A single dose of pegylated leridistim significantly improves neutrophil recovery in sublethally irradiated rhesus macaques.

Leridistim, a member of the myelopoietin family of dual receptor agonists that binds interleukin-3 and G-CSF receptors, has been shown to enhance hematopoietic activity in rhesus monkeys above that observed with either cytokine alone or in combination. This study demonstrated the ability of a pegylated form of leridistim (peg-leridistim), administered s.c., as a single- or two-dose regimen separated by 4 or 7 days, to significantly improve neutrophil recovery following radiation-induced myelosuppression. Rhesus macaques were total body x-irradiated (250 kVp, TBI) to 600 cGy. Following TBI, two groups received peg-leridistim (n = 5) or leridistim (n = 4) at a dose of 600 microg/kg on day 1, while two other groups (both n = 4) received peg-leridistim at 200 microg/kg on day 1 and day 4, or day 1 and day 7. The irradiation controls (n = 7) received 0.1% autologous serum for 18 days. All peg-leridistim treatment schedules significantly improved all neutrophil-related parameters following TBI as compared with nontreated controls and were equivalent in effect when compared among themselves. Administration of a single high dose or two separate lower doses of peg-leridistim significantly improved neutrophil regeneration, in a manner equal to that of conventional daily or abbreviated every-other-day administration of leridistim in this nonhuman primate model of severe myelosuppression.

A comparison of three Rorschach diagnostic systems and use of the Hand Test for detecting multiple personality disorder in outpatients.

Eleven individuals diagnosed with multiple personality disorder (MPD) on the basis of clinical observation by experienced therapists plus elevated scores on the Dissociative Experiences Scale (DES; Bernstein & Putnam, 1986) were administered the Rorschach Inkblot Test and the Hand Test. Results from the sample (n = 11) and a matched control group (N = 22) were analyzed and discussed in accordance with previous Rorschach diagnostic systems. The Wagner Signs diagnosed 91% (n = 10) of the MPD cases in this outpatient sample, with no false positives. The Labott Signs were found to have no utility, and the Barach Signs, when they occurred, seemed to be diagnostic of MPD but yielded a high rate of false negatives. Hand Test results were analyzed and found to be possibly diagnostic of MPD. Tentative criteria were proposed for its use as an additional tool for diagnosing MPD.

Purification and characterization of microbially expressed neomycin phosphotransferase II (NPTII) protein and its equivalence to the plant expressed protein.

The gene encoding neomycin phosphotransferase II (NPTII) has been used routinely as a selectable marker in the production of genetically engineered crops. To facilitate the safety assessment of this protein, the same coding sequence used for plant transformation was introduced into Escherichia coli to produce gram quantities of this protein. A unique, simple, rapid and efficient purification method was developed to purify thirty grams of NPTII protein. The microbially produced NPTII was shown to be chemically and functionally equivalent to the NPTII protein expressed in and purified from genetically engineered cotton seed, potato tubers and tomato fruit. Microbially produced and plant produced NPTII proteins have comparable molecular weights, immuno-reactivities, epitope structures, amino terminal amino acid sequences, biological activities and both lack glycosylation. Demonstrating the equivalence of NPTII protein from these sources establishes the validity of using the microbially produced NPTII to assess the safety of the NPTII protein produced in genetically engineered crops.

Comparison of performance, clinical chemistry, and carcass characteristics of finishing lambs treated with recombinant ovine or bovine somatotropins.

The variation in growth and carcass composition responses of lambs to somatotropin (ST) treatment may depend on the source of ST used as well as on other experimental conditions. In the present experiment, growth, carcass composition, and clinical chemistry responses to recombinantly produced ovine ST (oST) and two bovine ST (N-methionyl-bST[M-bST] and N-alanyl-bST[A-bST] were compared. Lambs weighing 42 kg were assigned to treatment groups of control (no injection) or 4 mg/d of M-bST, A-bST, or oST administered by s.c. injection for 6 wk. Growth rate was increased by an average of 30% and feed efficiency was improved by an average of 22% by ST treatment compared with control, and responses did not differ among ST. The IGF-I, insulin, and glucose concentrations were increased by 107, 700, and 53% compared with control, respectively, and did not differ among ST treatment groups. Urea nitrogen responses to A-bST and oST were transiently greater than those to M-bST. Although quality grade was not affected by treatment, an average .8-kg increase in weight of retail cuts was calculated from yield grade. Carcasses of ST-treated lambs were calculated to have 1.3 kg more muscle and 1.9 kg less fat. Although fat and muscle were affected more by oST than by M-bST on a percentage basis, they did not differ among treatment groups on a total weight basis. Thus, both bST variants and oST improved growth performance and carcass leanness. Decreased responses of some carcass variables to M-bST treatment may have been related to the presence of antibodies that were indicated by an increased number of positive responders in a relative bST binding assay.

High-level production of active HIV-1 protease in Escherichia coli.

High levels of active HIV-1 protease (PR) were produced in Escherichia coli, amounting to 8-10% of total cell protein. High production levels were achieved by altering the following parameters: (1) codon preference of the coding region, (2) A+T-richness at the 5' end of the coding region, and (3) promoter. To circumvent the toxicity of HIV-1 PR in E. coli, the gene was expressed as a fusion protein with two different proteolytic autocleavage sequences. In both the cases, the fusion protein could be cleaved in vivo to give an active molecule with the native sequence at the N terminus.

Synthetic peptide inhibitors of complement serine proteases--III. Significant increase in inhibitor potency provides further support for the functional equivalence hypothesis.

Synthetic peptides based on functionally equivalent (as defined by similar patterns of chemically equivalent amino acids) serine protease inhibitor (serpin) C-terminal sequences inhibit both classical and alternative pathways of complement activation. Inhibition was also found with hybrid peptides consisting of the cleavage site of one serpin (antithrombin III, alpha-1-antitrypsin, or antichymotrypsin) attached to the short and long functionally equivalent protease binding cores of the other two serpins. A hybrid peptide composed of the sequence at the site of cleavage of C4 by C1s attached to the long binding core of antithrombin III was selective in inhibiting the classical pathway with no effect on the alternative pathway at a concn of 10 microM. Extension of the functional equivalence hypothesis has produced inhibitors of complement activation named generic and generic +, whose sequences differ by 77% or 87%, respectively, from those of all three serpin sequences. A hybrid peptide composed of the antithrombin III cleavage site attached to the generic peptide is an inhibitor of complement activation at 500 nM, the most potent inhibitor found in this study.