human exposure through soil ingestion. Four soils (three ultramafic Brunisols with geogenic Ni and one Organic soil with anthropogenic Ni) were sieved into PS < 10 µm, 10-41 µm, 41-70 µm, 70-105 µm, 105-150 µm, and 150-250 µm, the [Ni]T for which ranged from 560 to 103000 mg/kg. Mass fraction-adjusted [Ni]IVBA (SBRC gastric) for each soil fraction was similar whether calculated for all particles <250 µm or <150 µm %NiIVBA ranged from 3% to 16% of [Ni]T and %NiABA (accumulated Ni in urine, kidneys, and small intestine of Sprague Dawley rats gavaged with a soil) ranged from 0% to 0.49%. The correlation between these two measurements was weak (R2 = 0.06). Multiple linear dose response relationships attributing variation in %NiABA to %NiIVBA plus soil physicochemical parameters known to influence trace element availability in soils were developed. As many soil properties measured in this study were highly correlated, ridge regression enabled a predictive relationship where the effect of each parameter was its true contribution to variation in %NiABA. Using a ridge constant (k) of 0.012, %NiABA could be predicted from %NiIVBA adjusted for soil absorptive entities (OrgC, and Fe oxides (negative coefficients)) and soil pH (positive coefficient). %NiABA predicted from this relationship was very close to 1:1 with the observed %NiABA except at the lowest observed values which were lower than predicted. This study shows that as the conditions increasingly favour soil Ni solubility, more of the Ni was bioavailable; this generalization was true regardless of particle size or soil origin.

Lessons Learned From Rapid Deployment of 100% Mortality Review for Patients With COVID-19 Across a Health System.

Mortality review is one approach to systematically examine delivery of care and identify areas for improvement. Health system leaders sought to ensure hospitals were adapting to the rapidly changing medical guidance for COVID-19 and delivering high-quality care. Thus, all patients with a COVID-19 diagnosis within the 6-hospital system who died between March and July 2020 were reviewed within 72 hours. Concerns for preventability advanced review to level 2 (content experts) or 3 (hospital leadership). Reviews included available autopsy and cardiac arrest data. Overall health system mortality for COVID-19 patient admissions was 12.5% and mortality for mechanically ventilated patients was 34.4%. Significant differences in mortality rates were observed among hospitals due to demographic variations in patient populations at hospitals. Mortality reviews resulted in the dissemination of evolving knowledge among sites using an electronic medical record order set, implementation of proning teams, and development of checklists for converting COVID-19 floors and units.

Derivation of a Ni bioaccessibility value for screening-level risk assessment of Ni substances in ingested materials including soils.

The objective of the present study was to derive a Ni bioaccessibility value for screening-level risk assessment of Ni substances in ingested materials including soils where multiple Ni substances are expected but not definitively identified. Broad ranges of Ni mass loading and dissolution time of a simple gastric assay were applied to pure Ni substances (removing the confounding factors of soil constituents on dissolution), thus broadening the applicability of the conclusions. The data were also used to support current knowledge of 'read across' for Ni substances. Release of Ni from pure manufactured Ni substances (Ni metal, NiO, NiSO4, Ni3S2, and NiS) was determined relative to Ni mass and substance surface area loading. Mass loadings ranged from 0.33 to 20.0 g Ni per L of 0.15 M HCl, and dissolution time ranged from 1 to 168 h. Proton exhaustion was indicated only at the highest loading (20 g/L) of NiO and Ni-M. Dissolution of substances other than NiSO4 was most likely limited by formation of intermediate products at the particle surface or particle agglomeration, impeding access to the principal Ni substance. The bioaccessibility of Ni for these substances was consistent with previously published data: substances other than NiSO4 were < 48% bioaccessible for a variety of gastric assays, which is much lower than all data for NiSO4, the usual reference substance. Thus, we suggest that Ni bioaccessibility data from gastric assays that are most relevant to human exposure can be relied upon to develop scientifically sound screening-level human health RA decisions for Ni contamination in soils and sediments in the absence of detailed Ni speciation.

Gastric bioaccessibility is a conservative measure of nickel bioavailability after oral exposure: Evidence from Ni-contaminated soil, pure Ni substances and Ni alloys.

Oral bioaccessibility (BAc) is a surrogate for the bioavailability (BAv) of a broad range of substances, reflecting the value that the approach offers for assessing oral exposure and risk. BAc is generally considered to have been validated as a proxy for oral BAv for the important soil contaminants Pb, Cd, and As. Here, using literature data for Ni BAc and BAv, we confirmed that Ni BAc (gastric only, with HCl mimicking stomach conditions) is a conservative measure of BAv for the oral exposure pathway. Measured oral BAv of Ni in soil was shown to be 50-100 times less than the simplest oral BAc estimates (%BAv = 0.012(%BAc) - 0.023 (r = 0.701, 95%CI [0.456, 0.847], n = 30)) in rats, demonstrating a significant conservatism for exposure assessment. The relationship between the oral BAv and BAc of nickel sulfate hexahydrate (NSHH) was comparable to that of soil, with measured oral BAv of NSHH (1.94%) being a small fraction of NSHH gastric BAc (91.1%). BAc and BAv reflect the underlying Ni speciation of the sample, with the bioaccessible leaching limits being represented by the highly soluble Ni salts and the poorly soluble Ni monoxide, and the environmental (e.g. soil properties) or gastric (e.g. food present) conditions. BAc has potential utility for chemical classification purposes because pure Ni substances can be grouped by %BAc values(using standardized methodologies for the relevant exposure routes), these groupings reflecting the underlying chemistry and speciation of the samples of substances tested here, with 0.008% %BAc for alloys (SS304, SS316, Inconel, Monel), <1% in green NiO and Ni metal massives, 0.9-23.6% for Ni powders, 9.8-22.7% for Ni sulfides, 26.3-29.6% for black oxidic Ni, and 82-91% for the soluble Ni salts. Oral BAc provides realistic yet conservative estimates of BAv for the hazard classification and risk assessment of Ni substances.

Modeling phytoremediation of aged soil Ni from anthropogenic deposition using Alyssum murale.

Several field-scale phytoextraction scenarios were created in a greenhouse study to investigate the feasibility of using Alyssum murale, to remediate three types of industrially Ni-contaminated soil (heavy clay, sand, organic muck) from Port Colborne, Ontario. The observed distribution of Ni mass between soil and aboveground vegetation was used in STELLA modeling software to predict timelines for the target soil Ni concentration, namely 1200 mg Ni/kg. Alyssum murale grown in sand would have a relatively constant pool of Ni available for plant uptake, which would not be the case for plants grown in organic muck and heavy clay. The maximum Ni extraction (%, plant Ni mass/soil Ni mass) was achieved in A. murale grown in unfertilized clay soil at the higher irrigation rate. Using these data, the STELLA model predicted that 246 years would be required to reduce soil Ni concentration in the most efficient combination of treatments to the remediation target. In addition, hypothetical A. murale Ni extraction in plant-soil systems optimized by manipulating soil chemistry and physical attributes, were modeled. The most optimized A. murale plant-soil systems for Ni extraction would require 9 years to achieve the same reduction, and it is not clear that this optimization can be achieved in the field. This study showed that phytoremediation using A. murale is not likely a time-sensitive approach for these soils.

Selected Veterinary Concerns of Geriatric Rats, Mice, Hamsters, and Gerbils.

Improved husbandry and better knowledge of exotic pets have led to a gradual increase in the life span of pets, such as rats, mice, hamsters, and gerbils. Much of the information on these senior patients is derived from the laboratory animal studies and anecdotal practitioner information. Although the small size of some of the patients makes blood collection problematic for hematology and organ function testing, the advent of polymerase chain reaction testing and other molecular diagnostics is allowing practitioners to test for specific etiologies with the small biologic samples available. Radiology and ultrasonography also are valuable diagnostic modalities.

Towards an exposure narrative for metals and arsenic in historically contaminated Ni refinery soils: Relationships between speciation, bioavailability, and bioaccessibility.

Archived soils contaminated with Ni, Cu, Co, and As from legacy operations of a nickel refinery at Port Colborne, Ontario, Canada were speciated using mineral liberation analysis. Four Ni minerals were identified as fingerprint compounds of the historical refinery emissions. Cu and Co were present in solid solution in these minerals due to their presence in the refinery's feed. The highest concentrations of Ni, Cu, Co, and As in these soils were 18,553, 1915, 196, and 79mg/kg, respectively, these elevated contaminant concentrations attesting to the importance of incidental soil ingestion to the oral exposure pathway in Port Colborne. The in vitro gastric bioaccessibility (BAc) was determined for these contaminants, as was in vivo oral bioavailability (BAv), using a mass balance approach in male Sprague-Dawley rats. In spite of the elevated soil concentrations of Cu, the BAv of this physiologically important metal could not be distinguished from that in commercial rat chow, suggesting low potential for exposure. Co and As also had low apparent BAv (<2%). For Ni, baseline oral BAv of naturally sourced dietary Ni was found to be approximately 2%, as was the oral BAv of Ni from nickel sulfate hexahydrate. The mass balances of NiSO4·6H2O were fully accounted-for in urine and feces after a single gavage dose, indicating little to no organ incorporation from this highly soluble salt. Therefore, the urinary estimates of Ni BAv for these soils were assumed to represent true BAv despite variable fecal recoveries. The high Ni concentrations enabled BAc-BAv relationships to be developed for these contaminated soils. For absolute bioavailability (ABA) and relative bioavailability (RBA) the relationships were: ABA=0.0116(BAc)-0.0479 and RBA=0.5542(BAc)-2.2817. These findings will advance the development of robust exposure narratives for soil metal contamination in Port Colborne and elsewhere.

Bioaccessibility estimates by gastric SBRC method to determine relationships to bioavailability of nickel in ultramafic soils.

Frameworks for human health risk assessment often include the opportunity to correct the estimate of exposure for bioavailability, which could be predicted from bioaccessibility. Lead and As are the only metallic elements for which bioavailability and bioaccessibility have been correlated across a spectrum of mineralogy and particle types. The objective of the present study is to correlate in vivo bioavailability with ex vivo bioaccessibility for elevated Ni in soils of ultramafic origin and explore attribution of any variation in this correlation to mineralogical characterization of the Ni. Ultramafic soils were field collected in British Columbia, CA. Rietveld quantitative X-ray diffraction was used for the characterization and quantification of crystalline materials containing Ni. Bioaccessible Ni was determined using the in vitro method developed by the Solubility/Bioaccessibility Research Consortium. Bioavailable Ni was determined by gavage dose of the soils to Sprague-Dawley rats. Urine and feces were collected every 24 h. At the end of 72 h, the animals were humanely sacrificed using carbon dioxide as per the approved animal care protocol. All organs were harvested, washed and preserved. Fecal elimination of gavaged Ni ranged from 35 to 95% including positive control. Relative bioavailability (RBA) ranged from 5 to 18%. In vitro bioaccessibility (IVBA) of soil Ni ranged from 0 to 17%; IVBA explained 86% of the variation in RBA. Normalizing both axes to soil olivine accounted for an additional 10% of the variation in RBA. For risk assessment of Ni contaminated soils, IVBA would be a useful and cost effective tool in estimating exposure of mammals through ingestion of soil particles, with some additional benefit of considering Ni mineralogy.

Comparison of nickel speciation in workplace aerosol samples using sequential extraction analysis and X-ray absorption near-edge structure spectroscopy.

There is a pressing need to further develop speciation knowledge of Ni workplace aerosols as the Zatka sequential extraction method used until now to speciate workplace Ni exposures has limitations. Here we compare the Zatka and XANES methods and evaluate XANES spectroscopy as a more appropriate and accurate technique for identifying nickel species in workplace aerosols. XANES spectroscopy is capable of identifying unique Ni species in the unaltered samples. Our findings indicate some significant departures in speciation assignment between the Zatka and XANES methods. In particular, the Zatka method can overestimate the soluble Ni fraction and it may underestimate the sulphidic and metallic fractions in some samples. Of particular importance, XANES is able to identify component sulphidic species. This information can lead to more accurate exposure matrices and more refined epidemiological analysis of respiratory cancer causation in sulphidic Ni processing.

Mycotoxins in South African foods: a case study on aflatoxin M1 in milk.

The contamination of cow's milk at the farm level with aflatoxin M1 was investigated in South Africa. Samples of feeds, forage, maize and milk were taken at nine dairy farms, and at the same time samples of the processed milk (retail milk) were collected from the respective dairies to which the farms delivered their milk. The feeds were analysed for aflatoxin B1 and the milk samples for aflatoxin M1 using high performance liquid chromatography (HPLC) and fluorescence detection. All milk samples from the dairy farms were positive for aflatoxin M1, ranging from 0.02 µg/l to 1.5 µg/l. Retail milk was also frequently contaminated with AFM1, at levels of 0.01-3.1 µg/l. High AFB1 levels in feed materials on the farms supplying the raw milk indicate that various sources account for this contamination frequency in milk.

Natural multi-occurrence of mycotoxins in rice from Niger State, Nigeria.

Twenty-one rice samples from field (ten), store (six) and market (five) from the traditional rice-growing areas of Niger State, Nigeria were analysed for aflatoxins (AFs), ochratoxin A (OTA), zearalenone (ZEA), deoxynivalenol (DON), fumonisin B(1) (FB(1)) and B(2) (FB(2)), and patulin (PAT) by thin-layer chromatography (TLC) and high-performance liquid chromatography (HPLC) respectively. T-2 toxin was determined using TLC only. AFs were detected in all samples, at total AF concentrations of 28-372 µg/kg. OTA was found in 66.7% of the samples, also at high concentrations (134-341 µg/kg) that have to be considered as critical levels in aspects of nephrotoxicity. ZEA (53.4%), DON (23.8), FB(1) (14.3%) and FB(2) (4.8%) were also found in rice, although at relatively low levels. T-2 toxin was qualitatively detected by TLC in only one sample. Co-contamination with AFs, OTA, and ZEA was very common, and up to five mycotoxins were detected in a single sample. The high AF and OTA levels as found in rice in this study are regarded as unsafe, and multi-occurrences of mycotoxins in the rice samples with possible additive or synergistic toxic effects in consumers raise concern with respect to public health.

In vitro estimates of bioaccessible nickel in field-contaminated soils, and comparison with in vivo measurement of bioavailability and identification of mineralogy.

This study determined nickel (Ni) bioaccessibility in weathered smelter-contaminated soils, separately for particle-sized fractions using two in vitro methods: simulated gastrointestinal digestion (PBET) and PBET followed by absorption by Caco-2 cells. Relative bioavailability of Ni in soils was determined in vivo using rats, validating in vitro estimates; a mineralogical basis of variation in bioavailability/bioaccessibility among soils was explored. In vitro assays identified the same difference in bioaccessibility for Ni among particle size fractions. PBET estimates were more precise, thus likely to be more useful in discerning differences among soils. In vivo bioavailability for Ni was below limit of detection for the small soil particles, and 31% and 56% for the larger particles. The relative bioavailability calculated from this work suggests that risk from ingesting Ni-contaminated soils could be overestimated by between 2- and 50-fold if the estimates of exposure are not adjusted for the lower bioavailability of weathered Ni originating from smelter emissions. The overestimation that would occur by using total Ni is greatest for the particle size that is most likely to adhere to the hands of children, demonstrating the importance of particle-size separation of soils for bioavailability determination and risk assessment.

The African Fusarium/maize disease.

There is a general but rather vague feeling that the use of maize (corn) as a staple foodstuff by black rural Africans is somehow a factor in, or is linked to, chronic disease found in these populations. An attempt is made in this review to consider the evidence for this connection and to identify what is actually the root of the problem. The main thrust of the argument to explain this perception is that maize is routinely contaminated with fungi and of these Fusarium verticillioides is found in maize throughout the world. Evidence is presented to this effect and, further, of the mycotoxins found in maize, the fumonisins are the most common and at the highest levels. Various animal chronic diseases arising from the consumption of contaminated maize are reviewed and possible human conditions listed, in some cases related to the known animal ones. A brief overview of the complicated cellular mechanisms of fumonisin B1 is given and it is concluded that the prime suspect in what might be called "the maize disease" can be attributed to the ingestion of this mycotoxin, sometimes in combination with other synergist mycotoxins and other disease factors, such as smoking and drinking.

The cytotoxic effect of fumonisin B1 and ochratoxin A on human and pig lymphocytes using the Methyl Thiazol Tetrazolium (MTT) assay.

Lymphocytes cell obtained from healthy human donors and pigs were exposed to fumonisin B1 (FB1) and ochratoxin A (OTA), which have been found to be immunosuppressive, carcinogenic and mutagenic, to ascertain their single and combined cytotoxic effects with time and to assess the suitability of animal lymphocytes as test agents in comparison to human cells. The main objectives of this work were to assess the use of animal lymphocytes, particularly pig lymphocytes, for their use in the Methyl Thiazol Tetrazolium (MTT) cytotoxicity test, making them more accessible to animal research-based institutes in comparison to human lymphocytes previously used, and to study the cytotoxic and synergism or antagonistic effects of FB1 and OTA. The MTT assay, which measures cell viability and proliferation based on reduction of MTT to a blue dye, also used the addition of phytohaemagglutinin (PHA) to stimulate the blood cells. The results showed a progressive decrease in lymphocytes viability with time of exposure to the toxins. It was also noted that FB1, as compared to OTA, had a lower cytotoxicity on both human and pig lymphocytes cells. In addition, when the two mycotoxins were combined, a synergistic decrease of cell viability in both human and pig lymphocytes was observed, with pig lymphocytes showing a greater sensitivity. This study has shown that the MTT assay can be used for the determination of cytotoxicity of mycotoxins using animal, and in particular pig, lymphocytes, which eliminates the use of human donors and other cell cultures.

The incidence of fungi and mycotoxins in South African barley and barley products.

Barley is grown as a crop in South Africa, mainly for use in the barley beer (lager) industry, particularly in the production of barley malt. This investigation was done to find out what fungal infection and mycotoxin contamination this barley and the malt contained. The survey, done in 2005, not only covered barley and malt but also the final product in the form of several brands of beer purchased from retail outlets. Analysis was done using a range of methodologies including chromatography, immunoaffinity/fluorimetry and cytotoxicity testing. The results show that barley, which also includes barley sold directly to the public, and malt, contain various fungi, sometimes at high incidence and also a range of mycotoxins which persisted through, although at low levels, to beer. The malt showed a different pattern of micro flora, as compared to the barley, which indicates infection during the malting process, which is not uncommon. Interestingly, fumonisin B1 was found in some of the samples, as well as the beer, although in the latter case these were at very low levels. Recent studies have shown that the immunoaffinity/fluorimetry method can give false positives for the fumonisins, so these results were confirmed by high performance liquid chromatography. As is the case with other cereals grown in South Africa, there is the concern that the public is exposed to mycotoxins on a regular basis through the consumption of products made from them.

Kinetics of the oxidoreductase involved in the conversion of O-methylsterigmatocystin to aflatoxin B1.

Among the enzymatic steps in the aflatoxin biosynthetic pathway, the conversion of O-methylsterigmatocystin (OMST) to the potent environmental carcinogen aflatoxin B1 (AFB1), has been proposed to be catalysed by an oxidoreductase (OR) that requires a cytochrome P-450 type of oxidoreductase activity. This enzyme displays relative specificity towards OMST homologues in fungal whole cells. These studies were extended to the action of a cell-free enzyme system (CFES), on five OMST homologues, with a view to establish the kinetics. In the current study a CFES, containing an oxidoreductase, was derived from a blocked mutant of Aspergillus parasiticus (Wh1-11-105). The key experimental steps involved rapid concentration and efficient dialysis by membrane filtration to remove small biomolecules (MW<10,000), co-factors, primary and secondary metabolites. The kinetic parameters of the enzyme-substrate reactions indicated that the reaction follows a Michealis-Menten kinetics and OR activity decreased in the order: O-butylsterigmatocystin>O-propylsterigmatocystin>O-ethylsterigmatocystin>O-methylsterigmatocystin>O-acetylsterigmatocystin>O-benzoylsterigmatocystin. The 7-O-alkyl homologues were the best substrate for the CFES, thereby substantially supporting that the 7-O-methyl group of OMST is preferred for OR catalytic activity in the absence of any other alkylating groups in vitro. The Km was calculated as 5.65 microM for this CFES and varied marginally among the OMST homologues studied.

The incidence of fungi and mycotoxins in South Africa wheat and wheat-based products.

This investigation was undertaken to survey the fungal and mycotoxin contamination of South African wheat ranging from that growing in the field to processed wheat products. Samples of wheat were taken from various growing areas in South Africa and screened for fungi and mycotoxins, using a range of methodologies, including chromatography, immunoaffinity/fluorimetry, and cytotoxicity testing. Similar samples were taken from supermarkets and retail outlets in South Africa and analyzed in a similar manner. The result showed that a range of fungi and mycotoxins could be detected in wheat in all these sample types. The major fungal contaminants were Fusarium spp. and their attendant mycotoxins, in particular deoxynivalenol, which is in keeping with the observations made in the rest of the world. An interesting observation was that samples of wheat taken from the field with heavy Fusarium contamination were contaminated with fumonisin B1, which is not normally associated with this crop. Of more concern were the low but persistent levels of mycotoxins and fungi in wheat-based products sold directly to the public.

The frequency of cytochrome P450 2E1 polymorphisms in Black South Africans.

Polymorphisms in the promoter region of the Cytochrome P4502E1 (CYP2E1) gene reportedly modify the metabolic activity of CYP2E1 enzyme, and have been associated with increased susceptibility to squamous cell carcinoma (SCC) of the oesophagus in high prevalence areas such as China. To assess the frequency of these polymorphisms in Black South Africans, a population with a high incidence of oesophageal SCC, this study examined genomic DNA from 331 subjects for restriction fragment length polymorphisms in the CYP2E1 (RsaI and PstI digestion). The frequency of the CYP2E1 c1/c1 and c1/c3 genotypes was 95% and 5% respectively. The frequency of the CYP2E1 allele distribution was found to be markedly different between Chinese and South African populations; hence it is important to place racial differences into consideration when proposing allelic variants as genetic markers for cancer.