A comparison of mass balance, pharmacokinetics and disposition of [14C(U)]- and [125I]recombinant human interleukin-2 in cynomolgus monkeys.

Recombinant human interleukin-2 (rHuIL-2) has been metabolically labeled with 14C amino acids in Escherichia coli and affinity purified on a rHuIL-2 receptor affinity column. The radiolabeled molecule had a specific radioactivity of 238 dpm/unit and the identical amino acid sequence and biological activity as unlabeled rHuIL-2. In this study, we used this labeled [14C(U)]rHuIL-2 and commercially available [125I]rHuIL-2 (identical in sequence to the [14C(U)]rHuIL-2) to compare the mass balance, pharmacokinetics, and disposition in cynomolgus monkeys. After a single intravenous bolus dose of 4 x 10(5) units/kg, serum samples were collected for 7 days and examined for biological activity, total radioactivity, and by molecular size exclusion chromatography. Urine and feces were analyzed for total radioactivity. When analyzed for biological activity, both [14C(U)]- and [125I]rHuIL-2 exhibited the following pharmacokinetic parameters: terminal elimination half-life of 1-2 hr, AUC0-infinity ranged from 2005 to 4659 units x hr/ml, clearance was 90-200 ml/hr/kg, and volume of distribution ranged from 103 to 163 ml/kg. Comparison of the pharmacokinetic profiles of the two radiolabels were very different from bioactivity, in that the elimination half-lives for radioactivity were approximately 8 days and 10 hr for [14C(U)]- and [125I]rHuIL-2, respectively. We conclude that the [14C(U)]rHuIL-2 was metabolized to constituent amino acids and recycled into newly synthesized proteins from our size exclusion chromatography studies.(ABSTRACT TRUNCATED AT 250 WORDS)

Ultraviolet-B (290-320 nm)-irradiation inhibits epidermal growth-factor binding to mammalian cells.

Mitogens, such as polypeptide growth factors and phorbol ester tumor promoters, act by binding to specific receptors and inducing a pleiotropic response in cultured mammalian cells, which results in the induction of cellular proliferation. An early effect of such agents is the inhibition of binding of epidermal growth factor (EGF) to its receptor. Ultraviolet radiation has also been shown to induce a proliferative response in vivo and in vitro and to act as a tumor promoter in animal skin. We, therefore, examined the effect of ultraviolet radiation (UVB - 290-320 nm) on EGF binding to cells in culture. We found that UVB (100-300 J/m2) induced a rapid, dose-dependent inhibition of EGF binding in a mouse fibroblast cell line, which resulted from a decrease in both number and affinity of binding sites. Phosphorylation of the EGF receptor by protein kinase C (PKC) is not likely to be the mechanism for inhibition, since UVB treatment did not result in PKC activation or modulation of phorbol diester binding.

Ultraviolet radiation alters choline phospholipid metabolism in human keratinocytes.

Ultraviolet radiation B (UVB-290-320 nm) induces inflammation and hyperproliferation in human epidermis. This response is associated with the recovery from irradiated skin of inflammatory mediators derived from membrane phospholipids. We have previously reported that UVB stimulates the production of such mediators by human keratinocytes (HK) in culture. In these studies we examined the effect of UVB on the metabolism of choline containing phospholipids in HK prelabeled with [3H] choline. UVB (400-1600J/m2) stimulated a dose dependent release of [3H] choline from HK within minutes of irradiation. Examination of media extracts by paper chromatography revealed that the released [3H] choline was predominately in the form of glycerophosphorylcholine. Examination of label remaining in membranes of cells after irradiation by acid precipitation and HPLC revealed that the origin of the released [3H] choline was the membrane phosphatidylcholine/lysophosphatidylcholine. These data support a concept of UVB stimulation of both a phospholipase A (1 or 2) and a lysophospholipase. These UVB induced alterations of HK membrane phospholipid metabolism likely have profound effects on UVB-induced inflammation and control of cell growth in human skin.

Effect of strategic anthelmintic treatment and pasture management on productivity and control of nematode parasites in weaner-yearling beef cattle.

Three groups of 17 beef calves were used to evaluate effects of strategic anthelmintic treatment on safe (group 1) and contaminated (group 2) pasture in comparison with minimal treatment at weaning and contaminated pasture (group 3). The investigation extended from weaning in November 1982 to the following August. Results of faecal egg counts, herbage larval counts, plasma pepsinogen and tracer calf worm counts in autumn and spring indicated minimal levels of infection on safe pastures provided in November and April (group 1). A decided weight gain advantage for group 1 was achieved from November to April, but the rate of gain was not consistent after April and transfer to the second safe pasture. Final average weights in late August were: group 1, 368 kg; group 2, 336 kg; group 3, 262 kg. All were significantly different (P less than 0.05). Worm counts from representative yearlings in September revealed low to moderate levels of Ostertagia ostertagi in group 1. In contrast group 2 cattle had large, almost exclusively O ostertagi infections; group 3 cattle had exceedingly high levels of Trichostrongylus axei infection and moderate to high levels of O ostertagi. Marginal evidence of type 2 ostertagiasis was observed in individual animals of group 2 and group 3.

Ultraviolet radiation acts as an independent mitogen for normal human melanocytes in culture.

Identification of growth factors for normal human melanocytes has been significantly aided by the recent development of in vitro culture systems for this cell. Utilizing such a system, we studied the effect of ultraviolet radiation (UVR) on both melanocyte growth and melanization by incorporation of 3H-thymidine and 3H-L-dihydroxyphenylalanine (3H-DOPA), respectively. 3H-thymidine incorporation was found to be significantly stimulated during the first 24 h following a single irradiation. 3H-DOPA incorporation was stimulated after a delay of 2 days postirradiation. Whereas UVR has long been known to induce melanocyte proliferation in vivo, these studies show that UVR can act as a mitogenic stimulus for this cell independent of the cutaneous environment. UVR can thus be added to a growing list of growth factors for epidermal pigment cells and is the only physical agent conclusively shown to act as a mitogen. Included in this list are substances that act via stimulation of the CAMP-kinase or protein kinase systems such as cholera toxin and phorbol esters. UVR is postulated to induce melanocyte proliferation by modulation of these second messenger pathways. With recent evidence linking growth factors, oncogenes and malignant transformation, this study supports the association between UVR exposure and the development of malignant melanoma, and suggests mechanisms whereby UVR may contribute to malignant transformation of this cell.

Benoxaprofen photosensitization of phospholipase activation in mammalian cells in culture.

Benoxaprofen (BPF) induces a clinical photosensitivity which resembles idiopathic solar urticaria. This response is mediated by degranulation of mast cells. Since mast-cell degranulation is known to be modulated by phospholipid alteration, we examined the effect of BPF and ultraviolet radiation (UV) on phospholipid metabolism of C3H 10 T1/2 cells in culture. BPF + UVB (280-320 nm) induced release of [3H]arachidonic acid (AA) but did not alter the release of [3H]choline from prelabelled cells. This suggests that BPF photosensitizes phospholipase A2 activation. Such activation probably represents an integral step in the mechanism of BPF photosensitization of mast-cell degranulation.

The effect of 12-O-tetradecanoylphorbol-13-acetate (TPA) on phospholipid metabolism of human epidermal keratinocytes in culture.

The tumor promoter 12-O-tetradecanoylphorbol-13-acetate (TPA) stimulated the release of [3H]choline from prelabelled membrane phospholipids of cultured keratinocytes obtained from normal human skin. In contrast, TPA in the concentration range of 10(-12) to 10(-6) g/ml failed to induce deacylation of [3H]arachidonic acid or stimulate [3H]prostaglandin production in prelabelled keratinocytes. In addition, TPA did not induce [3H]choline incorporation into the membrane phospholipids of these cells. The previously reported inability of TPA to stimulate a proliferative response in these cell cultures may be related to the resistance of these cells to TPA-induced alterations of arachidonate metabolism.

Efficacy of fenbendazole against inhibited larvae of Ostertagia ostertagi in yearling cattle.

Efficacy of fenbendazole, at doses of 7.5 and 10.0 mg/kg of body weight, against inhibited early 4th-stage larvae of Ostertagia ostertagi and other nematodes of the abomasum and intestinal tract, was investigated in naturally infected yearling heifers in late May 1982. In Louisiana, this is near the end of the period (March to May) in which maximal numbers of inhibition-prone larvae are acquired. The mean numbers of O ostertagi in 10 untreated control cattle were: adults, 4,880; developing 4th-stage larvae, 12,546; and inhibited early 4th-stage larvae, 167,931. At the 7.5 mg/kg dose level (10% liquid suspension) in 10 cattle, percentage reduction of O ostertagi in comparison with controls was: adults, 95.7%; developing 4th stages, 91.1%; and inhibited 4th stage, 55.0%. Percentage reductions of other genera were as follows: abomasum--Trichostrongylus axei, 99.6%; Haemonchus sp, 95.1%; intestinal tract--Cooperia spp, 97.8%; Trichostrongylus colubriformis, 100.0%; and Oesophagostomum radiatum 4th stage and adults, 100.0%. At the 10.0 mg/kg dose (10% liquid suspension) in 11 cattle, the percentage reduction of O ostertagi in comparison with controls was: adults, 98.6%; developing 4th stages, 92.9%; and inhibited 4th stage, 80.0%. Percentage reductions of other genera were: abomasum--T axei, 99.9%; Haemonchus sp, 98.8%; intestinal tract--Cooperia spp, 99.3%; T colubriformis, 100.0%; and Oes radiatum 4th stage and adults, 100.0%. Variability of efficacy against inhibited larvae was observed, particularly at the 7.5 mg/kg dose; at this dose, 7 of the 10 heifers in the group yielded in excess of 54,000 surviving larvae.(ABSTRACT TRUNCATED AT 250 WORDS)