The self in transformation: the passage from a two- to a three-dimensional internal world.

This paper seeks to describe some of the steps in the process of transformation from a two-dimensional to a three-dimensional inner world as experienced by a patient who had essentially used up her remaining vital resources. An important hurdle had first to be crossed when the analyst was required to demonstrate her capacity to survive with her in a two-dimensional claustrum, in a solitary confinement. Here, an oppositional defensive system had been created in which positive and negative personifications of vital but withdrawn energies ensured that she was locked within an interior world that sought to bar entry to anyone else, and where the possibility of the transformation of her self was disallowed. The patient had to accept the risk of breaching the system of self-defence before the analyst was allowed a mutual position within her interior world. A series of dreams tracks this transformation internally and in relation to the analyst.

Selective inhibition of N-formylpeptide-induced neutrophil activation by carbamate-modified peptide analogues.

Stimulation of the leukocyte N-formylpeptide receptor (FPR) induces chemotaxis, cell adhesion, free radical release, and degranulation, responses associated with infection and inflammation. Under conditions where continuous activation of the receptor prevails, neutrophil-dependent tissue damage ensues. Antagonists of the FPR have potential for use as diagnostic and therapeutic agents. Hence, we have synthesized and evaluated a series of amino-terminal carbamate analogues of the peptide Met-Leu-Phe (MLF) in order to determine the structural requirements for imparting agonist or antagonist activity at the human neutrophil FPR. Peptides were evaluated in three in vitro assays: receptor binding, superoxide anion release, and cell adhesion. Unbranched carbamates (methoxycarbonyl, ethoxycarbonyl, and n-butyloxycarbonyl) resulted in agonist activity, whereas branched carbamates (iso-butyloxycarbonyl, tert-butyloxycarbonyl, and benzyloxycarbonyl) were antagonists. The peptide antagonists were more potent inhibitors of superoxide anion release than cell adhesion by 4-7-fold. When iso-butyloxycarbonyl-MLF (i-Boc-MLF) was further modified at the carboxy terminus with Lys, antagonist potency was retained but without functional selectivity. Further C-terminal modification with the radionuclide linker diethylenetriaminepentaacetic acid did not alter the potency of i-Boc-MLFK. These results indicate that the switch from agonist to antagonist activity can be achieved by modifying the overall size and shape of the amino-terminal group; that modifications at both the amino and carboxy termini can alter the functional selectivity of the peptide; and that modifications can be tolerated at the carboxy terminus to allow for development of an antagonist for diagnostic applications.

[99mTc]tricine: a useful precursor complex for the radiolabeling of hydrazinonicotinate protein conjugates.

The stannous reduction of [99mTc]pertechnetate in the presence of tricine results in the formation of the new labeling precursor complex [[99m-Tc]tricine. This complex has improved efficacy for the 99mTc labeling of hydrazinonicotinate-modified IgG compared to [99mTc]glucoheptonate. FAB mass spectral analysis of the product formed by the reaction of [TcOCl4](-1) with tricine indicates the formation of [TcO(tricine x 2H)2](-1).

Inhibition of chemotactic peptide-induced neutrophil adhesion to vascular endothelium by cAMP modulators.

An early event associated with neutrophil-dependent tissue damage involves the adhesion of neutrophils to the vascular endothelium and the subsequent release of oxygen-derived free radicals and granule constituents. Elevations in intracellular cAMP are known to inhibit free radical release but not lysosomal enzyme release. The role of cAMP in FMLP-induced neutrophil adhesion was examined in this study by using an in vitro model of neutrophil-endothelial cell adhesion. FMLP stimulated a time- and concentration-dependent increase in human neutrophil adhesion to HUVEC. FMLP-mediated adhesion was inhibited by a diverse group of cAMP modulators: forskolin, isoproterenol, phosphodiesterase IV inhibitors (rolipram and Ro 20-1724), but not phosphodiesterase III inhibitors (milrinone and bemoradan). Endogenous adenosine has previously been shown to mediate FMLP-induced increases in cAMP enhanced in the presence of Ro 20-1724. In this study, adenosine deaminase prevented the inhibitory effects observed with rolipram and Ro 20-1724, implicating endogenous adenosine as a co-modulator of inhibition. FMLP stimulated neutrophil shape change and the surface expression of the beta 2 integrins CD11b/CD18 and CD11a/CD18. Both these responses were inhibited by rolipram but not bemoradan. With the use of 4,4'-diisothiocyanostilbene-2,2'disulfonic acid, we showed that mobilization of the intracellular pool of CD11b/CD18 paralleled adhesion. We conclude that neutrophil-endothelial cell adhesion is attenuated by elevating neutrophil intracellular cAMP and that inhibition of neutrophil CD11b/CD18 surface expression by cAMP accounts for this observed inhibition of adhesion.

Imaging of focal sites of inflammation in rhesus monkeys with 99mTc-labeled human polyclonal IgG.

The biological behavior of human polyclonal immunoglobulin G (IgG) radiolabeled with 99mTc via a nicotinyl hydrazine derivative, was evaluated in Rhesus monkeys. 99mTc-IgG and 111In-MACROSCINT DTPA-IgG were co-administered to Rhesus monkeys with focal sites of sterile inflammation and scintillation camera images were acquired at 6 and 19 h after injection. The biodistribution of the two antibody preparations were similar, however, small differences were detected: 99mTc-IgG > 111In-IgG in spleen and lung; 99mTc-IgG in bone and skeletal muscle. A linear correlation of the tissue-to-blood ratios of 99mTc- and 111In-labeled IgG was observed at both times (r2 > 0.98). The slopes of the regression lines were not significantly different from unity: 6 h-0.982 +/- 0.018; 19 h 1.0334 +/- 0.0226. Also, at both 6 and 19 h after injection, the target-to-background ratios (T/B) for the sites of inflammation were remarkably similar for 111In and 99mTc. These studies establish that human polyclonal IgG labeled with 99mTc via a nicotinyl hydrazine modified intermediate is equivalent to 111In-MACROSCINT DTPA-IgG for imaging focal sites of inflammation in monkeys.

Localization of indium-111-immunoglobulin G, technetium-99m-immunoglobulin G and indium-111-labeled white blood cells at sites of acute bacterial infection in rabbits.

Biodistribution and infection imaging properties of 111In-DTPA-IgG, 99mTc-hydrazino nicotinamide-IgG and 111In-WBC were compared in rabbits with E. coli infection. Groups of six rabbits were injected with 10 mCi of 99mTc-IgG plus 0.5 mCi of 111In-IgG or 1 mCi of 99mTc-IgG plus 0.05 mCi of 111In-WBC. At 4-5 and 18-20 hr, dual photon scintigrams were acquired. At both times, the distributions of 99mTc and 111In-IgG were nearly identical. The sites of infection were well visualized with all three radiopharmaceuticals. In the early images, the target-to-background ratios (T/B) for 111In and 99mTc-IgG determined by ROI analysis were 1.95 +/- 0.26 and 2.57 +/- 0.38 (p = NS). In the delayed images, the T/B ratios increased (p < 0.01) to 3.56 +/- 0.49 and 4.90 +/- 0.98. At both times, the T/B ratios for 111In-WBC were higher (p < 0.01); 4.17 +/- 0.78 at 4-5 hr and 8.52 +/- 1.52 at 18-20 hr. These results indicate that all three agents yield excellent images of infection sites. Although 111In-WBC had higher T/B ratios, the ease of preparation of the radiolabeled proteins makes them attractive alternatives for infection imaging.

A model of gastric emptying in cats shows solid emptying is promoted by MK-329: a CCK antagonist.

The effect of MK-329, a potent, orally active, nonpeptidal cholecystokinin (CCK) antagonist, was measured on the gastric emptying rate of a solid meal in cats. External scintigraphy of cats that had been fed a meal of technetium-99m-(99mTc) labeled rabbit liver and light cream allowed the measurement of the emptying rates of either the liquid or solid portion of a meal under physiologic conditions. In cats, liquids emptied 2.6 times faster than solids [163 +/- 11 min vs. 62 +/- 3 min (mean +/- s.e.)]. At 3 or 10 mg/kg p.o., MK-329 gastric emptying was significantly accelerated, with the mean half-time of emptying being decreased by 34 +/- 11% (mean +/- s.e.) of the control half-times (p less than 0.02). Using only responders (five of six animals), mean half-time was decreased by 55 +/- 4% of the control half-times. CCK is important in regulating the emptying of solid food from the stomach because the CCK antagonist MK-329 accelerates that emptying.

A survey of staphylococci isolated from the laboratory gerbil.

A microbiological survey of the Mongolian gerbil, Meriones unguiculatus, revealed coagulase-negative staphylococci to be common inhabitants of representative animals derived from three different breeding colonies. The nasal area was most often culture positive, and Staphylococcus xylosus was a predominant species. S. xylosus was the only organism cultured from nasal dermatitis. These organisms were found to be susceptible in vitro to the majority of the antimicrobial agents tested. This survey indicates that the possible role of S. xylosus as an opportunistic pathogen warrants further investigation.

(+)-3-[123I]Iodo-MK-801: synthesis and characterization of binding to the N-methyl-D-aspartate receptor complex.

Synthetic methods have been established for preparing high specific activity (+)-3-[123I]Iodo-MK-801 in high radiochemical yield. The binding of the radiotracer to rat cortical membranes has been examined to assess its potential use as an in vivo imaging agent for the N-methyl-D-aspartate (NMDA) receptor-ion channel complex. Under the conditions of the assay, specific (+)-3-[123I]Iodo-MK-801 binding to membrane homogenates represented greater than 95% of the total binding. Several structurally distinct, noncompetitive NMDA receptor antagonists inhibited binding with potencies in accordance with their reported inhibitory activity at the receptor complex. The concentration of (+/-)-3-Iodo-MK-801 required to inhibit 50% of (+)-3-[123I]Iodo-MK-801 binding (IC50) was 3.4 nM when using a low ionic strength assay buffer and 5.5 nM in a physiological buffer. In a thoroughly washed membrane preparation, (+)-3-[123I]Iodo-MK-801 binding was enhanced by L-glutamate and glycine at concentrations known to activate the NMDA receptor. The results indicate that (+)-3-[123I]Iodo-MK-801 specifically labels the NMDA receptor complex in rat brain membranes and the retention of high affinity under near physiological assay conditions suggests that it may be useful as a SPECT imaging agent for the receptor in vivo.

Estradiol distribution and penetration in rat skin after topical application, studied by high resolution autoradiography.

Transdermal pathways and targets in the skin for estradiol were investigated using dry-mount autoradiography. 3H-estradiol-17 beta was applied at doses of 30.1 pmol, 120.4 pmol and 301 pmol/cm2 to shaved rat skin in the dorsal neck region. Vehicles were DMSO, ethylene glycol or sesame oil. After 2 h of topical treatment with 30.1 pmol 3H-estradiol x cm-2 dissolved in DMSO a distinct cellular distribution was apparent. Target cells with concentrations of radioactivity were found in epidermis, sebaceous glands, dermal papillae of hair and fibroblasts. After treatment with 120.4 and 301 pmol/cm2, a penetration gradient of radioactivity was recognizable however it masked specific cellular and subcellular uptake. The stratum corneum accumulated and retained radioactivity, apparently forming a depot for the hormone. Strong concentration and retention of the hormone was conspicuous in sebaceous glands for more than 24 h, suggesting that sebaceous glands serve as a second storage site for the hormone. In all autoradiograms two penetration pathways to the dermis were visible: one through the stratum corneum and epidermis, the other through the hair canals and hair sheaths.