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Biomedical subjects

M F Smith

Publications and source records attributed to M F Smith.

At least 19 recordsLinked to original sources
PubMed1992

Results of intraoperative 5-fluorouracil supplementation on trabeculectomy for open-angle glaucoma.

The success rate of filtration surgery has been increased by the postoperative subconjunctival injection of 5-fluorouracil, a potent antimetabolite. However, the optimal route of administration has not been determined. Trabeculectomy was performed on one eye each of 14 patients. Topical 5-fluorouracil was applied intraoperatively (50 mg/ml for five minutes) and subconjunctival 5-fluorouracil was injected postoperatively (an average of 5.8 injections) (mean total dose, 29 mg). Seven of the 14 eyes had primary open-angle glaucoma, and seven eyes had open-angle glaucoma with either uveitis, aphakia, or previous failed trabeculectomy. Mean preoperative intraocular pressure was 24.7 mm Hg during treatment with an average of three antiglaucoma medications, and mean final intraocular pressure was 11.9 mm Hg during treatment with an average of 0.2 medication. Thirteen of 14 eyes (93%) had final intraocular pressure of 18 mm Hg or less. Mean follow-up was 6.4 months (range, four to nine months). No remarkable complications occurred. Visual acuity remained stable in 13 of 14 eyes (93%). Intraoperative 5-fluorouracil may be a helpful adjunct in achieving low final intraocular pressure after trabeculectomy.

Administration, Topical↗
PubMed1992

Human IL-1 receptor antagonist promoter. Cell type-specific activity and identification of regulatory regions.

To study the molecular mechanisms involved in transcriptional regulation of the human IL-1R antagonist (IL-1ra) we have isolated 1680-bp of 5'-flanking region DNA from the IL-1ra gene. This region of DNA was sequenced and cloned into the luciferase expression vector pA3Luc (pRA-1680.Luc) for use in gene transfer studies aimed at determining the cis-acting DNA elements required for IL-1ra expression. Sequence analysis of the IL-1ra promoter revealed a TATAA box at -26, with consensus sequences for possible NF-kB-, NFIL-1 beta A-, AP-1-, and CRE-binding sites located further upstream. When transfected into a variety of human and murine cell lines, the cloned IL-1ra promoter was preferentially active in those cell lines in which expression of the endogenous IL-1ra gene could be detected. The cloned promoter and the endogenous IL-1ra promoter utilized the same transcriptional start site. This promoter activity was LPS-inducible in the RAW 264.7 murine macrophage cell line. In the human monocytic cell line U937, IL-1ra promoter activity was inducible by LPS or PMA treatment, but the combination of LPS and PMA led to the greatest increase in promoter activity, identical to the pattern of expression of endogenous IL-1ra mRNA as detected by polymerase chain reaction analysis. A series of 5'-truncated promoter constructs having a common 3'-end at +27 were created to map potential cis-acting transcriptional elements important for full IL-1ra promoter activity. Removal of sequences between -294 and -148 led to a greater than 90% decrease in both unstimulated and LPS-induced promoter activity; further deletion to -85 led to an almost complete abrogation of promoter activity. These studies demonstrate that the cloned IL-1ra promoter behaves in a manner consistent with that of the endogenous gene. Two regions within the IL-1ra promoter are identified which are required for full promoter activity.

Animals↗
PubMed1992

Quantitation of molecular densities by cryo-electron microscopy. Determination of the radial density distribution of tobacco mosaic virus.

We have determined the absolute mass and radial scattering density distribution of tobacco mosaic virus in the frozen-hydrated state by energy-filtered low-dose bright-field transmission electron microscopy. The absolute magnitude of electron scattering from tobacco mosaic virus in 150 nm of ice was within 3.0% of that predicted, with inelastic scattering accounting for approximately 80% of the scattering contrast. In order to test the accuracy of the radial reconstruction, a computer model of tobacco mosaic virus was built from the atomic co-ordinates assuming uniform solvent density. The validity of the model was confirmed by comparison of X-ray scattering and predictions of the model (R factor = 0.05). First-order corrections for the microscope contrast transfer function were necessary and sufficient for conversion of the cryo-electron microscopy images into accurate representations of the mass density. At 1.9 nm resolution the compensated reconstruction and model had density peaks of similar magnitude at 2.4, 4.2, 6.0 and 7.8 nm radius and a central hole of 2 nm radius. Equatorial Fourier transforms of the corrected electron images were in excellent agreement with predictions of the model (R factor = 0.12). Thus, the uniform solvent approximation was adequate at 1.9 nm resolution to describe quantitatively X-ray scattering in liquid water and electron imaging in vitreous ice. This is the first demonstration that cryo-electron microscopy images can be used to quantitate the absolute mass, mass per unit length and internal density distributions of proteins and nucleic acids.

Computer Simulation↗
PubMed1992

Comparison of the double-plate Molteno drainage implant with the Schocket procedure.

A randomized, prospective study was undertaken to compare the results of the double-plate Molteno implant with those of the anterior chamber tube shunt to encircling band (ACTSEB) procedure, described by Schocket, for eyes with complicated glaucoma. One surgeon performed all the procedures. For statistical analysis there were 21 eyes in the ACTSEB group and 19 eyes in the Molteno group, with at least 6 months of follow-up (mean follow-up, 26 months; range, 6 to 49 months). Eyes with aphakia/pseudophakia, uveitis, previous penetrating keratoplasty, previous unsuccessful trabeculectomy, and neovascular glaucoma were evenly distributed between the two groups. Mean preoperative intraocular pressures in the ACTSEB and Molteno groups were 35.2 and 34.6 mm Hg, respectively. With the exclusion of one phthisical eye in each group final mean intraocular pressure in the ACTSEB group was 15.1 mm Hg, with a mean of 0.43 medications, and 14.4 mm Hg, with a mean of 0.95 medications, in the Molteno group. Seven (33%) of 21 eyes and 13 (68%) of 19 eyes in the ACTSEB and Molteno groups, respectively, were receiving additional antiglaucoma medication at the most recent visit (P = .027). Final visual acuity was stable (within one Snellen line of baseline visual acuity) or improved in 10 (48%) of 21 eyes in the ACTSEB group compared with 13 (68%) of 19 eyes in the Molteno group. Complications or inadequate pressure control requiring further surgical intervention were seen in nine (47%) of 19 eyes in the Molteno group and 12 (57%) of 21 eyes in the ACTSEB group.(ABSTRACT TRUNCATED AT 250 WORDS)

Drainage↗
PubMed1992

Homozygous acute intermittent porphyria: compound heterozygosity for adjacent base transitions in the same codon of the porphobilinogen deaminase gene.

A sister and brother with severe porphobilinogen (PBG) deaminase deficiency are described. Each of their parents carries a different mutation for acute intermittent porphyria and the children are homozygous for the PBG-deaminase deficiency that causes this disorder. Both are compound heterozygotes for adjacent base transitions in the same codon in exon 10 of the PBG deaminase gene.

Base Sequence↗
PubMed1992

Intraoperative and post operative treatment with 5-fluorouracil and mitomycin-c: long term effects in vivo on subconjunctival and scleral fibroblasts.

Rabbits undergoing full thickness glaucoma filtering surgery were exposed to one of 4 treatments. Group 1 received intraoperative distilled water, group 2 received intraoperative mitomycin-c 0.2 mg/ml for 5 minutes, group 3 received 5 post operative injections of 5-Fluorouracil (5-FU) 5 mg in 0.1 ml, and group 4 received intraoperative 5-FU 50 mg/ml followed by 5 post operative injections of 5-FU. 30 days after the operation tissue biopsies were taken from the subconjunctival and scleral tissue at the treated area and 90 and 180 degrees from the treated area. The biopsies were then placed in tissue culture media and the outgrowths quantitated. The fibroblast outgrowths from all areas did not differ significantly from each other except for the outgrowths from the areas directly treated with mitomycin 0.2 mg/ml which were significantly smaller. In addition then cells were morphologically abnormal although there were foci of normal cells which appeared to be growing from localised areas in the tissue biopsies. The outgrowths from the areas 90 and 180 degrees from the treated area were normal. Intraoperative treatments with mitomycin-c result in long term inhibition of fibroblast proliferation limited to the treated area, when compared with intraoperative and postoperative treatment with 5-FU. Failure of filtration surgery in eyes treated with intraoperative mitomycin may in part be due to unaffected cells reproliferating.

Animals↗
PubMed1992

Quantitative energy-filtered electron microscopy of biological molecules in ice.

The theoretical and experimental bases for quantitative electron microscopy of frozen-hydrated specimens are described, with special considerations of energy filtration to improve the images. The elastic and inelastic scattering from molecules in vacuum and in ice are calculated, and simple methods to approximate scattering are introduced. Multiple scattering calculations are used to describe the scattering from vitreous ice and to predict the characteristics of images of frozen-hydrated molecules as a function of ice thickness and accelerating voltage. Energy filtration is predicted to improve image contrast and signal-to-noise ratio. Experimental values for the inelastic scattering of ice, the energy spectrum of thick ice, and the contrast of biological specimens are determined. The principles of compensation for the contrast transfer function are presented. Tobacco mosaic virus is used to quantify the accuracy of interpreting image intensities to derive the absolute mass, mass per unit length, and internal mass densities of biological molecules. It is shown that compensation for the contrast transfer function is necessary and sufficient to convert the images into accurate representations of molecular density. At a resolution of 2 nm, the radial density reconstructions of tobacco mosaic virus are in quantitative agreement with the atomic model derived from X-ray results.

Cryopreservation↗
PubMed1992

Three-dimensional photon detection kernels and their application to SPECT reconstruction.

In single photon emission computed tomography (SPECT), three-dimensional photon detection kernels characterize the probabilities that photons emitted by radio-isotopes in different parts of the source region will be detected at particular projection pixels of the projection images. Monte Carlo modelling is used to study these kernels for the case of parallel hole collimators. The use of vectorized Monte Carlo computer code speeds the modelling computations. The contributions of direct and scattered photons to projection data in a transverse plane from neighbouring planes are significant for the case of uniform activity within a water-filled cylinder. A reconstruction method using the 3D kernels is proposed in which projection measurements in three adjacent planes are used simultaneously to estimate the source activity of the center plane. This multiple slice method accounts for the fact that photons detected in a given transverse plane may have originated in other transverse planes with different activity distributions. The matrix equations for image reconstruction are solved using generalized matrix inverses. The new method shows compensation for 3D photon detection effects when applied to projection data from a numerical simulation and a cardiac phantom experiment. Quantitation for the numerical study is improved compared with results from a single slice reconstruction method.

Heart↗
PubMed1992

Mitochondrial DNA-like sequence in the nuclear genome of an akodontine rodent.

Initial amplification and sequencing of a 366-bp fragment of the cytochrome b gene by a conserved primer pair (MVZ 03 and MVZ 04) revealed a nonfunctional copy of the gene with two deletions (one of which is 17 bp in length and the other of which is 3 bp in length) in Chroeomys jelskii, a South American akodontine rodent. By means of an alternative primer to MVZ 03--namely, MVZ 05--from the region of the tRNA for glutamic acid, a functional copy of cytochrome b was subsequently amplified. Both primer pairs amplify functional sequence when applied to purified mitochondrial DNA (mtDNA). Restriction-endonuclease digestion of purified mtDNA from C. jelskii did not reveal any additional sets of bands that would suggest heteroplasmy in the mitochondrial genome. When probed with both functional and nonfunctional gene fragments, MboI restriction digests revealed the same pattern, providing further evidence that the nonfunctional copy must be located in the nucleus. Observed differences in the mitochondrial and nuclear sequences from two populations are consistent with a faster rate of change in mtDNA than in nuclear DNA.

Animals↗
PubMed1992

Luteal function after intrauterine infusion of recombinant bovine interferon-alpha I1 into postpartum beef cows expected to have short or normal luteal phases.

This study was conducted to determine whether intrauterine infusion of recombinant bovine interferon-alpha I1 (rboIFN-alpha I1), which has 70% sequence identity to bovine trophoblast protein-1, will prevent regression of corpora lutea anticipated to have a short lifespan. Twenty-six beef cows in good body condition were allotted to four treatment groups at parturition in a 2 x 2 factorial design. Treatments were: group 1, saline; group 2, rboIFN-alpha I1; group 3, norgestomet-saline; and group 4, norgestomet-rboIFN-alpha I1. Norgestomet implants were inserted on days 21-24 postpartum and removed 9 days later (before injection of human chorionic gonadotrophin (hCG)). Ovulation was induced 30 to 33 days postpartum with 5000 or 10,000 iu hCG. Groups 1 (n = 7) and 3 (n = 5) were given intrauterine infusions (rectocervical approach) twice daily with saline on days 1-12 or 13-24 after hCG injection, respectively. Cows allotted to groups 2 (n = 8) and 4 (n = 6) were given intrauterine infusions (rectocervical approach) of 2 mg rboIFN-alpha I1 twice daily on days 1-12 or 13-24 after hCG injection, respectively. Treatment with both norgestomet and rboIFN-alpha I1 delayed (P less than 0.01) luteolysis. Lengths of luteal phases (days; mean +/- SEM) were 8.4 +/- 0.7 (group 1, saline), 14.1 +/- 1.0 (group 2, rboIFN-alpha I1), 18.6 +/- 1.3 (group 3, norgestomet-saline) and 20.8 +/- 1.2 (group 4, norgestomet-rboIFN-alpha I1). Concentration of progesterone in serum was similar among all groups the first 6 days following hCG-induced ovulation, but differed (P less than 0.01) thereafter.(ABSTRACT TRUNCATED AT 250 WORDS)

Animals↗
PubMed1992

Comparative validity of two hearing loss screening questionnaires.

BACKGROUND: Hearing loss is one of the most common of all physical impairments, but physicians seldom screen adults for it, and patients often overlook or deny hearing problems. This study was designed to validate the use of two self-administered hearing loss questionnaires. METHODS: Two self-administered screening questionnaires and a hearing screening evaluation using the Welch Allyn Audioscope 3 instrument were given to 409 consecutive family practice patients over the age of 18 years. Correlational, discriminate, sensitivity, and specificity analyses were conducted on the data. RESULTS: Neither of the existing questionnaires was clinically sensitive enough to be recommended for use. A new tool based on a discriminate function analysis of the existing questionnaires was developed. In contrast, the audioscope proved to be a sensitive screening tool. Of those patients who were identified, 88% did not follow recommendations to obtain further evaluation. CONCLUSIONS: Existing self-administered questionnaires cannot be recommended for use. A controlled clinical study using the newly derived questionnaire, the Smith Hearing Screening, should be conducted.

Adult↗
PubMed1991

IL-1 receptor antagonist and IL-1 beta production in human monocytes are regulated differently.

Human monocytes may synthesize simultaneously both the agonist IL-1 beta and a specific receptor antagonist of IL-1 (IL-1ra). These studies examined whether monocyte production of these two structurally related cytokines was regulated differently. IL-1ra and IL-1 beta protein levels in cell supernatants and lysates were measured with specific ELISA. Relative steady-state mRNA levels, relative transcriptional rates as determined by the nuclear run-on technique, and mRNA stability were all assessed using specific cDNA probes. Monocytes were stimulated with LPS alone, adherent IgG, or both LPS and adherent IgG. Monocytes stimulated with LPS produced near equivalent amounts of IL-1ra and IL-1 beta proteins over 22 h; relative steady-state mRNA levels paralleled the protein levels. In addition, LPS-induced monocytes exhibited enhanced rates of transcription for both IL-1ra and IL-1 beta, in comparison to adherent control cells without LPS. mRNA half-lives in LPS-induced monocytes also were similar for IL-1ra and IL-1 beta. Monocytes cultured on adherent IgG exhibited a low level of IL-1 beta transcription with an absence of protein production. In contrast, adherent IgG led to a high and prolonged rate of IL-1ra protein production. Furthermore, monocytes cultured on adherent IgG exhibited a specific induction of IL-1ra transcription and a marked prolongation in IL-1ra mRNA stability. However, LPS in a high concentration, 1 microgram/ml, reversed the IgG induction of IL-1ra production by decreasing both transcriptional rate and mRNA stability. These results indicate that production of IL-1ra by human monocytes is characterized by different patterns of regulation in comparison with IL-1 beta. LPS induces production of both proteins whereas adherent IgG stimulates only IL-1ra production. The effects of IgG and LPS on induction of IL-1ra production in human monocytes are mediated at both transcriptional and post-transcriptional levels.

Humans↗
PubMed1991

Effect of constant infusion of oxytocin on luteal lifespan and oxytocin-induced release of prostaglandin F2 alpha in heifers.

Two experiments were conducted to determine whether constant infusion of oxytocin would prolong the luteal phase and inhibit uterine prostaglandin F2 alpha (PGF2 alpha) secretion in heifers. In Experiment 1, twelve heifers, treated with saline (SAL) or oxytocin (OXY) via jugular cannulae infusions (INF) or osmotic minipumps (OMP), were allotted at estrus into four treatment groups (n = 3). Treatments were: SAL-INF, SAL-OMP, OXY-INF and OXY-OMP. Physiological saline or oxytocin was given from Days 10 to 23 (Day 0 = estrus) of the estrous cycle. Method of treatment (jugular cannula infusion or osmotic minipump) had no effect (P greater than 0.05) on estrous cycle length or pattern of secretion of progesterone; therefore, data were pooled. Estrous cycle lengths were extended (P less than 0.01) for heifers which received oxytocin (25.3 +/- 0.4 d) compared to saline (20.5 +/- 0.4 d). Luteolysis did not occur in oxytocin-treated heifers until after treatment ceased. Experiment 2 was designed and conducted identically to Experiment 1 with the addition of a "challenge" injection of oxytocin (100 IU oxytocin, i.v.) given on Day 16 of the estrous cycle. Treatment of heifers with oxytocin extended (P less than 0.05) estrous cycle length by an average of 3 d compared to heifers treated with saline. The "challenge" injection induced (P less than 0.05) secretion of PGF2 alpha (as measured by the stable PGF2 alpha metabolite, 15-keto-13,14-dihydro-PGF2 alpha) in saline-treated but not oxytocin-treated heifers. In both Experiment 1 and 2, serum concentrations of FSH were elevated (P less than 0.05) in oxytocin-treated heifers. No increase was observed for LH or prolactin. The rise in estradiol-17 beta at luteolysis was not affected (P greater than 0.10) by treatment. In summary, constant infusion of oxytocin extended luteal lifespan, prolonged secretion of progesterone, and inhibited oxytocin-induced secretion of PGF2 alpha. Constant infusion of oxytocin did not affect serum concentrations of estradiol-17 beta, LH or prolactin; however, serum concentrations of FSH were elevated during the oxytocin treatment period.

Animals↗
PubMed1991

Problems identified by family caregivers in counseling.

Content analysis of the problems that caregiving daughters and daughters-in-law (N = 51) elected to work on during counseling yielded seven categories: improving coping skills, meeting elder's care needs, responding to family issues, concern over the caregiver-recipient relationship, eliciting formal and informal supports, feelings of inadequacy and guilt, and planning for elder's future. Two global themes of maintaining control and increasing responsibility within the caregiver role were also evident. Case examples illustrate problems, and practice implications are considered.

Counseling↗