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Biomedical subjects

K A Smith

Publications and source records attributed to K A Smith.

At least 37 records · Page 2Linked to original sources
PubMed2000

PU.1 inhibits GATA-1 function and erythroid differentiation by blocking GATA-1 DNA binding.

The lineage-specific transcription factors GATA-1 and PU.1 can physically interact to inhibit each other's function, but the mechanism of repression of GATA-1 function by PU.1 has not been elucidated. Both the N terminus and the C terminus of PU.1 can physically interact with the C-terminal zinc finger of GATA-1. It is demonstrated that the PU.1 N terminus, but not the C terminus, is required for inhibiting GATA-1 function. Induced overexpression of PU.1 in K562 erythroleukemia cells blocks hemin-induced erythroid differentiation. In this system, PU.1 does not affect the expression of GATA-1 messenger RNA, protein, or nuclear localization. However, GATA-1 DNA binding decreases dramatically. By means of electrophoretic mobility shift assays with purified proteins, it is demonstrated that the N-terminal 70 amino acids of PU.1 can specifically block GATA-1 DNA binding. In addition, PU.1 had a similar effect in the G1ER cell line, in which the GATA-1 null erythroid cell line G1E has been transduced with a GATA-1-estrogen receptor fusion gene, which is directly dependent on induction of the GATA-1 fusion protein to effect erythroid maturation. Consistent with in vitro binding assays, overexpression of PU.1 blocked DNA binding of the GATA-1 fusion protein as well as GATA-1-mediated erythroid differentiation of these G1ER cells. These results demonstrate a novel mechanism by which function of a lineage-specific transcription factor is inhibited by another lineage-restricted factor through direct protein-protein interactions. These findings contribute to understanding how protein-protein interactions participate in hematopoietic differentiation and leukemogenesis. (Blood. 2000;96:2641-2648)

Biological Transport↗
PubMed2000

Inhibitory effect of cystic fibrosis sputum on adenovirus-mediated gene transfer in cultured epithelial cells.

Effective gene transfer to the airway epithelial cells of individuals with cystic fibrosis (CF) requires gene therapy vectors to effectively penetrate the mucous lining of the airways of these patients. In this study, we examined the effects of the aqueous sol fraction of sputum recovered from CF patients (CF sol) on adenovirus (Ad)-mediated gene transfer to cultured epithelial cells. Sputum collected from patients with CF was separated into aqueous sol and gel fractions by ultracentrifugation and the sol fraction from different individuals was pooled. To determine if CF sol affects Ad-mediated transfection, Fisher rat thyroid (FRT) epithelial cells or normal human bronchial epithelial (NHBE) cells were infected with an Ad encoding beta-galactosidase (Ad2/betagal-2) in the presence or absence of the pooled CF sol. Transfection efficiency was determined by measuring beta-Gal activity. CF sol significantly inhibited Ad2-mediated gene transfer in a dose-dependent manner when the vector was incubated with CF sol prior to exposure to the cells. In contrast, preincubation of the cells with the sol was without effect. The inhibition of Ad-mediated gene transfer by CF sol was not related to its low pH, was abrogated by preadsorption with an Ad2 serotype vector, and was neutralized by heat treatment, but was not affected by treatment with protease inhibitors. Analysis of CF sol fractions from seven different individuals with CF showed inhibition of Ad-mediated gene transfer in four of the seven samples tested and, further, the inhibitory effect was correlated with the presence of Ad-specific antibodies. We conclude that preexisting adenovirus-specific antibodies present in some of the patient samples were the predominant factor inhibiting Ad-mediated gene transfer.

Adenoviridae↗
PubMed2000

Gamma knife radiosurgery for trigeminal neuralgia: the initial experience of The Barrow Neurological Institute.

PURPOSE: To assess the efficacy and complications of Gamma Knife radiosurgery for trigeminal neuralgia. METHODS AND MATERIALS: The Barrow Neurological Institute (BNI) Gamma Knife facility has been operational since March 17, 1997. A total of 557 patients have been treated, 89 for trigeminal neuralgia (TN). This report includes the first 54 TN patients with follow-up exceeding 3 months. Patients were treated with Gamma Knife stereotactic radiosurgery (RS) in uniform fashion according to two sequential protocols. The first 41 patients received 35 Gy prescribed to the 50% isodose via a single 4-mm isocenter targeting the ipsilateral trigeminal nerve adjacent to the pons. The dose was increased to 40 Gy for the remaining 13 patients; however, the other parameters were unvaried. Outcome was evaluated by each patient using a standardized questionnaire. Pain before and after RS was scored as level I-IV per our newly-developed BNI pain intensity scoring criteria (I: no pain; II: occasional pain, not requiring medication; III: some pain, controlled with medication; IV: some pain, not controlled with medication; V: severe pain/no pain relief). Complications, limited to mild facial numbness, were similarly graded by a BNI scoring system. RESULTS: Among our 54 TN patients, 52 experienced pain relief, BNI score I in 19 (35%), II in 3 (6%), III in 26 (48%), and IV in 4 (7%). Two patients (4%) reported no relief (BNI score V). Median follow-up was 12 months (range 3-28). Median time to onset of pain relief was 15 days (range 0-192), and to maximal relief 63 days (range 0-253). Seventeen (31%) noted immediate improvement (</= 24 h). Prior to RS, all patients were on pharmacologic therapy felt to be optimal or maximal. Twenty-two (41%) were able to stop medications entirely (BNI score I or II). Another 16 (30%), with BNI Score III relief, decreased medication intake by at least 50%. Patients with classical TN pain symptoms were more likely to stop medications than those with atypical features, 49% (21 of 43) versus 9% (1 of 11). This difference was significant at p = 0.040. Statistically, the finding most predictive for pain relief was new facial numbness following RS. Each of the 5 patients with new numbness after RS developed BNI score I relief, contrasting with 35% for the 49 patients with no new numbness (p = 0.019). Complications have been limited to delayed, mild facial sensory loss. Before RS, 17 patients had numbness from prior invasive procedures, none of whom reported a worse numbness score after treatment. Thirty-seven patients had no facial numbness at the time of RS, of whom 5 developed facial hypesthesia. Each rated this as "mild, not bothersome." There have been no other sequellae. CONCLUSION: RS is an effective treatment, and is the least invasive nonpharmacologic therapy for TN. It carries a small risk of mild facial hypesthesia, a side effect which, somewhat ironically, may be desirable, because it appears to correlate favorably with an excellent pain response. We currently include radiosurgery among the appropriate options for TN patients who have failed optimal medical management, with or without prior invasive neurosurgical procedures. We present here BNI scoring systems for pain intensity and facial numbness. These have proved simple and reliable, have facilitated data collection, rendered analysis more objective, and improved our ability to discuss results with patients and colleagues.

Adult↗
PubMed2000

Immunogene therapy for murine melanoma using recombinant adenoviral vectors expressing melanoma-associated antigens.

Adenoviral vectors expressing tumor-associated antigens can be used to evoke a specific immune response and inhibit tumor growth. In this study, we tested the efficacy of adenoviral vectors encoding human gp100 (Ad2/hugp100), murine gp100 (Ad2/mugp100), or murine TRP-2 (Ad2/muTRP-2) for their ability to elicit a specific cellular immune response and inhibit the growth of B16 melanoma tumor cells in the mouse. C57BL/6 mice were immunized with Ad2/hugp100, Ad2/mugp100, or Ad2/muTRP-2 either 2 weeks prior to B16-F10 tumor challenge (prophylactic treatment) or 3 days after tumor challenge (active treatment). Ad2/hugp100 and Ad2/muTRP-2 administered to two or more intradermal (i.d.) sites inhibited subsequent subcutaneous tumor growth in > or = 80% of the mice and elicited an antigen-specific cytotoxic T lymphocyte response, whereas other administration routes were not as effective. Ad2/mugp100 administered to two i.d. sites did not inhibit tumor growth or provoke cellular immunity. Immunization was less effective with active treatment where tumor growth was not significantly inhibited by a single dose of either Ad2/muTRP-2 or Ad2/hugp100. However, increasing the number of intradermal immunization sites and the number of doses resulted in progressive improvements in protection from tumor growth in the active treatment model. In conclusion, breaking host tolerance to elicit protective immunity by using adenoviral vectors expressing melanoma-associated antigens is dependent upon the choice of antigen, the site of administration, and the number of doses. These observations provide insights into the clinical applicability of adenoviral vaccines for immunotherapy of malignant diseases.

Adenoviridae↗
PubMed2000

Experimental analysis of chaotic neural network models for combinatorial optimization under a unifying framework.

The aim of this paper is to study both the theoretical and experimental properties of chaotic neural network (CNN) models for solving combinatorial optimization problems. Previously we have proposed a unifying framework which encompasses the three main model types, namely, Chen and Aihara's chaotic simulated annealing (CSA) with decaying self-coupling, Wang and Smith's CSA with decaying timestep, and the Hopfield network with chaotic noise. Each of these models can be represented as a special case under the framework for certain conditions. This paper combines the framework with experimental results to provide new insights into the effect of the chaotic neurodynamics of each model. By solving the N-queen problem of various sizes with computer simulations, the CNN models are compared in different parameter spaces, with optimization performance measured in terms of feasibility, efficiency, robustness and scalability. Furthermore, characteristic chaotic neurodynamics crucial to effective optimization are identified, together with a guide to choosing the corresponding model parameters.

Algorithms↗
PubMed2000

Modelling a loss event: effect of imagined bereavement on the hypothalamic-pituitary-adrenal axis.

BACKGROUND: Loss events are the stressors most closely associated with the onset of depressive illnesses. The acute cortisol response to loss has been little studied although it could be an important mediator of the effects of environmental stress on psychological state. METHODS: The salivary cortisol response to an established negative mood induction procedure involving music and an imagined bereavement was measured in 30 healthy volunteers. RESULTS: Considerable but transient mood lowering in response to the negative mood induction was associated with a small increase in cortisol output over 30 min. CONCLUSIONS: This procedure has some potential as a tool to investigate individual differences in the neuroendocrine response to loss events, but this is limited. There remains a need for laboratory models of relevant psychosocial stressors in mood disorders research.

Adult↗
PubMed2000

Provocative use testing of methyldibromo glutaronitrile in a cosmetic shampoo.

Methyldibromo glutaronitrile (1,2-dibromo-2,4-dicyanobutane, MDGN) is a component of Euxyl K400, a broad-spectrum preservative increasingly used in cosmetics over the last 10 years. Contact allergy to MDGN is not uncommon, with leave-on cosmetics and moistened toilet tissue being the most important sources. Rinse-off products have rarely been reported to elicit allergic reactions in patients sensitized to MDGN and shampoos have only been implicated in hairdressers. To determine whether subjects sensitized to MDGN would react in a provocative use test with a shampoo containing the preservative, we asked 12 pre-sensitized subjects to use a shampoo containing 0.02% MDGN for a period of 9-13 weeks. During this time, none of the subjects showed any skin reactions indicative of contact allergy. The results of this study provide evidence to support that a shampoo product containing 0.02% MDGN may safely be used by most individuals who are presensitized to MDGN.

Adolescent↗
PubMed2000

Autoantibodies to the flavoprotein subunit of succinate dehydrogenase: analysis of specificity in autoimmune thyroid disease.

OBJECTIVE: Thyroid-associated ophthalmopathy is a progressive eye disorder affecting the extraocular muscle and orbital connective tissue and is considered to have an autoimmune aetiology. A recent study reported a close relationship between serum antibodies against the flavoprotein subunit of succinate dehyhdrogenase (SDHFp) and active thyroid-associated ophthalmopathy involving eye muscle damage. The aim of the present study was to develop a sensitive and quantitative radiobinding assay for the detection of antibodies to the flavoprotein subunit of succinate dehydrogenase and to use this to determine the distribution of antibodies in different patient groups. DESIGN AND PATIENTS: Serum samples from the following patient groups were analysed: 20 systemic lupus erythematosus; 20 Addison's disease; 26 autoimmune hypothyroidism; 28 Graves' hyperthyroidism; 12 pretibial myxoedema; 25 thyroid-associated ophthalmopathy. Sera from 20 healthy subjects were used as controls. [35S]-labelled succinate dehydrogenase flavoprotein was produced in an in vitro transcription-translation system and subsequently used in immunoprecipitation experiments with sera from patient and control groups to test for the presence of antibodies to the flavoprotein. RESULTS: Succinate dehydrogenase flavoprotein antibodies were detected in five of the 20 (25%) patients with Addison's disease, six of the 20 (30%) with systemic lupus erythematosus, five of the 26 (19%) with autoimmune hypothyroidsm, six of the 28 (21%) with Graves' hyperthyroidism, two of the 12 (17%) with pretibial myxoedema and three of the 25 (12%) with thyroid-associated ophthalmopathy. The frequencies of flavoprotein antibodies were significantly greater than controls (P-value < 0.05) for patients with systemic lupus erythematosus (P = 0.02), but not for patients with either Addison's disease (P = 0.05), pretibial myxoedema (P = 0.13), Graves' hyperthyroidism (P = 0.07), autoimmune hypothyroidism (P = 0.06) or thyroid-associated ophthalmopathy (P = 0.24). For the patients with thyroid-associated ophthalmopathy, the frequency of SDHFp antibodies did not appear to be related to the length of time from diagnosis: the group containing samples taken less than one year from diagnosis showed no increased frequency of SDHFp antibodies when compared to controls (P = 0.10), with three of the 18 (17%) patients being positive. With respect to seven patients with thyroid-associated ophthalmopathy diagnosed for more than a year, SDHFp antibodies were not detected in any of their serum samples. In addition, the clinical severity of the disease, as recorded by the NOSPECS classification, did not correlate with the frequency of SDHFp antibodies: P = 0.13, 0.33 and 0.38, respectively, for patients with Grade II, III and IV ophthalmopathy. Similar results were also found in the case of patients with pretibial myxoedema and eye disease: P = 0.06 for patients with Grade III ophthalmopathy and, SDHFp antibodies were not detected in any of the sera taken from patients with Grade IV ophthalmopathy. In addition, no association was found between disease duration and the frequency of antibodies to the flavoprotein in this patient group. CONCLUSIONS: Our results indicate that succinate dehydrogenase flavoprotein antibodies are not a suitable marker for thyroid-associated ophthalmopathy, at least with the assay system used, as they can be found in patients who do not have eye disease and therefore lack the disease specificity required of a diagnostic tool.

Addison Disease↗
PubMed2000

Impaired regulation of brain serotonin function during dieting in women recovered from depression.

BACKGROUND: Amino acid mixtures that lower brain availability of the serotonin (5-HT) precursor tryptophan produce acute depressive relapse in women with a history of major depression. Dieting also lowers brain tryptophan availability, but its effects on brain 5-HT function in recovered depressed women have not been studied. AIM: To test the hypothesis that women with a history of major depression would show impaired regulation of brain 5-HT function during a period of dieting-induced tryptophan depletion. METHOD: Women with and without a history of major depression were placed on a daily 1000 kcal (approximately 4200 kj) diet for three weeks. Before the diet and in the final week we measured fasting plasma tryptophan levels and the prolactin response to an intravenous tryptophan challenge. RESULTS: Dieting lowered plasma tryptophan levels equivalently in women with and without a history of depression. In women without a history of depression, dieting also increased the prolactin response to tryptophan. This increase did not occur in women with a history of depression. CONCLUSIONS: Women with a history of depression showed impaired regulation of brain 5-HT function in response to dieting.

Adult↗
PubMed2000

Self-evaluation maintenance in a larger social context.

Two studies extended Tesser's (1988) Self-evaluation Maintenance (SEM) model to include two close others and to examine the implications of intermediate performances on self-definition. Both demonstrated that when outperformed by both close others, participants rated activities low in self-relevance. When participants outperformed both close others, relevance was moderate or high. When participants were superior to one close other, but inferior to the other, relevance was low or moderate. When participants were uncertain about relative performances, and when there was disagreement over relative performances among the two close others and participants, ratings of relevance were attenuated as a means of protecting participants' self-evaluations. Results are discussed in terms of the importance of extending the SEM model to the larger social network.

Attitude↗
PubMed2000

Do-not-resuscitate orders in the operating room: required reconsideration.

Advances in medical technology are giving health care providers the tools with which to keep patients alive for longer and longer periods of time. However, in our struggle to keep patients alive, we must not forget that the patient is the one who controls his or her own destiny. Advance directives and do-not-resuscitate orders are becoming more commonplace as patients strive to make their wishes known to all who care for them. The use of such orders in the operating room has historically been a hotbed of controversy because it appears contrary to the necessity of surgery in the first place. Surgeons, anesthetists, operating room nurses, and others all have concerns regarding this issue, and they will be discussed here. The answer to the dilemma lies in a policy of "required reconsideration" to examine all factors of the do-not-resuscitate order and its applicability to the situation at hand.

Advance Directives↗
PubMed1999

Human cord blood progenitors sustain thymic T-cell development and a novel form of angiogenesis.

There is growing interest in using human umbilical cord blood (CB) for allogeneic bone marrow transplantation (BMT), particularly in children. Thus, CB has been identified as a rich source of hematopoietic progenitors of the erythroid, myeloid, and B-cell lineages. Whether CB blood cells engrafting in the BM space also comprise T-cell progenitors capable of trafficking to the thymus and reconstituting a functional thymopoiesis in young recipients is presently unknown. Here, we show that CB progenitors, engrafted in the BM of immunodeficient mice, sustain human thymopoiesis by generating circulating T-cell progenitors capable of homing to and developing within a human thymic graft. Surprisingly, development of CB stem cells in this in vivo model extended to elements of the endothelial cell lineage, which contributed to the revascularization of transplants and wound healing. These results demonstrate that human CB stem cell transplantation can reconstitute thymic-dependent T-cell lymphopoiesis and show a novel role of CB-derived hematopoietic stem cells in angiogenesis.

Animals↗
PubMed1999

Cytokine response gene 6 induces p21 and regulates both cell growth and arrest.

Cytokine response gene #6 (CR6), cloned from interleukin 2-stimulated T lymphocytes, is homologous to GADD45 and MyD118, genes which promote cell cycle arrest and apoptosis. To determine how this gene family could possibly mediate both cell survival/proliferation and cell cycle arrest/death, transfectants were generated so that the genes could be expressed ectopically, independently from their normal inducing agents. In cycling retinoblastoma protein-negative (pRb-) cells, ectopic CR6 expression blocked G2/M transition, but did not prevent G1/S transition so that endoreduplication resulted. By comparison, when CR6, GADD45, and MyD118 genes were expressed ectopically in proliferating pRb+ cells, either G1/S or G2/M transition was effectively blocked, so that there was no endoreduplication. Consistent with these findings, in proliferating pRb-cells, ectopic expression of CR6 promoted the expression of both G1 and G2/M cyclins. By comparison, in pRb+ cells, the expression of G1 cyclins was increased, while expression of the mitotic cyclins was decreased. However, in pRb+ cells, cyclin-dependent kinase activities associated with both G1 and G2/M cyclins were decreased. Moreover, ectopic expression of all three genes resulted in the expression of the CKI, p21, both in pRb- and pRb+ cells. The physiologic induction of CR6 expression by IL2 in quiescent normal human T cells occurs transiently in the first half of G1, coordinately with the expression of p21. Therefore, this gene family regulates G1 and G2, and promotes either cell growth or arrest by a common mechanism.

Carrier Proteins↗
PubMed1999

PU.1 and the granulocyte- and macrophage colony-stimulating factor receptors play distinct roles in late-stage myeloid cell differentiation.

PU.1 is a hematopoietic cell-specific ets family transcription factor. Gene disruption of PU.1 results in a cell autonomous defect in hematopoietic progenitor cells that manifests as abnormal myeloid and B-lymphoid development. Of the myeloid lineages, no mature macrophages develop, and the neutrophils that develop are aberrantly and incompletely matured. One of the documented abnormalities of PU. 1 null (deficient) hematopoietic cells is a failure to express receptors for granulocyte colony-stimulating factor (G-CSF), granulocyte-macrophage (GM)-CSF, and M-CSF. To elucidate the roles of the myeloid growth factor receptors in myeloid cell differentiation, and to distinguish their role from that of PU.1, we have restored expression of the G- and M-CSF receptors in PU.1-deficient cells using retroviral vectors. We have similarly expressed PU.1 in these cells. Whereas expression of growth factor receptors merely allows a PU.1-deficient cell line to survive and grow in the relevant growth factor, expression of PU.1 enables the development of F4/80(+), Mac-1(+)/CD11b(+) macrophages, expression of gp91(phox) and generation of superoxide, and expression of secondary granule genes for neutrophil collagenase and gelatinase. These studies reinforce the idea that availability of PU.1 is crucial for normal myeloid development and clarify some of the molecular events in developing neutrophils and macrophages that are critically dependent on PU.1.

Animals↗
PubMed1999

Regulators of G protein signaling exhibit distinct patterns of gene expression and target G protein specificity in human lymphocytes.

The newly recognized regulators of G protein signaling (RGS) attenuate heterotrimeric G protein signaling pathways. We have cloned an IL-2-induced gene from human T cells, cytokine-responsive gene 1, which encodes a member of the RGS family, RGS16. The RGS16 protein binds Gialpha and Gqalpha proteins present in T cells, and inhibits Gi- and Gq-mediated signaling pathways. By comparison, the mitogen-induced RGS2 inhibits Gq but not Gi signaling. Moreover, the two RGS genes exhibit marked differences in expression patterns. The IL-2-induced expression of the RGS16 gene in T cells is suppressed by elevated cAMP, whereas the RGS2 gene shows a reciprocal pattern of regulation by these stimuli. Because the mitogen and cytokine receptors that trigger expression of RGS2 and RGS16 in T cells do not activate heterotrimeric G proteins, these RGS proteins and the G proteins that they regulate may play a heretofore unrecognized role in T cell functional responses to Ag and cytokine activation.

Cells, Cultured↗
PubMed1999

Transduction of human CD34+ cells that mediate long-term engraftment of NOD/SCID mice by HIV vectors.

Efficient gene transfer into human hematopoietic stem cells (HSCs) is an important goal in the study of the hematopoietic system as well as for gene therapy of hematopoietic disorders. A lentiviral vector based on the human immunodeficiency virus (HIV) was able to transduce human CD34+ cells capable of stable, long-term reconstitution of nonobese diabetic/severe combined immunodeficient (NOD/SCID) mice. High-efficiency transduction occurred in the absence of cytokine stimulation and resulted in transgene expression in multiple lineages of human hematopoietic cells for up to 22 weeks after transplantation.

Animals↗