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Biomedical subjects

S E Pratt

Publications and source records attributed to S E Pratt.

At least 19 recordsLinked to original sources
PubMed2002

Expression of multidrug resistance-associated protein 2 (MRP2) in normal human tissues and carcinomas using tissue microarrays.

AIMS: Using tissue microarrays, this study analysed the expression of the multidrug resistance protein, MRP2, by immunohistochemistry with two different MRP2 antibodies. This is the first study to address the expression of MRP2 in various common human neoplasms. METHODS AND RESULTS: Immunohistochemistry was performed on zinc formalin-fixed tissue to evaluate normal tissues and carcinomas using two antibodies against MRP2 (EAG5, a polyclonal antibody, and M2-lll-6, a monoclonal antibody). Immunostaining was localized in neoplastic cells mainly on the cell membrane with M2-lll-6 and cell membrane and cytoplasm with EAG5. In normal tissues MRP2 was expressed in liver, gastrointestinal tract, and kidney tubular epithelial cells with both antibodies. MRP2 was seen in nine of 22 renal cell carcinomas, eight of 13 gastric carcinomas, 25 of 49 breast carcinomas, 14 of 32 lung carcinomas, 39 of 50 colon carcinomas, and 16 of 17 ovarian carcinomas. There was < 10% variability between the two antibodies. MRP2 expression was highest in moderate to poorly differentiated tumours from colon, lung, gastric, and ovarian carcinomas and in grade 2 and 3 breast and renal carcinomas. CONCLUSION: The expression of MRP2 in many solid human tumours indicates that inherent drug resistance may play an important role as a biomarker for predictive chemotherapy treatment.

Antibodies, Monoclonal↗
PubMed1994

Differential glycosylation of the glucose transporter coincides with enhanced sugar transport in respiration deficient cells.

Sugar transport regulation was characterized in terms of the expression and subcellular distribution of the glucose transporter (GT) in the V79 hamster fibroblast cell line and in a respiration deficient mutant (G14) of the V79 cell line. Comparison of GT content in V79 and G14 cells and cell fractions revealed that the 3-fold elevation in basal sugar transport observed in the G14 cell line did not coincide with any significant difference in either the whole cell or plasma membrane GT content when compared to the V79 parental cell line. Determination of delta-antibody binding to intact cell monolayers supported the finding that the two cell lines demonstrate equivalent plasma membrane GT content. Further, D-glucose inhibitable cytochalasin B binding to total cell membranes indicates that additional, unrecognized GT isoforms do not occur in either cell line. A higher average molecular weight GT was detected in the G14 cell line, and treatment of GT enriched preparations with endoglycosidase F established that the G14 cell line exhibits a hyperglycosylated form of the same core GT protein expressed in V79. These results suggest that an enhancement of the intrinsic activity of the GT expressed in G14 is responsible for its increased sugar transport capabilities and this may be related to differences in GT post-translational processing.

Animals↗
PubMed1994

Insulin-like growth factor binding protein 3 (IGF-BP3) inhibits estrogen-stimulated breast cancer cell proliferation.

We have recently demonstrated that exposure of MCF7 cells to antiestrogens in vitro results in both accumulation of IGF-BP3 in media and reduced mitogenic responsivity to IGFs (Cancer Res. 53:5193-5198). We show here that MCF7 cell proliferation in steroid-stripped, serum containing media is significantly attenuated by rhIGF-BP3 (p < 0.05), with a maximal 40% inhibition of serum stimulated growth achieved by 6.25nM IGF-BP3. 10(-10) M estradiol (E2) significantly stimulated MCF7 proliferation, and co-incubation of estrogen containing cultures with 50nM IGF-BP3 resulted in significant attenuation of the estrogen-stimulated proliferation ([3H]thymidine incorporation: E2, 147 +/- 18% of control; E2 +/- IGF-BP3, 111 +/- 18% of control, p < 0.05). These results demonstrate antagonism of steroid stimulated proliferation by an IGF binding protein and are compatible with the hypothesis that antiestrogen-induced accumulation of IGF-BP3 in the conditioned media of MCF7 cells contributes to the cytostatic action of these drugs.

Breast Neoplasms↗
PubMed1994

Evidence that modulation of glucose transporter intrinsic activity is the mechanism involved in the allose-mediated depression of hexose transport in mammalian cells.

In serum starved V79 Chinese hamster lung fibroblast cells, replacement of D-glucose with D-allose resulted in a significant 38 +/- 18% (P < 0.05) reduction of 2-deoxy-D-glucose (2-DG) transport. Similarly, in a respiration-deficient mutant cell line (V79-G14), which has elevated 2-DG transport activity, D-allose reduced 2-DG transport by 59 +/- 18% (P < 0.05). [3H]D-allose uptake by V79 cells occurred slowly and was not inhibited by cytochalasin B, suggesting diffusion as the mode of D-allose entry. Western blot analysis using a rabbit polyclonal antibody to the human erythrocyte glucose transporter (GT) demonstrated that, in both cell lines, GT content and GT subcellular distribution were not significantly different in D-glucose vs. D-allose-treated cells. delta-Antibody, which has been shown to bind to exofacial epitopes of the GT (Harrison et al., 1990, J. Biol. Chem., 265:5793-5801), did not demonstrate any differences in surface binding to D-glucose vs. D-allose-treated intact V79 cells. D-allose treatment of 3T3 fibroblasts resulted in a similar decrease (72%) of 2-DG transport, however D-allose had no apparent effect on basal sugar transport in 3T3 adipocytes. These results suggest that D-allose reduces sugar transport through a modulation of the intrinsic activity of the GT, and that D-allose may act in a tissue-specific manner.

3T3 Cells↗
PubMed1993

Estrogen and antiestrogen modulation of MCF7 human breast cancer cell proliferation is associated with specific alterations in accumulation of insulin-like growth factor-binding proteins in conditioned media.

Many neoplastic cell lines secrete insulin-like growth factor binding proteins (IGFBPs). The physiological role of these proteins is incompletely characterized; under various conditions IGFBPs have been observed to either enhance or inhibit the biological activity of insulin-like growth factors. MCF7 human breast cancer cells are known to be mitogenically responsive to insulin-like growth factors and estrogens, to secrete several IGFBPs, including BP-2, BP-4 and BP-5, and to be growth inhibited by antiestrogens. We report here that the pure antiestrogen ICI 182,780 and, to a lesser extent, the commonly used drug tamoxifen significantly increase levels of a M(r) 43,000-46,000 IGFBP (BP-3) and significantly reduce levels of a M(r) 24,000 IGFBP (BP-4) in the conditioned medium of MCF7 cells. Effects of estradiol and antiestrogens on M(r) 30,000 and M(r) 36,000 IGFBPs are also described. The effects of ICI 182,780 on IGFBPs in the conditioned medium of MCF7 cells may contribute to the remarkable ability of this compound to attenuate insulin-like growth factor I stimulated MCF7 cell proliferation.

Antineoplastic Agents↗
PubMed1987

The role of sulfhydryl groups in permitting transformation and DNA binding of the glucocorticoid receptor.

Treatment of rat liver cytosol containing temperature-transformed, [3H]dexamethasone-bound receptors at 0 degree C with the sulfhydryl-modifying reagent methyl methanethiosulfonate (MMTS) inhibits the DNA-binding activity of the receptor, and DNA-binding activity is restored after addition of dithiothreitol (DTT). When cytosol containing untransformed receptors is heated at 25 degrees C in the presence of MMTS, the 90-kDa heat shock protein dissociates from the receptor in the same manner as in the absence of MMTS, and the receptor will bind to DNA-cellulose if DTT is added subsequently at 0 degree C. These observations are consistent with the conclusion of Bodwell et al. (Bodwell, J. E., Holbrook. N. J. and Munck, A. (1984) Biochemistry 23, 1392-1398) that sulfhydryl moieties on the receptor are absolutely required for the receptor to bind to DNA, and they show that the sulfhydryl-modifying reagent does not inhibit the temperature-mediated dissociation of the heteromeric receptor complex that accompanies transformation to the DNA-binding state. When steroid-receptor complexes that are prebound to DNA-cellulose are exposed to MMTS, the steroid rapidly dissociates, but the receptor remains bound to DNA. Thus, the presence of steroid is not required for the receptor to remain bound to DNA in a high affinity manner. Treatment of cytosol containing transformed glucocorticoid-receptor complexes at 0 degrees C with 20 mM hydrogen peroxide also inactivates the DNA-binding activity of the receptor. The peroxide-induced inactivation is reversed by DTT. Incubation of rat liver cytosol containing untransformed glucocorticoid-receptor complexes at 25 degrees C with hydrogen peroxide prevents their transformation to the DNA-binding form as shown by their inability to bind to DNA-cellulose after addition of DTT. The presence of peroxide during heating of the cytosol also prevents dissociation of the receptor complex as assayed both by reduction in sedimentation value of the receptor and by dissociation of the 90-kDa heat shock protein from the steroid-binding protein. These results strongly suggest that critical sulfur moieties in the receptor complex must be in a reduced form for the temperature-mediated dissociation of the receptor to occur.

Animals↗
PubMed1987

The heat-stable cytosolic factor that promotes glucocorticoid receptor binding to DNA is neither thioredoxin nor ribonuclease.

Treatment of rat liver cytosol containing temperature-transformed [3H]dexamethasone-bound receptors at 0 degree C with the sulfhydryl modifying reagent methyl methanethiosulfonate (MMTS) inhibits the DNA-binding activity of the receptor, and DNA-binding activity is restored after addition of dithiothreitol (DTT). However, transformed receptors that are treated with MMTS and then separated from low Mr components of cytosol by passage through a column of Sephadex G-50 have very little DNA-binding activity when DTT is added to regenerate sulfhydryl moities. The receptors will bind to DNA if whole liver cytosol or boiled liver cytosol is added in addition to DTT. The effect of boiled cytosol is mimicked by purified rat thioredoxin or bovine RNase A in a manner that does not reflect the reducing activity of the former or the catalytic activity of the latter. This suggests that the reported ability of each of these heat-stable peptides to stimulate DNA binding by glucocorticoid receptors is not a biologically relevant action. We suggest that stimulation of DNA binding of partially purified receptors by boiled cytosol does not constitute a reconstitution of a complete cytosolic system in which the dissociated receptor must associate with a specific heat-stable accessory protein required for DNA binding, as has been suggested in the "two-step" model of receptor transformation recently proposed by Schmidt et al. (Schmidt T.J., Miller-Diener, A., Webb M.L. and Litwack G. (1985) J. biol. Chem. 260, 16255-16262).

Animals↗
PubMed1986

An X-autosome fusion chromosome of Caenorhabditis elegans.

The translocation mnT12(IV;X) is a fusion of holocentric chromosomes IV and X, the breakpoints occurring near the left end of IV and the right end of X. Animals homozygous for mnT12 are viable and fertile; they contain five pairs of chromosomes rather than the normal set of six pairs. The mnT12 chromosome is larger than all wild-type chromosomes and thus identifies linkage groups IV and X cytologically. Hermaphrodites heterozygous for mnT12 show high frequency meiotic nondisjunction both between mnT12 and the X chromosome, which results in a high incidence of male self progeny (27% compared to the wild-type incidence of 0.2%), and between mnT12 and chromosome IV, which results in a high incidence of self progeny essentially trisomic for chromosome IV (karyotype IV/mnT12/mnT12). The viability of chromosome IV trisomics has been confirmed by constructing animals trisomic for only normal copies of chromosome IV; these animals are morphologically wild type. Meiotic chromosome disjunction in mnT12 homozygotes appears to be normal, although the frequency of recombination between markers that are normally X-linked is significantly reduced. Males of genotype IV/mnT12/0 are fertile. They can be thought of as having a neo-X(mnT12) neo-Y(normal IV) karyotype since it is possible to maintain a male-hermaphrodite stock of C. elegans consisting of such males and hermaphrodites carrying two neo-X chromosomes and no neo-Y; the organism is thus converted from an XO:XX type of sex determination to an XY:XX system.

Animals↗
PubMed1986

Targeting errors and reduced oculomotor range following ablations of the superior colliculus or pretectum/thalamus.

The physiology of the superior colliculus (SC) implicates it in the visual control of eye movements. In the primate, acute inactivation of the superior colliculus delays the onset of a visually guided saccade, slows its velocity, and shortens its amplitude. Previous research leaves uncertain whether other oculomotor disorders which sometimes follow ablation of this structure are due to tectal pathology, to neural damage surrounding the tectum, or to both causes. In this study, 7 cynomolgus monkeys received SC ablations. In 3 others, control lesions were placed in the pretectal/posterior thalamic region. Both procedures produced a qualitatively similar syndrome of 4 oculomotor changes. Reaction time to initiate saccades to visual targets was slowed. Secondly, the surgery constricted the normal ocular range. At the worst, movement was confined to a radius of 10-12 degrees of primary gaze. The monkeys displayed two kinds of inaccuracies when attempting to foveate stationary visual targets within their surviving ocular range. Saccadic amplitude was reduced, causing the monkeys' initial attempt to fall short of foveating the target. If the target remained lighted there then ensued a series of stepwise corrective saccades toward it. The corrective saccades ceased with the eyes still at a position short of the target. Eye position remained in error for the duration of the trial. The final position was independent of the target's retinal position or the vector of the motor command needed to acquire the target. Rather, the error was related to the angular position of the target about the head ( = desired eye position). The syndrome appeared qualitatively similar whether resulting from tectal or the more rostral pretectal/diencephalic ablation. When occurring along the horizontal axis, the deficits appeared to require damage to the superior colliculus, perhaps combined with pathology of some other structure. The same syndrome along the vertical axis was better correlated with pretectal/diencephalic pathology. Invasion of these areas together with invasion of the transthalamic axons from the frontal eye fields is interpreted as the critical pathology responsible for the syndrome. A similar oculomotor trajectory can be modelled by supposing a loss in the gain of the signal which conveys the target's retinal position, combined with one other fault in the circuit: either a loss in gain of the eye position signal, or the signal representing the target's position in craniocentric coordinates.

Animals↗
PubMed1984

Evaluation of fenbendazole as an extended anthelmintic treatment regimen for swine.

Fenbendazole was given in the feed to swine at a cumulative dosage of 9 mg/kg of body weight over a period of 3, 6, and 12 days to compare efficacy. Four treatment groups of ten 2- to 3-month-old pigs each, with a mean of 15 kg of body weight per group, received 3 mg of fenbendazole/kg/day for 3 days, 1.5 mg/kg/day for 6 days, 0.75 mg/kg/day for 12 days, and no medication. Medicated feed was scheduled so that all treated pigs reached the last day of treatment on the same day, thus making the time between the last treatment and necropsy equal for all groups. Ascaris suum and Trichuris suis were the target species, their presence before treatment being determined by fecal egg counts and at necropsy by worm counts. At necropsy, 9 control pigs were infected with A suum (mean of 18.0 worms/pig), and all control pigs had T suis infection (mean of 36.5 worms/pig). All 3 treatment schedules were 100% effective in removal of A suum; and for T suis, the 3-day regimen was 100% effective, the 6-day regimen, 99.2%, and the 12-day regimen, 91.0%.

Administration, Oral↗
PubMed1984

Anthelmintic effect of febantel/praziquantel paste in dogs and cats.

A paste formulation of 3.4% febantel and 0.34% praziquantel was given in soft food at a rate of 0.29 g/kg of body weight, once daily for 3 days to dogs and cats. Eleven of 12 dogs were naturally infected with Ancylostoma caninum, 10 with Trichuris vulpis, 2 with Toxocara canis, 5 with Taenia sp, and 3 with Dirofilaria immitis. Ten of 11 cats were naturally infected with Toxocara cati, 2 with Ancylostoma tubaeforme, 3 with Taenia sp, 5 with Capillaria aerophila, and 1 with Paragonimus kellicotti. Efficacy was 100% for the removal of A caninum, T canis, and Taenia sp and 99.9% in clearance of T vulpis in dogs, and 100% in removal of A tubaeforme, T cati, and Taenia sp in cats. In dogs, Dirofilaria appeared unaffected, and in cats, there was no apparent effect on Capillaria or on Paragonimus. The paste formulation given in soft food was well tolerated and without observable adverse effects.

Animals↗
PubMed1982

Effect of febantel against Ancylostoma caninum and Trichuris vulpis infections in dogs.

Febantel was administered as a 3.4% paste to 45 dogs infected with 1 or more nematode species, which included Ancylostoma caninum and Trichuris vulpis. Dosage levels of 5, 10, or 15 mg/kg of body weight daily for 3 consecutive days for a total of 15, 30, or 45 mg/kg were administered. At 15 mg/kg, all dogs were completely cleared of all nematode and cestode infections, including A caninum, T vulpis, Uncinaria stenocephala, Toxocara canis, Toxascaris leonina, Taenia sp, and Dipylidium caninum, except for 1 Physaloptera sp adult. The 2 other dosage levels produced somewhat lower efficacy, with the 5 mg/kg being the least effective. The paste appeared palatable to the dogs and there were no apparent adverse effects.

Ancylostomiasis↗
PubMed1981

Prevalence of Dirofilaria immitis and Dipetalonema reconditum infections in Missouri dogs.

The prevalence of Dirofilaria immitis and Dipetalonema reconditum infections in Missouri dogs was investigated by means of a statewide veterinary mail survey and a study of shelter dogs from central Missouri and Mississippi River communities. The veterinary survey revealed microfilariae of D immitis in 447 of 11,823 (3.8%) dogs, microfilariae of D reconditum in 14 (0.1%), and microfilariae of both species in 8 (less than 0.1%). Of 493 shelter dogs examined, using the modified Knott test, 23 (4.7%) were positive for D immitis, 23 (4.7%) for D reconditum, and 2 (0.4%) for both helminths. The prevalence of infection with both D immitis (9.4%) and D reconditum (6.3%) was higher in shelter dogs from the Mississippi River communities than in those from central Missouri (1.7% for D immitis, 3.7% for D reconditum). The microfilariae of D immitis in shelter dogs ranged in length from 255 to 343 micrometer (mean, 314 micrometer) and in width from 5.0 to 7.5 micrometer (mean, 6.5 micrometer); those of D reconditum ranged in length from 238 to 295 micrometer (mean, 257 micrometer) and in width from 3.2 to 6.5 micrometer (mean, 4.9 micrometer). Buttonhook tails were seen in 41% of D reconditum. In the final analysis, the shape of the anterior end was considered the most reliable criterion for differentiation.

Animals↗
PubMed1981

Relative efficacies of pyrantel tartrate and pyrantel citrate against Oesophagostomum sp in swine.

The relative efficacies of pyrantel tartrate and of pyrantel citrate against Oesophagostomum sp in swine were evaluated in a controlled-critical study and the efficacy of pyrantel citrate in a field trial. In the controlled-critical study, pigs naturally infected with Oesophagostomum dentatum were either not treated or were treated with pyrantel citrate or pyrantel tartrate at a dosage of 510 mg of free pyrantel base/kg of feed. Six days later, the pigs were necropsied, adult O dentatum was recovered and counted, and fecal samples were examined for helminth eggs. The efficacies of pyrantel citrate and pyrantel tartrate were each 100% based on fecal egg counts and numbers of adults at necropsy. The field trial was conducted in a similar manner except that pyrantel citrate only was tested against a control group on the basis of fecal egg counts made both at the beginning and at the termination of the trial. In this study pyrantel citrate was found to reduce Oesophagostomum sp egg counts by 89.4%.

Animals↗
PubMed1979

Dose titration of oxfendazole against common nematodes of swine.

Oxfendazole, a benzimidazole carbamate, was administered as a 0.5% feed additive to 88 pigs naturally infected with two to four nematode species. Dose rates of 1.5 mg/kg of body weight, 3.0 mg/kg, 3.75 mg/kg, 4.5 mg/kg, 6.75 mg/kg, or 9.0 mg/kg were 100% efficacious against Oesophagostomum dentatum and 99.2% to 100% effective against Ascaris suum. Dose rates of 4.5 mg/kg, 6.75 mg/kg, and 9.0 mg/kg were 92.7%, 98.9%, and 99.5% effective, respectively, against mixed populations of Metastrongylus apri and M pudendotectus. Results were variable with Trichuris suis infections. Efficacy was based on the number of nematodes recovered at necropsy. Palatability and acceptability of the feed additive were good, and adverse reactions following administration were not observed.

Animals↗