ALBERT

Notes: Abstract Supercritical fluid extraction (SFE) is a technique for sample preparation that is quickly replacing Soxhlet extraction and ultrasonic extraction. This manuscript introduces two real-world applications of SFE that serve as the basis of undergraduate laboratory experiments: the analysis of caffeine in ground coffee and nicotine in cigarette tobacco. The extracts are analyzed with gas chromatography-mass spectrometry (GC-MS). The experiments are presented in a discovery fashion that enables students to develop a theoretical understanding of the role of intermolecular forces in the extraction process. The focus on theoretical aspects of SFE makes these experiments appropriate for analytical and physical chemistry labs.

Notes: Abstract Monte Carlo simulations of fatigue in welded joints have been performed using an approach which combines a model for initiation with a multiple surface crack propagation model. The results have been compared with experiments on T-plate welded joints in which initiation and propagation of surface cracks were monitored using potential drop techniques. Predictions of initiation life using a local strain approach were conservative. Despite this underprediction of initiation life, predictions of total fatigue life were very good as a consequence of accurate simulations of propagation life and the fact that initiation represented on average only 12 to 22% of total life, depending on stress level. The initiation model considered variability due to the local weld toe angle and radius, and material strain–life behaviour. The only variability considered in the propagation analysis was the position and timing of initiation events, which leads to variability in coalescence. The underprediction of the variability in propagation and total lives was attributed to the underprediction of initiation life and the fact that out-of-plane coalescence effects were ignored.

Notes: Abstract Cerium can be determined spectrofluorimetrically (λem 350 nm, λex 260 nm) based on the relatively intense native fluorescence of the cerium(III) aquo-ion. The main potential interference in the analysis of steel from iron(III), cerium(IV) and chromium(VI) are removed by use of a carrier solution containing 2.5% w/v hydroxylammonium chloride. The slight residual interference from iron(II) can be corrected by a matrix matching factor linearly related to the amount of iron present. The calibration graphs are linear over the range 0–7 μg ml−1 based on 250 μl injection volumes. The sampling rate was 30 h−1. The relative standard deviation was 2.0% (n=5) at 3 μg ml−1 cerium. The system has been applied to the determination of cerium in carbon or low alloy steels.

Notes: Abstract. In previous studies we found that the calciotropic hormone 1,25-dihydroxyvitamin D3 [1,25(OH)2D3] augments the action of either prostaglandin E1 (PGE1) or NaF to induce differentiation of human promyelocytic HL-60 cells, a process that features increased generation of nitric oxide (NO) via up-regulation of inducible nitric oxide synthase (iNOS). We have now examined the short-term interaction of 1,25(OH)2D3 with phorbol 12-myristate 13-acetate (PMA) and dimethylsulfoxide (DMSO) in these cells. PMA (100 nM) alone generally up-regulated several classical indices of macrophagic differentiation and stimulated cellular production of interleukin (IL)-1α, IL-6, tumor-necrosis factor (TNF)-α, PGE2, and NO. Increased generation of NO primarily resulted from increased expression of cellular iNOS. When 1,25(OH)2D3 (10 nM) was added to PMA treatments, most PMA-induced changes, particularly its effects to up-regulate iNOS-dependent NO production and change cell morphology, were multiplicatively augmented. In contrast, DMSO (1.3%) alone, an inducer of granulocytic differentiation, increased cytokine production, but failed to stimulate NO production or induce iNOS. In contrast to its striking interaction with PMA, 1,25(OH)2D3 could not augment DMSO's differentiative effects. Changes in cellular cytokine production were eliminated as the driving force in HL-60 differentiation when specific neutralizing antibodies failed to produce any attenuation of iNOS up-regulation or of the shifts in cell morphology. However, indomethacin (30 μM) blocked the synergistic interaction between 1,25(OH)2D3+ PMA to shift cell morphology and stimulate NO production. Subsequently adding PGE2 (1 ng/ml) to indomethacin-treated cells restored the ability of 1,25(OH)2D3+ PMA to interactively increase cellular NO production, but failed to fully replicate the strong shift in cell morphology typical of PMA + 1,25(OH)2D3 treatments. Our findings suggest that interaction between 1,25(OH)2D3 and PMA to induce macrophagic differentiation increases iNOS-dependent NO production by a mechanism involving a cyclooxygenase product(s), possibly PGE2.

Notes: Abstract Human promyelocytic HL-60 cells can be induced by biochemical agents to differentiate in vitro towards divergent types of myelomonocytic cells. It has been reported that prostaglandin E1 (PGE1) can induce granulocytic differentiation and that 1,25-dihydroxyvitamin D3 (1,25(OH)2D3) can induce monocytic differentiation. We have now examined the effects of these compounds, both alone and in combination, on HL-60 cell differentiation. PGE1 (1 μg/ml) or 1,25(OH)2D3 (10 nM) each inhibited cell proliferation over 48–96 hours of treatment, but combined treatment with both agents was necessary to produce a strong inhibition. The percentage of HL-60 cells that can reduce nitroblue tetrazolium (NBT) (a characteristic index of early monocytic or granulocytic differentiation) increased 13-fold within 72 hours of PGE1 treatment, and 1,25(OH)2D3 produced a fivefold stimulation. However, combined treatment (PGE1 plus 1,25(OH)2D3) produced a dramatic 35-fold increase. HL-60 cells did not produce significant levels of nitric oxide (NO) before 48 hours in culture, and treatment with PGE1 or 1,25(OH)2D3 did not significantly increase cellular NO elaboration over control levels. However, combined treatment produced a striking 12-fold increase over control levels. Similarly, combined treatment was necessary to obtain the maximal time-dependent stimulation of cellular lactate dehydrogenase (LDH) activity (a marker of granulocytic differentiation) as well as acid phosphatase (ACP) activity. During this same period of time, PGE1, but not 1,25(OH)2D3, markedly stimulated cellular claboration of interleukin (IL)-1α, IL-6, and tumor necrosis factor (TNF)-α, and 1,25(OH)2D3 cotreatment strongly augmented these effects. Thus, combined treatment with 1,25(OH)2D3 plus PGE1 generally augmented the apparent conversion of these cells, producing synergistic (multiplicative) or additive effects. Furthermore, PGE1 induced within 48 hours the more general phenotypic changes classically associated with the differentiation of these cells: increased expression of chloroacetate esterase (ChAE) (a granulocytic marker), decreases in the nuclear/cytoplasmic ratio (characteristic of development beyond the promyelocyte/myelocyte stage), and major alterations in morphology from floating spherical cells to loosely adherent, elliptical polygons. 1,25(OH)2D3 had little effect itself on most of these parameters, but augmented the morphological changes induced by PGE1 treatment. Within 48 hours, the ability of these cells to reduce the tetrazolium salt WST-1, a general measure of cellular metabolic activity, was increased by PGE1, but not by 1,25(OH)2D3; however, the combination of 1,25(OH)2D3 and PGE1 again produced the strongest stimulation. Similarly, only PGE1 significantly reduced intracellular ATP levels, but combined treatments produced a more pronounced decrease. In summary, our findings suggest that PGE1, not 1,25(OH)2D3, is sufficient to promote rapid in vitro differentiation of HL-60 cells along the granulocytic pathway; however, the PGE1-induced conversion of these cells is markedly augmented by cotreatment with 1,25(OH)2D3. In addition, these converted HL-60 cells preferentially utilize the glycolytic pathway, rather than the citric acid cycle, for production of ATP, a metabolic characteristic that resembles that described for mature granulocytes.

Notes: Abstract Schönbein’s correspondence with Berzelius (1836–1847) and with Faraday (1836–1862) are shown to be excellent accessible sources for his changing views as to the nature of ozone. Two of Schönbein’s qualitative tests for ozone, namely the reaction with neutral potassium iodide solution and that with indigo have current quantitative application. Andrew’s quantitative studies of the ozone reaction with potassium iodide (1856) and his work with Tait on the volumetric gas phase decomposition of ozone (1860) confirmed the allotropic nature of ozone but did not yield the structure which was first proposed by Odling (1861). Modern spectroscopic studies owe their origin to Hartley’s studies of 1881.

Notes: Abstract  Levels of cadmium, copper, aluminium, iron, manganese and zinc (Cd, Cu, Al, Fe, Mn and Zn) in deep-sea sediments and sediment-feeding holothurians are reported. Profiles of metals were found to vary with depth in the sediment. Holothurian foregut metal values are generally closer to surficial sediment levels, whereas body wall levels, with the exception of Cd and Cu, were generally lower. Holothurians are presented as potential biomonitors of metal concentrations in the deep-sea floor environment.

Notes: Abstract The Experimental Watershed Liming Study (EWLS) was initiated to evaluate the application of CaCO3 to a forested watershed in an effort to mitigate the acidification of surface water. The objective of the EWLS was to assess the response of the Woods Lake watershed to an experimental addition of CaCO3. During October 1989, 6.89 Mg CaCO3/ha was applied by helicopter to two subcatchments comprising about 50% (102.5 ha) of the watershed area. The EWLS involved individual investigations of the response of soil and soil water chemistry, forest and wetland vegetation, soil microbial processes, wetland, stream and lake chemistry, and phytoplankton and fish to the CaCO3 treatment. In addition, the Integrated Lake/Watershed Acidification (ILWAS) model was applied to the site to evaluate model performance and duration of the treatment. The results of these studies are detailed in this volume. The purposes of this introduction and synthesis paper are to: 1) present the overall design of the EWLS, 2) discuss the linkages between the individual studies that comprise the EWLS, and 3) summarize the response of the lakewater chemistry to watershed addition of CaCO3 and compare these results to previous studies of direct lake addition. An analysis of lake chemistry revealed the watershed treatment resulted in a gradual change in pH, acid neutralizing capacity (ANC) and Ca2+ in the water column. This pattern was in contrast to direct lake additions of CaCO3 which were characterized by abrupt changes following base addition and subsequent rapid reacidification. Over the three-year study period, the supply of ANC to drainage waters was largely derived from dissolution of CaCO3 in wetlands. Relatively little dissolution of CaCO3 occurred in freely draining upland soils. The watershed treatment had only minor effects on forest vegetation. The watershed treatment eliminated the episodic acidification of streamwater and the near-shore region of the lake during snowmelt, a phenomenon that occurred during direct lake treatments. Positive ANC water in the near-shore area may improve chemical conditions for fish reproduction, and allow for the development of a viable fish population. The watershed CaCO3 treatment also decreased the transport of Al from the watershed to the lake, and increased the concentrations of dissolved organic carbon (DOC) and dissolved silica (H4SiO4) in stream and lakewater. The watershed treatment appeared to enhance soil nitrification, increasing concentrations of NO3 − in soilwater and surface waters. However, the acidity associated with this NO3 − release was small compared to the increase in ANC due to CaCO3 addition and did not alter the acid-base status of Woods Lake. Acid neutralizing capacity (ANC) budgets for 12-month periods before and after the watershed treatment showed that the lake shifted from a large source of ANC to a minor source due to retention of SO4 2−, NO3 −, Al and the elevated inputs of Ca2+ associated with the watershed CaCO3 application. In contrast to the direct lake treatments, Ca2+ inputs from the watershed application were largely transported from the lake.