Preparation of 125I-thyroxine and 125I-triiodothyronine of high specific activity for the radioimmunoassay of serum total T4 and T3.

A simple and inexpensive procedure for the preparation of high specific activity 125I radioiodinated triiodothyronine and thyroxine is described. The specific activities achieved are approximately 350 Ci/g for 125I-T4, and either 200 Ci/g for 125I-T3 depending on the reagents employed. The high specific activity 125I-T4 and 125I-T3 are suitable for the routine radioimmunoassay of serum total T3 and T4. When properly stored they have a self life of at least seven weeks.     

Mechanism-based pharmacokinetic-pharmacodynamic modeling of 5-HT1A receptor agonists: estimation of in vivo affinity and intrinsic efficacy on body temperature in rats

The pharmacokinetic-pharmacodynamic (PK-PD) correlations of seven prototypical 5-HT(1A) agonists were analyzed on the basis of a recently proposed semi-mechanistic PK-PD model for the effect on body temperature. The resulting concentration-effect relationships were subsequently analyzed on the basis of the operational model of agonism to estimate the operational affinity (pK(A)) and efficacy (log tau) at the 5-HT(1A) receptor in vivo. The values obtained in this manner were compared with estimates of the affinity (pK(i)) and intrinsic efficacy (log[agonist ratio]) in a receptor-binding assay. Between 5-HT(1A) agonists wide differences in in vivo affinity and efficacy were observed, with values of the pK(A) ranging from 5.67 for flesinoxan to 8.63 for WAY-100,635 [N-(2-(4-(2-methoxyphenyl)-1-piperazinyl)ethyl)-N-2-pyridinyl-cyclohexanecarboxamide] and of the log tau ranging from -1.27 for WAY-100,135 [N-(1,1-dimethylethyl)-4-(2-methoxyphenyl)-alpha-phenyl-1-piperazine-propanamide] to 0.62 for R-(+)-8-hydroxy-2-[di-n-propylamino)tetralin. Poor correlations were observed between the in vivo receptor affinity (pK(A)) and the affinity estimates in the in vitro receptor binding assay (pK(i); r(2) = 0.55, P > 0.05), which could in part be explained by differences in blood-brain distribution. In contrast, a highly significant correlation was observed between the efficacy parameters in vivo (log tau) and in vitro (log [agonist ratio]; r(2) = 0.76, P < 0.05). Thus by combining the previously proposed semi-mechanistic PK-PD model for the effect on body temperature with the operational model of agonism, a full mechanistic PK-PD model for 5-HT(1A) receptor agonists has been obtained, which is highly predictive of the in vivo intrinsic efficacy.     

A monoclonal antibody detecting a new human T cell antigen, HuLy-m2

HuLy-m2 is a new T cell antigen detected by a monoclonal antibody. The antibody (anti-HuLy-m2) reacts with 85% of human T cells, 20% B cells, and 50% null cells in peripheral blood. The antibody is cytotoxic, binds protein A, and is of the IgG2a subclass. HuLy-m2 antigen consists of a glycoprotein dimer Mr 37,000 subunit structure, in which sialic acid and carbohydrate play some role in antigenicity. Capping studies showed the HuLy-m2 antigen to be distinct from the previously described OKT-3, 4, 8, HuLy-m1 (OKT11), and HuLy-m3 antigens.     

Characterization of activated lymphocyte-tumor cell adhesion

This study demonstrates the variable expression of ICAM-1 and leukocyte function antigen-3 (LFA-3) on four tumor cell lines (COLO526, K562, Daudi, and HT-29). In addition, phorbol ester (PMA) activation of lymphocytes modulated LFA-1 from a uniform to a clustered surface distribution; whereas after treatment with high levels of Mg2+ ions, the unique epitope for high-affinity LFA-1 was identified using clone Mab24. Using a flow cytometric adhesion assay it was demonstrated that PMA-activated lymphocytes formed conjugates with COLO526 and Daudi, and that these conjugates were inhibited by anti-CD2 with varying inhibition by LFA-1 clones MHM24 and 25.3.1. When lymphocytes were induced to express the high-affinity form of LFA-1, conjugates were identified with COLO526, K562, and Daudi and these conjugates were sensitive to the presence of both CD2 and LFA-1 antibodies. Further studies using confocal microscopy confirmed significant adhesion between peripheral blood lymphocytes pretreated with either PMA or high levels of Mg2+ and the adherent cell line COLO526. In conclusion, this unique study has demonstrated for the first time the important role of the active form of LFA-1 on the lymphocyte cell surface for conjugate formation with an ICAM-1-expressing tumor cell; also, two pathways of cell signaling were identified for conjugate formation to occur.     

Effect of Hand Positioning on DXA Total and Regional Bone and Body Composition Parameters,Precision Error,and Least Significant Change

Dual-energy X-ray absorptiometry (DXA) body composition measurements are performed in both clinical and research settings for estimations of total and regional fat mass, lean tissue mass, and bone mineral content. Subject positioning influences precision and positioning instructions vary between manufacturers. The aim of the study was to determine the effect of hand position and scan mode on regional and total body bone and body composition parameters and determine protocol-specific body composition precision errors. Thirty-eight healthy subjects (men; mean age: 27.1?±?12.1?yr) received 4 consecutive total body GE-Lunar iDXA (enCORE v 15.0) scans with re-positioning, and scan mode was dependent on body size. Twenty-three subjects received scans in standard mode and 15 received scans in thick scan modes. Two scans per subject were conducted with subject hands prone and 2 with hands mid-prone. The precision error (root mean squared standard deviation; percentage coefficient of variation) and least significant change for each protocol were determined using the International Society for Clinical Densitometry calculator. Hands placed in the mid-prone position increased arm bone mineral density (BMD) (standard mode: 0.185?g*cm^?2, thick mode: 0.265?g*cm^?2; p?<?0.05), total body BMD (standard mode: 0.051?g*cm^?2, thick mode: 0.069?g*cm^?2; p?<?0.001), and total body BMD Z-score (standard mode: 0.5. thick mode: 0.7; p?<?0.001). This was due to reductions in bone area and bone mineral content. In standard mode, hands mid-prone reduced fat mass (0.05?kg, p?<?0.05) and increased lean mass (0.11?kg, p?<?0.05). There were no differences in body composition for thick mode scans. Hands mid-prone reduced lean mass precision error at the arms, trunk, and total body (p?<?0.01). DXA clinical and research centers are advised to maintain consistency in their hand positioning and scan mode protocols, and consideration should be given to the hand positioning used for reference data. As a best practice recommendation, published DXA-based studies and reports for clinic-based total body assessments should ensure that subject positioning is fully described.     

Distribution of ω-Conotoxin GVIA binding sites in teleost cerebellar and electrosensory neurons

The distribution of ω-Conotoxin GVIA (CgTx) binding sites was used to localize putative N-type Ca²⁺ channels in an electrosensory cerebellar lobule, the eminentia granularis pars posterior, and in the electrosensory lateral line lobe of a gymnotiform teleost (Apteronotus leptorhynchus). The binding sites for CgTx revealed by an anti-CgTx antibody had a consistent distribution on somatic and dendritic membranes of specific cell types in both structures. The distribution of CgTx binding was unaffected by co-incubation with nifedipine or Aga Toxin IVA, blocking agents for L- and P-type Ca²⁺ channels, respectively. Incubation with CgTx in the presence of varying levels of extracellular Ca²⁺ altered the number but not the cell types exhibiting immunolabel. A punctate immunolabel was detected on somatic membranes of granule and stellate cell interneurons in both the eminentia granularis pars posterior and the electrosensory lateral line lobe. Punctate CgTx binding sites were also present on spherical cell somata and on the large presynaptic terminals of primary afferents that terminate on spherical cells in the electrosensory lateral line lobe. No label was detected in association with distal dendritic membranes of any cell class, or with parallel fibers in the respective molecular layers. Binding sites for CgTx in the eminentia granularis are consistent with the established role for N-type Ca²⁺ channels in cell migrations, an activity which is known to persist in this layer in adult Apteronotus. The distribution of labeled stellate cells with respect to topographic maps in the electrosensory lateral line lobe further suggest that N-type Ca²⁺ channels are expressed in relation to functional activity across these sensory maps. © 1996 Wiley-Liss, Inc.