Established Laboratory Procedures Research Articles

Assessment of the benefits and risks of sewage sludge application as soil amendment for agriculture in Eswatini

Sewage sludge is an important soil conditioner, and source of nutrients with a potential for use in agriculture. However, such benefit needs to be weighed against the risks due to the presence of heavy metals, and other substances that may endanger human health, plants, soil and the ecosystem. This research had the objective of evaluating the agricultural potential of sewage sludge together with the risks that may be present in use. Samples of sewage sludge from seven wastewater treatment plants in Eswatini were analyzed for selected physical and chemical parameters, and heavy metal concentrations using commonly established laboratory procedures. The analysis results indicated that, sludge samples exhibited high organic matter content, cation exchange capacity, macro nutrients such as nitrogen and phosphorus and micro nutrients (trace metal elements) needed for plant growth. Anaerobically digested sludge samples showed higher carbon to nitrogen ratio because of biomass loss in the form of methane and carbon dioxide. The heavy metal concentrations are all within safe limits except the sludge from Matsapha area that had levels of chromium, and nickel above regulatory limits. With respect to their heavy metal contents, most of the sludge samples would qualify as Class A sludge ready to be used as organic fertilizer for agriculture without regular monitoring of heavy metals in the soil to which they are applied. The study results indicated the importance of sewage sludge for their agricultural potential through supplementation of both micro and macro nutrients needed for plant growth, for improving the soil properties such as water holding capacity and permeability, through the increase of organic matter content, and retention of nutrients due to the high cation exchange capacity. All these benefits are realized with sewage sludge being a lower cost alternative to commercial fertilizers. Moreover, the low concentration of heavy metals present in the sludge presents lower risk that may arise in the course of utilizing the sludge as agricultural supplement. Key words: Agriculture, fertilizer, heavy metals, nutrient, sewage, sludge, nitrogen, phosphorous

Investigatory study on the qualities of locally grown Rice brands sold in main markets of Abeokuta, South-Western Nigeria

Qualities of some selected rice varieties sold in the main markets of Abeokuta, the capital of Ogun State where rice is mostly grown and consumed in the south-western part of Nigeria was investigated. Rice (Aroso, Mokwa, Royal niger, Orlam and Ofada rice) were evaluated for their physical, chemical properties and sensory qualities using established laboratory procedures. The average width of the rice ranged from “0.261 to 0.268 mm”, “0.260 to 0.265 mm, “0.260 to 0.262 mm”, “0.269 to 0.270 mm”, and 0.271 to 0.275 mm” respectively. The rice average lengths ranged between “0.346 to 0.350 mm”, “0.365 to 0.368 mm”, “0.356 to 0.359 mm”, “0.357 to 0.357 mm” and “0.365 to 0.379 mm” respectively. The physical appearance of the rice investigated ranged from good to very good except for mokwa and orlam rice which shows fair colours. Chemical properties of the rice samples ranged from 10.21 to 11.97%, 0.83 to 2.62, 1.89 to 2.16%, 0.24 to 0.37%, 5.50 to 7.56%, 77.89 to 79.60%, 59.90 to 65.17%, 12.15 to 14.61% and 85.39 to 87.85% for moisture, total ash, fat, crude fibre, crude protein, carbohydrate, starch, amylose and amylopectin contents respectively. The cooking properties ranged from 2 to 7.5, 230 to 250% and 5.49 to 5.64 in length and 0.689 to 1.337mm for volume expansion ratio, water uptake and elongation ratio respectively. This study revealed that Aroso and Orlam rice were of excellent in physical, chemical, sensory qualities and are both recommended for consumption. This research finding serves as a basis of choice for consumers and it also reveals the areas for improvement in terms of qualities of the local rice investigated.

Effect of short-term grazing exclusion on herbage species composition, dry matter productivity, and chemical composition of subtropical grasslands

Abstract Grazing exclusion (GE) is a useful management technique for restoring degraded grasslands. The herbage mass productivity and chemical makeup in the grazing-excluded subtropical grassland environment has, however, received little attention. A subtropical riverine grassland was selected to determine the effect of GE on herbage mass productivity and chemical composition in Nepal. In three successive harvesting times from September to November 2020, the herbage was sampled along the six randomly selected transects of 100 m length and at two treatments (GA: Grazing-allowed and GE: Grazing-excluded plots) at three different times of harvest from a 1,000-ha grassland. A total of 108 herbage cut samples were collected from the individual 1 m × 1 m quadrats at the three harvests, respectively, from the GA and GE plots. Fences were used to maintain the GE plots to avoid grazing to prevent the vegetation altered by grazing. Day before herbage sampling, the functional groups, cover-abundance within the sampling quadrats were investigated. By cutting the fresh herbage 5 cm above the ground and subjecting it to oven drying for laboratory examination, the herbage mass productivity within each quadrat was measured. Using established laboratory procedures, the chemical analysis of herbage was evaluated for its proximate, fibre, and mineral contents. The results of the study demonstrated that GE significantly increased grass species than other-forbs, other-graminoids, and legumes, respectively, and increased dry matter productivity, which could be seen by an increase in leaf stem ratio, tiller productivity, increased coarseness (fibrous content), total ash, calcium (Ca), and phosphorus (P), but with a decreased nonstructural carbohydrate, and the concentrations of ether extract and crude protein. Research results also confirmed that GE increases herbages’ fibrousness and productivity, though the herbage quality, intake, and digestibility decline. It further demonstrates that grazing is a crucial biological component for maintaining pasture quality in subtropical grasslands and that managing grasslands through livestock grazing would make grasslands more stable and nutrient-enriched. The findings of this study can be useful in the long-term monitoring of grazing livestock in the subtropical grasslands when considering further investigations with the multiple factors in future.

E076 To investigate the mechanisms of action of novel compounds ex vivo in cells obtained from the inflammatory synovium of patients with rheumatoid arthritis

Abstract Background/Aims To investigate the ex vivo effect of JAK1/2 inhibitor baricitinib on the expression of pro-inflammatory mediators in normal and RA diseased fibroblasts stimulated with oncostatin M (OSM) as well as in cells derived from RA inflamed synovial membrane cells (RA SMC). Methods RA SMC and fibroblast like synoviocytes (RA FLS) were isolated from synovial membranes of patients undergoing surgery using enzymatic digestion and cell propagation. Normal fibroblast-like synoviocytes (FLS) and normal human skin fibroblasts (HSF) were purchased from commercial sources. All fibroblasts were cultured in media supplemented foetal bovine serum and antibiotics in incubators using established laboratory procedures, and serum-starved prior to stimulation with OSM with or without baricitinib. RA SMC were cultured in the presence of baricitinib without OSM as they spontaneously release pro-inflammatory cytokines. Levels of mRNA and proteins of the various inflammatory cytokines (IL-6, IL-8, MCP-1, CCL5 and IP-10) were determined by qPCR, ELISA and MSD. Western blot was used to measure levels of phospho-JAK specific components. Results qPCR analysis found that baricitinib inhibited OSM induced IP-10 and MCP-1 mRNA expression but not RANTES mRNA expression in normal human skin fibroblasts and RA FLS. In normal FLS however, baricitinib was found to inhibit RANTES and MCP-1 mRNA expression, but not IP-10 mRNA expression. Expression levels of IL-8 were too low for accurate conclusions to be drawn in all cell types apart from the RA SMCs, and in this case baricitinib did not inhibit IL-8 expression. ELISA results showed that baricitinib inhibited OSM induced MCP-1 expression in all cell types investigated. OSM induced IL-6 expression was inhibited in RA FLS but no other cell type. Conclusion Baricitinib inhibits OSM stimulated pro-inflammatory cytokine production through JAK1, 2. It has a varying inhibitory effect on the production of different cytokines: it is shown to particularly inhibit MCP-1. Baricitinib was found to have no effect on the production of spontaneously released pro-inflammatory cytokines in RA mixed cell populations. Disclosure M. Turner: None.

P191 To investigate the mechanisms of action of novel compounds ex vivo in cells obtained from the inflammatory synovium of patients with rheumatoid arthritis

Abstract Background/Aims To investigate the ex vivo effect of JAK1/2 inhibitor baricitinib on the expression of pro-inflammatory mediators in normal and RA diseased fibroblasts stimulated with oncostatin M (OSM) as well as in cells derived from RA inflamed synovial membrane cells (RA SMC). Methods RA SMC and fibroblast like synoviocytes (RA FLS) were isolated from synovial membranes of patients undergoing surgery using enzymatic digestion and cell propagation. Normal fibroblast-like synoviocytes (FLS) and normal human skin fibroblasts (HSF) were purchased from commercial sources. All fibroblasts were cultured in media supplemented foetal bovine serum and antibiotics in incubators using established laboratory procedures, and serum-starved prior to stimulation with OSM with or without baricitinib. RA SMC were cultured in the presence of baricitinib without OSM as they spontaneously release pro-inflammatory cytokines. Levels of mRNA and proteins of the various inflammatory cytokines (IL-6, IL-8, MCP-1, CCL5 and IP-10) were determined by qPCR, ELISA and MSD. Western blot was used to measure levels of phospho-JAK specific components. Results qPCR analysis found that baricitinib inhibited OSM induced IP-10 and MCP-1 mRNA expression but not RANTES mRNA expression in normal human skin fibroblasts and RA FLS. In normal FLS however, baricitinib was found to inhibit RANTES and MCP-1 mRNA expression, but not IP-10 mRNA expression. Expression levels of IL-8 were too low for accurate conclusions to be drawn in all cell types apart from the RA SMCs, and in this case baricitinib did not inhibit IL-8 expression. ELISA results showed that baricitinib inhibited OSM induced MCP-1 expression in all cell types investigated. OSM induced IL-6 expression was inhibited in RA FLS but no other cell type. Conclusion Baricitinib inhibits OSM stimulated pro-inflammatory cytokine production through JAK1, 2. It has a varying inhibitory effect on the production of different cytokines: it is shown to particularly inhibit MCP-1. Baricitinib was found to have no effect on the production of spontaneously released pro-inflammatory cytokines in RA mixed cell populations. Disclosure M.P. Turner: None.

A Comparative Evaluation of Rain Water Obtained from Corrugated Roofing Sheets within Awka Metropolis, Anambra State

The objective of this study was to evaluate comparatively the effects of different brands of corrugated roofing sheets on rainwater utilized within Awka metropolis. Different physicochemical parameters were assessed in the rain water harvested using established standard laboratory procedures. The rain water was harvested directly (control) and also collected after passing through the different brands of corrugated roofing sheets (samples). The results showed no appreciable effects on the physical appearance, observable odour, or taste of the water samples compared to the control sample. However, at 30 °C, there were minor changes in the pH. The amount of suspended solids (SS) in mg/100ml, total solids (TS), and total dissolved solids (TDS) found in each of the samples, including the control sample, did not vary. This showed that the corrugated roofing sheet has no significant effect on the SS, TS, and TDS of the rain water harvested within the study area. Iron, zinc, and chloride ions followed a similar trend as SS, with Fe and Zn occurring at non-detectable limits of the instrumentation, while the levels of Cl- ion (3mg/100ml) did not vary in all the samples, including the control sample. The results revealed that corrugated roofing sheets affect the total alkalinity and methyl orange alkalinity of the harvested rain water in the study area greatly but had no impact on their physical appearance.

The Role of Electrochemistry and Mineralogy in the Geotechnical Behavior of Salinized Soils

The Atterberg limits are essential information and the first step in soil classification for geotechnical purposes. Established laboratoryprocedures use distilled water in the plasticity and liquid limits determination. However, saline solutions frequently interact with soilsin the construction environment through fluid percolation processes. This work aims to understand the variation of the geotechnicalbehavior of two standard materials with different mineralogical compositions (kaolinitic and smectitic) when affected by NaCl ionicsolutions in different concentrations. The purpose is to simulate different soils in environments with the presence of saline solutions.This paper reports an experimental program in which a kaolinite-rich and a smectite-rich material received NaCl solutions in threedifferent concentrations (0.6 %, 3.5 %, and 15.0 %) and had their Atterberg limits determined under these conditions. Additionally,non-contaminated samples of both materials have had their limits measured using distilled water. Physical characterization testsincluded hygroscopic moisture, grain size distribution, grain density, plastic limit (PL), and liquid limit (LL). These data allowed thedetermination of the Skempton activity index (AI), plasticity index (PI), consistency index (CI), classification of soils in the UnifiedSoil Classification System (USCS), and in the Highway Research Board (HRB) with the group index (GI). Mineralogy was determinedby X-ray diffraction and physical chemistry by measuring pH in H2O and KCl, determining the ΔpH, the point of zero-charge (PZC),and the surface electrical potential (Ψo). The results show that the pH values rise with increasing salinity, while ΔpH, PZC, Ψo, LL,AI, PI, GI decrease with increasing salinity. The PL decreases with the increase in salinity for smectite and increases for kaolinite. TheUSCS and HRB demonstrate that the materials start to behave as fewer plastic materials with increased salinity. It is concluded that thevariations in the physicochemical parameters of the environment control and modify the geotechnical behavior of the fine-grained soils.

A numerical study on the coupled hydro-mechanical behaviour of compacted bentonite

Understanding the mechanical behaviour of compacted bentonite upon re-saturation is of outmost importance in most designs of nuclear waste disposal repositories. The behaviour of bentonite is characterized by its stress-path dependency and it is typically interpreted on the basis of its microstructural interactions. Up to now, effective stress-based models have had limited success in reproducing consistently the main responses. Here a recently proposed model for the modelling of volumetric behaviour of compacted bentonites is extended to triaxial stress states. The model is formulated using a conventional effective stress expression and the degree of saturation. Because these two variables are directly related to the water retention, a suitable formulation for bentonites is used. The resulting equations are characterised by a high degree of hydro- mechanical couplings and a low number of material parameters, which can be obtained on the basis of well- established laboratory procedures. In order to demonstrate its capabilities, the model is used to simulate the behaviour of MX-80 bentonite for several stress paths under oedometric conditions. The emphasis is put on the process of parameter determination. The predictive capabilities of the model are also highlighted.

Distinct Trajectories of Cortisol Response to Prolonged Acute Stress Are Linked to Affective Responses and Hippocampal Gray Matter Volume in Healthy Females.

The development of robust laboratory procedures for acute stress induction over the last decades has greatly advanced our understanding of stress responses in humans and their underlying neurobiological mechanisms. Nevertheless, attempts to uncover linear relationships among endocrine, neural, and affective responses to stress have generally yielded inconsistent results. Here, 79 healthy females completed a well established laboratory procedure of acute stress induction that was modified to prolong its effect. Endocrinological and subjective affect assessments revealed stress-induced increases in cortisol release and negative affect that persisted 65 and 100 min after stress onset, respectively, confirming a relatively prolonged acute stress induction. Applying latent class linear mixed modeling on individuals' patterns of cortisol responses identified three distinct trajectories of cortisol response: the hyper-response (n = 10), moderate-response (n = 21), and mild-response (n = 48) groups. Notably, whereas all three groups exhibited a significant stress-induced increase in cortisol release and negative affect, the hyper-response and mild-response groups both reported more negative affect relative to the moderate-response group. Structural MRI revealed no group differences in hippocampal and amygdala volumes, yet a continuous measure of cortisol response (area under the curve) showed that high and low levels of stress-induced cortisol release were associated with less hippocampal gray matter volume compared with moderate cortisol release. Together, these results suggest that distinct trajectories of cortisol response to prolonged acute stress among healthy females may not be captured by conventional linear analyses; instead, quadratic relations may better describe links between cortisol response to stress and affective responses, as well as hippocampal structural variability.SIGNIFICANCE STATEMENT Despite substantial research, it is unclear whether and how individual neuroendocrine stress response patterns are linked to affective responses to stress and structural variability in neuroendocrine regulatory brain regions. By applying latent class linear mixed modeling on individuals' patterns of cortisol responses to a prolonged acute stressor, we identified three distinct trajectories of cortisol response. Relative to the group showing a moderate cortisol response, groups characterized by hyper and mild cortisol response were both associated with more negative affect. Moreover, a continuous measure of cortisol response showed that high and low levels of stress-induced cortisol release correlated with reduced hippocampal gray matter volume. Given that neuroendocrine stress responses are conceptualized as biomarkers of stress susceptibility, these insights may have clinical implications.

Validation of single photon absorptiometry for on-farm measurement of density and mineral content of tail bone in cattle

A validation study examined the accuracy of a purpose-built single photon absorptiometry (SPA) instrument for making on-farm in vivo measurements of bone mineral density (BMD) in tail bones of cattle. In vivo measurements were made at the proximal end of the ninth coccygeal vertebra (Cy9) in steers of two age groups (each n = 10) in adequate or low phosphorus status. The tails of the steers were then resected and the BMD of the Cy9 bone was measured in the laboratory with SPA on the resected tails and then with established laboratory procedures on defleshed bone. Specific gravity and ash density were measured on the isolated Cy9 vertebrae and on 5-mm2 dorso-ventral cores of bone cut from each defleshed Cy9. Calculated BMD determined by SPA required a measure of tail bone thickness and this was estimated as a fraction of total tail thickness. Actual tail bone thickness was also measured on the isolated Cy9 vertebrae. The accuracy of measurement of BMD by SPA was evaluated by comparison with the ash density of the bone cores measured in the laboratory. In vivo SPA measurements of BMD were closely correlated with laboratory measurements of core ash density (r = 0.92). Ash density and specific gravity of cores, and all SPA measures of BMD, were affected by phosphorus status of the steers, but the effect of steer age was only significant (P < 0.05) for steers in adequate phosphorus status. The accuracy of SPA to determine BMD of tail bone may be improved by reducing error associated with in vivo estimation of tail bone thickness, and also by adjusting for displacement of soft tissue by bone mineral. In conclusion a purpose-built SPA instrument could be used to make on-farm sequential non-invasive in vivo measurements of the BMD of tailbone in cattle with accuracy acceptable for many animal studies.

Replacing liquid fyrite with digital to improve <i>in vitro</i> fertilization laboratory quality control procedures

In clinical in vitro fertilization (IVF), optimal culture conditions are required for production of high quality embryos and for achieving high pregnancy rates. Cell culture systems require vigilant attention to quality control and quality assurance, and upgrades to equipment and procedures require strenuous deliberation. During a 2-week maintenance period, we undertook an extensive analysis of incubator carbon dioxide (CO2) monitoring and the effect on culture media pH by comparing our traditional liquid Fyrite instruments to a certified and calibrated digital CO2 analyzer. The digital analyzer produced consistently lower CO2 readings and significantly greater precision than the liquid Fyrite. Media pH measurements showed significant variation depending on CO2 calibration device; however pH remained within manufacturers’ specifications. After superior performance by the digital analyzer, we incorporated this device into the incubator calibration and daily quality control procedures. A retrospective comparison of overall lab performance before and after this equipment switch demonstrated improved clinical pregnancy and implantation rates. This report illustrates the necessary caution when altering established laboratory procedures and equipment while highlighting the benefits of judiciously updating techniques and equipment in a laboratory setting that is often stubborn to change pre-existing, ingrained methodology.

Defining an absolute reference frame for ‘clumped’ isotope studies of CO 2

We present a revised approach for standardizing and reporting analyses of multiply substituted isotopologues of CO 2 (i.e., ‘clumped’ isotopic species, especially the mass-47 isotopologues). Our approach standardizes such data to an absolute reference frame based on theoretical predictions of the abundances of multiply-substituted isotopologues in gaseous CO 2 at thermodynamic equilibrium. This reference frame is preferred over an inter-laboratory calibration of carbonates because it enables all laboratories measuring mass 47 CO 2 to use a common scale that is tied directly to theoretical predictions of clumping in CO 2, regardless of the laboratory’s primary research field (carbonate thermometry or CO 2 biogeochemistry); it explicitly accounts for mass spectrometric artifacts rather than convolving (and potentially confusing) them with chemical fractionations associated with sample preparation; and it is based on a thermodynamic equilibrium that can be experimentally established in any suitably equipped laboratory using commonly available materials. By analyzing CO 2 gases that have been subjected to established laboratory procedures known to promote isotopic equilibrium (i.e., heated gases and water-equilibrated CO 2), and by reference to thermodynamic predictions of equilibrium isotopic distributions, it is possible to construct an empirical transfer function that is applicable to data with unknown clumped isotope signatures. This transfer function empirically accounts for the fragmentation and recombination reactions that occur in electron impact ionization sources and other mass spectrometric artifacts. We describe the protocol necessary to construct such a reference frame, the method for converting gases with unknown clumped isotope compositions to this reference frame, and suggest a protocol for ensuring that all reported isotopic compositions (e.g., Δ 47 values; Eiler and Schauble, 2004; Eiler, 2007) can be compared among different laboratories and instruments, independent of laboratory-specific analytical or methodological differences. We then discuss the use of intra-laboratory secondary reference frames (e.g., based on carbonate standards) that can be more easily used to track the evolution of each laboratory’s empirical transfer function. Finally, we show inter-laboratory reproducibility on the order of ±0.010 (1 σ) for four carbonate standards, and present revised paleotemperature scales that should be used to convert carbonate clumped isotope signatures to temperature when using the absolute reference frame described here. Even when using the reference frame, small discrepancies remain between two previously published synthetic carbonate calibrations. We discuss possible reasons for these discrepancies, and highlight the need for additional low temperature (<15 °C) synthetic carbonate experiments.

Demonstration That CobG, the Monooxygenase Associated with the Ring Contraction Process of the Aerobic Cobalamin (Vitamin B12) Biosynthetic Pathway, Contains an Fe-S Center and a Mononuclear Non-heme Iron Center

The ring contraction process that occurs during cobalamin (vitamin B(12)) biosynthesis is mediated via the action of two enzymes, CobG and CobJ. The first of these generates a tertiary alcohol at the C-20 position of precorrin-3A by functioning as a monooxygenase, a reaction that also forms a gamma lactone with the acetic acid side chain on ring A. The product, precorrin-3B, is then acted upon by CobJ, which methylates at the C-17 position and promotes ring contraction of the macrocycle by catalyzing a masked pinacol rearrangement. Here, we report the characterization of CobG enzymes from Pseudomonas denitrificans and Brucella melitensis. We show that both contain a [4Fe-4S] center as well as a mononuclear non-heme iron. Although both enzymes are active in vivo, the P. denitrificans enzyme was found to be inactive in vitro. Further analysis of this enzyme revealed that the mononuclear non-heme iron was not reducible, and it was concluded that it is rapidly inactivated once it is released from the bacterial cell. In contrast, the B. melitensis enzyme was found to be fully active in vitro and the mononuclear non-heme iron was reducible by dithionite. The reduced mononuclear non-heme was able to react with the oxygen analogue NO, but only in the presence of the substrate precorrin-3A. The cysteine residues responsible for binding the Fe-S center were identified by site-directed mutagenesis. A mechanism for CobG is presented.

Evaluation of analytical methods and workflow performance of the Architect ci8200 integrated serum/plasma analyzer system

Background The Architect ci8200 is an integrated serum analyzer for photometric, electrochemical and immunological assays. Several assays of each category and the workflow performance of the system were compared with established laboratory procedures in two laboratories. Methods Measurements were compared with the ELECSYS 2010 (Roche Diagnostics) for CEA, PSA, FPSA, AFP, folate, vitamin B12, with the CENTAUR (Bayer) for TSH, T4, FT4, FSH and Estradiol, with the LIAISON (DiaSorin) for TSH, FT4 and FT3, with the Behring Nephelometer BN II (Dade-Behring) for ferritin, and with the INTEGRA 800 (Roche Diagnostics), and the AU640 (Olympus) for clinical chemistry assays. Workflow studies were performed to compare times of analysis required for defined analytical workloads. Results The coefficients of variation (CVs) for within-run imprecision were between 3% and 6% for CEA, PSA, FPSA, AFP and ferritin, and between 3% and 11% for TSH, FT4, FT3, folate and vitamin B12. The CVs for day-to-day imprecision for immunoassays were between 3% and 10%, except for vitamin B12 (CVs 11–13%) and FT4 (CV 10% –13%). For clinical chemistry tests corresponding CVs for within-run imprecision were < 1%, except for HDL, triglyceride, creatinine, ALT, LD and lipase (CVs < 2%) and bicarbonate (CV 3%–6%) and magnesium (CV < 3%). The CVs for day-to-day imprecision for clinical chemistry tests were < 1%, except for sodium, CO 2, magnesium, phosphorus, glucose, uric acid, HDL, triglyceride, ALT, AST CK, lipase with CVs < 6% and for CO 2 < 11%. Dilutional linearity testing of seven immunoassays and five clinical chemistry analytes resulted in recovery rates of 90–110%. Correlation studies with 15 immunoassays and 25 clinical chemistry tests showed acceptable agreements with established methods. Work flow analyses demonstrated a net gain in time of analysis up to 109 min depending on the size of the sample batch analyzed with the Architect ci8200 as the main analyzer as compared to the currently installed routine laboratory equipment. Median turn-around times were 7 and 30 min for chemistry assays and immunoassays, respectively, when ordered as STAT analyses, and 18 min when chemistry assays were ordered as routine determinations. Conclusions Assays on the Architect ci8200 performed well, fulfilling quality control requirements as defined for instance by German quality control guidelines (RiliBÄK). Method comparisons showed acceptable agreements with established assays. Workflow studies using the Architect ci8200 documented shorter times of analyses as compared with the conventionally established laboratory routine demonstrating the potential of integrated chemistry/immunoassay analyzers to provide faster and more efficient performance.

Use of Steel Slag Leach Beds for the Treatment of Acid Mine Drainage

Steel slag from the Waylite steel-making plant in Bethlehem, Pennsylvania was leached with acidic mine drainage (AMD) of a known quality using an established laboratory procedure. Leaching continued for 60 cycles and leachates were collected after each cycle. Results indicated that the slag was very effective at neutralizing acidity. The AMD/slag leachates contained higher average concentrations of Ba, V, Mn, Cr, As, Ag, and Se and lower average concentrations of Sb, Fe, Zn, Be, Cd, Tl, Ni, Al, Cu, and Pb than the untreated AMD. Based on these tests, slag leach beds were constructed at the abandoned McCarty mine site in Preston County, West Virginia. The leach beds were constructed as slag check dams below limestone-lined settling basins. Acid water was captured in limestone channels and directed into basins to leach through the slag dams and discharge into a tributary of Beaver Creek. Since installation in October 2000, the system has been consistently producing net alkaline, pH 9 water. The treated water is still net alkaline and has a neutral pH after it encounters several other acidic seeps downstream.

Monitoring urea levels during haemodialysis with a pulsed-flow chemiluminescence analyser

We have developed a rapid and robust method for the determination of urea in spent haemodialysis fluid as a measure of the efficiency of haemodialysis treatments. A novel flow analysis instrument (which generates a pulsed solution flow) was coupled with a chemiluminescence detection system, based on the oxidation of urea with hypobromite. The ‘pulsed-flow chemiluminescence analyser’ exhibited high precision (1.6% relative standard deviation (R.S.D.) for a 1×10 −5 M urea standard, n=10) and good limit of detection (9×10 −7 M, S/N=3) as a result of the rapid and reproducible mixing of small volumes of reagent and sample at the point of detection. The proposed chemiluminescence technique and an established urease-based laboratory procedure were compared, and showed a very similar trend for the change in urea concentration during a typical haemodialysis treatment. The relative chemiluminescence response from the oxidation of species with similar structure has revealed the inherent selectivity of the light producing pathway, but a positive interference was obtained from protein when this technique was applied to the determination of urea in serum samples. Arginine was identified as the predominant source of this interference.

A nuclear magnetic resonance study of Antarctic sea ice brine diffusivity

We have measured the diffusive motion of water molecules in the brine inclusions of Antarctic sea ice using a specially constructed nuclear magnetic resonance (NMR) apparatus. The method relies on the use of pulsed magnetic field gradients in precise analogy to well established laboratory procedures. One version of the apparatus utilised core samples extracted from the ice sheet which were subsequently analysed on site while a later version utilised a probehead which was inserted into the ice sheet, thus minimising any sample perturbation. The diffusive motion of water molecules in the brine inclusions is found to be strongly anisotropic, and, over short length scales, exhibits a rapidity greatly in excess of that expected for thermal equilibrium Brownian behaviour, an effect which we attribute to convective transport.

Predicting exchangeable cation distributions in soil by using exchange coefficients and solution activity ratios

A simple model which predicts the distribution of surface-applied chemicals in soil following leaching is described. The applied cation is considered to reach equilibrium between the solution and exchange phases prior to moving into the underlying soil layer. The distribution between the solution and exchange phases is defined by the Gapon relationship. The Gapon exchange coefficient is derived from exchange isotherms determined using a batch technique. Movement of the applied cation through successive layers and cation exchange with the soil continues until the activity ratio in solution equals the equilibrium activity ratio determined from the exchange isotherm. The applied cation remaining in solution is then considered to move 'freely'. A number of physical and chemical processes have been omitted in developing the model to retain simplicity and to ensure that the required parameters can readily be obtained using simple and well established laboratory procedures. In particular, only a binary cation exchange system is considered, and it is assumed that the rate of exchange is instantaneous. The inputs to the model are the amount of cation added, the Gapon exchange coefficient and corresponding solution activity ratio, the equilibrium activity ratio, the initial exchangeable and solution cation concentrations, and the average volumetric moisture content of soil during leaching. The simple model satisfactorily predicted the distribution of exchangeable K+ and Mg2+, and percentage of applied K+ or Mg2+ leaching 'freely' in the soil solution in homogeneous columns of soil having varying texture, clay mineralogy, and initial cation composition.

Histomorphometry of fresh frozen iliac crest bone biopsies

Although methods are well established for the rapid histological preparation of fresh frozen soft tissues, they remain inadequate for the preservation of hard tissues. A recent technique for sectioning fresh frozen bone, in which sections can be prepared within an hour, has been applied to undecalcified human iliac crest bone biopsies. Quantitative analysis has shown that the static remodeling variables deduced from frozen sections closely resemble those derived from plastic-embedded sections prepared for routine use by established laboratory procedures. It is concluded that the rapid method is reliable for immediate diagnostic and histomorphometric purposes.

A simple and rapid method to initiate Trypanosoma brucei brucei and T. brucei evansi bloodstream form cultures

Initiation of cultures of bloodstream forms of trypanosomes from the Trypanozoon subgenus is an established laboratory procedure. The trypanosomes are usually separated from the blood cells of the donor animal by centrifugation on a density gradient (Hirumi et al., 1977) or by differential centrifugation (Hill et al., 1978; Brun et al., 1981; Zweygarth et al., 1982; Baltz et al., 1985). Isolation on an anion exchange column (Lanham and Godfrey, 1970) has been little used for this purpose. We describe a simple and rapid method for the initiation of trypanosome bloodstream form cultures from infected host blood, avoiding centrifugation and anion exchange column procedures. Two T.b. brucei stocks, CP 271 and CP 547, and two T.b. evansi stocks, CP 1134 and CP 2087, were used. Stock CP 271 was isolated in 1980 from a goat in Matuga/Kenya, stock CP 547 was isolated in 1985 from a naturally infected bovine in Jilib/Somalia. The two T.b. evansi stocks were both isolated from camels, stock CP 1134 in 1979 in Kulal/Kenya, stock CP 2087 in 1980 in the Sudan.