Search PubMed⌕ Search

Biomedical subjects

S A Howard

Publications and source records attributed to S A Howard.

At least 19 recordsLinked to original sources
PubMed2005

Effects of celecoxib in human retinoblastoma cell lines and in a transgenic murine model of retinoblastoma.

BACKGROUND/AIM: Celecoxib, a cyclooxygenase-2 inhibitor and antiangiogenic agent, has demonstrated potent anticancer effects in preclinical studies and in human clinical trials. To evaluate the potential utility of this agent in the treatment of retinoblastoma, the authors investigated the effects of celecoxib in retinoblastoma cell lines and in a murine model of this disease. METHODS: Growth inhibitory effects of celecoxib were evaluated in Y79 and Weri-RB1 human retinoblastoma cell lines by WST-1 cell proliferation assay. For animal study, two groups of 24, 8 week old LHbeta-TAg transgenic mice were treated with celecoxib (250 mg/kg, orally once a day) or vehicle control, 5 days/week for 6 weeks. Mice were sacrificed on day 43. Enucleated eyes were serially sectioned and ocular tumour burden was quantified by histopathological analysis. RESULTS: Celecoxib did not inhibit proliferation of Y79 or Weri-RB1 cells, even at concentrations far exceeding clinically achievable levels. No significant difference in ocular tumour burden between celecoxib treated and control mice (p=0.73) was found. CONCLUSION: Celecoxib was ineffective at inhibiting proliferation of retinoblastoma cells in vitro and was ineffective at controlling retinoblastoma tumour growth in a murine model of this disease. On the basis of these findings, oral celecoxib therapy is unlikely to have clinical utility in the treatment of retinoblastoma.

Angiogenesis Inhibitors↗
PubMed2001

Mechanisms of estrogenic protection against gp120-induced neurotoxicity.

gp120, an HIV coat glycoprotein that may play a role in AIDS-related dementia complex (ADC), induces neuronal toxicity characterized by NMDA receptor activation, accumulation of intracellular calcium, and downstream degenerative events including generation of reactive oxygen species and lipid peroxidation. We have previously demonstrated estrogenic protection against gp120 neurotoxicity in primary hippocampal cultures. We here characterize the mechanism of protection by blocking the classical cytosolic estrogen receptors and by measuring oxidative end points including accumulation of extracellular superoxide and lipid peroxidation. Despite blocking ERalpha and ERbeta with 1 microM tamoxifen, we do not see a decrease in the protection afforded by 100 nM 17 beta-estradiol against 200 pM gp120. Additionally, 17alpha-estradiol, which does not activate estrogen receptors, protects to the same extent as 17beta-estradiol. 17beta-Estradiol does, however, decrease gp120-induced lipid peroxidation and accumulation of superoxide. Together the data suggest an antioxidant mechanism of estrogen protection that is independent of receptor binding.

Animals↗
PubMed1999

Glucocorticoid modulation of gp120-induced effects on calcium-dependent degenerative events in primary hippocampal and cortical cultures.

The HIV coat protein gp120 has been implicated in damaging the nervous system and may play a role in AIDS-related dementia complex. The glycoprotein triggers the release of a glutamatergic agent from infected microglia and macrophages, causing NMDA receptor- and calcium-dependent excitotoxic damage to neurons. We have previously shown that glucocorticoids, the adrenal steroids secreted during stress, worsen gp120 neurotoxicity and calcium mobilization in various brain regions. This study explores events down-stream of gp120-induced calcium mobilization, specifically, generation of reactive oxygen species (ROS) and subsequent lipid peroxidation, destruction of the cytoskeleton through spectrin proteolysis, and the glucocorticoid modulation of these events in primary hippocampal cultures. We observe that 200 pM gp120 causes a significant accumulation of ROS, including superoxide, and of lipid peroxidation. Counter to our predictions, pretreatment with the glucocorticoid corticosterone (CORT) did not worsen the effects of gp120 on ROS accumulation, but did increase lipid peroxidation. We also observed that neither gp120 alone nor gp120 plus CORT caused detectable proteolysis of the cytoskeletal protein spectrin, whose breakdown has been shown to be a damaging consequence of calcium excess in other models of necrotic neuronal injury.

Animals↗
PubMed1998

The relative effectiveness of human plasma glutathione peroxidase as a catalyst for the reduction of hydroperoxides by glutathione.

To reveal clues to the function of human plasma glutathione peroxidase (GPx), we investigated its catalytic effectiveness with a variety of hydroperoxides. Comparisons of hydroperoxides as substrates for plasma GPx based on the ratio of Vmax/Km were blocked by the limited solubility of the organic hydroperoxides, which prevented kinetic saturation of the enzyme at the chosen glutathione concentration. Therefore, we compared the hydroperoxides by the fold increase in the apparent first-order rate constants of their reactions with glutathione owing to catalysis by plasma GPx. The reductions of aromatic and small hydrophobic hydroperoxides (cumene hydroperoxide, t-amyl hydroperoxide, t-butyl hydroperoxide, paramenthane hydroperoxide) were better catalyzed by plasma GPx than were reductions of the more "physiological" substrates (linoleic acid hydroperoxide, hydrogen peroxide, peroxidized plasma lipids, and oxidized cholesterol).

Catalysis↗
PubMed1998

Energy dependency of glucocorticoid exacerbation of gp120 neurotoxicity.

The HIV envelope glycoprotein, gp120, a well documented neurotoxin, may be involved in AIDS-related dementia complex. gp120 works through an NMDA receptor- and calcium-dependent mechanism to damage neurons. We have previously demonstrated that both natural and synthetic glucocorticoids (GCs) exacerbate gp120-induced neurotoxicity and calcium mobilization in hippocampal mixed cultures. GCs, steroid hormones secreted during stress, are now shown to work in conjunction with gp120 to decrease ATP levels and to work synergistically with gp120 to decrease the mitochondrial potential in hippocampal cultures. Furthermore, energy supplementation blocked the ability of GCs to worsen gp120's effects on neuronal survival and calcium mobilization. A GC-induced reduction in glucose transport in hippocampal neurons, as previously documented, may contribute to this energetic dependency. These results may have clinical significance, considering the common treatment of severe cases of Pneumocystis carinii pneumonia, typical of HIV infection, with large doses of synthetic GCs.

Adenosine Triphosphate↗
PubMed1996

Endocrine modulators of necrotic neuron death.

In recent years, there has been extraordinary progress in understanding the cellular and molecular cascades that mediate neuron death following necrotic insults. With this knowledge has come the recognition of ways in which these cascades can be modulated by extrinsic factors, altering the likelihood of subsequent neuron death. In this review, we consider the ability of a variety of hormones to modulate necrotic death cascades. Specifically, we will examine the ability of the stress hormones glucocorticoids and corticotropin-releasing factor, of thyroid hormone, and of pre-ischemic exposure to catecholamines to augment necrotic neuron death. In contrast, estrogen, insulin and postischemic exposure to catecholamines appear to decrease necrotic neuron death. We review the heterogeneous mechanisms that are likely to mediate these hormone effects, some possible clinical implications and the therapeutic potentials of these findings.

Animals↗
PubMed1992

Purification of a Trypanosoma cruzi trypomastigote 60-kilodalton surface glycoprotein that primes and activates murine lymphocytes.

We have purified a glycoprotein with a relative molecular mass of 60 kDa and present on the surface of Trypanosoma cruzi trypomastigotes and studied its ability to prime and stimulate the proliferation of murine spleen cells. T. cruzi trypomastigote membrane proteins were separated by preparative isoelectrofocusing. A trypomastigote 60-kDa surface protein with an isoelectric point of 4.2 was enriched by chromatofocusing and was readily purified in native form to homogeneity by gel filtration on a Superose column by use of a fast protein liquid chromatography system. Biotinylated wheat germ agglutinin, Ricinus communis agglutinin, and Datura stramonium agglutinin bound to blots containing the purified trypomastigote 60-kDa surface protein, indicating that this protein was glycosylated. The purified trypomastigote 60-kDa glycoprotein was recognized by antibodies produced during human infection, and immunoglobulin G against the purified glycoprotein immunoprecipitated a biotinylated 60-kDa molecule from the surface of trypomastigotes but not epimastigotes. Specific immunoglobulin G against the 60-kDa glycoprotein also increased the uptake of trypomastigotes and promoted parasite killing by macrophages. The purified 60-kDa glycoprotein was able to specifically activate primed lymphocytes, since there was a significant increase in [3H]thymidine incorporation by spleen cells obtained from CBA mice primed with this glycoprotein, with respect to control values. Furthermore, the 60-kDa glycoprotein did not stimulate unprimed spleen cells, indicating that the lymphoproliferation induced by this glycoprotein was specific and was not due to polyclonal activation. Our findings indicate that this T. cruzi trypomastigote 60-kDa surface glycoprotein primes and activates lymphocytes, which could lead to a beneficial immune response in the host.

Animals↗
PubMed1983

Dissolution profiles for finely divided drug suspensions.

A suspension of micronized prednisolone acetate was separated into four fractions by the technique of centrifugal elutriation. Data showed that each fraction had a narrow particle size. The dissolution experiments were carried out under sink conditions (less than 10% of saturation concentration) in a dissolution apparatus with a rotating filter assembly and a continuous circulation of filtered fluid samples through a recording spectrophotometer. The dissolution profile was highly reproducible and substantially different for each fraction. As expected, fractions with the smallest and largest particles showed the fastest and slowest dissolution, respectively. Almost the entire dissolution profiles for four small particle size fractions can be satisfactorily described by the Higuchi-Hiestand model with the dissolution rate constant, K, in the range of 1.5-2.0 X 10(-9)cm2/sec. This is approximately 3.5 times greater than the value for K calculated on the basis of reported reasonable values for diffusion coefficient, density, and solubility.

Models, Theoretical↗
PubMed1982

Oral immunization against influenza.

An anti influenza vaccine was administered by oral route, as capsules, to 24 volunteers. The presence of IgAs in nasal secretions led us to the fact that some lymphocytes of the small intestine were the precursors of circulating IgAs, and, while proliferating, they settled on mucous sites at a distance.

Administration, Oral↗
PubMed1979

Hydrodynamic characterization of a spin-filter dissolution device.

The spin-filter dissolution device was characterized using a two-dimensional convective diffusion model. Experimental model testing involved analysis of dissolution rates from nondisintegrating salicylic acid disks. The disks were prepared as double-layer tablets, with an ethylcellulose layer as a nondissolving surface. For each dissolution run, the disk was positioned so that the dissolving salicylic acid surface was parallel to the flow of the circulating fluid. Experimental variables included the stirring speed, the tablet radius, and the distance of the tablet from the stirring source. At the farthest distance from the stirring source, the average numerical exponents for stirring speed and tablet radius were 0.58 and 1.54, respectively, which compare favorably with the values of 0.50 and 1.50 from the model. When the dissolving salicylic acid surface was positioned closer to the stirring source, the numberical exponent for the stirring speed increased significantly, while the average numerical exponent for the tablet radius was lowered to 1.07, indicating a change is dissolution mechanism as a function of distance from the stirring source. These data indicate that dissolution rates are not necessarily proportional to surface area as predicted by the Nernst equation and that distance from the stirring source is significant.

Chemistry, Pharmaceutical↗
PubMed1979

Suspending agent effects on steroid suspension dissolution profiles.

Dissolution profiles and particle-size analyses were determined for two lots of prednisolone acetate. The effects of common suspending agents on dissolution and particle-size distributions of these suspensions also were investigated. Lot-to-lot variation in the prednisolone acetate dissolution rate was observed and was apparently related to the percentage of fine particles within the distribution. Carboxymethylcellulose sodium inhibition of prednisolone acetate dissolution occurred with only one lot of raw material and seemed to be related to aggregation of the fine particles. Hydroxypropyl methylcellulose inhibited both prednisolone acetate lots and was observed with or without small particle aggregation. The dissolution variations observed have important implications in suspension formulation.

Excipients↗
PubMed1979

Tablet position and basket type effects in spin-filter dissolution device.

The effects of stirring and basket placement on tablet dissolution using the previously developed Shah spin-filter device were investigated. Visualization of flow and dissolution patterns was possible by testing nondisintegrating colored tablets. Dissolution experiments were conducted on nondisintegrating double-layered tablets containing salicylic acid as the dissolving layer and ethylcellulose as an inert nondissolving layer. Visual observations revealed that color was drawn more rapidly from the tablet face resting on the bottom of the basket. Dissolution data from multilayered tablets revealed that when the salicylic acid face was resting on the bottom of the basket, the dissolution was appreciably more rapid than when it was facing up in the basket. This phenomenon was found for several stirring speeds.

Diffusion↗
PubMed1978

Separation of multisized drug suspensions into narrow distributions by centrifugal elutriation.

A centrifugal elutriator rotor was used to separate suspensions of micronized hydrocortisone acetate and prednisolone acetate. Up to five distinct particle fractions could beisolated from a single parent suspension, and reasonable reproducibility was exhibited between various elutriator runs. Arithmetic means and associated standard deivations, based on volume, were calculated for each isolated fraction. These data demonstrate the narrowness and uniqueness of each fraction. The particle-size data show an apparent log-normal distribution for each isolate. The uniqueness of each fraction also was demonstrated by dissolution experiments where Fraction 1, the smallest fraction of prednisolone acetate, dissolved much more rapidly than Fraction 4, the largest fraction. The substantial difference in dissolution behavior of these two fractions of prednisolone acetate powder was biopharmaceutically significant.

Centrifugation↗