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Biomedical subjects

H Browning

Publications and source records attributed to H Browning.

11 recordsLinked to original sources

Tea2p is a kinesin-like protein required to generate polarized growth in fission yeast.

Cytoplasmic microtubules are critical for establishing and maintaining cell shape and polarity. Our investigations of kinesin-like proteins (klps) and morphological mutants in the fission yeast Schizosaccharomyces pombe have identified a kinesin-like gene, tea2(+), that is required for cells to generate proper polarized growth. Cells deleted for this gene are often bent during exponential growth and initiate growth from improper sites as they exit stationary phase. They have a reduced cytoplasmic microtubule network and display severe morphological defects in genetic backgrounds that produce long cells. The tip-specific marker, Tea1p, is mislocalized in both tea2-1 and tea2Delta cells, indicating that Tea2p function is necessary for proper localization of Tea1p. Tea2p is localized to the tips of the cell and in a punctate pattern within the cell, often coincident with the ends of cytoplasmic microtubules. These results suggest that this kinesin promotes microtubule growth, possibly through interactions with the microtubule end, and that it is important for establishing and maintaining polarized growth along the long axis of the cell.

Amino Acid Sequence↗

Effect of pumice prophylaxis on the bond strength of orthodontic brackets.

Pumice prophylaxis has long been accepted as a prerequisite for achieving adequate enamel etching during orthodontic bonding procedures. Three methods were used in this study to examine the effects of pumice prophylaxis on the bond strength of orthodontic brackets: (1) shear bond strength of brackets that were bonded to extracted premolars after surface preparation procedures, which either included or did not include prior pumice prophylaxis, was evaluated; (2) scanning electron microscopy (SEM) was used to examine the surface characteristics of teeth that had been etched with and without prior pumice prophylaxis; and (3) rate of bracket failure in patients who had had brackets bonded with and without prior pumice prophylaxis was recorded during an average treatment time of 18 months. No significant differences were noted in bond strength, general etched enamel surface characteristics, or bracket retention rates. Some specific differences, however, were noted on SEM in localized areas of the etched enamel surfaces, although these did not appear to affect the bond strength or bracket retention rates ultimately attained.

Acid Etching, Dental↗

Isolation of Enterobacteriaceae from the mouth and potential association with malodor.

Bad breath is a common phenomenon, usually the result of bacterial metabolism in the oral cavity. It is generally accepted that Gram-negative bacteria are responsible for this problem, largely through degradation of proteinaceous substances. In initial experiments, screening of malodorous isolates following outgrowth of samples obtained from saliva, periodontal pockets, and the tongue dorsum yielded enterobacterial isolates. Clinical studies were conducted to examine the prevalence of such bacteria in four different populations: orthodontic patients, malodor clinic patients, complete-denture wearers, and a healthy young population. The prevalence of Enterobacteriaceae in the oral cavities of the denture-wearing population was very high (48.0%) as compared with the other groups: 27.1% in the malodor clinic patients, 16.4% in the normal population, and 13% among orthodontic patients. Isolates of Klebsiella and Enterobacter emitted foul odors in vitro which resembled bad breath, with concomitant production of volatile sulfides and cadaverine, both compounds related to bad breath. When incubated on a sterile denture, enterobacterial isolates produced typical denture foul odor. Isolates exhibited cell-surface hydrophobic properties when tested for adhesion to acryl and aggregation with ammonium sulphate. The results, taken together, suggest that Klebsiella and related Enterobacteriaceae may play a role in denture malodor.

Adolescent↗

Macrorestriction analysis of Caenorhabditis elegans genomic DNA.

The usefulness of genomic physical maps is greatly enhanced by linkage of the physical map with the genetic map. We describe a "macrorestriction mapping" procedure for Caenorhabditis elegans that we have applied to this endeavor. High molecular weight, genomic DNA is digested with infrequently cutting restriction enzymes and size-fractionated by pulsed field gel electrophoresis. Southern blots of the gels are probed with clones from the C. elegans physical map. This procedure allows the construction of restriction maps covering several hundred kilobases and the detection of polymorphic restriction fragments using probes that map several hundred kilobases away. We describe several applications of this technique. (1) We determined that the amount of DNA in a previously uncloned region is < 220 kb. (2) We mapped the mes-1 gene to a cosmid, by detecting polymorphic restriction fragments associated with a deletion allele of the gene. The 25-kb deletion was initially detected using as a probe sequences located approximately 400 kb away from the gene. (3) We mapped the molecular endpoint of the deficiency hDf6, and determined that three spontaneously derived duplications in the unc-38-dpy-5 region have very complex molecular structures, containing internal rearrangements and deletions.

Animals↗

A sperm-supplied factor required for embryogenesis in C. elegans.

The paternal-effect embryonic-lethal gene, spe-11, is required for normal development of early C. elegans embryos. Spe-11 embryos fail to complete meiosis, form a weak eggshell, fail to orient properly the first mitotic spindle, and fail to undergo cytokinesis. Here we report cloning and sequencing of the spe-11 gene, which encodes a novel protein. As predicted by the paternal-effect mutant phenotype, the gene is expressed during spermatogenesis but is not detectable in females undergoing oogenesis, and the protein is present in mature sperm. To investigate whether SPE-11's essential function is during spermatogenesis or whether sperm-delivered SPE-11 functions in the newly fertilized embryo, we engineered animals to supply SPE-11 to the embryo through the oocyte rather than through the sperm. We found that maternal expression is sufficient for embryonic viability. This result demonstrates that SPE-11 is not required during spermatogenesis, and suggests that SPE-11 is a sperm-supplied factor that participates directly in development of the early embryo. In contrast to the many known maternal factors required for embryogenesis, SPE-11 is the first paternally contributed factor to be genetically identified and molecularly characterized.

Amino Acid Sequence↗

Ceramic bracket fracture resistance to second order arch wire activations.

The fracture resistance of ceramic brackets to orthodontic activations has been incorrectly estimated by previous investigations that have reported second order loads in terms of force magnitudes rather than moments. Because force magnitudes alone do not reflect the influence of distance from the site of force application on total load, it is impossible to apply previous results to actual clinical situations. The purpose of this study was to determine the average moments (in gram-millimeters) necessary to fracture various ceramic brackets subjected to second order tipping activations and compare them with actual clinical loads. Central and lateral incisor ceramic brackets from seven manufacturers were subjected to mesial-distal tipping arch wire activations at two speeds of load application with a testing apparatus designed for that purpose. Significant differences in fracturability among the brackets of various manufacturers and between central and lateral incisor brackets were found. There were no differences related to the speed of load application. Once the influence of bracket width was considered, the differences in fracture resistance between central and lateral incisor brackets were no longer apparent. Second order activations required to fracture the ceramic brackets in this study were all much greater than measured clinical orthodontic loads. It is unlikely that second order arch wire activations are a significant cause of ceramic bracket failure.

Ceramics↗

A novel, quantitative bioassay for type I interferon using a recombinant indicator cell line.

We describe a specific and quantitative novel assay for biologically active human type I interferon (IFN), the MxR assay. It is based on a Vero cell line containing multiple copies of a hybrid gene consisting of the murine Mx promoter, which is responsive to type I IFN, linked to the human growth hormone (hGH) transcription unit. Exposure of this cell line to IFN-alpha or -beta for 12-48 hours results in the production of hGH that is measured by a commercially available radio-immune assay. The response to IFN-alpha is dose-dependent between 3 and 1000 units/ml. There is no response to TNF, IL-1 and a number of other cytokines and growth factors, and only a negligible response to IFN-gamma.

Animals↗

Transport of the murine Mx protein into the nucleus is dependent on a basic carboxy-terminal sequence.

Cytoplasmic microinjection of murine Mx mRNA synthesized in vitro or nuclear microinjection of Mx cDNA under the control of a constitutive promoter into murine Mx- cells led to the accumulation of Mx protein in the nucleus and inhibited the replication of influenza virus but not of vesicular stomatitis virus (VSV). Similar results were also found with dog, rat, chicken, and monkey cells. A human lung fibroblast cell line (A549) was exceptional in that Mx protein was located predominantly in the cytoplasm and showed antiviral activity. Truncation of the 19 last residues of murine Mx protein almost completely abolished accumulation of Mx protein in the nucleus; however the activity against influenza virus was at least partially retained. The truncated region contains a segment rich in basic amino acids, similar to that reported for several nuclear location signals.

Amino Acid Sequence↗

Embryonal nephroma in a wapiti.

Ebryonal nephroma was diagnosed in a young, free-ranging female wapiti (Cervus canadensis) from the San Juan National Forest in southwestern Colorado. Metastases were found in the lung.

Animals↗