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Determination of methylmercury in fish samples using GC/AA and sodium tetraethylborate derivatization.

A simple technique is described for the rapid determination of methylmercury in fish tissue. Following simple dissolution in methanolic KOH solution, aqueous phase ethylation by derivatization with NaB(C2H5)4, cryogenic trapping on a packed chromatographic column, and GC separation, volatile mercury species are detected by atomic absorption spectrometry. Absolute detection limits are 4 pg of Hg for CH3Hg+ and 75 pg of Hg for labile Hg2+. Concentration detection limits for this optimized procedure are 4 ng of Hg for CH3Hg+ and 75 ng of Hg for labile Hg2+ per gram of pulverized dried fish tissue. Analysis of standard reference materials demonstrates the accuracy, precision, and reproducibility of the analytical method.

Animals↗

Target-promoted alkylation of DNA.

Inducible and selective alkylation of DNA was accomplished under neutral conditions by use of a silyl-protected phenol that served as a precursor for a highly reactive quinone methide. As expected, addition of fluoride triggered reaction of a model compound, 3-(tert-butyldimethylsiloxy)-4-[(p-nitrophenoxy)methyl]benzamide, and its oligodeoxynucleotide conjugate. Surprisingly, the silyl phenol was also specifically yet more slowly activated by the environment of duplex DNA in the absence of fluoride. This alternative process was associated with the hybridization of probe and target strands, and single-stranded DNA was unable to induce a similar activation. Therefore, DNA appears to effect its own alkylation by promoting the formation of an electrophilic and nondiffusible intermediate.

Alkylation↗

A[Bi(3)Ti(4)O(13)] and A[Bi(3)PbTi(5)O(16)] (A = K, Cs): New n = 4 and n = 5 Members of the Layered Perovskite Series, A[A'(n)()(-)(1)B(n)()O(3)(n)()(+1)], and Their Hydrates.

We describe the synthesis and structural characterization of new layered bismuth titanates, A[Bi(3)Ti(4)O(13)] and A[Bi(3)PbTi(5)O(16)] for A = K, Cs, corresponding to n = 4 and 5 members of the Dion-Jacobson series of layered perovskites of the general formula, A[A'(n)()(-)(1)B(n)()O(3)(n)()(+1)]. These materials have been prepared by solid state reaction of the constituents containing excess alkali, which is required to suppress the formation of competitive Aurivillius phases. Unlike the isostructural niobates and niobium titanates of the same series, the new phases reported here are spontaneously hydrated-a feature which could make them potentially useful as photocatalysts for water splitting reaction. On hydration of the potassium compounds, the c axis expands by ca. 2 Å and loses its doubling [for example, the tetragonal lattice parameters of K[Bi(3)Ti(4)O(13)] and its dihydrate are respectively a = 3.900(1) Å, c = 37.57(2) Å; a = 3.885(1) Å, c = 20.82(4) Å]; surprisingly, the cesium analogues do not show a similar change on hydration.

Journal Article↗

(39)K NMR of solid potassium salts at 21 T: effect of quadrupolar and chemical shift tensors.

39K Solid State NMR spectra (static and magic angle spinning (MAS)) on a set of potassium salts measured at 21.14 T show that the chemical shift range for K(+) ions in diamagnetic salts is well in excess of 100 ppm contrary to previous assumptions that it was quite small. Inequivalent potassium sites in crystals can be resolved through differences in chemical shifts, with chemically similar sites showing differences of over 10 ppm. The quadrupolar coupling constants obtained from MAS and solid echo experiments on powders cover the range from zero for potassium in cubic environments in halides to over 3 MHz for the highly asymmetric sites in K2CO3. Although the quadrupolar effects generally dominate the 39K spectra, in several instances, we have observed subtle but significant contributions of chemical shift anisotropy with values up to 45 ppm, a first such observation. Careful analysis of static and MAS spectra allows the observation of the various chemical shift and quadrupole coupling tensor components as well as their relative orientations, thereby demonstrating that high-field 39K NMR spectroscopy in the solid state has a substantial sensitivity to the local environment with parameters that will be of considerable value in materials characterization and electronic structure studies.

Anisotropy↗

Cyclosporin A-sensitive cytochrome c release and activation of external pathway of NADH oxidation in liver mitochondria due to pore opening by acidification of phosphate-containing incubation medium.

Acidification of a high phosphate incubation medium from pH 7.4 to 6.5 promotes increase in rates of succinate oxidation and exogenous NADH oxidation via external (rotenone-and myxothiazol-resistant) pathway by factors 2 and 2.3 respectively. Cyclosporin A prevents these effects. To measure the cytochrome c release, mitochondrial cytochrome c concentration was calculated from absorption spectrum of alpha-band of cytochromes c + c1. The cytochrome c release is shown to be equal to 27 +/- 4%, 40 +/- 12%, 70 +/- 5% at pH 7.4, 7.0, 6.5, respectively, the last value being reduced by cyclosporin A to 10 +/- 3%. Immunoblot method gives the similar results. It is concluded that acidification of the high phosphate medium induces release of a large part of the cytochrome c pool from liver mitochondria due to opening the Ca(2+)-dependent cyclosporin A-sensitive permeability transition pore and subsequent high amplitude swelling.

Animals↗

Relation between cavity design and marginal adaptation in a machine-milled ceramic restorative system.

The purpose of the current study was to evaluate the influence of inclination of the cusp as well as convergence/divergence of the cavity wall on marginal adaptation in a machine-milled ceramic restorative system. Sixty inlay specimens were prepared from a pre-fabricated feldspathic porcelain (Vitablocs Mark 2) by means of a computer integrated manufacturing system (Cerec 2). Specimens were divided into 12 groups according to original cavity preparations consisting of four inclinations of the cusp (0 degree, 15 degrees, 30 degrees and 45 degrees) and three types of cavity form (tapered box, regular box, and undercut box). The restorations were cemented with a dual-activated luting agent (Clapearl DC) and cut into two halves with a low speed cutting saw, after which adaptation at the occlusal surface of each inlay specimen was measured with a microscope. The results showed that the inclination of the cusp did not affect adaptation of the restorations (P > 0.05), whereas the inlay specimens inserted into undercut cavity preparation exhibited significantly poor adaptation as compared with those inserted into regular and tapered cavities in two inclinations of the cusp (0 degree and 15 degrees; P < 0.05). The experimental results suggest that either regular or tapered box form cavity preparation is preferable to the undercut box form to improve occlusal marginal adaptation of the Cerec 2 ceramic restorative system.

Aluminum Silicates↗

In vitro reconstitution of osmoregulated expression of proU of Escherichia coli.

Osmoregulated expression of proU has been reconstituted in a cell-free system. proU encodes an osmotically inducible, high-affinity transport system for the osmoprotectant glycine betaine in Escherichia coli. Previously, a proU-lacZ fusion gene had been cloned, resulting in plasmid pOS3. In vivo osmoregulation of this extrachromosomal proU-lacZ fusion gene at low copy number showed that the plasmid-encoded fusion contained all the necessary sequences in cis for correctly receiving osmoregulatory signals during induction by osmotic stress and repression by glycine betaine. Using a cell-free (S-30) extract, plasmid pOS3 was then used to program protein synthesis in vitro. The ionic compound potassium glutamate specifically stimulated proU-lacZ expression in a concentration-dependent manner. Potassium acetate also induced some proU expression, but other salts were ineffective, thereby ruling out ionic strength as the stimulatory signal. High concentrations of sucrose, trehalose, or glycine betaine did not induce proU expression in vitro either, eliminating osmolarity per se as the stimulus. Reconstitution in a cell-free system rules out osmoregulatory mechanisms that depend on turgor, trans-membrane signaling, or trans-acting regulators synthesized after osmotic upshock.

Chromosomes, Bacterial↗

Time courses of plasma magnesium concentrations and urinary magnesium excretion in cows subjected to acute changes in potassium intake.

Hypomagnesaemic tetany in cows develops occasionally after an acute increase in K intake such as can occur when cows are transferred to spring grass. There is evidence that under these conditions plasma Mg concentrations are only transiently decreased. In this study the questions addressed were whether the plasma Mg concentration, indeed adapts to a high K intake as only dietary variable, and whether urinary Mg excretion is associated with this adaptation. Dry cows were fed rations containing either 26 or 50 g K/kg dm, the extra K being in the form of KHCO3. When the cows were acutely transferred from the low to the high K ration, plasma Mg concentrations fell slightly, but significantly from 0.86 to 0.76 mmol/l within five days, but rose again to 0.80 mmol/l after another 23 days, this rise being also statistically significant. None of the animals developed tetany. The decrease in plasma Mg concentration in individual animals after five days on the high-K ration ranged from 6 to 21%. The time course of urinary Mg excretion resembled that of plasma Mg concentration; minimum Mg excretion was seen after four to six days on the high-K ration with a subsequent increase thereafter. To explain the transient lowering of plasma Mg concentration, it is suggested that the K-induced decrease in Mg status caused a delayed increase in the carrier-mediated component of Mg absorption, which in turn caused an increase in urinary Mg excretion. When the cows were acutely switched from the high to the low K ration, plasma Mg concentration and urinary Mg excretion rose, but no transient changes were seen.

Adaptation, Biological↗

Intracellular transport of phosphatidylcholine to the plasma membrane.

We have used pulse-chase labeling of Chinese hamster ovary cells with choline followed by plasma membrane isolation on cationic beads to study the transport of phosphatidylcholine from the endoplasmic reticulum to the plasma membrane. We have found that the process is rapid (t1/2 [25 degrees C] = 2 min) and not affected by energy poisons or by cytochalasin B, colchicine, monensin, or carbonyl cyanide p-chlorophenylhydrazone. Cooling cells to 0 degree C effectively stops the transport process. The intracellular transport of phosphatidylcholine is distinct in several ways from the intracellular transport of cholesterol (Kaplan, M. R., and R. D. Simoni, 1985, J. Cell. Biol., 101:446-453).

Animals↗

Transport of cholesterol from the endoplasmic reticulum to the plasma membrane.

We have studied the transport of newly synthesized cholesterol from the endoplasmic reticulum to the plasma membrane in Chinese hamster ovary cells using a cell fractionation assay. We found that transport is dependent on metabolic energy, but that the maintenance of the high differential concentration of cholesterol in the plasma membrane is not an energy-requiring process. We have tested a variety of inhibitors for their effect on cholesterol transport and found that cytochalasin B, colchicine, monensin, cycloheximide, and NH4Cl did not have any effect. The cholesterol transport process shows a sharp temperature dependence; it ceases at 15 degrees C, whereas cholesterol synthesis continues. When synthesis occurs at 15 degrees C, the newly synthesized cholesterol accumulates in the endoplasmic reticulum and in a low density, lipid-rich vesicle fraction. These results suggest that cholesterol is transported via a vesicular system.

Animals↗

Dietary electrolyte balance for broiler chickens exposed to thermoneutral or heat-stress environments.

Ross male broiler chicks (n = 480) on new litter were used in a randomized block design with two blocks (environmental rooms) and four treatments having four replicate pens (1.0 x 2.5 m; 15 chicks) each to evaluate dietary electrolyte balance (DEB; P < 0.05). Two rooms were 1) thermoneutral (Weeks 1 through 6, with decreasing maximum from 32 to 25 degrees C and minimum from 28 to 19 degrees C; relative humidity 49 to 58%) and 2) cyclic daily heat stress (Weeks 1 and 2, thermoneutral; Weeks 2 through 6, maximum temperatures 35, 35, 33, and 33 degrees C, respectively; and minimum temperatures 23, 20, 19, and 19 degrees C, respectively; relative humidity 51 to 54%). The DEB treatments (0, 140, 240, or 340 mEq Na + K - Cl/kg) had NaHCO3 plus NH4Cl, or KHCO3, or both added to corn-soybean meal mash basal diets with 0.30% salt (NaCl). In the thermoneutral room, DEB 240 increased 42-d weight gain and 44-d lymphocyte percentage and decreased heterophil percentage and heterophil to lymphocyte ratio compared to the DEB 40 treatment. The DEB 240 diets had 0.35 and 0.35% Na and 0.37% and 0.29% Cl in starter (0.75% K) and grower (0.67% K) diets, respectively. No DEB treatment differences were found in the heat stress room. For combined rooms, 42-d feed intake was higher for DEB 240 than for DEB 40. The 21-d weight gain was higher for DEB 240 than for DEB 40 or 140; and 21-d feed/gain was lower for DEB 40 than for DEB 340. The predicted maximum point of inflection for 21- and 42-d weight gains were DEB 250 and 201, with highest 42-d feed intake at 220.

Ammonium Chloride↗

Potassium iodate and its comparison to potassium iodide as a blocker of 131I uptake by the thyroid in rats.

Potassium iodide is the preferred thyroid blocker for personnel handling radioiodine and is recommended as a prophylaxis for the population in the near-field of a nuclear reactor which would be likely to be exposed to radioiodine in an accidental breach of containment. However, in hot and humid climates, this hygroscopic chemical has a poor shelf life due to hydrolytic loss of iodine vapors. On the other hand, another iodine-rich salt, potassium iodate (KIO3), is quite stable and has a much longer shelf life. The present study compares potassium iodide and KIO3 as thyroid blockers and examines the appropriate time at which they should be administered in case of radioiodine exposure. Either of the two were given in recommended dosage (100 mg stable iodine per 70 kg body weight) at -2, 0, +2, +4, +6, and +8 h after administration of tracer quantities of radioiodine (131I) to age-, weight-, and sex-matched rats. 131I uptake in thyroid was measured 24 h after its administration in the experimental animals and compared with placebo administered controls. Results suggest that KIO3 is as effective a thyroid blocking agent as potassium iodide. In comparison to controls, 24-h thyroid uptake of 131I can be substantially reduced if potassium iodide or KIO3 is given to the animals within 2-4 h after exposure to 131I. Another noteworthy observation is that KIO3 is effective even at 8 h when administered at twice the usual dosage in comparison to the single dose, which does not show appreciable thyroid blocking properties after 8 h.

Animals↗

KMo5O13 and KNb(1.76)Sb(3.24)O13.

The title compounds, potassium pentamolybdenum oxide, KMo(5)O(13), and potassium niobate antimonate or potassium niobium antimony oxide (1/1.76/3.24), KNb(1.76)Sb(3.24)O(13), were synthesized by solid-state reactions and are isomorphous in space group Cmcm. The structure of the Mo complex has three unique Mo atoms and consists of MoO(6) octahedra sharing edges to form Mo(2)O(6) pairs and Mo(3)O(9) triplets, which, in turn, form layers by sharing corners. These layers are linked together in the [100] direction, yielding a three-dimensional network similar to that of KSb(5)O(13). This framework delimits interconnected tunnels, running approximately along the [110] and [-110] directions, in which K(+) ions are located. In the isomorphous KNb(1.76)Sb(3.24)O(13) structure, one of the Mo sites in KMo(5)O(13) is replaced by Sb and the other two Mo sites have been replaced by Nb/Sb.

Journal Article↗

Generating isomorphous heavy-atom derivatives by a quick-soak method. Part I: test cases.

Screening for heavy-atom derivatives remains a time-consuming and cumbersome process that often results in non-isomorphous derivatives whose phases cannot be combined. Using lysozyme and FcgammaRIII receptor crystals as test cases, an improved soaking method for the generation of conventional heavy-atom derivatives has been developed. The method is based on soaking crystals in heavy-atom compounds for a very brief time at near-saturation concentrations. Compared with the current heavy-atom soaking method, which often takes days to achieve a derivatization, the quick-soak method completes a derivatization within 10 min to 2 h. The bound heavy-atom sites display higher peak heights from quick soaks than from overnight soaks in all cases tested. The quick-soak derivatives also preserved native-like diffraction resolution and data quality that was better than the prolonged-soak derivatives. Furthermore, derivatives generated by brief soaks are more isomorphous to the native than those generated by overnight soaks. Short soaks not only increase the likelihood of success in heavy-atom screening by reducing the pitfalls associated with prolonged soaks, such as lack of isomorphism and overall lattice disorder, but also have the potential to transform a time-consuming derivative screening into an 'on-the-fly' real-time derivatization process.

Animals↗

The iodide perchlorate discharge test in women with previous post-partum thyroiditis: relationship to sonographic appearance and thyroid function.

BACKGROUND: Post-partum thyroid disease occurs in 50% of anti-thyroid peroxidase (TPO) antibody positive women (detected at 16 weeks' gestation) and is characterized by a transient episode of hyper, hypo or hyper-hypothyroidism. In approximately 20% of these women the hypothyroidism is permanent. However, the extent of long-term thyroid dysfunction, possibly mediated by immune attack, in those anti-TPO Ab + ve women who have had only transient or no thyroid dysfunction during the postpartum period is not clear. OBJECTIVE: We have therefore studied the frequency of iodide organification defects by iodide perchlorate discharge testing, and of thyroid morphological abnormalities by ultrasound scanning in euthyroid women following their episode of post-partum thyroiditis (PPT). DESIGN: The study group comprised 17 women with previous PPT (PPT + ve) and 12 women who had positive anti-TPO antibodies during pregnancy but who did not develop PPT (PPT - ve). Women were studied 15-47 months following their episode of PPT. RESULTS: Iodide perchlorate discharge tests were positive (more than 10% discharge) in 7 (41%) PPT + ve and 5 (42%) PPT-ve subjects (P = NS). Morphological abnormalities on thyroid ultrasound were detected in 7 of 14 (50%) PPT + ve and 7 of 9 (77%) PPT - ve subjects (P = NS). There was a strong association between abnormalities of iodide organification and morphology: of 11 subjects with positive iodide perchlorate discharge tests, 10 had abnormal (positive) ultrasound scans; of 12 subjects with negative iodide perchlorate discharge tests 8 had negative ultrasound scans (P = 0.013, Fisher's exact test). CONCLUSIONS: Long-term subtle defects of thyroid function and morphology are common in women with anti-TPO antibodies in pregnancy, whether or not they develop post-partum thyroiditis. The clinical significance of these findings is unclear but a continuing thyroid pathological process is suggested.

Adult↗