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Biomedical subjects

J Keller

Publications and source records attributed to J Keller.

At least 163 records · Page 9Linked to original sources

Effects of gene dosage and sequence modification on the frequency and timing of transposition of the maize element Activator (Ac) in tobacco.

The effect of Ac copy number on the frequency and timing of germinal transposition in tobacco was investigated using the streptomycin phosphotransferase gene (SPT) as an excision marker. The activity of one and two copies of the element was compared by selecting heterozygous and homozygous progeny of transformants carrying single SPT::Ac inserts. It was observed that increasing gene copy not only increases the transposition frequency, but also occasionally alters the timing of transposition such that earlier events are obtained. The result is that some homozygous plants generate multiple streptomycin resistant progeny carrying the same transposed Ac (trAc) element. We have also investigated the effect of modification of the sequence in the region around 82 bp downstream of the polyadenylation site and 177 bp from the 3' end of the element on germinal excision frequencies. Alteration of three bases to create a Bgl II site at this location caused a minor decrease in germinal excision events, but insertion of four bases to create a Cla I site caused a 10-fold decrease in the transposition activity of the Ac element.

Agrobacterium tumefaciens↗

McDonald cerclage under pudendal nerve block.

OBJECTIVE: The purpose of our study was to evaluate McDonald cerclage placement with pudendal anesthesia. Patient pain and hospital charges were analyzed and compared with a control group. STUDY DESIGN: This was a prospective, nonrandomized study. Pain was evaluated with a visual analog scale. Two groups were compared with the Student t test. RESULTS: Of 69 patients, 20 had cerclage with pudendal anesthesia; 49 served as control and had cerclage with regional anesthesia. There were no statistical differences in pain between the two groups or complications from pudendal anesthesia, and there was a large cost savings. CONCLUSION: All 20 patients had McDonald cerclage placed successfully with pudendal anesthesia. McDonald cerclage can be easily and safely placed under pudendal anesthesia, resulting in cost savings and improved use of time and resources for the patient and her physician.

Adult↗

An index of scientific quality for health reports in the lay press.

Although the quality of health reporting has been criticized for being unscientific, evaluations of health care reporting have been limited by the lack of a reliable and credible measure of scientific quality. We developed an index of scientific quality (ISQ) for health-related news reports and tested its reliability and sensibility. Items were generated from a survey of the literature and experts in research methodology. Items that were unclear, confusing or discriminated poorly between articles of high and low scientific quality were revised or deleted in an iterative process wherein potential criteria were independently applied to samples of 5 to 15 articles by 6 raters. To test the reliability of the final criteria 60 articles were drawn from three sampling frames: newspapers, magazines, and professional journals. Articles were intentionally selected to obtain a wide range of quality and topics. Two categories of raters were used: research assistants and physicians with research training. All 6 raters assessed all 60 articles. The sensibility of the index was tested by a questionnaire with 13 items related to face validity and content validity as well as other aspects of sensibility. The questionnaire was completed by 20 researchers and 13 health and science writers. The final ISQ includes 7 items that address the extent to which a report allows readers to draw conclusions about the applicability, validity and practical importance of the information that is reported. Chance corrected agreement (kappa) among all 6 raters for overall scientific quality was 0.62 (SE 0.02). The index was found to be sensible with only one major problem, the need for judgment in making ratings. While some degree of subjectivity appears to be inevitable in rating the scientific quality of health reports, the ISQ is acceptable reliable and credible and should be useful for evaluating and improving the scientific quality of health reporting.

Evaluation Studies as Topic↗

Soluble forms of the human T cell receptor CD4 are efficiently expressed by Streptomyces lividans.

We have developed a new gene expression and secretion system for Streptomyces lividans and used it to produce soluble forms of a human T-cell receptor CD4 at levels greater than 300 mg/l. The system uses the transcription, translation and secretion signals of the serine protease inhibitor gene STI-II which is naturally produced by S. longisporus. Using these signals, soluble derivatives of CD4 were secreted directly into the culture supernatant as correctly processed soluble, biologically active proteins. High level expression of the CD4 proteins depended on the transcription initiation signal, the amino acid sequence surrounding the signal peptide cleavage site and temporally controlled protease activities. We discuss these results in the context of the potential of this system for producing other eukaryotic proteins in Streptomyces.

Amino Acid Sequence↗

Dentin formation in miniature pigs with special reference to indomethacin and orthodontic treatment.

The rate of dentin mineralization and the influence of indomethacin on the dentin mineralization rate during orthodontic treatment was determined in miniature pigs by intravital labeling with tetracycline. The results demonstrated that the dentin mineralization rate in the control animals was 3.8 microns/day, a rate corresponding to that of human teeth. Both indomethacin and orthodontics had an effect on the dentin mineralization rate, indomethacin reducing and orthodontic forces increasing it. In combination, the two factors neutralized each other.

Animals↗

Age differences in the breast cancer management of rural women.

This article describes the effect of age on the pattern of access to breast cancer care among rural women diagnosed in 1986-1989. Cases were identified by the Illinois State Cancer Registry and information on breast cancer management was obtained through review of hospital records and physician survey. Case follow-back was more than 99% complete. State-of-the-art breast cancer management was defined by the Physician Data Query (PDQ) and included diagnostic evaluation, prognostic evaluation, and stage-specific definitive treatment. The data did not indicate age differences in bilateral diagnostic mammography, performance of hormone receptor assays, radiation therapy, and stage-appropriate treatment. Tumor staging and axillary node dissection did differ by age with significant negative trends in the multivariate model. Also, women younger than 55 years were the most likely to receive separate diagnostic biopsy, limited surgery, and chemotherapy, whereas women 55 to 74 years were most likely to receive hormone therapy. Although rural populations may experience barriers to some aspects of state-of-the-art breast cancer management, age does not always exacerbate them.

Age Factors↗

Transforming growth factor-beta enhances fracture healing in rabbit tibiae.

The ability of exogenous Transforming Growth Factor-beta (TGF-beta) to stimulate bone formation in fracture healing was investigated. TGF-beta was continuously applied in doses of 1 and 10 micrograms/day for 6 weeks to 2 groups of adult rabbits with unilateral plated mid-tibial osteotomies. A group receiving solvent without TGF-beta served as control. Fracture healing was evaluated by mechanical tests, bone morphometry and bone densitometry. Increased maximal bending strength and callus formation were demonstrated in the groups receiving TGF-beta. TGF-beta had no effect on bending-stiffness, bone mineral content, cortical thickness or haversian canal diameter. We conclude that local application of exogenous TGF-beta may enhance fracture healing in rabbits.

Animals↗

Effect of local prostaglandin E2 on fracture callus in rabbits.

We investigated the effect of local infusion with prostaglandin E2 (PGE2) in doses of 0.0003 to 4.0 mg/hour per kg body weight for 6 weeks on a plated unilateral osteotomy in rabbits. PGE2 caused a dose-dependent stimulation of callus formation. Total bone mineral content increased, although the mineral content per volume of the callus was reduced. In another experiment, PGE2 was infused either in the first half or in the second half of the healing period. No effect of PGE2 infusion could be observed in the first half of the 6-week healing period, whereas PGE2 infusion during the second half caused callus stimulation.

Animals↗

Indomethacin for prevention of heterotopic ossification. A randomized controlled study in 41 hip arthroplasties.

The effect of indomethacin 25 mg 3 times daily during the first 2 postoperative weeks in preventing heterotopic bone formation after cemented total hip arthroplasty was investigated in a randomized, double-blind and placebo-controlled clinical trial on 57 patients. 16 patients were secondarily excluded, leaving 19 patients in the indomethacin group and 22 patients in the placebo group. Evaluated from the 3-month radiographs, 18/19 indomethacin patients developed either no or only the milder Grade 1 ossification. In contrast, 11/22 placebo patients developed Grade 2 or 3 ossifications. Our observations favor indomethacin prophylaxis for 2 weeks in cemented arthroplasty of the hip.

Aged↗

Megestrol acetate in cancer anorexia and weight loss.

High-dose megestrol acetate has been associated with increased appetite and weight. To examine the effects of high-dose megestrol acetate in the treatment of anorexia and weight loss in patients with advanced hormone-insensitive malignant lesions, a randomized double-blind placebo-controlled trial was conducted. Patients receiving megestrol acetate for 1 month reported a significant improvement in appetite and adequacy of food intake compared with those receiving placebo. A three-item scale measuring appetite, adequacy of food intake, and concern about weight revealed a higher improvement with megestrol acetate than with placebo. Patients who worsened while receiving placebo had similar favorable changes after the cross over to megestrol acetate. These data indicate that megestrol acetate may improve appetite and food intake in patients with advanced cancer.

Adult↗

Effect of local prostaglandin E2 on periosteum and muscle in rabbits.

We assessed the target tissue for the stimulatory effect of prostaglandin E2 (PGE2) on bone formation previously observed during fracture healing. PGE2 was infused into tibial periosteal tissue in the right leg of 7 rabbits and into the anterior tibial muscle in the right leg of 7 other rabbits for 6 weeks. Solvent solution was infused into the left leg. PGE2 infusion at the periosteum caused the formation of primitive woven bone with large amounts of connective tissue; solvent infusion caused small amounts of normal periosteal bone formation. In the neighboring cortical bone, remodeling was increased after PGE2 infusion compared to solvent infusion. In the muscle, PGE2 infusion caused the formation of connective tissue with small amounts of woven bone. Thus, the major effects of PGE2 infusion at the site of the periosteum was the formation of primitive woven bone and in muscles the formation of connective tissue.

Animals↗

Germinal and somatic activity of the maize element Activator (Ac) in Arabidopsis.

We have investigated the germinal and somatic activity of the maize Activator (Ac) element in Arabidopsis with the objective of developing an efficient transposon-based system for gene isolation in that plant. Transposition activity was assayed with a chimeric marker that consists of the cauliflower mosaic virus 35S promoter and a bacterial streptomycin phosphotransferase gene (SPT). Somatic activity was detected in seedlings germinated on plates containing streptomycin as green-resistant sectors against a background of white-sensitive cells. Germinal excisions resulted in fully green seedlings. The transposition frequency was extremely low when a single copy of the transposon was present, but appeared to increase with an increase in Ac copy number. Plants that were selected as variegated produced an increased number of green progeny. The methylation state of the Ac elements in lines with either low or high levels of excision was assessed by restriction analysis. No difference was found between these lines, indicating that the degree of methylation did not contribute to the level of Ac activity. Germinal excision events were analyzed molecularly and shown to carry reinserted transposons in about 50% of the cases. In several instances, streptomycin-resistant siblings carried the same transposed Ac element, indicating that excision had occurred prior to meiosis in the parent. We discuss parameters that need to be considered to optimize the use of Ac as a transposon tag in Arabidopsis.

DNA↗

Identification and characterization of novel low-temperature-inducible promoters of Escherichia coli.

Escherichia coli promoters that are more active at low temperature (15 to 20 degrees C) than at 37 degrees C were identified by using the transposon Tn5-lac to generate promoter fusions expressing beta-galactosidase (beta-Gal). Tn5-lac insertions that resulted in low-temperature-regulated beta-Gal expression were isolated by selecting kanamycin-resistant mutants capable of growth on lactose minimal medium at 15 degrees C but which grew poorly at 37 degrees C on this medium. Seven independent mutants were selected for further studies. In one such strain, designated WQ11, a temperature shift from 37 degrees C to either 20 or 15 degrees C resulted in a 15- to 24-fold induction of beta-Gal expression. Extended growth at 20 or 15 degrees C resulted in 36- to 42-fold-higher beta-Gal expression over that of cells grown at 37 degrees C. Treatment of WQ11 with streptomycin, reported to induce a response similar to heat shock, failed to induce beta-Gal expression. In contrast, treatment with either chloramphenicol or tetracycline, which mimics a cold shock response, resulted in a fourfold induction of beta-Gal expression in strain WQ11. Hfr genetic mapping studies complemented by physical mapping indicated that in at least three mutants (WQ3, WQ6, and WQ11), Tn5-lac insertions mapped at unique sites where no known cold shock genes have been reported. The Tn5-lac insertions of these mutants mapped to 81, 12, and 34 min on the E. coli chromosome, respectively. The cold-inducible promoters from two of the mutants (WQ3 and WQ11) were cloned and sequenced, and their temperature regulation was examined. Comparison of the nucleotide sequences of these two promoters with the regulatory elements of other known cold shock genes identified the sequence CCAAT as a putative conserved motif.

Anti-Bacterial Agents↗

Phenotypes of murine leukemia virus-induced tumors: influence of 3' viral coding sequences.

Murine leukemia viruses (MuLVs) induce leukemias and lymphomas in mice. We have used fluorescence-activated cell sorter analysis to determine the hematopoietic phenotypes of tumor cells induced by a number of MuLVs. Tumor cells induced by ecotropic Moloney, amphotropic 4070A, and 10A1 MuLVs and by two chimeric MuLVs, Mo(4070A) and Mo(10A1), were examined with antibodies to 13 lineage-specific cell surface markers found on myeloid cell, T-cell, and B-cell lineages. The chimeric Mo(4070A) and Mo(10A1) MuLVs, consisting of Moloney MuLV with the carboxy half of the Pol region and nearly all of the Env region of 4070A and 10A1, respectively, were constructed to examine the possible influence of these sequences on Moloney MuLV-induced tumor cell phenotypes. In some instances, these phenotypic analyses were supplemented by Southern blot analysis for lymphoid cell-specific genomic DNA rearrangements at the immunoglobulin heavy-chain, the T-cell receptor gamma, and the T-cell receptor beta loci. The results of our analysis showed that Moloney MuLV, 4070A, Mo(4070A), and Mo(10A1) induced mostly T-cell tumors. Moloney MuLV and Mo(4070A) induced a wide variety of T-cell phenotypes, ranging from immature to mature phenotypes, while 4070A induced mostly prothymocyte and double-negative (CD4- CD8-) T-cell tumors. The tumor phenotypes obtained with 10A1 and Mo(10A1) were each less variable than those obtained with the other MuLVs tested. 10A1 uniformly induced a tumor consisting of lineage marker-negative cells that lack lymphoid cell-specific DNA rearrangements and histologically appear to be early undifferentiated erythroid cell-like precursors. The Mo(10A1) chimera consistently induced an intermediate T-cell tumor. The chimeric constructions demonstrated that while 4070A 3' pol and env sequences apparently did not influence the observed tumor cell phenotypes, the 10A1 half of pol and env had a strong effect on the phenotypes induced by Mo(10A1) that resulted in a phenotypic consistency not seen with other viruses. This result implicates 10A1 env in an active role in the tumorigenic process.

Animals↗

Controlled trial of respiratory muscle training in chronic airflow limitation.

BACKGROUND: Whether respiratory muscle training is of benefit to patients with chronic airflow limitation is controversial. The objective of the study was to determine the effect of resistance breathing training on physiological and functional measures in patients with chronic airflow obstruction. METHODS: The design was a randomised, double blind, controlled trial with a six month follow up. Eighty two patients with a forced expiratory volume in one second (FEV1) of less than 70% predicted, and an FEV1/vital capacity ratio of less than 0.7, were randomised to receive training for 10 minutes five times daily with progressively larger resistances through a resistive breathing device (PFLEX) as tolerated or to a sham device which gave minimal resistance. The main outcome measures, respiratory muscle strength and endurance, a progressive exercise test, a six minute walk test and physical and emotional function (chronic respiratory questionnaire) were assessed at monthly intervals. Patients in both groups were also randomised to wear or not wear nose clips during their training. RESULTS: No significant differences were observed between treatment and control groups, with or without nose clips, for any of the outcomes. Confidence intervals on the difference between treatments were narrow, excluding clinically important difference in any major outcome. CONCLUSION: This training regimen fails to strengthen respiratory muscles or improve exercise or functional capacity in patients with chronic airflow limitation.

Aged↗

Are race and sex differences in lung function explained by frame size? The CARDIA Study.

Using the CARDIA cohort of 20- to 32-yr-old black and white men and women, FVC and FEV1 were standardized for standing height, sitting height, leg height, elbow breadth, and biacromial diameter in such a way that the standardized lung function showed minimal statistical dependence on these measures of frame size. Race and sex differences in lung function have been reported even after adjustment for height; however, these differences might depend on aspects of frame size other than height. We found that within this age group height2 provided robust standardization for FVC and FEV1 for all race and sex strata of the population. Height explained approximately 40% of the variance of FVC and FEV1 in whites, 30% in black women, and 20% in black men. In black men only, standardization for the combination of sitting height, leg height, elbow breadth, and biacromial diameter improved explained variance to nearly 40% for FVC and nearly 30% for FEV1. After standardization for height, FVC and FEV1 were found to be 14 to 19% higher in whites than in blacks, and in men than in women. Standardization of FVC and FEV1 for sitting height, leg height, elbow breadth, and biacromial diameter combined reduced these differences to 13-16%. Thus, race and sex differences in lung function exist even after detailed adjustment for frame size.

Adult↗