Search PubMed⌕ Search

Biomedical subjects

R E Steele

Publications and source records attributed to R E Steele.

At least 37 records · Page 2Linked to original sources
PubMed1995

Single step purification of biologically active recombinant rat basic fibroblast growth factor by immobilized metal affinity chromatography.

The construction and use of a plasmid which allows the expression and single step purification of recombinant rat basic fibroblast growth factor (bFGF) is described. A cDNA encoding rat bFGF was subcloned into the expression plasmid pQE-9 (Qiagen) in such a way that the bFGF which is produced from the resulting construct contains 6 histidine residues near the amino terminus. The resulting plasmid, pQE-9-bFGF, was expressed in the E. coli strain M15[pREP4] and the 6 x His-bFGF was purified to homogeneity from the soluble fraction of the bacterial cell lysate in a single step by affinity chromatography on a nickel chelate resin. About 5 mg of 6 x His-bFGF was obtained from the soluble fraction from one liter of bacterial cell culture. Testing of the 6 x His-bFGF in a PC12 cell differentiation assay showed that its activity was comparable to the activities for native bFGF and recombinant bFGF purified by multistep methods.

Amino Acid Sequence↗
PubMed1995

Prevention of spontaneous tumours in female rats by fadrozole hydrochloride, an aromatase inhibitor.

Mammary tumours are oestrogen dependent in female Sprague-Dawley rats and in a significant proportion of women, so pharmacological treatment to inhibit oestrogen production is a valuable therapeutic measure to prevent or slow the progression of disease. Here we show that a non-steroidal aromatase inhibitor, which competitively inhibits the conversion of androstenedione to oestrone, prevents the development of both benign and malignant spontaneous mammary neoplasms in female Sprague-Dawley ats. It also slows the spontaneous development of pituitary pars distalis adenomas in female rats, and reduces the incidence of spontaneous hepatocellular tumours in male and female rats.

Animals↗
PubMed1994

The cosmetic outcome of the scar formation after cesarean section: percutaneous or intracutaneous suture?

Three methods of skin closure after cesarean section were tested and compared in a prospective trial. Eighty-nine (82.5%) appeared for follow-up investigation 4-5 months after delivery. The mean scar width was significantly narrower after phannenstiel incision compared with percutaneous nylon sutures after lower midline incision, 3.1 mm versus 11.3 mm. Intracutaneous continuous PDS-suture after lower midline incision produced scars 40% narrower than with percutaneous sutures, 6.8 versus 11.3 mm. 37.5% of the women sutured intracutaneously complained over persistent secretion after discharge from the hospital. Some of these complaints were probably due to the introduction of new materials and skin closure technique. None consulted a physician with their complaints, i.e. the complications were sub-clinical. Exclusion of the women with persistent secretions from the material led to significantly narrower scars compared with percutaneous closure, 4.5 versus 11.1. Thus, even better results can be expected as experience with the technique increases. Observer and patient satisfaction with the cosmetic outcome were measured independently on a 'Lasa-line'. Their opinions coincided; the order of acceptability was worst with percutaneous wound closure after lower midline incision, next best with intracutaneous closure after lower midline incision, and best after phannenstiel incision with intracutaneous closure.

Adult↗
PubMed1994

Identification of a gene encoding a novel protein-tyrosine kinase containing SH2 domains and ankyrin-like repeats.

Using the polymerase chain reaction with primers corresponding to conserved regions in the kinase domain of protein-tyrosine kinases, we amplified segments of several protein-tyrosine kinase genes from Hydra vulgaris, a member of the ancient metazoan phylum Cnidaria. Characterization of cDNA clones for one of these genes, HTK16, revealed that it encodes a non-receptor protein-tyrosine kinase with two SH2 domains but no SH3 domain. In this regard the predicted HTK16 protein resembles two mammalian non-receptor protein-tyrosine kinases, the products of the ZAP-70 and syk genes. However, the HTK16 protein contains five ankyrin-like repeats, a structural motif which has not previously been found in protein-tyrosine kinases. The HTK16 protein also contains a potential tyrosine phosphorylation site in its carboxyl-terminal tail which resembles the phosphorylation site in members of the src family. RNA hybridization analysis indicates that the HTK16 gene is expressed in epithelial cells, cells which also express the Hydra homologue of the src protein. Our finding of the HTK16 gene in Hydra indicates that diversification of genes encoding non-receptor protein-tyrosine kinases was a very early event in metazoan evolution.

Amino Acid Sequence↗
PubMed1993

Alternative splicing of a neural-specific Src mRNA (Src+) is a rapid and protein synthesis-independent response to neural induction in Xenopus laevis.

Alternative splicing is used by the src protooncogene to derive two gene products, pp60c-src and pp60+. pp60+ is expressed exclusively in neurons, whereas pp60c-src is expressed in many tissues. The specific role of the src+ gene product in neurons is not yet understood. Expression of src+ mRNA begins early in neural development and is restricted to the neural plate in Xenopus embryos. Using a reverse transcription/polymerase chain reaction assay, we demonstrate that expression of src+ mRNA is dependent on neural induction. Src+ mRNA is expressed in animal cap ectoderm which is cocultured with mesoderm tissue, but not in ectoderm cultured alone. The protein kinase C (PKC) activator 12-O-tetradecanoylphorbol-13-acetate (TPA) induces src+ mRNA expression in cultured stage 10+ animal cap ectoderm. This induction is rapid, with src+ mRNA being detected within 1 hr of TPA addition. Only dorsal ectoderm tissue is competent to express src+ mRNA upon TPA treatment, and stage 10+ ectoderm tissue becomes unresponsive to TPA after 4 hr in culture. TPA-dependent induction of src+ mRNA in ectoderm tissue is not blocked by cycloheximide and therefore is not dependent on protein synthesis.

Alternative Splicing↗
PubMed1993

Expression of Cnox-2, a HOM/HOX gene, is suppressed during head formation in hydra.

Using PCR, six genes of the HOM/HOX class have been identified in hydra. One of them, Cnox-2, has been sequenced and resembles the Deformed gene of Drosophila. As shown previously, the expression pattern of Cnox-2 suggested that it may be involved in axial patterning as it was strongly expressed in the epithelial cells of the body column and foot, but only weakly expressed in the head. In addition, expression of Cnox-2 decreased sharply in body column tissue as it was converted into head tissue, thereby providing further support for a role in hydra pattern formation. To examine this possibility further, additional manipulations were carried out which convert body column tissue into either head or foot tissue. When body column tissue was converted into head tissue by regeneration, transplantation, or during budding, Cnox-2 expression was sharply reduced, indicating that expression of this gene is suppressed during head formation. When body column tissue was converted into foot tissue by regeneration or Li+ treatment, the changes in Cnox-2 expression patterns indicated that Cnox-2 had no direct role in foot formation. Surprisingly, however, the observed changes in Cnox-2 expression patterns indicate that following bisection, both head and foot regeneration processes are initiated at the injured edge of the bisected animal.

Animals↗
PubMed1993

A molecular snapshot of the metazoan 'Eve'.

A description of the molecular make-up of the ancestral multicellular animal is emerging from the growing availability of molecular biological and biochemical data gleaned from the study of modern members of ancient groups of animals. We use the distributions of classes of transcription factors, signal transduction systems and other molecular innovations among metazoan phyla to infer some of the characteristics of the first animals.

Animals↗
PubMed1993

Expression of Cnox-2, a HOM/HOX homeobox gene in hydra, is correlated with axial pattern formation.

Cnox-2 is a HOM/HOX homeobox gene that we have identified in the simple metazoan Hydra vulgaris (Cnidaria: Hydrozoa). Cnox-2 is most closely related to anterior members of the Antennapedia gene complex from Drosophila, with the greatest similarity to Deformed. The Cnox-2 protein is expressed in the epithelial cells of adult hydra polyps in a region-specific pattern along the body axis, at a low level in the head and at a high level in the body column and the foot. The expression pattern of Cnox-2 is consistent with a role in axial pattern formation. Alteration of hydra axial patterning by treatment with diacylglycerol (DAG) results in an increase of head activation down the body column and in a coordinate reduction of Cnox-2 expression in epithelial cells in 'head-like' regions. These results suggest that Cnox-2 expression is negatively regulated by a signaling pathway acting through protein kinase C (PKC), and that the varying levels of expression of Cnox-2 along the body axis have the potential to result in differential gene expression which is important for hydra pattern formation.

Amino Acid Sequence↗
PubMed1992

Reduction of cardiovascular and thyroxine-suppressing activities of L-T3 by liver targeting with cholic acid.

This study was designed to determine whether the conjugation product of L-T3 with cholic acid would result in a liver-targeted compound (CGH 509A) with hypocholesterolemic (HC) activity significantly dissociable from cardiovascular (CV) and thyroxine-suppressing (TS) effects normally observed with thyroid hormone. Evaluation of HC activity in lipemic rats showed that CGH 509A was 6 times less potent than L-T3 with ED25 values estimated at 150 and 25 nmol/kg, respectively. CV function measured as changes in atrial rate, atrial tension and heart weight was determined in euthyroid rats. CGH 509A was at least 64 times less cardio-stimulant than L-T3 with minimum effective doses estimated at 2350 and 37 nmol/kg, respectively. TS activity was assessed in euthyroid rats as the potency of any compound to reduce plasma T4 levels. CGH 509A was 50 times less potent than-L-T3 with ED50 values estimated at 900 and 18 nmol/kg, respectively. From these results, it is clear that, while L-T3 was equally potent on HC, CV and TS activities, the HC potency of CGH 509A was at least 15 and 6 times greater than its CV and TS potencies, respectively.

Animals↗
PubMed1992

Effects of coaxial CO2/Nd:YAG irradiation on periodontal wound healing.

This study evaluated a coaxial CO2/Nd:YAG laser used during periodontal surgery in mongrel dogs for the purpose of ablating the osseous portions of root surfaces to increase reattachment of soft tissue. Periodontal defects were produced with ligatures of orthodontic wire and elastics. The left mandibular osseous root surfaces were irradiated with a defocused, CW laser beam [power density (PD) = 42.46 W/cm2, energy density (ED) = 424.63 J/cm2]. The left maxillary osseous root surfaces were treated with a beam at twice the energy density [PD = 84.93 W/cm2, ED = 849.3 J/cm2]. The mandibular right quadrant received conventional surgery and the maxillary right quadrant served as the untreated control. Fourteen days after treatment, facial/lingual attached gingivae were scored for soft tissue adherence to bone. Laser irradiation at low ED did not improve soft tissue attachment when compared to conventional treatment. Higher ED irradiation produced significantly less attachment and increased tissue necrosis.

Aluminum Silicates↗
PubMed1992

Identification and developmental expression of Src+ mRNAs in Xenopus laevis.

Alternative splicing of src mRNA has been demonstrated in several vertebrate species to yield a neuron-specific form of src protein termed pp60+. The function of pp60+ is unknown. The early developmental expression pattern of src+ RNA has not been previously examined. We have identified and characterized src+ transcripts corresponding to the two src genes in Xenopus laevis using a reverse transcription/polymerase chain reaction (RT/PCR) method. Both Xenopus pp60+ proteins have a 5-amino-acid insert in contrast to the 6-amino-acid insert in fish, birds, and mammals. Src+ mRNA first appears in neural plate stage Xenopus embryos, after neural induction signaling events but prior to neural differentiation. Analysis of dissected neural plate stage embryos showed that src+ mRNAs are localized to the neural plate. These findings suggest that pp60+ may play a role in elaboration of neuron structure.

Amino Acid Sequence↗
PubMed1992

Evolution of the technique of combination therapy for staghorn calculi: a decreasing role for extracorporeal shock wave lithotripsy.

Percutaneous nephrostolithotomy with or without extracorporeal shock wave lithotripsy (ESWL) has been extensively used in the management of staghorn calculi, with stone-free rates varying from 23 to 86%. Explanations for the variability of such results include differences in stone burden, differences in percutaneous techniques and an overreliance on ESWL. The results and changing trends in the relative roles of percutaneous nephrostolithotomy and ESWL in the management of staghorn calculi at the Methodist Hospital of Indiana were examined. We reviewed 343 cases of staghorn calculi (partial and complete) with adequate followup data, which were managed with initial percutaneous nephrostolithotomy. Cases were divided into 2 groups: group 1 (252 cases) from 1984 to 1987 when access was performed by a radiologist and group 2 (91 cases) from 1988 to 1990 when access was performed by a urologist. Although stones in group 2 tended to be larger, a decreasing dependence on ESWL was noted (64.7% in group 1 compared to 35.2% in group 2, p less than 0.001). Despite the larger stone size in group 2, stone-free rates were similar (83.3% in group 1, 86.8% in group 2). In groups 1 and 2 percutaneous nephrostolithotomy alone achieved stone-free rates of 91% and 91.5%, respectively. The stone-free rate with the combination approach was 79.1% and 78.1% in groups 1 and 2, respectively. Technical refinements with percutaneous nephrostolithotomy in group 2 include accurate and carefully selected accesses (superior pole in 36.3%) for best approach to the stone, multiple accesses (13.3%) and improved skills in flexible nephroscopy. Blood transfusion has not been required in group 2 compared with 11.1% in group 1. Mean hospital stay was 12.4 days in group 1 and 10.3 days in group 2 (percutaneous nephrostolithotomy alone, 7.2 days). With careful attention to percutaneous nephrostolithotomy techniques, complex renal stones can be successfully managed endourologically, reducing the need for combination ESWL. Complications previously associated with percutaneous nephrostolithotomy have decreased.

Combined Modality Therapy↗
PubMed1992

Staghorn calculi: analysis of treatment results between initial percutaneous nephrostolithotomy and extracorporeal shock wave lithotripsy monotherapy with reference to surface area.

Treatment recommendations and results reported for the management of staghorn calculi are highly variable. In an attempt to provide a more objective means to compare treatment results for staghorn renal calculi, stone burden as measured by stone surface area was used. Stone surface area was determined by computer analysis. A total of 380 cases of staghorn calculi treated at the same institution was evaluated. Treatment consisted of initial percutaneous nephrostolithotomy with or without extracorporeal shock wave lithotripsy (ESWL*) in 298 cases and ESWL monotherapy in 82. When considered as a group, the overall stone-free rate for initial percutaneous nephrostolithotomy (mean surface area 1,378.3 mm.2) was 84.2% compared to 51.2% (p less than 0.0001) for ESWL monotherapy (mean surface area 693.4 mm.2). For staghorn calculi smaller than 500 mm.2 a stone-free rate of 94.4% was achieved in the percutaneous nephrostolithotomy with or without ESWL group compared to 63.2% for ESWL monotherapy (p = 0.0214). For calculi of 501 to 1,000 mm.2 the stone-free rates were 86% and 45.7%, respectively (p less than 0.0001). When stone surface area exceeded 1,000 mm.2 the stone-free rate for percutaneous nephrostolithotomy with or without ESWL was 82.4% but it was only 22.2% for ESWL monotherapy (p = 0.0002). Overall, when adjusted for stone surface area the odds of being stone-free were more than 8 times higher for initial percutaneous nephrostolithotomy versus ESWL monotherapy (odds ratio = 8.36, p less than 0.0001). While percutaneous nephrostolithotomy with or without ESWL appears to be the procedure of choice for most staghorn stones, ESWL monotherapy may have a role for some stones smaller than 500 mm.2. In 12 such cases associated with a nondilated renal collecting system (mean surface area 380.5 mm.2) a stone-free rate of 91.7% was achieved. The number of procedures required to complete therapy was higher in the initial percutaneous nephrostolithotomy group (2.8 versus 2.1, p less than 0.0001). Although complications were more common in the ESWL monotherapy group (manifested as obstruction in 30.5%), bleeding requiring blood transfusion was more frequent in the initial percutaneous nephrostolithotomy group (9.4%).

Adolescent↗
PubMed1992

Biochemical and cytological changes associated with expression of deregulated pp60src in Xenopus oocytes.

We have examined the effects of Xenopus pp60c-src with constitutive kinase activity on the morphology and maturation of Xenopus laevis oocytes. When RNA encoding this deregulated variant was injected into stage VI oocytes, we observed a gross alteration in the cortex of the oocyte. This alteration involved aggregation of pigment and invagination of the cortex in a large area proximal to the site of injection. This phenomenon was not seen in oocytes injected with RNA encoding wild-type pp60c-src. We have correlated this phenomenon with the tyrosine phosphorylation of 84- and 100-kDa proteins. These phosphorylated proteins colocalized with the alteration in the oocyte cortex when assayed by both biochemical and immunocytochemical methods. Neither the pigment aggregation nor phosphorylation of the 84- and 100-kDa proteins was observed in oocytes expressing a nonmyristoylated version of the deregulated pp60c-src. Expression of deregulated Xenopus fyn, a src-family member, resulted in a phenotype similar to that seen with deregulated src. However, in the fyn-injected oocytes, many more proteins were phosphorylated on tyrosine than in the src-injected oocytes. Progesterone stimulation of oocytes expressing deregulated pp60c-src resulted in an increase in the number of tyrosine-phosphorylated proteins. This change may represent the response of pp60src to the resumption of the cell cycle in maturing oocytes. These data suggest that the oocyte may be a particularly useful system for investigating the role of pp60c-src in the regulation of cytoskeletal structure and in the regulation of events associated with the cell cycle.

3T3 Cells↗
PubMed1992

Structural organization of a src gene from Xenopus laevis.

By sequence analysis of genomic clones, the exon-intron structure of one of the two src genes from Xenopus laevis has been determined. The coding region of the gene is interrupted by 10 introns whose locations are identical to the introns in the coding regions of the src genes of human and chicken. The 5' untranslated region is contained on a separate exon with no sequence conservation relative to the corresponding region of the chicken gene. The 5' untranslated region of the Xenopus gene contains a G + C-rich stem-loop sequence and two ATG triplets. A 1.4-kb fragment containing the 5' untranslated region and sequences upstream of it acts as a promoter when introduced in the correct orientation into X. laevis cell lines. The DNA sequence of this fragment lacks the typical arrangement of TATA and CCAAT sequences but contains the ATGCAAAT octamer sequence and a (TA)39 sequence.

Amino Acid Sequence↗
PubMed1991

Enamel surface roughness and dental pulp response to coaxial carbon dioxide-neodymium: YAG laser irradiation.

The purpose of this study was to evaluate the effect of a coaxial carbon dioxide/neodymium:yttrium aluminium garnet laser beam on enamel surface roughness and the dental pulps of mongrel dogs. In four dogs, four maxillary left posterior teeth were irradiated at 16 cm source-tooth distances. Two teeth were irradiated with 16 W CO2/16 W Nd:YAG and the remaining two with 16 W CO2/40 W Nd:YAG. Two maxillary right teeth were untreated controls. In addition, mandibular premolars were irradiated at the same distance and power levels, extracted, and analysed for surface roughness. Significant differences in surface roughness were found between control samples and either power level, but not between enamel surfaces at the two power levels. Maxillary teeth were removed at 10 days postoperatively, sectioned and stained (H & E). The reaction of pulpal cells to irradiation was scored. Data analysis revealed statistically significant differences between the control and lower power Nd:YAG groups and between the control and higher power Nd:YAG groups. The difference in pulpal response between both laser groups approached significance.

Aluminum Silicates↗