Optimisation of plutonium separations using TEVA cartridges and ICP-MS/MS analysis for applicability to large-scale studies in tropical soils.

The analysis of plutonium (Pu) in soil samples can inform the understanding of soil erosion processes globally. However, there are specific challenges associated for analysis in tropical soils and so an optimal analytical methodology ensuring best sensitivity is critical. This method aimed to demonstrate the feasibility of sample preparation and analysis of Pu isotopes in African soils, considering the environmental and cost implications applicable to low-resource laboratories. The separation procedure builds upon previous work using TEVA columns, further demonstrating their usefulness for the reduction of uranium (U) interference in ICP-MS analysis with enhanced selectivity for Pu. Here several steps were optimised to enhance Pu recovery, reducing method blank concentration, and improving the separation efficiency through the determination of the elution profiles of U and Pu. The elimination of the complexing agent in the eluent, increased the spike recovery by improving matrix tolerance of the plasma, and simplified the separation procedure, improving throughput by 20%. The subsequent method was validated through the analysis of Certified Reference Material IAEA-384, where high accuracy and improved precision of measurement were demonstrated (measured value 114 ± 12 versus certified value 108 ± 13 Bq kg-1). Optimisation of the column separation, along with the analysis of the samples using O2 gas in ICP-MS/MS mode to mass shift Pu isotopes away from interfering molecular U ions provided a simple, robust, and cost-effective method with low achievable method detection limits of 0.18 pg kg-1 239+240Pu, applicable to the detection of ultra-trace fallout Pu in African soils.

Artisanal gold mining in Kakamega and Vihiga counties, Kenya: potential human exposure and health risk.

Artisanal and small-scale gold mining (ASGM) represents 20% of gold supply and 90% of gold mining workforce globally, which operates in highly informal setups. Pollutants from mined ores and chemicals introduced during gold processing pose occupational and inadvertent health risks to the extent that has not been well elucidated in Africa. Trace and major elements were analysed using inductively coupled plasma mass spectrometry in soil, sediment and water samples from 19 ASGM villages in Kakamega and Vihiga counties. Associated health risks for residents and ASGM workers were assessed. This paper focuses on As, Cd, Cr, Hg, Ni and Pb for which 96% of soil samples from mining and ore processing sites had As concentrations up to 7937 times higher than the US EPA 12 mg kg-1 standard for residential soils. Soil Cr, Hg and Ni concentrations in 98%, 49% and 68% of the samples exceeded respective USEPA and CCME standards, with 1-72% bioaccessibility. Twenty-five percentage of community drinking water sources were higher than the WHO 10 µg L-1 drinking water guideline. Pollution indices indicated significant enrichment and pollution of soils, sediment and water in decreasing order of As > Cr > Hg > Ni > Pb > Cd. The study revealed increased risks of non-cancer health effects (98.6) and cancer in adults (4.93 × 10-2) and children (1.75 × 10-1). The findings will help environment managers and public health authorities better understand the potential health risks in ASGM and support evidence-based interventions in ASGM processes, industrial hygiene and formulation of public health policy to protect residents and ASGM workers' health in Kenya.

Method development to characterise elephant tail hairs by LA-ICP-MS to reflect changes in elemental chemistry.

This paper evaluated analytical methods used to generate time-series data from elephant tail hairs, which can be used to reflect changing exposure to environmental geochemistry. Elephant tail hairs were analysed by three methods sequentially, each providing data to inform subsequent analysis. Scanning Electron Microscopy (SEM) and X-ray Microanalysis visually showed the structure of the hair, specific structures such as tubules, and the mineral crusting around the edge of the hair, informing targeting of subsequent analysis by Laser Ablation-Inductively Coupled Plasma Mass Spectrometry (LA-ICP-MS). LA-ICP-MS generated time-series data which informed sectioning of the tail hairs for subsequent quantitative analysis for potentially toxic elements and micronutrients using Inductively Coupled Plasma Mass Spectrometry (ICP-MS) of dissolved tail hairs. This novel approach to characterise the tail hair enabled time-series analysis to reflect changes in environmental exposure which may result from seasonal or geochemical spatial variation and could inform elephant movement patterns. The seasonal change between wet and dry seasons was reflected down the length of the hair. Correlations were seen between LA-ICP-MS data and ICP-MS data in several elements including Mg, P, Ca, Fe, Na, Mn and U. This study provided time-series data for the analysis of elephant tail hairs by evaluating analytical challenges to obtaining quantitative data, such as improving protocols to ensure removal of extraneous material, determining where to section the tail hairs to best reflect environmental changes/exposure and ensuring representative analyses. A protocol was established to determine mineral status across a 12-18 month time period utilizing single elephant tail hairs.

Assessment of bioaccessibility and health risk of mercury within soil of artisanal gold mine sites, Niger, North-central part of Nigeria.

The occurrence of mercury (Hg) in the environment globally has been linked largely to its use for gold processing. In this research, ore samples, agricultural soil and mine wastes were taken within the vicinity of an artisanal gold mine and processing sites in Niger state, a north-central part of Nigeria to determine Hg contamination in the environment and estimate the potential hazard to health. The values of Hg measured in ore, agricultural soil and mine wastes ranged between 0.03 and 5.9, 0.002 and 5.57 and 0.19 and 20.99 mg/kg, respectively, with the majority of samples observed above the crustal average values of 0.003 mg/kg. All of the samples were 100 times greater than the USEPA residential soil screening level of 0.0023 mg/kg, but were lower than comparable mine sites within the same region. Contamination indices were used to demonstrate the potential exposure to Hg contamination in the study area which ranged from a medium to high level of contamination. Average daily dose and hazard quotient (HQ) were calculated for adults and children in the study area and decreased in the following order: ADDvapour > ADDingestion > ADDdermal > ADDinhalation. The non-carcinogenic health risk index (HI) of Hg calculated for children and adults in the study area was children: 7.42, 2.19, 1.49 and adults: 4.45, 1.26, 1.19, for mine wastes, agricultural soil and ore, respectively. All of these values were higher than a considered safe level (= 1) and therefore showed that Hg posed a serious non-carcinogenic HI for both adults and children exposed to the soil in the study area. The bioaccessible fraction as a measure of ingestion for Hg was generally < 13% across all sample matrices, suggesting a low bioaccessibility. An HQ incorporating bioaccessible data (BHQ) ranged between 0.000005 and 4.06 with a mean value of 0.62. Values for the BHQ were still > 1, threshold limit in some samples and showed that Hg could present a risk to health via ingestion, although further research is required to assess dermal and inhalation bioaccessibility to assess fully the risk to residents. However, the values were lower than the non-carcinogenic health risk index, which is assumed to be overestimated.

Assessment of chromium species dynamics in root solutions using isotope tracers.

BACKGROUND: Chromium (Cr) exists in the environment in two chemical forms; CrIII is an essential micronutrient for glucose and lipid metabolism, whereas CrVI is toxic and a recognised carcinogen through inhalation. Numerous studies have attempted to evaluate their transfer mechanisms from soil and solution media into plants, usually with respect to the hyperaccumulation, detoxification and tolerance of the plant to CrVI. METHODS: Isotopically enriched species of Cr, added as 50CrIII and 53CrVI, were used to investigate transfer from solution into the root systems of Spinacia oleracea. In addition the effect of sulphate (SO42-), as a competitor for CrVI uptake, was investigated. Separation of 50CrIII and 53CrVI was undertaken using HPLC-ICP-QQQ following isolation of root solutions using freeze/thaw centrifugation. RESULTS: Irrespective of supplied CrVI concentration (250, 500 or 1000 µg L-1), the dominant species in both apoplastic (routed through cell wall and intercellular space as a passive mechanism) and symplastic (routed through cytoplasm as an active mechanism) root solutions was CrIII. There was evidence for CrVI reduction in the rhizosphere prior to uptake as an additional detoxification mechanism. Sulphate promoted uptake of CrVI through the active pathway, although increases in SO42- concentration did not yield a proportional increase in Cr symplastic solution concentration; CrIII was also the dominant species in these root solutions. CONCLUSION: The results indicate that Spinacia oleracea plants can effectively reduce CrVI to CrIII and that the uptake pathways for both CrIII and CrVI are more complex than previously reported. Further work is required to understand the physiological processes that result in the reduction of CrVI prior to, and during, uptake. The efficacy of sulphate to augment existing agricultural management strategies, such as liming and organic reincorporation, also requires further investigation to establish suitable application rates and applicability to other environmental contaminants.

Arsenic speciation in the bracket fungus Fomitopsis betulina from contaminated and pristine sites.

Uptake, distribution and speciation of arsenic (As) were determined in the bracket fungus Fomitopsis betulina (previously Piptoporus betulinus), commonly known as the birch polypore, collected from a woodland adjacent to a highly contaminated former mine in the Southwest UK and at an uncontaminated site in Quebec, Canada, with no past or present mining activity. The fruiting body was divided into cap, centre and pores representing the top, middle and underside to identify trends in the distribution and transformation of As. Total As, determined by inductively coupled plasma-mass spectrometry (ICP-MS), was approximately tenfold higher in the mushroom from the contaminated compared to the uncontaminated site. Overall, accumulation of As was low relative to values reported for some soil-dwelling species, with maximum levels of 1.6 mg/kg at the contaminated site. Arsenic speciation was performed on aqueous extracts via both anion and cation high-performance liquid chromatography-ICP-MS (HPLC-ICP-MS) and on whole dried samples using X-ray absorption near edge structure (XANES) analysis. Seven As species were detected in F. betulina from the contaminated site by HPLC-ICP-MS: arsenite (AsIII), arsenate (AsV), dimethylarsinate (DMAV), methylarsonate (MAV), trimethylarsine oxide (TMAO), tetramethylarsonium ion (Tetra) and trace levels of arsenobetaine (AB). The same As species were observed at the uncontaminated site with the exception of TMAO and Tetra. Arsenic species were localized throughout the fruiting body at the contaminated site, with the cap and pores containing a majority of AsV, only the cap containing TMAO, and the pores containing higher concentrations of DMAV and MAV as well as tetra and a trace of AB. XANES analysis demonstrated that the predominant form of As at the contaminated site was inorganic AsIII coordinated with sulphur or oxygen and AsV coordinated with oxygen. This is the first account of arsenic speciation in F. betulina or any fungi of the family Fomitopsidaceae.

Environmental geochemistry and cancer: a pertinent global health problem requiring interdisciplinary collaboration.

Primary prevention is a key strategy to reducing the global burden of cancer, a disease responsible for ~ 9.6 million deaths per year and predicted to top 13 million by 2030. The role of environmental geochemistry in the aetiology of many cancers-as well as other non-communicable diseases-should not be understated, particularly in low- and middle-income countries where 70% of global cancer deaths occur and reliance on local geochemistry for drinking water and subsistence crops is still widespread. This article is an expansion of a series of presentations and discussions held at the 34th International Conference of the Society for Environmental Geochemistry and Health in Livingstone, Zambia, on the value of effective collaborations between environmental geochemists and cancer epidemiologists. Key technical aspects of each field are presented, in addition to a case study of the extraordinarily high incidence rates of oesophageal cancer in the East African Rift Valley, which may have a geochemical contribution. The potential merit of veterinary studies for investigating common geochemical risk factors between human and animal disease is also highlighted.

Fertilizer management and soil type influence grain zinc and iron concentration under contrasting smallholder cropping systems in Zimbabwe.

Micronutrient deficiencies remain prevalent in food systems of southern Africa, although advances in biofortification through crop breeding and agronomy provide opportunities to address these. We determined baseline soil availability of zinc (Zn) and iron (Fe) and the effects of soil type and farmer management on extractable soil Zn and Fe and subsequent concentration in cereal and legume grains under two contrasting agro-ecologies in Zimbabwe. Soil and crop surveys were conducted in Hwedza and Mutasa Districts of Zimbabwe in 2015-16 on 350 locations over different soil types. Fields with different levels of productivity (designated as "most" and "least" productive fields) were sampled using an inherited hierarchical randomized sampling design. Grain Zn and Fe concentration in maize (Zea mays), sorghum (Sorghum bicolor), finger millet (Eleusine coracana) and cowpea (Vigna unguiculata) were generally insufficient for adequate human nutrition. A Linear Mixed Effects (LME) model revealed that diethylene triamine penta-acetic acid- (DTPA) extractable soil Zn concentration and grain Zn concentration were affected primarily by field productivity level. DTPA-extractable soil Zn concentration was more than two-fold greater on the most productive fields (mean 0.8 mg kg-1) than on the least productive fields, with mean grain Zn concentration of 25.2 mg grain Zn kg-1 which was 13% greater than seen on the least productive fields.  An interaction effect of field productivity level and total soil Zn concentration on DTPA-extractable soil Zn concentration suggests potential contribution of organic matter management to unlocking unavailable forms of soil Zn. DTPA-extractable soil Fe and grain Fe concentration were primarily affected by soil type and crop type, respectively. The LME modelling approach revealed additional soil geochemical covariates affected DTPA-extractable soil Zn and Fe concentration and grain Zn and Fe concentration within Districts. Future studies can therefore be powered to detect their roles at wider spatial scales for sustainable management of crop Zn and Fe nutrition.

Intra-household agreement of urinary elemental concentrations in Tanzania and Kenya: potential surrogates in case-control studies.

Element deficiencies and excesses play important roles in non-communicable disease aetiology. When investigating their roles in epidemiologic studies without prospective designs, reverse-causality limits the utility of transient biomarkers in cases. This study aimed to investigate whether surrogate participants may provide viable proxies by assessing concentration correlations within households. We obtained spot urine samples from 245 Tanzanian and Kenyan adults (including 101 household pairs) to investigate intra-household correlations of urinary elements (As, Ba, Ca, Cd, Co, Cs, Cu, Fe, Li, Mn, Mo, Ni, Pb, Rb, S, Se, Sr, Tl, V and Zn) and concentrations (also available for: Bi, Ce, Sb, Sn and U) relative to external population-levels and health-based values. Moderate-strong correlations were observed for As (r = 0.65), Cs (r = 0.67), Li (r = 0.56), Mo (r = 0.57), Se (r = 0.68) and Tl (r = 0.67). Remaining correlations were <0.41. Median Se concentrations in Tanzania (29 µg/L) and Kenya (24 µg/L) were low relative to 5738 Canadians (59 µg/L). Exceedances (of reference 95th percentiles) were observed for: Co, Mn, Mo, Ni and U. Compared to health-based values, exceedances were present for As, Co, Mo and Se but deficiencies were also present for Mo and Se. For well correlated elements, household members in East African settings provide feasible surrogate cases to investigate element deficiencies/excesses in relation to non-communicable diseases.

Chromium speciation in foodstuffs: A review.

Numerous critical reviews have evaluated exposure to toxic and carcinogenic hexavalent chromium (Cr(VI)) from a number of pathways; including workplace air, cement and packaging materials. The contribution of foodstuffs to dietary Cr(VI) has been increasingly under investigation, however no summary of this work has been carried out. The objective of this article is to review the last twenty years of chromium speciation research in foodstuffs. Alkaline extraction, used for chromium speciation in other solids, is the most widely-reported procedure. Previous measurement of Cr(VI) in foodstuffs is questionable due to the reducing power of organic matter and antioxidants, leading to the development of speciated isotope dilution mass spectrometry (SIDMS) techniques to monitor interconversions. Evaluation of the genotoxicity of trivalent chromium (Cr(III)), which acts through a different pathway to that of Cr(VI), requires reconsideration towards measurement of Cr(III), which is present at higher concentrations in foodstuffs following reduction of the more-bioavailable Cr(VI).

Spatial distribution and human health risk assessment of mercury in street dust resulting from various land-use in Ahvaz, Iran.

Mercury as a toxic element and its associated health hazard has been an important topic of research for urban pollution for many years. In this paper, the spatial distribution, pollution assessment, and health risk associated with Hg in roadside dust 96 street dust samples, representing differing land-uses, have been investigated. Land-uses included residential areas (RA), industrial areas (IA), public gardens (PG), roadside areas (RS), and suburban areas (SA) in the city of Ahvaz, Iran were investigated. Compared with other cities, the concentration of Hg in Ahvaz was considerably higher with a mean value of 2.53 mg kg-1, ranging from 0.02 to 8.75 mg kg-1. Residential areas exhibited higher Hg in street dust than other areas, as demonstrated by spatial mapping illustrating hot spots associated with old urban areas with high residential density, high volume traffic of roadside areas, and industrial districts: including oil-drilling activities, steel smelting-related industries, and small industrial towns around Ahvaz. However, Hg concentrations in street dust near to the public gardens (PG) and suburban areas (SA) were not at elevated levels compared other land-uses investigated in this study. A health risk assessment model of non-carcinogenic effects was evaluated for both children and adults. The HQ values also revealed that the main exposures route for children and adults decreased as follows: vapour > ingestion > dermal contact > inhalation. The hazard index (HI) in each area is less than the safe level (HI ≤ 1) for children and adults, but higher for children. The HI value decreases as the following order: RS > IA > RA > SA > PG, which indicates potentially serious health hazards for children in the study areas.

Optimisation and quality control of cell processing for autologous stem cell transplantation.

Clinical practice and the technology of cell processing for autologous stem cell transplantation has continued to evolve over the last two decades and merits review of current quality control expectations. The external regulatory era has improved quality and safety standards but there is still variable practice, with specific risks illuminated by a number of clinical incidents. Viable CD34+ cell assays may fail to indicate significant losses in progenitor function during storage, particularly after cryopreservation, and there is a need to develop an alternative, real time functional assay to replace colony assays. The ultimate guide to potency and successful cell processing for haematopoietic progenitor cell products is prompt and reproducible engraftment and close monitoring is essential for safety and quality control.

Re-evaluation of progenitor thresholds and expectations for haematopoietic recovery based on an analysis of 810 autologous transplants: Implications for quality assurance.

Haematological engraftment was assessed in 804 autologous transplants. Neutrophil recovery occurred in over 99% within 14 d but platelet recovery was delayed beyond this time in 14·8%. Time to recovery was dependent on the progenitor cell dose infused. The minimum CD34+ cell threshold adopted in this study (2 × 106 /kg) was safe although recovery was faster with a dose >5 × 106 /kg. CD34+ cell doses of between 1 and 2 × 106 /kg were also acceptable if either the granulocyte-macrophage colony-forming cell dose exceeded 2 × 105 /kg or this dose was due to splitting a higher yield harvest. Prompt neutrophil recovery affords important quality assurance for laboratory processing.

Post-thaw viability of cryopreserved peripheral blood stem cells (PBSC) does not guarantee functional activity: important implications for quality assurance of stem cell transplant programmes.

Standard quality assurance (QA) of cryopreserved peripheral blood stem cells (PBSC) uses post-thaw viable CD34(+) cell counts. In 2013, concerns arose at Great Ormond Street Hospital (GOSH) about 8 patients with delayed engraftment following myeloablative chemotherapy with cryopreserved cell rescue, despite adequate post-thaw viable cell counts in all cases. Root cause analysis was undertaken; investigations suggested the freeze process itself was a contributing factor to suboptimal engraftment. Experiments were undertaken in which a single PBSC product was divided into three and cryopreserved in parallel using a control-rate freezer (CRF) or passive freezing method (-80°C freezer) at GOSH, or the same passive freezing at another laboratory. Viable CD34(+) counts were equivalent and adequate in each. Granulocyte-monocyte colony-forming unit assays demonstrated colonies from the products cryopreserved using passive freezing (both laboratories), but no colonies from products cryopreserved using the CRF. The CRF was shown to be operating within manufacturer's specifications with freeze-profile within acceptable limits. This experience has important implications for quality assurance for all transplant programmes, particularly those using cryopreserved products. The failure of post-thaw viable CD34(+) counts, the most widely used routine QA test available, to ensure PBSC function is of great concern and should prompt reassessment of protocols and QA procedures.

Assessing urinary flow rate, creatinine, osmolality and other hydration adjustment methods for urinary biomonitoring using NHANES arsenic, iodine, lead and cadmium data.

BACKGROUND: There are numerous methods for adjusting measured concentrations of urinary biomarkers for hydration variation. Few studies use objective criteria to quantify the relative performance of these methods. Our aim was to compare the performance of existing methods for adjusting urinary biomarkers for hydration variation. METHODS: Creatinine, osmolality, excretion rate (ER), bodyweight adjusted ER (ERBW) and empirical analyte-specific urinary flow rate (UFR) adjustment methods on spot urinary concentrations of lead (Pb), cadmium (Cd), non-arsenobetaine arsenic (As(IMM)) and iodine (I) from the US National Health and Nutrition Examination Survey (NHANES) (2009-2010 and 2011-2012) were evaluated. The data were divided into a training dataset (n = 1,723) from which empirical adjustment coefficients were derived and a testing dataset (n = 428) on which quantification of the performance of the adjustment methods was done by calculating, primarily, the correlation of the adjusted parameter with UFR, with lower correlations indicating better performance and, secondarily, the correlation of the adjusted parameters with blood analyte concentrations (Pb and Cd), with higher correlations indicating better performance. RESULTS: Overall performance across analytes was better for Osmolality and UFR based methods. Excretion rate and ERBW consistently performed worse, often no better than unadjusted concentrations. CONCLUSIONS: Osmolality adjustment of urinary biomonitoring data provides for more robust adjustment than either creatinine based or ER or ERBW methods, the latter two of which tend to overcompensate for UFR. Modified UFR methods perform significantly better than all but osmolality in removing hydration variation, but depend on the accuracy of UFR calculations. Hydration adjustment performance is analyte-specific and further research is needed to establish a robust and consistent framework.

Africa's Oesophageal Cancer Corridor: Geographic Variations in Incidence Correlate with Certain Micronutrient Deficiencies.

BACKGROUND: The aetiology of Africa's easterly-lying corridor of squamous cell oesophageal cancer is poorly understood. Micronutrient deficiencies have been implicated in this cancer in other areas of the world, but their role in Africa is unclear. Without prospective cohorts, timely insights can instead be gained through ecological studies. METHODS: Across Africa we assessed associations between a country's oesophageal cancer incidence rate and food balance sheet-derived estimates of mean national dietary supplies of 7 nutrients: calcium (Ca), copper (Cu), iron (Fe), iodine (I), magnesium (Mg), selenium (Se) and zinc (Zn). We included 32 countries which had estimates of dietary nutrient supplies and of better-quality GLOBCAN 2012 cancer incidence rates. Bayesian hierarchical Poisson lognormal models were used to estimate incidence rate ratios for oesophageal cancer associated with each nutrient, adjusted for age, gender, energy intake, phytate, smoking and alcohol consumption, as well as their 95% posterior credible intervals (CI). Adult dietary deficiencies were quantified using an estimated average requirements (EAR) cut-point approach. RESULTS: Adjusted incidence rate ratios for oesophageal cancer associated with a doubling of mean nutrient supply were: for Fe 0.49 (95% CI: 0.29-0.82); Mg 0.58 (0.31-1.08); Se 0.40 (0.18-0.90); and Zn 0.29 (0.11-0.74). There were no associations with Ca, Cu and I. Mean national nutrient supplies exceeded adult EARs for Mg and Fe in most countries. For Se, mean supplies were less than EARs (both sexes) in 7 of the 10 highest oesophageal cancer ranking countries, compared to 23% of remaining countries. For Zn, mean supplies were less than the male EARs in 8 of these 10 highest ranking countries compared to in 36% of other countries. CONCLUSIONS: Ecological associations are consistent with the potential role of Se and/or Zn deficiencies in squamous cell oesophageal cancer in Africa. Individual-level analytical studies are needed to elucidate their causal role in this setting.

A high prevalence of zinc- but not iron-deficiency among women in rural Malawi: a cross-sectional study.

BACKGROUND: Zinc deficiency is often associated with nutritional iron deficiency (ID), and may be exacerbated by low selenium status. AIM: To investigate risk of iron and zinc deficiency in women with contrasting selenium status. METHODS: In a cross-sectional study, 1-day diet composites and blood samples were collected from self-selected Malawian women aged 18-50 years from low- (Zombwe) (n=60) and high-plant-available soil selenium (Mikalango) (n=60) districts. Diets were analyzed for trace elements and blood for biomarkers. RESULTS: Zinc deficiency (>90 %) was greater than ID anemia (6 %), or ID (5 %), attributed to diets low in zinc (median 5.7 mg/day) with high phytate:zinc molar ratios (20.0), but high in iron (21.0 mg/day) from soil contaminant iron. Zombwe compared to Mikalango women had lower (p<0.05) intakes of selenium (6.5 vs. 55.3 µg/day), zinc (4.8 vs. 6.4 mg/day), iron (16.6 vs. 29.6 mg/day), lower plasma selenium (0.72 vs. 1.60 µmol/L), and higher body iron (5.3 vs. 3.8 mg/kg), although plasma zinc was similar (8.60 vs. 8.87 µmol/L). Body iron and plasma zinc were positive determinants of hemoglobin. CONCLUSION: Risk of zinc deficiency was higher than ID and was shown not to be associated with selenium status. Plasma zinc was almost as important as body iron as a hemoglobin determinant.

Engraftment defect of cytokine-cultured adult human mobilized CD34(+) cells is related to reduced adhesion to bone marrow niche elements.

In vitro exposure of haematopoietic stem and progenitor cells (HSPC) to cytokines in expansion or gene therapy protocols reduces homing and engraftment in vivo. We have previously reported that this is related in part to altered tissue specificity of short-term homing, leading to loss of cells in non-haematopoietic tissues. Here we demonstrate that defective engraftment persists when cultured HSPC are transplanted by intrabone injection. Changes in engraftment function occur within 24 h of cytokine exposure, and are evident when engraftment is analysed solely in the injected bone. A novel ex vivo model of the bone marrow was developed, in which the attachment of infused HSPC in rodent long bones is reduced following culture with cytokines. Finally, cultured HSPC demonstrated reduced adhesion to N-cadherin, osteopontin and vascular cell-adhesion molecule-1, ligands present in bone marrow niches. These changes in adhesive function occur rapidly, and are not related to downregulation of the relevant receptors. Our findings suggest that cytokine exposure of adult human HSPC results in altered adhesion within bone marrow niches, further leading to reduced engraftment potential in vivo.

Peripheral blood stem cell yield in 400 normal donors mobilised with granulocyte colony-stimulating factor (G-CSF): impact of age, sex, donor weight and type of G-CSF used.

Mobilised peripheral blood is now the main source of stem cells collected from normal donors. We report our experience of mobilising and collecting 400 normal healthy donors using standardised procedures and techniques. Target recipient doses were reached with one aphaeresis in 63% of donors and with two aphereses in 81% of donors. Approximately 2% of donors yielded such low progenitor values that they were termed 'poor mobilisers'. There were minor effects of donor age, weight and sex and where possible, larger male donors under the age of 55 years should be selected. Two forms of granulocyte colony-stimulating factor (G-CSF) were used at the same dose and no significant difference was seen in the yield of CD34+ cells collected/l of blood processed. However, a greater number of granulocyte-macrophage colony-forming cells were harvested using lenograstim (glycosylated G-CSF) compared with filgrastim (non-glycosylated G-CSF; P = 0.002). CD34+ cell yields were also measured halfway through the aphaeresis procedure. This was found to be highly predictive of final yield and facilitated distribution of the stem cell product to other centres. The observation that CD34+ yields did not decline in the second half compared with the first half of aphaeresis suggests that the circulating cell numbers are not static.