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PubMed1985

Chromatographic separation of nontransformed and transformed glucocorticoid-receptor complexes from rat liver cytosol. Use of tungstate in the purification, inactivation, and resolution of receptor forms.

Aliquots of rat liver cytosol glucocorticoid-receptor complexes (GRc) were transformed by an incubation with 8-10 mM ATP at 0 degrees C and were compared with those transformed by an exposure to 23 degrees C. The extent of receptor transformation was measured by chromatography of the samples over columns of DEAE-Sephacel. The ATP-transformed complexes, like those which were heat-transformed, exhibited lower affinity for the positively charged ion-exchange resin and were eluted with 0.12 M KCl (peak-I): the nontransformed complexes appeared to possess higher affinity and required 0.21 M KCl (peak II) for their elution. As expected, the receptor in the peak-I exhibited the DNA-cellulose binding capacity and sedimented as 4S in sucrose gradients. Peak II contained an 8-9S glucocorticoid receptor (GR) form that showed reduced affinity for DNA-cellulose. Presence of sodium tungstate (5 mM) prevented both heat and ATP transformation of the GRc resulting in the elution of the complexes in the region of nontransformed receptors. When parallel experiments were performed, binding of the cytosol GRc to rat liver nuclei or DNA-cellulose was seen to increase 10-15 fold upon transformation by heat or ATP: tungstate treatment blocked this process completely. The transformed and nontransformed GRc were also differentially fractionated by (NH4)2SO4: tungstate-treated (nontransformed) receptor required higher salt concentration and was precipitated at 55% saturation. In addition, the GRc could be extracted from DNA-cellulose by an incubation of the affinity resin with sodium tungstate resulting in approximately 500-fold purification of the receptor with a 30% yield. These studies show that the nontransformed, and the heat-, salt-, and ATP-transformed GRc from the rat liver cytosol can be separated chromatographically, and that the use of tungstate facilitates the resolution of these different receptor forms. In addition, extraction of the receptor from DNA-cellulose by tungstate provides another new and efficient method of partial receptor purification.

Adenosine Triphosphate↗

PubMed2002

Thermoluminescence of PbWO4 irradiated with UV light.

PbWO4 single crystals were grown by the Czochralski method in argon and air. The glow curves induced by UV light and the absorption spectra of PbWO4 annealed at various temperatures were measured. The glow curves of PbWO4 were strongly dependent on the growing atmospheres. The activation energies of the 110 K peak of PbWO4 grown in argon and the 122 K peak of PbWO4 grown in air were calculated to be 0.23 eV and 0.29 eV, respectively. The ratio of the 2.76 eV emission band to the 2.48 eV or 2.26 eV emission bands of the PbWO4 grown in air was smaller than that of the PbWO4 grown in argon. The glow curve of PbWO4 grown in argon was similar to that of PbWO4 grown in air when the annealing temperatures were increased.

Air↗

PubMed2006

Defects creation under UV irradiation of PbWO4 crystals.

A systematic study of photothermally stimulated defects creation processes is carried out by the thermally stimulated luminescence (TSL) method for a large number of undoped and doped PbWO4 crystals under irradiation at 80-180 K in the 3.4-4.3 eV energy range. The activation energy Ea for the regular exciton state disintegration is found to be approximately 0.1 eV. For defect-related states disintegration, Ea varies in the crystals studied from 0.03 to 0.36 eV. The origin of the defect-related states is discussed. The conclusion is made that not only a release of charge carriers but also charge transfer processes take place under UV irradiation of PbWO4 crystals.

Crystallization↗

PubMed1989

Lack of HPA-23 antiviral activity in HIV-infected patients without AIDS.

A randomized study of 12 treated patients and seven controls was conducted in order to evaluate HPA-23 anti-HIV activity in HIV-infected patients. The antiviral activity was assessed by determining HIV p24 antigenemia. A persistence or even increase in antigenemia was shown in treated patients and thrombocytopenia was observed in nine out of the 12 patients. This suggests that HPA-23 should not be used in anti-HIV therapy.

Antimony↗

PubMed1997

Xanthine oxidase mediates myocardial injury after hepatoenteric ischemia-reperfusion.

OBJECTIVES: To determine if myocardial injury results from hepatoenteric ischemia-reperfusion. We also proposed to determine if this remote heart injury is mediated by a xanthine oxidase-dependent mechanism. DESIGN: Randomized, controlled animal study. SETTING: University-based animal research facility. SUBJECTS: Thirty-six New Zealand white male rabbits, weighing 1.8 to 3 kg. INTERVENTIONS: Anesthetized rabbits were randomly assigned to one of four groups (n = 9 per group): a) a sham-operated group; b) a sham-operated group pretreated with sodium tungstate (xanthine oxidase inactivator); c) an aorta occlusion group; and d) an aorta occlusion group pretreated with sodium tungstate. Descending thoracic aorta occlusion was maintained for 40 mins with a 4-Fr Fogarty embolectomy catheter, followed by 2 hrs of reperfusion. MEASUREMENTS AND MAIN RESULTS: Myocardial injury, manifested by increased circulating creatine kinase-MB fraction activity, was significantly associated with aortic occlusion and reperfusion (p < .05). Sodium tungstate pretreatment significantly (p < .05) reduced circulating and myocardial xanthine oxidase activity. Xanthine oxidase inactivation by sodium tungstate significantly decreased circulating creatine kinase-MB fraction activity after hepatoenteric ischemia-reperfusion (p < .05). Finally, circulating creatine kinase-MB fraction activity was significantly associated with circulating xanthine oxidase activity (r2 = .85; p < .001). CONCLUSIONS: We conclude that remote myocardial injury is caused by hepatoenteric ischemia-reperfusion. The pathoetiology of this myocardial injury involves a xanthine oxidase-dependent mechanism.

Animals↗

PubMed2000

WENDI: an improved neutron rem meter.

Neutron rem meters are routinely used for real-time field measurements of neutron dose equivalent where neutron spectra are unknown or poorly characterized. These meters are designed so that their response per unit fluence approximates an appropriate fluence-to-dose conversion function. Typically, a polyethylene moderator assembly surrounds a thermal neutron detector, such as a BF3 counter tube. Internal absorbers may also be used to further fine-tune the detector response to the shape of the desired fluence conversion function. Historical designs suffer from a number of limitations. Accuracy for some designs is poor at intermediate energies (50 keV-250 keV) critical for nuclear power plant dosimetry. The well-known Andersson-Braun design suffers from angular dependence because of its lack of spherical symmetry. Furthermore, all models using a pure polyethylene moderator have no useful high-energy response, which makes them inaccurate around high-energy accelerator facilities. This paper describes two new neutron rem meter designs with improved accuracy over the energy range from thermal to 5 GeV. The Wide Energy Neutron Detection Instrument (WENDI) makes use of both neutron generation and absorption to contour the detector response function. Tungsten or tungsten carbide (WC) powder is added to a polyethylene moderator with the expressed purpose of generating spallation neutrons in tungsten nuclei and thus enhance the high-energy response of the meter beyond 8 MeV. Tungsten's absorption resonance structure below several keV was also found to be useful in contouring the meter's response function. The WENDI rem meters were designed and optimized using the Los Alamos Monte Carlo codes MCNP, MCNPX, and LAHET. A first generation prototype (WENDI-I) was built in 1995 and its testing was completed in 1996. This design placed a BF3 counter in the center of a spherical moderator assembly, whose outer shell consisted of 30% by weight WC in a matrix of polyethylene. A borated silicone rubber (5% boron by weight) absorber covered an inner polyethylene sphere to control the meter's response at intermediate energies. A second generation design (WENDI-II) was finalized and tested in 1999. It further extended the high-energy response beyond 20 MeV, increased sensitivity, and greatly facilitated the manufacturing process. A 3He counter tube is located in the center of a cylindrical polyethylene moderator assembly. Tungsten powder surrounds the counter tube at an inner radius of 4 cm and performs the double duty of neutron generation above 8 MeV and absorption below several keV. WENDI-II is suitable for field use as a portable rem meter in a variety of work place environments, and has been recently commercialized under license by Eberline Instruments, Inc. and Ludlum Measurements, Inc. Sensitivity is about a factor of 12 higher than that of the Hankins Modified Sphere (Eberline NRD meter) in a bare 252Cf field. Additionally, the energy response for WENDI-II closely follows the contour of the Ambient Dose Equivalent per unit fluence function [H'(10)/phi] above 0.1 MeV. Its energy response at 500 MeV is approximately 15 times higher than that of the Hankins and Andersson-Braun meters. Measurements of the energy and directional response of the improved meter are presented and the measured response function is shown to agree closely with the predictions of the Monte Carlo simulations in the range from 0.144 MeV to 19 MeV.

Borates↗

PubMed2001

Effect of somatostatin on cholecystokinin-induced amylase release in rat pancreatic acini.

The effect of somatostatin on cholecystokinin-induced amylase release was investigated in isolated rat pancreatic acini. Acini were isolated by enzymatic digestion and incubated in a HEPES buffered Ringer's solution with testing reagents for 30 minutes at 37 degrees C. The activity of released amylase, cAMP, and inositol phosphate formation were measured. Intracellular calcium concentration ([Ca2+]i) was also checked. Somatostatin 14 and octreotide, a somatostatin analog, inhibited CCK-stimulated amylase release in a concentration-dependent manner. The inhibitory effect of octreotide on CCK-induced amylase release was not shown when the acini were treated with 8-Br-cAMP, irrespective of the presence of IBMX. Forskolin potentiated CCK-induced amylase release and this effect was blocked by octreotide treatment; although CCK-8 (3 x 10(-11) M) failed to stimulate cAMP formation, octreotide significantly inhibited basal cAMP formation in the acini. The increase of [Ca2+]i in response to CCK was inhibited by octreotide. However, CCK-induced inositol phosphate formation was not changed by 10(-9) M octreotide. Octreotide had no effect on CCK-stimulated tyrosine phosphorylation, and tyrosine phosphatase inhibitors (NaF and Na2WO4) did not influence the effect of octreotide on CCK-induced amylase release. From these results, we conclude that octreotide inhibits CCK-induced amylase release by inhibiting basal cAMP formation and decreasing the [Ca2+]i stimulated by CCK.

1-Methyl-3-isobutylxanthine↗

PubMed2004

Long-term beneficial effects of vanadate, tungstate, and molybdate on insulin secretion and function of cultured beta cells.

The ultratrace elements vanadate, tungstate, and molybdate exhibit significant antihyperglycemic effects in both type 1 and 2 diabetic animals, but possible effects on the function of pancreatic beta cells are understudied. In the present study, clonal BRIN BD11 cells were cultured for 3 days with each ultratrace element to establish doses lacking detrimental effects on viable beta cell mass. Vanadate treatment (4 micromol/L) had no effect on cellular insulin content but improved glucose-induced insulin secretory responsiveness. However, insulin secretion mediated by PKA and PKC activation was desensitized in vanadate-treated cells. Culture with tungstate (300 micromol/L) and molybdate (1 mmol/L) increased cellular insulin content and enhanced basal insulin release and the responsiveness to glucose and a wide range of other secretagogues. These observations suggest significant effects of ultratrace elements on pancreatic beta cells that may contribute to their antihyperglycemic action.

Animals↗

PubMed2005

A novel method for creating endodontic access preparations through all-ceramic restorations: air abrasion and its effect relative to diamond and carbide bur use.

Access through porcelain restorations is a technically delicate and stressful procedure. Although this is a common dilemma in endodontics, little research has explored alternatives in cutting through porcelain. The purpose of this study was to compare the use of a carbide bur plus water, diamond bur plus water, and air abrasion to access through porcelain. All-ceramic samples were accessed using the different techniques. Samples were evaluated using two transillumination methods, white light, and fluorescent liquid penetrant described by the American Society for Testing and Materials. Edge chipping, microcracking, and catastrophic fracture of porcelain caused by the techniques were statistically compared. Fluorescent liquid penetrant was a more sensitive method for microcrack detection. There were significant differences between the preparation techniques. Air abrasion was significantly less destructive, and caused no catastrophic fractures, edge chipping or microcracks. Preparation by air abrasion took longer to complete.

Air Abrasion, Dental↗

PubMed2005

Hard metal lung disease: high resolution CT and histologic correlation of the initial findings and demonstration of interval improvement.

Hard metal lung disease (HMLD) is a rare form of interstitial lung disease caused by sensitization to cobalt. It occurs in patients exposed to hard metal (tungsten carbide) and in diamond workers who use cobalt-containing polishes. The chest radiograph may be normal or show a nodular, reticulonodular, or reticular pattern. The high-resolution CT findings have been described in a small number of patients. Reported abnormalities include patchy lobular ground glass opacities, consolidation, reticulation, centrilobular nodularity, and, occasionally, honeycombing.We describe the high-resolution CT findings in a patient with pathologically proven HMLD who demonstrated interval improvement in the ground glass opacities and persistence of the centrilobular nodules following cessation of exposure to hard metal and treatment with corticosteroids. To our knowledge, this is the first described case of HMLD with follow-up CT examination.

Adult↗

PubMed1976

Effect of ammonium 5-tungsto-2-antimoniate on encephalomyocarditis and vesicular stomatitis virus infections in mice.

Ammonium 5-tungsto-2-antimoniate (HPA 23) protected micr partially or completely against two strains of encephalomyocarditis (EMC) virus and one strain of vesicular stomatitis (VSV) virus. The best protective effect was obtained with EMC strain VR 129 and VSV when a single i.p. injection of HPA 23 was administered shortly before virus inoculation. Mice protected by HPA 23 against EMC strain VR129 had virus titres in the blood and brain similar to those in untreated mice. A synergism between interferon and HPA 23 was observed in mice infected with EMC VR129. Our results demonstrate the in vivo activity of HPA 23 against two lethal viral infections and suggest that, at least in mice infected with EMC, death may not be related solely to virus multiplication.

Animals↗

PubMed1978

Inhibition of rabies virus in vitro by the ammonium-5-tungsto-2-antimoniate.

In vitro multiplication of rabies virus was inhibited by a condensed mineral ion, ammonium-5-tungsto-2-antimoniate (HPA 23). The inhibitory effect was evaluated by two different methods, plaque reduction and one step virus growth. Plaquing showed 50% inhibition with 4.5 microgram/ml of HPA 23 and complete inhibition with 12.5 microgram/ml. A reduction of two logs in virus yield was obtained in BHK21C13S cells in suspension treated with 50 microgram/ml of HPA 23. Inhibition also occurred when treatment with HPA 23 was started 18 to 24 h after infection in the plaque assay but no effect was seen when HPA 23 was added 48 h after virus inoculation. All these inhibitory effects of HPA 23 on rabies virus multiplication were observed at non cytotoxic doses. Therefore HPA 23 contrasts with other antiviral drugs which do not inhibit rabies virus multiplication without affecting the viability of cells.

Antimony↗

PubMed1984

Modulation by the polyoxotungstate HPA-23 of Epstein-Barr virus early antigen expression in Raji cells treated with iododeoxyuridine.

The polyoxotungstate HPA-23 was found to exert a differential effect on the induction of Epstein-Barr virus early antigen in Raji cells induced with 5-iodo-2'-deoxyuridine (IUdR). Thus, treatment of Raji cells concomitantly with IUdR and HPA-23 inhibited early antigen expression, and the extent of inhibition was proportional to the duration of treatment with HPA-23. In contrast, pretreatment of Raji cells with HPA-23 prior to induction with IUdR stimulated early antigen expression in exponentially multiplying but not in stationary-phase cells. HPA-23 alone had no effect on early antigen expression in Raji cells. Activation of the latent Epstein-Barr virus genome by IUdR is dependent upon incorporation of the thymidine analogue into cellular DNA during the S-phase of the cell cycle. Synthesis of Epstein-Barr virus DNA also takes place during S-phase, suggesting a possible participation in this process of cellular DNA polymerase alpha which is thought to be responsible for cellular DNA replication and the activity of which increases several-fold during S-phase. Treatment of Raji cells with HPA-23 caused a marked decrease in DNA polymerase alpha activity, which could result in an inhibition of IUdR incorporation leading to the observed reduction of early antigen expression in cells treated concomitantly with IUdR and HPA-23.

Antigens, Viral↗

PubMed2000

Auxin-induced ethylene triggers abscisic acid biosynthesis and growth inhibition.

The growth-inhibiting effects of indole-3-acetic acid (IAA) at high concentration and the synthetic auxins 7-chloro-3-methyl-8-quinolinecarboxylic acid (quinmerac), 2-methoxy-3,6-dichlorobenzoic acid (dicamba), 4-amino-3,6, 6-trichloropicolinic acid (picloram), and naphthalene acetic acid, were investigated in cleavers (Galium aparine). When plants were root treated with 0.5 mM IAA, shoot epinasty and inhibition of root and shoot growth developed during 24 h. Concomitantly, 1-aminocyclopropane-1-carboxylic acid (ACC) synthase activity, and ACC and ethylene production were transiently stimulated in the shoot tissue within 2 h, followed by increases in immunoreactive (+)-abscisic acid (ABA) and its precursor xanthoxal (xanthoxin) after 5 h. After 24 h of treatment, levels of xanthoxal and ABA were elevated up to 2- and 24-fold, relative to control, respectively. In plants treated with IAA, 7-chloro-3-methyl-8-quinolinecarboxylic acid, naphthalene acetic acid, 2-methoxy-3,6-dichlorobenzoic acid, and 4-amino-3,6,6-trichloropicolinic acid, levels of ethylene, ACC, and ABA increased in close correlation with inhibition of shoot growth. Aminoethoxyvinyl-glycine and cobalt ions, which inhibit ethylene synthesis, decreased ABA accumulation and growth inhibition, whereas the ethylene-releasing ethephon promoted ABA levels and growth inhibition. In accordance, tomato mutants defective in ethylene perception (never ripe) did not produce the xanthoxal and ABA increases and growth inhibition induced by auxins in wild-type plants. This suggests that auxin-stimulated ethylene triggers ABA accumulation and the consequent growth inhibition. Reduced catabolism most probably did not contribute to ABA increase, as indicated by immunoanalyses of ABA degradation and conjugation products in shoot tissue and by pulse experiments with [(3)H]-ABA in cell suspensions of G. aparine. In contrast, studies using inhibitors of ABA biosynthesis (fluridone, naproxen, and tungstate), ABA-deficient tomato mutants (notabilis, flacca, and sitiens), and quantification of xanthophylls indicate that ABA biosynthesis is influenced, probably through stimulated cleavage of xanthophylls to xanthoxal in shoot tissue.

Abscisic Acid↗

PubMed1989

Role of nitrate and nitrite in the induction of nitrite reductase in leaves of barley seedlings.

The role of NO3- and NO2- in the induction of nitrite reductase (NiR) activity in detached leaves of 8-day-old barley (Hordeum vulgare L.) seedlings was investigated. Barley leaves contained 6 to 8 micromoles NO2-/gram fresh weight x hour of endogenous NiR activity when grown in N-free solutions. Supply of both NO2- and NO3- induced the enzyme activity above the endogenous levels (5 and 10 times, respectively at 10 millimolar NO2- and NO3- over a 24 hour period). In NO3(-)-supplied leaves, NiR induction occurred at an ambient NO3- concentration of as low as 0.05 millimolar; however, no NiR induction was found in leaves supplied with NO2- until the ambient NO2- concentration was 0.5 millimolar. Nitrate accumulated in NO2(-)-fed leaves. The amount of NO3- accumulating in NO2(-)-fed leaves induced similar levels of NiR as did equivalent amounts of NO3- accumulating in NO3(-)-fed leaves. Induction of NiR in NO2(-)-fed leaves was not seen until NO3- was detectable (30 nanomoles/gram fresh weight) in the leaves. The internal concentrations of NO3-, irrespective of N source, were highly correlated with the levels of NiR induced. When the reduction of NO3- to NO2- was inhibited by WO4(2-), the induction of NiR was inhibited only partially. The results indicate that in barley leaves in NiR is induced by NO3- directly, i.e. without being reduced to NO2-, and that absorbed NO2- induces the enzyme activity indirectly after being oxidized to NO3- within the leaf.

Enzyme Induction↗

PubMed2006

Crystallization and halide phasing of the C-terminal domain of human KIN17.

Here, the crystallization and initial phasing of the C-terminal domain of human KIN17, a 45 kDa protein mainly expressed in response to ionizing radiation and overexpressed in certain tumour cell lines, are reported. Crystals diffracting to 1.4 A resolution were obtained from 10% ethylene glycol, 27% PEG 6000, 500 mM LiCl and 100 mM sodium acetate pH 6.3 in space group P2(1)2(1)2(1), with unit-cell parameters a = 45.75, b = 46.31, c = 60.80 A and one molecule in the asymmetric unit. Since this domain has a basic pI, heavy-atom derivatives were obtained by soaking the crystals with negatively charged ions such as tungstate and iodine. The replacement of LiCl by KI in the cryosolution allowed the determination of phases from iodide ions to give an interpretable electron-density map.

Chlorides↗

PubMed1985

Biochemical studies on a novel vanadate- and molybdate-sensitive acid phosphatase from human epidermis.

A novel vanadate- and molybdate-sensitive human skin epidermal acid phosphatase was purified and characterized. The enzyme was extracted from epidermal sheets with a 0.1% Triton X-100 solution buffered at pH 7.0. The purification procedure consisted of molecular permeation chromatography on Sephadex G-200 followed by chromatography on hydroxylapatite using an ammonium sulfate gradient. The molecular weight of the enzyme was 82,000 and the isoelectric point was at pH 5.6. At the optimum pH (5.1) the enzyme hydrolyzed most rapidly 1-naphthyl phosphate (Km = 0.28 mM) and 4-nitrophenyl phosphate (Km = 0.28 mM). In general, the best substrates had an aromatic leaving group. Fluoride (Ki = 39 microM; noncompetitive) and phosphate (competitive) inhibited by binding to different binding sites of the enzyme. The most potent inhibitors were vanadate (Ki = 1.9 X 10(-6)M), tungstate (Ki = 1.4 X 10(-7)M), and molybdate (Ki = 2.0 X 10(-9)M). Chemical modification and kinetic experiments suggested that the activity of the enzyme is based on imidazole, tyrosyl, and carboxyl groups. Benzoyl peroxide was a relatively potent inhibitor (Ki = 5.0 X 10(-5)M; noncompetitive). This enzyme resembled the prostatic acid phosphatase with regard to substrate specificity, inhibition characteristics, and functional groups.

Acid Phosphatase↗

PubMed1991

Biological responses of isolated macrophages to cobalt metal and tungsten carbide-cobalt powders.

A previous study from this laboratory, using morphological and biochemical (LDH release) parameters, has shown that tungsten carbide-cobalt dust exhibits a greater cytotoxicity toward isolated macrophages than cobalt metal powder alone. The present study extends this comparison by examining additional biological parameters. Glucose uptake and superoxide anion production by isolated macrophages were significantly more depressed by the tungsten carbide-cobalt mixture (WC-Co) than by cobalt alone (Co) while pure tungsten carbide (WC) had no effect or even stimulated the cells. For glucose-6-phosphate dehydrogenase and cell-associated plasminogen activator (PA) activities, no difference between Co and WC-Co dusts was observed. These observations add further evidence to our previous findings regarding the different biological reactivity of cobalt metal alone or mixed with tungsten carbide.

Animals↗