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J S Wiley

Publications and source records attributed to J S Wiley.

At least 55 records · Page 3Linked to original sources
PubMed1993

The ATP4- receptor-operated channel (P2Z class) of human lymphocytes allows Ba2+ and ethidium+ uptake: inhibition of fluxes by suramin.

It is now recognized that extracellular ATP can open a receptor-operated ion channel in a variety of cell types. In human lymphocytes this P2Z purinergic channel conducts Na+, K+, Rb+, Li+, and Ca2+ but its permeability to larger cations is not known. Fluorometric measurements were used to show that ATP4- induced the entry of Sr2+ (87 Da) and Ba2+ (137 Da) into human lymphocytes loaded with fura-2. Flow cytometry was used to show that ATP4- induced the entry of ethidium+ (314 Da) but that the larger propidium2+ cation (414 Da) was excluded. ATP(4-)-induced entry of both Ba2+ and ethidium+ showed features previously demonstrated for smaller cation permeants: (i) inhibition by amiloride analogs, (ii) sigmoid dependence of flux on ATP concentrations, and (iii) inhibition by extracellular Na+ ions. Specific inhibitors of L-type voltage-gated Ca2+ channels (nisoldipine and diltiazem) had no effect on ATP(4-)-induced Ba2+ influx. Suramin and reactive blue 2, which are recognized antagonists of ATP-operated purinergic receptors in other tissues, inhibited ATP-induced uptake of ethidium+ in lymphocytes with K1/2 of 61 and 69 microM, respectively. However, hexamethylene amiloride was a more potent inhibitor of ATP-induced ethidium+ uptake with a K1/2 of 13 microM. These data show that the ATP4- receptor-operated channel of human lymphocytes allows influx of cations as large as Ba2+ or ethidium+ and that this influx is inhibited by suramin and reactive blue 2.

Adenosine Triphosphate↗
PubMed1993

The problem of partial competition in the quantitative characterization of interactions by competitive binding assays.

Binding expressions are derived and analytical procedures developed for the quantitative characterization of inhibitor binding that is only partially competitive with the interaction between an acceptor and the ligand that is being monitored. Two such situations are considered: (i) that in which the partial competition reflects binding of inhibitor to fewer acceptor sites than available to ligand; and (ii) that in which the partial competition reflects binding of inhibitor to acceptor sites in addition to those occupiable by ligand. The potential efficacy of the suggested analyses is then explored by their application to simulated data that span the likely range of experimental behavior. Quantitative analysis of the displacement of [3H]nitrobenzylthioinosine from cultured leukemic cells by an adduct of 5'-S-(2-amino-ethyl)-N6-(4-nitrobenzyl)-5'-thioadenosine with fluorescein-5-isothiocyanate is used to establish that the cells possess 6% fewer sites (150,000 cf. 159,000 sites/cell) for the fluorescent adduct than for the tritiated ligand, and that the binding is 10-fold weaker (binding constant of 0.28 cf. 2.8 nM-1). Corresponding analysis of results obtained with bovine aorta endothelial cells indicates that a 3-fold weaker interaction (binding constant of 1.1 cf. 3.3 nM-1) occurs between the fluorescent adduct and 79% of the cell sites accessible to the tritiated ligand. The present analytical procedures extend the utility of competitive binding assays for the quantitative screening of potential inhibitors by removing the inherent limitation of existing analyses that all acceptor sites be accessible to both the competing solute and the indicator ligand.

Adenosine↗
PubMed1993

Flow cytometric quantitation of nucleoside transporter sites on human leukemic cells.

Quantitation of equilibrative, nitrobenzylthioinosine (NBMPR) sensitive (es) nucleoside transporters on blast cells isolated from patients with acute myeloblastic leukemia is useful in predicting intracellular accumulation of the antileukemic nucleoside drug, cytosine arabinoside. We previously reported the synthesis of a fluorescein-labeled ligand for the es nucleoside transporter, 5-(SAENTA-x2)-fluorescein. This paper reports the synthesis of 5-(SAENTA-x8)-fluorescein in which the linkage between fluorescein and nucleoside ligand has been increased from 2 atoms to 8 atoms. This new ligand had a sixfold increase in affinity (Kd 0.9 +/- 0.1 nM) as well as an 86% increase in the cell associated fluorescence output compared to its prototype 5-(SAENTA-x2)-fluorescein. The fluorescence signal arising from 5-(SAENTA-x8)-fluorescein specifically bound to freshly isolated and cultured leukemic myeloblasts was converted to molecules of equivalent soluble fluorescein (MESF) using standardized fluorescein microbeads and compared with the number of es nucleoside transporter sites assayed concurrently by [3H]NBMPR equilibrium binding analysis. A high correlation between the two assays was observed (r = 0.98), which enabled the cell-bound fluorescence output of 5-(SAENTA-x8)-fluorescein to be expressed in numbers of es nucleoside transporter sites per cell. The improved properties of 5-(SAENTA-x8)-fluorescein over those of its prototype molecule make it a suitable reagent for flow cytometric quantitation of nucleoside transporter expression on leukemic cells isolated from patient samples.

Adenosine↗
PubMed1993

The effects of ruminally undegradable protein, propionic acid, and monensin on puberty and pregnancy in beef heifers.

Two hundred ten spring-born English crossbred beef heifers were used in two experiments in consecutive years using a randomized complete block design. The objectives were to determine the effects of increasing dietary intake of ruminally undegradable protein or propionic acid on the occurrence of puberty and to elucidate the mechanism by which monensin decreases age of puberty in beef heifers. Diets were fed for 120 d beginning in mid-November and consisted of mature grass hay and barley straw with .45 kg.animal-1 x d-1 supplement (CON) formulated to obtain .4 kg.animal-1 x d-1 of BW gain; .5 kg of supplement similar to CON and containing an additional 250 g of ruminally undegradable intake protein supplied from blood and corn gluten meal (UIP); .45 kg of supplement similar to CON composition and containing 400 g.animal-1 x d-1 of a 50% water and 50% propionic acid mixture (PROP); or .45 kg of supplement similar to CON supplying 200 mg.animal-1 x d-1 of monensin (MON). The hay:straw ratio in each diet was adjusted biweekly, to maintain similar BW gains for heifers fed each dietary treatment. As planned, BW gains were similar (P > .1) for all heifers. The heifers fed MON were 21 d younger (P < .05) at first estrus than heifers fed UIP; CON and PROP were intermediate and similar to heifers fed MON and UIP. The heifers fed UIP were 7 to 8 kg heavier (P < .05) at puberty than CON- or PROP-treated heifers. The heifers fed PROP were 6 kg heavier (P < .1) at puberty than heifers fed MON. Heifers that received UIP required less (P < .05) TDN than did heifers that received CON or PROP to achieve similar gain. Pregnancy rate was similar for all treatment groups. Feeding UIP in excess of NRC CP requirements may improve energy utilization of heifers fed mature roughages but may delay the onset of puberty compared with heifers fed monensin. This study demonstrated that nutrient composition of the diet may influence age of puberty independent of growth rate.

Animal Feed↗
PubMed1992

The ATP4- receptor-operated ion channel of human lymphocytes: inhibition of ion fluxes by amiloride analogs and by extracellular sodium ions.

Extracellular ATP is known to increase the membrane permeability of a variety of cells. Addition of ATP to human leukemic lymphocytes loaded with the Ca2+ indicator, fura-2, induced a rise in cytosolic Ca2+ concentration which was attenuated or absent in NaCl media compared with KCl, choline Cl, or NMG Cl media. In contrast, anti-immunoglobulin antibody gave similar Ca2+ transients in NaCl and KCl media. A half-maximal inhibition of peak ATP-induced Ca2+ response was observed at 10-16 mM extracellular Na+. Basal 45Ca2+ influx into lymphocytes was stimulated 9.6-fold by ATP added to cells in KCl media, but the effect of ATP was greatly reduced for cells in NaCl media. Hexamethylene amiloride blocked 74% of the ATP-stimulated Ca45 uptake of cells in KCl media. Flow cytometry measurements of fluo-3-loaded cells confirmed that the ATP-induced rise in cytosolic Ca2+ was inhibited either by extracellular Na+ or by addition of hexamethylene amiloride. Extracellular ATP stimulated 86Rb efflux from lymphocytes 10-fold and this increment was inhibited by the amiloride analogs in a rank order of potency 5-(N-methyl-N-isobutyl)amiloride greater than 5-(N,N-hexamethylene)amiloride greater than 5-(N-ethyl-N-isopropyl)amiloride greater than amiloride. ATP-induced 86Rb efflux showed a sigmoid dependence on the concentration of ATP and Hill analysis gave K1/2 of 90 and 130 microM and n values of 2.5 and 2.5 for KCl and NaCl media, respectively. However, the maximal ATP-induced 86Rb efflux was 3-fold greater in KCl than in NaCl media. Raising extracellular Na+ from 10 to 100 mM increased ATP-induced Na+ influx from a mean of 2.0 to 3.7 nEq/10(7) cells/min, suggesting either saturability or self-inhibition by Na+ of its own influx. These data suggest that ATP opens a receptor-operated ion channel which allows increased Ca2+ and Na+ influx and Rb+ efflux and these fluxes are inhibited by extracellular Na+ ions as well as by the amiloride analogs.

Adenosine↗
PubMed1992

All-trans retinoic acid in the treatment of acute promyelocytic leukaemia.

All-trans-retinoic acid (ATRA) is known to induce differentiation of promyelocytes in vitro and also to induce remission of acute promyelocytic leukaemia in vivo. We treated 11 patients with poor prognosis acute promyelocytic leukaemia (APL) with ATRA and obtained seven complete and one partial remission. Remissions took one to three months to achieve and were associated with adverse effects including dry skin and bone pain. In eight patients the white cell count rose above 20 x 10(9)/L within the first ten days of retinoic acid treatment and this was associated with the development of pulmonary leukostasis in three patients which was fatal in one. Another two patients died of intracranial haemorrhage also within the first ten days. ATRA is a promising new agent in the induction therapy of this particular category of acute leukaemia.

Adult↗
PubMed1991

A new fluorescent probe for the equilibrative inhibitor-sensitive nucleoside transporter. 5'-S-(2-aminoethyl)-N6-(4-nitrobenzyl)-5'-thioadenosine (SAENTA)-chi 2-fluorescein.

The N6-(4-nitrobenzyl) derivative of adenosine is a tight-binding inhibitor of the equilibrative inhibitor-sensitive nucleoside transporter of mammalian cells. A fluorescent ligand for this transporter has been synthesized by allowing an adenosine analogue. 5'-S-(2-aminoethyl)-N6-(4-nitrobenzyl)-5'-thioadenosine (SAENTA), to react with fluorescein isothiocyanate. The purified adduct had a SAENTA/fluorescein molar ratio of 0.92:1 calculated from its absorption spectrum. The intensity of fluorescent emission from the SAENTA-chi 2-fluorescein adduct was 30% that of fluorescein isothiocyanate (chi 2 is the number of atoms in the linkage between fluorescein and SAENTA). SAENTA-chi 2-fluorescein inhibited the influx of nucleosides into cultured leukaemic cells with an IC50 (total concentration of inhibitor producing 50% inhibition) of 40 nM. The adduct inhibited the binding of [3H]nitrobenzylthioinosine ([3H]NBMPR) with half-maximal inhibition at 50-100 nM. Mass Law analysis of the competitive-binding data suggested the presence of two classes of sites for [3H]NBMPR binding, only one of which was accessible to SAENTA-chi 2-fluorescein. Flow cytometry was used to analyse equilibrium binding of SAENTA-chi 2-fluorescein to leukaemic cells and a Kd of 6 nM was obtained. SAENTA-chi 2-fluorescein is a high-affinity ligand for the equilibrative inhibitor-sensitive nucleoside transporter which allows rapid assessment of transport capacity by flow cytometry.

Adenosine↗
PubMed1991

Treatment of acute promyelocytic leukaemia relapsing after allogeneic bone marrow transplantation with all-trans-retinoic acid: suppression of the leukaemic clone.

We describe the clinical experience of a male patient with acute promyelocytic leukaemia, relapsing after sex-mismatched allogeneic bone marrow transplantation and treated with all-trans-retinoic acid. Resolution of the coagulopathy was observed by day 7 of therapy. A complete remission was achieved by day 47 after a period of pancytopenia, dysplastic myeloid maturation and bone marrow hypocellularity with necrosis and fibrosis. Serial cytogenetic analyses revealed a progressive loss of the male leukaemic clone [46XY,t(15;17)] and emergence of normal female (donor) cells [46XX] which became completely dominant with remission. Adverse effects of all-trans-retinoic acid included bone pain and a prominent leucocytosis requiring leukaphereses and hydroxyurea therapy. All-trans-retinoic acid can induce complete remission of recurrent acute promyelocytic leukaemia following bone marrow transplantation. The data suggest that remission is due to differentiation and suppression of the leukaemic clone while allowing repopulation of the marrow with non-malignant cells.

Adult↗
PubMed1991

Production from first-calf beef heifers fed a maintenance or low level of prepartum nutrition and ruminally undegradable or degradable protein postpartum.

Two experiments were conducted in consecutive years to determine the effects of prepartum nutrient level and postpartum ruminally undegraded protein intake on nutrient status, milk production, subsequent calf production, and reproductive performance of 126 crossbred, primiparous beef heifers. Prepartum treatments were low nutrient intake (LN) (approximately 2.5 kg of TDN, .5 kg of CP animal-1.d-1 and maintenance nutrient intake (MN) (5 kg of TDN, 1 kg of CP animal-1.d-1), which were fed for 75 d before parturition. Two postpartum protein supplements were formulated to provide 250 g/d of ruminally degradable protein (RD) and one to supply ruminally undegraded protein (UD) at 250 g/d of additional UD CP compared to the RD supplement. Cholesterol was lower (P less than .01) in heifers given UD than in heifers given RD. Blood urea nitrogen was higher (P less than .01) for UD-fed heifers than for RD-fed heifers and was higher in LN heifers (P less than .06) than in MN heifers. Milk production did not differ (P greater than .11) as a result of LN, MN, UD, or RD. Postpartum cow weight gain was greatest (P less than .01) for UD and LN heifers. The percentage of heifers bred during the first estrous cycle of the breeding season was greater (P less than .02) for UD than for RD. Overall, prepartum nutrition did not interact with postpartum protein supplement, nor did it have any effect on postpartum interval, whereas UD increased cow weight gain postpartum and reduced postpartum interval.

Animal Nutritional Physiological Phenomena↗
PubMed1991

The influence of timing and the addition of urea to supplements containing DL-methionine on ruminal fermentation and cow weight change in beef cows.

A study was conducted to determine an optimal time for supplementation of DL-methionine in relation to time of forage intake by mature British breed-type crossbred cows, and two other experiments were conducted to determine whether ruminal ammonia concentration limited changes in disappearance rates in situ and ruminal functions caused by supplements containing DL-methionine. Experiments 1 and 2 used 4 x 4 Latin square designs with four cows in each experiment. Treatments in Exp. 1 were no supplement (CON), DL-methionine at feeding (0800), DL-methionine 4 h after feeding (1200), and DL-methionine 7 h after feeding (1500). Treatments in Exp. 2 were .5 kg of beet pulp (CON), .5 kg of beet pulp plus 16.5 g of DL-methionine (MET), .5 kg of beet pulp plus 16.5 g of DL-methionine and 55 g of urea (METU), and .4 kg of soybean (SOY). Experiment 3 was conducted in a 326-ha pasture with treatments of no supplement (CON), .5 kg of beet pulp plus 12.5 g of DL-methionine and 30 g of urea (METU), and .4 of kg soybean meal (SOY). In Exp. 1, in situ NDF disappearance rates of cows supplemented at 1200 and 1500 were greater (P less than .01) than those of cows supplemented at 0800. In Exp. 2, ruminal ammonia concentration was greatest (P less than .001) for METU (5.2 mg/dl) and least for CON (1.5 mg/dl). Disappearance rates in situ for DM differed (P less than .01) among treatments; METU was similar (P greater than .2) to SOY but faster (P less than .01) than MET. Disappearance rates of NDF were greatest (P less than .001) for METU-supplemented cows. In Exp. 3, DM and NDF disappearance rates were faster (P less than .10) for SOY than for METU. Cow BW change was positive for METU and SOY, and decrease in condition score was least (P less than .10) for METU- and SOY-treated cows. These studies indicate favorable ruminal responses to DL-methionine supplementation; however, the response depended on time of supplementation and ruminal ammonia concentration.

Ammonia↗
PubMed1990

Extracellular ATP stimulates an amiloride-sensitive sodium influx in human lymphocytes.

Extracellular ATP has been shown to increase the Na+ permeability of human lymphocytes by 3 to 12-fold. The kinetics of this ATP-induced response were studied by measuring 22Na+ influx into chronic lymphocytic leukemic lymphocytes incubated in low-sodium media without divalent cations. ATP-stimulated uptake of 22Na-ions was linear over 4 min incubation and this influx component showed a sigmoid dependence on ATP concentration. Hill analysis yielded a K1/2 of 160 microM and a n value of 2.5. The nucleotide ATP-gamma-S (1-2 mM) gave 30% of the permeability increase produced by ATP, but UTP (2 mM) and dTTP (2 mM) had no effect on 22Na influx. The amiloride analogs 5-(N-ethyl-N-isopropyl) amiloride and 5-(N,N-hexamethylene) amiloride, which are potent inhibitors of Na(+)-H+ countertransport, abolished 72-95% of the ATP-stimulated 22Na+ influx. However, the involvement of Na(+)-H+ countertransport in the ATP-stimulated Na+ influx was excluded by three lines of evidence. Sodium influx was stimulated 7-fold by extracellular ATP but only 2.4-fold by hypertonic conditions which are known to activate Na(+)-H+ countertransport. Addition of ATP to lymphocytes produced no change in intracellular pH when these cells were suspended in isotonic NaCl media. Finally ATP caused a membrane depolarization of lymphocytes which is inconsistent with stimulation of electroneutral Na(+)-H+ exchange. These data suggest that ATP acts cooperatively to induce the formation of membrane channels which allow increased Na+ influx by a pathway which is partially inhibited by amiloride and its analogs.

Adenosine Triphosphate↗
PubMed1990

Saturation of intracellular cytosine arabinoside triphosphate accumulation in human leukemic blast cells.

Accumulation of cytosine arabinoside triphosphate (araCTP) from a range of cytosine arabinoside (araC) concentrations (1-50 microM) was measured during incubations of leukemic cells freshly isolated from patients with acute leukemia. In all but one patient, increments in extracellular araC above 10 microM did not increase intracellular araCTP levels. This maximal level of araCTP accumulation ranged from 254 to 1607 pmol/10(7) cells attained after 1 h incubation and did not correlate with either the number of nucleoside transporters on the cell membrane or the Vmax of araC phosphorylation in cell free extracts. Extremely low araCTP accumulation (103 pmol/10(7) cells/h at 50 microM araC) was observed in an AML patient with the unusual finding of micromyeloblasts. These cells also had very low numbers of nucleoside transport sites (less than 500 sites/cell) and were mitotically inactive. The unique feature of the myeloblasts from this patient was that intracellular araCTP accumulation showed a linear dependence on extracellular araC up to 50 microM with no evidence of saturation.

Acute Disease↗
PubMed1990

Substitution of DL-methionine for soybean meal as a winter supplement for gestating cows grazing native range.

A winter grazing study was conducted to determine whether DL-methionine could replace soybean meal as a N supplement for gestating beef cows. During two winters (Trial 1, n = 51; Trial 2, n = 60), crossbred beef cows grazed native foothill range. Three treatment groups were supplemented with either none (CON), DL-methionine (7.5 g Trial 1 and 9 g Trial 2) in .5 kg beet pulp carrier (BPM) or .4 kg soybean meal (SBM). Cows were supplemented individually every other day. Small differences were noted in cow BW, condition score and blood metabolites. Unsupplemented cows lost the greatest amount of BW (P less than .01) in both trials and lost more (P less than .05) condition during Trial 1 than cows fed BPM or SBM supplements. Blood samples were obtained on two consecutive days during each trial (45 d and 25 d prepartum) and analyzed for blood urea N, total bilirubin, creatinine, albumin, total protein and cholesterol. A treatment x day preparatum interaction (P less than .05) was noted for blood urea. Blood urea nitrogen declined as gestation length increased for CON and SBM cows, but blood urea of BPM-supplemented cows remained low and unchanged. In situ forage digestion was measured in 12 ruminally cannulated cows (four/treatment). In both trials, in situ rate of NDF disappearance was greater (P less than .05) for SBM than for BPM. In Trial 2, a treatment x sampling hour interaction was detected for purine concentration of whole ruminal contents; SBM maintained greater purine concentrations throughout the 48-h supplementation cycle than BPM did. Principal component analysis suggested that ruminal ammonia limited the microbial growth response to DL-methionine. Therefore, alternate-day supplementation of DL-methionine plus beet pulp did not effectively substitute for soybean meal in these trials.

Animal Feed↗
PubMed1989

Nucleoside transport and proliferative rate in human thymocytes and lymphocytes.

The thymus is a site of active T-lymphoid cell proliferation and DNA synthesis. In this study, the capacity of human thymocytes for nucleoside transport was assessed both by cytosine arabinoside influx and by equilibrium binding of nitrobenzylmercaptopurine riboside (NBMPR), a specific ligand for the equilibrative nucleoside transporter of leukocytes. The proportion of freshly isolated thymocytes synthesizing DNA was 8.6% +/- 2.1% (n = 12) by 3H-thymidine labeling index and 7.8% +/- 2.9% (n = 4) S-phase cells by flow cytometric analysis of DNA content. In comparison, both methods gave proliferation S-phase values less than 1% for peripheral blood lymphocytes (PBLs). Thymocytes expressed a high density of specific NBMPR binding sites (26,068 +/- 8,776 sites per cell, n = 12) as compared with PBLs (1,123 +/- 553 sites per cell, n = 8). The initial influx of cytosine arabinoside into thymocytes was 14-fold greater than into PBLs, and in both cell types the influx of nucleoside was totally inhibited by 0.5 mumol/L NBMPR, which is known to inhibit the major equilibrative nucleoside transporter in white blood cells. Depletion of mature CD3+ cells from the thymocyte preparation by anti-CD3 antibody left a residual population with both increased labeling index and up to twofold greater density of NBMPR binding sites. When PBLs were cultured for 48 hours with the T-cell mitogen phytohemagglutinin, a 40-fold increase in labeling index was observed, together with a 30-fold increase in the density of specific NBMPR binding sites. Thus, fresh thymocytes from human thymus are actively proliferating and express high densities of a functional nucleoside transporter. The more immature cells in the thymocyte population which are proliferating more actively have a greater density of nucleoside transporters than the whole population. In contrast, mitotically inactive PBLs-have few nucleoside transporters, but after mitogenic stimulation PBLs express large numbers of this transmembrane molecule.

Antigens, Differentiation, T-Lymphocyte↗
PubMed1989

Transport and metabolism of 1-beta-D-arabinofuranosylcytosine in human ovarian adenocarcinoma cells.

1-beta-D-Arabinofuranosylcytosine (araC) is an effective drug in the i.p. therapy of ovarian carcinoma but little is known of its transport and metabolism in this tumor. Influx of araC at 1 microM into cultured human ovarian carcinoma cells (CI 80-13S) was largely inhibited by nanomolar concentrations of the nucleoside transport inhibitor, nitrobenzylthioinosine, while the residual influx (approximately 10%) was inhibited only by micromolar concentrations of nitrobenzylthioinosine. There was a two fold greater density of specific [3H]nitrobenzylthioinosine binding to the nucleoside transporters on the ovarian than on cultured human leukemic cells (RC2a). Calculated turnover rates of the nucleoside transporter for 1 microM araC were 5-fold less in ovarian than in leukemic cells. The major metabolic product of araC was 1-beta-D-arabinofuranosylcytosine 5'-triphosphate (araCTP) which accumulated in the ovarian cells to levels half those achieved in the leukemic cells. AraC was the major product of araCTP degradation in ovarian cells consistent with a pathway (araCTP--------araCMP----araC) which is different from that previously found in leukemic cells (araCTP--------araCMP----araUMP----araU). Despite these differences, ovarian carcinoma cells show substantial accumulation of araCTP from extracellular araC.

Adenocarcinoma↗