Thrombalexins: Cell-Localized Inhibition of Thrombin and Its Effects in a Model of High-Risk Renal Transplantation.

Allograft transplantation into sensitized recipients with antidonor antibodies results in accelerated antibody-mediated rejection (AMR), complement activation, and graft thrombosis. We have developed a membrane-localizing technology of wide applicability that enables therapeutic agents, including anticoagulants, to bind to cell surfaces and protect the donor endothelium. We describe here how this technology has been applied to thrombin inhibitors to generate a novel class of drugs termed thrombalexins (TLNs). Using a rat model of hyperacute rejection, we investigated the potential of one such inhibitor (thrombalexin-1 [TLN-1]) to prevent acute antibody-mediated thrombosis in the donor organ. TLN-1 alone was able to reduce intragraft thrombosis and significantly delay rejection. The results confirm a pivotal role for thrombin in AMR in vivo. This approach targets donor organs rather than the recipient and is intended to be directly translatable to clinical use.

An antibody combination that targets activated T cells extends graft survival in sensitized recipients.

Memory T cells are the very essence of adaptive immunity with their rapid and efficient response to antigen rechallenge and long-term persistence. However, it is becoming increasingly evident that when primed with self or transplanted tissue, these cells play a key role in causing and perpetuating tissue damage. Furthermore, current treatments, which efficiently control the naive response, have limited effects on primed T cells. We have used a treatment based on a combination of antibodies specific for molecules expressed by activated T lymphocytes to selectively remove these cells. This approach, which we termed multi-hit therapy, leads to cumulative binding of antibodies to the target T cells and a striking prolongation of skin graft survival in presensitized recipients in a stringent skin transplant model. The findings are consistent with the depletion of graft-specific CD4+ and CD8+ T cells, although other modes of action, such as T-cell regulation and altered migration could play a role. In conclusion, our therapeutic strategy controls primed T cells which are a major driving force in the pathology of many autoimmune diseases and in transplant rejection.

Pharmacist-designed and -implemented pharmaceutical care plan for antipsychotic-induced movement disorders.

We developed a systematic approach to assess the presence, severity, and management of extrapyramidal symptoms (EPS) in patients treated with antipsychotics. Patients were evaluated by the Modified Simpson-Angus scale, Abnormal Involuntary Movement Scale, and Dyskinesia Identification System: Condensed User Scale. We completed 235 sets of evaluations in 83 patients. A pharmaceutical intervention was proposed in 54% (130) of evaluations, of which 82% (107) were accepted and followed. In 93% (99) evaluations in which a recommendation was followed, clinical outcome was positive. The most common intervention was reducing the dosage or discontinuing the antidyskinetic agent, most often an anticholinergic (55% of cases). Our results show that detailed monitoring of EPS in a clinical pharmacist-operated clinic promotes rational drug therapy, limits unnecessary drugs, and improves clinical outcome of patients with EPS.

Predominant role for C5b-9 in renal ischemia/reperfusion injury.

Previous work has indicated that complement is a mediator of ischemia/reperfusion (I/R) injury. To investigate the components of complement responsible for this effect, we examined a model of renal I/R injury in C3-, C4-, C5-, and C6-deficient mice. We occluded the renal arteries and veins (40-58 minutes) and, after reperfusion (0-72 hours), assessed renal structural and functional injury. C3-, C5-, and C6-deficient mice were protected from renal I/R injury, whereas C4-deficient mice were not protected. C6-deficient mice treated with antibody to block C5a generation showed no additional protection from I/R injury. Reconstitution with C6 alone restored the I/R injury in C6-deficient mice. Tubular epithelial cells were the main structures damaged by complement-mediated attack, and, in contrast, the renal vessels were spared. Neutrophil infiltration and myeloperoxidase activity were reduced in C-deficient mouse kidney, but by a similar extent in C3-deficient and C6-deficient mice. We conclude that the membrane attack complex of complement (in which C5 and C6 participate) may account for the effect of complement on mouse renal I/R injury. Neither C5a-mediated neutrophil infiltration nor the classic pathway, in which C4 participates, appears to contribute to I/R injury in this model. By contrast with other organs, such as the heart, the primary effect of complement in the ischemic area is on the parenchymal cell rather than the vascular endothelial cell. The membrane attack complex of complement is a potential target for prevention of I/R injury in this model.

Inhibition of expression of the Galalpha1-3Gal epitope on porcine cells using an intracellular single-chain antibody directed against alpha1,3galactosyltransferase.

The carbohydrate epitope Galalpha1-3Gal has been shown to be the major target of natural antibodies responsible for hyperacute rejection of porcine tissues transplanted into primates. We have sought to produce a phenotypic knockout of the alpha1, 3Galactosyltransferase enzyme that is responsible for generating this epitope, using an intracellular antibody approach. We have isolated high affinity anti-alpha1,3Galactosyltransferase single-chain antibodies from a semi-synthetic phage display library. Expression of a KDEL-tagged anti-alpha1,3Galactosyltransferase single-chain antibody in a porcine endothelial cell line resulted in the decreased expression of the Galalpha1-3Gal epitope and increased resistance to lysis by human serum.

Management of plant viral diseases through chemical control of insect vectors.

In this article we consider the role of epidemiological factors and transmission processes of insect-vectored viruses on the effectiveness of insecticides in a disease management program. We also discuss the use of insecticides within the broader framework of the chemical environment surrounding vectors, and how chemical-induced alterations in the behavior of vectors can influence transmission. Our analysis confirms the belief of Heathcote, who stated in 1973, that "no one method of control is likely to keep crops entirely free from virus infection and as many preventative measures should be taken as are economically justified."

Stimulus cancellation by macaques with unilateral frontal or parietal lesions.

Monkeys' spontaneous behaviors in cancelling a variety of visual and somatosensory stimuli were measured before, and acutely after, unilateral periarcuate (N = 16) and inferior parietal (n = 14) cortical removals. Postoperative behavior was analyzed for both severity of change from the preoperative baseline, and for the type of behavior (perceptual or premotor) affected by the lesion. Overall the two lesion groups could not be differentiated by severity or type of deficit. In two tasks, premotor deficits, manifest as extreme disuse of the hand contralateral to the lesion, were significantly worse in the parietal than the frontal group. In a third, the frontal group showed a greater perceptual deficit, manifest as marked preference for acting within ipsilesional space, than the parietal. In the three remaining tasks, premotor and perceptual deficits were equal in the two groups. These quantitative behavioral data suggest that deficits are more highly contingent upon task requirements than upon lesion sites. This in turn suggests that frontal and parietal association cortical fields each play multiple, and sometimes interchangeable, roles in the spatially directed attention and motor behavior of the monkey.

Identification of a whitefly species by genomic and behavioral studies.

An introduced whitefly species, responsible for over a half billion dollars in damage to U.S. agricultural production in 1991, is morphologically indistinguishable from Bemisia tabaci (Gennadius). However, with the use of polymerase chain reaction-based DNA differentiation tests, allozymic frequency analyses, crossing experiments, and mating behavior studies, the introduced whitefly is found to be a distinct species. Recognition of this new species, the silverleaf whitefly, is critical in the search for management options.