Adenovirus-mediated gene transfer into cold-preserved liver allografts: survival pattern and unresponsiveness following transduction with CTLA4Ig.

The immune response of liver transplant recipients was modulated via adenovirus-mediated transduction of the cold-preserved liver with sequences encoding CTLA4Ig. Transplanted allografts demonstrated rapid transient local expression and recombinant protein production shortly after revascularization, resulting in intact liver function, indefinite survival of the recipient, and the development of donor-specific unresponsiveness. Lymphocytic infiltration of the graft was mainly of the T helper 2 (Th2) subset and was not associated with injury to primary cellular targets of the alloimmune response. These findings demonstrate a successful outcome of a feasible and potentially clinically relevant system of gene delivery of sequences encoding proteins capable of inhibiting the alloimmune response.

Tissue-specific consequences of the anti-adenoviral immune response: implications for cardiac transplants.

The immune response to adenoviral vectors can induce inflammation and loss of transgene expression in transfected tissues. This would limit the use of adenovirus-mediated gene transfer in disease states in which long-term gene expression is required. While studying the effect of the anti-adenoviral immune response in transplantation, we found that transgene expression persisted in cardiac isografts transfected with an adenovirus encoding beta-galactosidase. Transfected grafts remained free of inflammation, despite the presence of an immune response to the vector. Thus, adenovirus-mediated gene transfer may have therapeutic value in cardiac transplantation and heart diseases. Furthermore, immunological limitations of adenoviral vectors for gene therapy are not universal for all tissue types.

The amino terminus of GLUT4 functions as an internalization motif but not an intracellular retention signal when substituted for the transferrin receptor cytoplasmic domain.

Previous studies have demonstrated that the amino-terminal cytoplasmic domain of GLUT4 contains a phenylalanine-based targeting motif that determines its steady state distribution between the surface and the interior of cells (Piper, R. C., C. Tai, P. Kuleza, S. Pang, D. Warnock, J. Baenziger, J. W. Slot, H. J. Geuze, C. Puri, and D. E. James. 1993. J. Cell Biol. 121:1221). To directly measure the effect that the GLUT4 amino terminus has on internalization and subsequent recycling back to the cell surface, we constructed chimeras in which this sequence was substituted for the amino-terminal cytoplasmic domain of the human transferrin receptor. The chimeras were stably transfected into Chinese hamster ovary cells and their endocytic behavior characterized. The GLUT4-transferrin receptor chimera was recycled back to the cell surface with a rate similar to the transferrin receptor, indicating that the GLUT4 sequence was not promoting intracellular retention of the chimera. The GLUT4-transferrin receptor chimera was internalized at half the rate of the transferrin receptor. Substitution of an alanine for phenylalanine at position 5 slowed internalization of the chimera by twofold, to a level characteristic of bulk membrane internalization. However, substitution of a tyrosine increased the rate of internalization to the level of the transferrin receptor. Neither of these substitutions significantly altered the rate at which the chimeras were recycled back to the cell surface. These results demonstrate that the major function of the GLUT4 amino-terminal domain is to promote the effective internalization of the protein from the cell surface, via a functional phenylalanine-based internalization motif, rather than retention of the transporter within intracellular structures.

The clinical anatomy of the cephalic vein in the deltopectoral triangle.

Identification and recognition of the cephalic vein in the deltopectoral triangle is of critical importance when considering emergency catheterization procedures. The aim of our study was to conduct a cadaveric study to access data regarding the topography and the distribution patterns of the cephalic vein as it relates to the deltopectoral triangle. One hundred formalin fixed cadavers were examined. The cephalic vein was found in 95% (190 right and left) specimens, while in the remaining 5% (10) the cephalic vein was absent. In 80% (152) of cases the cephalic vein was found emerging superficially in the lateral portion of the deltopectoral triangle. In 30% (52) of these 152 cases the cephalic vein received one tributary within the deltopectoral triangle, while in 70% (100) of the specimens it received two. In the remaining 20% (38) of cases the cephalic vein was located deep to the deltopectoral fascia and fat and did not emerge through the deltopectoral triangle but was identified medially to the coracobrachialis and inferior to the medial border of the deltoid. In addition, in 4 (0.2%) of the specimens the cephalic vein, after crossing the deltopectoral triangle, ascended anterior and superior to the clavicle to drain into the subclavian vein. In these specimens a collateral branch was observed to communicate between the cephalic and external jugular veins. In 65.2% (124) of the cases the cephalic vein traveled with the deltoid branch of the thoracoacromial trunk. The length of the cephalic vein within the deltopectoral triangle ranged from 3.5 cm to 8.2 cm with a mean of 4.8+/-0.7 cm. The morphometric analysis revealed a mean cephalic vein diameter of 0.8+/-0.1 cm with a range of 0.1 cm to 1.2 cm. The cephalic vein is relatively large and constant, usually allowing for easy cannulation.

Amelioration of collagen-induced arthritis by CD95 (Apo-1/Fas)-ligand gene transfer.

Both rheumatoid arthritis and animal models of autoimmune arthritis are characterized by hyperactivation of synovial cells and hyperplasia of the synovial membrane. The activated synovial cells produce inflammatory cytokines and degradative enzymes that lead to destruction of cartilage and bones. Effective treatment of arthritis may require elimination of most or all activated synovial cells. The death factor Fas/Apo-1 and its ligand (FasL) play pivotal roles in maintaining self-tolerance and immune privilege. Fas is expressed constitutively in most tissues, and is dramatically upregulated at the site of inflammation. In both rheumatoid arthritis and animal models of autoimmune arthritis, high levels of Fas are expressed on activated synovial cells and infiltrating leukocytes in the inflamed joints. Unlike Fas, however, the levels of FasL expressed in the arthritic joints are extremely low, and most activated synovial cells survive despite high levels of Fas expression. To upregulate FasL expression in the arthritic joints, we have generated a recombinant replication-defective adenovirus carrying FasL gene; injection of the FasL virus into inflamed joints conferred high levels of FasL expression, induced apoptosis of synovial cells, and ameliorated collagen-induced arthritis in DBA/1 mice. The Fas-ligand virus also inhibited production of interferon-gamma by collagen-specific T cells. Coadministration of Fas-immunoglobulin fusion protein with the Fas-ligand virus prevented these effects, demonstrating the specificity of the Fas-ligand virus. Thus, FasL gene transfer at the site of inflammation effectively ameliorates autoimmune disease.

Phase I trial of docetaxel with estramustine in androgen-independent prostate cancer.

PURPOSE: To evaluate the toxicity, efficacy, and pharmacokinetics of docetaxel when combined with oral estramustine and dexamethasone in a phase I study in patients with progressive metastatic androgen-independent prostate cancer. PATIENTS AND METHODS: Thirty-four men were stratified into minimally pretreated (MPT) and extensively pretreated (EPT) groups. Estramustine 280 mg PO tid was administered 1 hour before or 2 hours after meals on days 1 through 5, with escalated doses of docetaxel from 40 to 80 mg/m2 on day 2. Treatment was repeated every 21 days. RESULTS: Thirty-four patients were assessable for toxicity and 33 for response. In the MPT patients, dose-limiting myelosuppression was reached at 80 mg/m2, with six patients experiencing grade 3/4 granulocytopenia. In EPT patients, escalation above 70 mg/m2 was not attempted. Fourteen MPT (70%) and six EPT (50%) patients had a > or = 50% decline in serum PSA on two consecutive measurements taken at least 2 weeks apart. The overall 50% PSA response rate was 63% (95% confidence interval [CI], 28% to 81%). Of the 18 patients with bidimensionally measurable disease, five (28%; 95% CI, 11% to 54%) achieved a partial response. At the time of entry onto the study, 15 patients required narcotic analgesics for bone pain; after treatment, eight (53%) discontinued their pain medications. The area under the curve for docetaxel increased linearly from 40 to 70 mg/m2. At 80 mg/m2, the measured area under the curve was 8.37 (standard deviation, 0.724), which was significantly higher than the previously reported values. CONCLUSION: The recommended phase II dose of docetaxel combined with estramustine is 70 mg/m2 in MPT patients and 60 mg/m2 in EPT patients. This combination is active in men with androgen-independent prostate cancer.

Immunosuppression through blockade of CD28:B7-mediated costimulatory signals.

It is now well established that T cells require two signals for activation and effector function. The first signal is provided through the T cell receptor for antigen. The best-characterized pathway which provides the second, or costimulatory, signal is through the CD28 receptor on the surface of T cells. In vitro, ligation of the T cell receptor without a second signal induces a long-lived state of anergy in T cells. CD28 has two known ligands, B7-1 and B7-2, expressed on activated antigen-presenting cells. A soluble fusion protein called CTLA4Ig has been produced which binds B7-1 and B7-2 and acts as a competitive inhibitor of CD28. In vitro and in vivo studies with CTLA4Ig demonstrate that it is an extremely effective immunosuppressive agent in models of transplantation and autoimmunity. Mechanistic studies indicate that CTLA4Ig may work by partially inhibiting the expansion of antigen-reactive cells and inducing anergy in the residual population.

Phase I/II studies of docetaxel (Taxotere) combined with estramustine in men with hormone-refractory prostate cancer.

Evaluation of the combined regimen of estramustine and docetaxel (Taxotere; Rhône-Poulenc Rorer, Collegeville, PA) in men with hormone-refractory prostate cancer is in its early stages. While this combination is promising in terms of efficacy, adverse events associated with estramustine are a concern. Estramustine has been associated with side effects such as nausea, vomiting, edema, and serious vascular events. Reported here are the results of phase I and phase II trials in which 280 mg estramustine was given three times daily on days I to 5 in 21-day treatment cycles with docetaxel at varying doses. Data from patients evaluable thus far support the efficacy of this combination, both in chemotherapeutically naive patients and in those who have had prior therapy. A survival benefit from this combination appears achievable from these early studies. As significant antitumor activity can be achieved with docetaxel alone, future studies need to define the minimal dose of estramustine for this combination.

The job satisfaction-job performance relationship: a qualitative and quantitative review.

A qualitative and quantitative review of the relationship between job satisfaction and job performance is provided. The qualitative review is organized around 7 models that characterize past research on the relationship between job satisfaction and job performance. Although some models have received more support than have others, research has not provided conclusive confirmation or disconfirmation of any model, partly because of a lack of assimilation and integration in the literature. Research devoted to testing these models waned following 2 meta-analyses of the job satisfaction-job performance relationship. Because of limitations in these prior analyses and the misinterpretation of their findings, a new meta-analysis was conducted on 312 samples with a combined N of 54,417. The mean true correlation between overall job satisfaction and job performance was estimated to be .30. In light of these results and the qualitative review, an agenda for future research on the satisfaction-performance relationship is provided.

Synthesis and biological evaluation of antiplatelet 2-aminochromones.

The synthesis and biological evaluation of a series of antiplatelet 2-morpholinylchromones has been described. Modification of the C-7 phenylmethoxy group of 8-methyl-7-(phenylmethoxy)-2-(4-morpholinyl)-4H-1-benzopyran-4-one (2) has led to the discovery of a series of 7-[(amino-ethyl)oxy]-8-methyl derivatives which are potent inhibitors of ADP-induced platelet aggregation. Several members of this class proved active in preventing platelet-dependent thrombus formation in the dog, including 8-methyl-7-[2-(4-methyl-1-piperazinyl)ethoxy]-2-(4- morpholinyl)-4H-1-benzopyran-4-one (39) which was devoid of hemodynamic effects at the effective antithrombotic dose.

Utility of adenoviral-mediated Fas ligand gene transfer to modulate islet allograft survival.

BACKGROUND: One of the best-defined mechanisms for the induction of apoptosis involves signaling via the cell surface molecule Fas, after binding of Fas ligand. Expression of Fas ligand is tightly regulated, being expressed primarily by T cells after activation, where it serves as a self-regulatory mechanism for immune responses. Fas ligand has also been found to be expressed constitutively at sites of immune privilege such as the testes and the anterior chamber of the eye. Recently, co-transplantation of Fas ligand-transfected myoblasts in association with islet cell allografts was shown to prolong islet allograft survival but only rarely led to indefinite graft survival. Graft rejection was associated with loss of Fas ligand on the myoblasts, suggesting that direct expression of the transgene on the islets might be more effective. METHODS: A replication-defective adenoviral construct containing murine Fas ligand (Ad/MFL) was prepared by homologous recombination. NIH 3T3 cells, rodent splenocytes, and murine islets were infected with Ad/MFL and examined in vitro for functional murine Fas ligand expression. Survival of Ad/MFL-infected islets was subsequently evaluated in vivo in both syngeneic and allogeneic islet transplantation models. RESULTS: Cell lines and islet allografts transfected with Ad/MFL expressed a functional Fas ligand, capable of inducing apoptosis (confirmed by three distinct assays for DNA fragmentation) in Fas+ targets, but not in Fas- controls. Furthermore, Ad/MFL was able to modify allogeneic immune responses in vitro, as addition of this virus, but not a control adenovirus, significantly reduced proliferation in a mixed lymphocyte reaction. Surprisingly, however, transplantation of islet allografts transfected with Ad/MFL resulted in long-term allograft survival in only 1 of 30 recipients. Moreover, adenoviral-mediated Fas ligand gene transfer was complicated by transient, dose-dependent islet dysfunction, perhaps contributing to the lack of long-term engraftment. CONCLUSION: These data suggest that adenoviral-mediated Fas ligand expression may impair normal islet function in vivo, and indicate that alternative strategies for Fas ligand transgene delivery may be required in this setting.

Inhibition of human scleral fibroblast proliferation with heparin.

Proliferation of fibroblasts is a serious problem in ocular trauma and surgical wound healing. Depending on the location of the injury, the growth of fibroblasts can lead to different problems. In glaucoma filtering surgery, fibroblast proliferation may contribute to scar tissue formation and premature wound closure. Fibroblastic growth in proliferative vitreoretinopathy may lead to the formation of preretinal membranes, which can contract, causing retinal detachment. In an effort to find a more effective method of inhibiting ocular fibroblast proliferation, we have investigated the effect of heparin, a sulfated polysaccharide, on the proliferation of fibroblasts obtained from the sclera of donor eyes. Heparin inhibits the incorporation of 3H-thymidine in a dose-dependent manner in the presence of fetal bovine serum (FBS). This inhibition is partially reversed by endothelial cell growth factor (ECGF). The heparin antagonist protamine sulfate causes a reversal of heparin inhibition and, in some instances, a significant increase in 3H-thymidine incorporation compared to serum controls. Heparin was equally effective in inhibiting cell proliferation in control and heparin-protamine sulfate-pretreated medium. These results were apparently unrelated to a direct toxic effect on cells, as a Trypan Blue exclusion assay showed no significant difference in viability when heparin treated cells were compared to control cells. Direct cell counts showed that heparin was effective in inhibiting cell proliferation over a long time period, but only if it was reinstilled every 2 days. Heparin treatment shows promise as a method for controlling fibroblast proliferation in the eye.

The age context of performance-evaluation decisions.

Organizational demography contends that demographic characteristics of individuals, examined at individual, dyadic, group, and organizational levels of analysis, exert significant effects on organizational processes. The purpose of this article was to test the contextual effects created by the interaction of work-group age composition and supervisor age on supervisor evaluations of subordinate performance. Two competing models of age demography were tested. The similarity model predicts that supervisors similar in age to the work group they supervise will issue generally higher performance ratings. The dissimilarity model, developed in this article, predicts the opposite. Support was indicated for the dissimilarity model. Implications of the results are discussed.

Relationship of core self-evaluations to goal setting, motivation, and performance.

A newly developed personality taxonomy suggests that self-esteem, locus of control, generalized self-efficacy, and neuroticism form a broad personality trait termed core self-evaluations. The authors hypothesized that this broad trait is related to motivation and performance. To test this hypothesis, 3 studies were conducted. Study 1 showed that the 4 dispositions loaded on 1 higher order factor. Study 2 demonstrated that the higher order trait was related to task motivation and performance in a laboratory setting. Study 3 showed that the core trait was related to task activity, productivity as measured by sales volume, and the rated performance of insurance agents. Results also revealed that the core self-evaluations trait was related to goal-setting behavior. In addition, when the 4 core traits were investigated as 1 nomological network, they proved to be more consistent predictors of job behaviors than when used in isolation.

Relationship of core self-evaluations traits--self-esteem, generalized self-efficacy, locus of control, and emotional stability--with job satisfaction and job performance: a meta-analysis.

This article presents meta-analytic results of the relationship of 4 traits--self-esteem, generalized self-efficacy, locus of control, and emotional stability (low neuroticism) with job satisfaction and job performance. With respect to job satisfaction, the estimated true score correlations were .26 for self-esteem, .45 for generalized self-efficacy, .32 for internal locus of control, and .24 for emotional stability. With respect to job performance, the correlations were .26 for self-esteem, .23 for generalized self-efficacy, .22 for internal locus of control, and .19 for emotional stability. In total, the results based on 274 correlations suggest that these traits are among the best dispositional predictors of job satisfaction and job performance. T. A. Judge, E. A. Locke. and C. C. Durham's (1997) theory of core self-evaluations is used as a framework for discussing similarities between the 4 traits and their relationships to satisfaction and performance.

Interviewers' perceptions of persons-organization fit and organizational selection decisions.

A model of person-organization fit and organizational hiring decisions is developed and tested, using data from 38 interviewers making hiring decisions about 93 applicants. Results suggest that interviewers can assess applicant-organization values congruence with significant levels of accuracy and that interviewers compare their perceptions of applicants' values with their organizations' values to assess person-organization fit. Results also suggested that interviewers' subjective person-organization fit assessments have large effects on their hiring recommendations relative to competing applicant characteristics, and that interviewers' hiring recommendations directly affect organizations' hiring decisions (e.g., job offers).

Relationship between conscientiousness and learning in employee training: mediating influences of self-deception and self-efficacy.

A field study of 97 employees tested a model of the mediating influences of self-deception and task-specific self-efficacy in the relationship between conscientiousness and learning. The setting was an introductory Windows 3.1 software training course. Findings indicated that, as hypothesized, self-deception and self-efficacy mediated the relationship between conscientiousness and learning. Specifically, conscientiousness was positively related to self-deception, which was negatively related to learning, and conscientiousness was positively related to self-efficacy, which was positively related to learning. In addition, 4 alternative models were estimated. The results of the tests of the 4 alternative models were not supported by the data, further substantiating the validity of the hypothesized model.

Five-factor model of personality and transformational leadership.

This study linked traits from the 5-factor model of personality (the Big 5) to transformational leadership behavior. Neuroticism, Extraversion, Openness to Experience, and Agreeableness were hypothesized to predict transformational leadership. Results based on 14 samples of leaders from over 200 organizations revealed that Extraversion and Agreeableness positively predicted transformational leadership; Openness to Experience was positively correlated with transformational leadership, but its effect disappeared once the influence of the other traits was controlled. Neuroticism and Conscientiousness were unrelated to transformational leadership. Results further indicated that specific facets of the Big 5 traits predicted transformational leadership less well than the general constructs. Finally, transformational leadership behavior predicted a number of outcomes reflecting leader effectiveness, controlling for the effect of transactional leadership.

Personality and job satisfaction: the mediating role of job characteristics.

This study tested a model of the relationship between core self-evaluations, intrinsic job characteristics, and job satisfaction. Core self-evaluations was assumed to be a broad personality concept manifested in 4 specific traits: self-esteem, generalized self-efficacy, locus of control, and low neuroticism. The model hypothesized that both subjective (perceived) job characteristics and job complexity mediate the relationship between core self-evaluations and job satisfaction. Two studies were conducted to test the model. Results from Study 1 supported the hypothesized model but also suggested that alternative models fit the data well. Results from Study 2 revealed that core self-evaluations measured in childhood and in early adulthood were linked to job satisfaction measured in middle adulthood. Furthermore, in Study 2 job complexity mediated part of the relationship between both assessments of core self-evaluations and job satisfaction.

Job and life attitudes of male executives.

Despite executives' important positions in organizations, their attitudes have not received much research attention. To remedy this deficiency, the authors tested a hypothesized model of executive attitudes involving job satisfaction, life satisfaction, job stress, and work-family conflict. Using data from a large, representative sample of male executives (because of the small number of female executives in the study, the analyses were confined to men), the authors obtained LISREL results indicating support for the overall model and the specific relationships within the model. These results--the first to simultaneously consider job satisfaction, life satisfaction, job stress, and work-family conflict--constitute the most comprehensive evidence to date on executive attitudes. The meaning and contributions of the findings are discussed.