Microencapsulated chlorpyrifos distribution in loamy sand and silty clay loam soils when applied with a sub-slab injector for subterranean termite control.
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Biomedical subjects
Publications and source records attributed to R W Davis.
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Centromere binding factor 1 (Cbf1p or CP1) binds to the CDEI region of Saccharomyces cerevisiae centromeres and is a member of the basic helix-loop-helix (bHLH) class of proteins. Deletion of the gene encoding Cbf1p results in an increased frequency of chromosome loss, hypersensitivity to low levels of microtubule disrupting drugs (such as thiabendazole and benomyl) and methionine auxotrophy. By polymerase chain reaction-based random mutagenesis of the CBF1 gene we have obtained a number of mutant alleles that make full-length protein with impaired function. The mutations in these alleles are clustered in or just downstream from the bHLH domain. Among the alleles obtained was a class that was more compromised for transcriptional activation and a class that was more compromised for chromosome loss and thiabendazole hypersensitivity. These results indicate that at least some aspects of the role of Cbf1p in chromosome segregation and transcriptional activation are distinct. In contrast, increased chromosome loss and thiabendazole hypersensitivity were not separated in any of the alleles, suggesting that these phenotypes reflect the same mechanistic defect. These observations are consistent with a model that suggests that one role of Cbf1p in chromosome segregation may be to improve the efficiency with which contact between the kinetochore and spindle microtubules is established or maintained.
The HAT4 gene from the plant Arabidopsis thaliana encodes a homeo domain protein that contains a leucine zipper motif. Homeo domain-leucine zipper (HD-Zip) proteins have not been found in animal systems, suggesting that HAT4 may define a new family of transcription factors that regulate higher plant development. To explore this possibility, functional studies of HAT4 were carried out in yeast and in transgenic plants. Point mutants of HAT4 isolated in yeast define functionally critical residues within the HD-Zip domain, many of which correspond to highly conserved positions in known homeo domains and leucine zippers. Transgenic plants bearing constructs that alter HAT4 expression exhibit a series of interesting developmental phenotypes, including changes in morphology and developmental rate. Thus, the HAT4 gene of Arabidopsis encodes an HD-Zip protein that functions as a novel developmental regulator.
Plants distinguish among the pollen grains that land on the stigma, permitting only compatible pollen to fertilize egg cells. To investigate these cell-cell interactions, Arabidopsis mutations that affect pollen-pistil communication were isolated. A male-sterile mutation that disrupts pollen-pistil interactions by eliminating the extracellular pollen coat (tryphine) is described here. Stigma cells that contact the mutant pollen produce callose, a carbohydrate synthesized in response to foreign pollen. The mutant pollen fails to germinate because it does not absorb water from the stigma, yet germinates in vitro, indicating it is viable. The defect is also conditional; high humidity results in pollen hydration and successful fertilization. Analysis of mature, mutant pollen indicated that it is deficient in long-chain lipids and has none of the lipoidic tryphine normally present on its surface. Immature mutant pollen grains have aberrant tryphine that disappears during pollen development. The sterile plants also lack stem waxes, and pollen from other wax-defective (eceriferum) mutants with reduced fertility has few of the lipid droplets normally present in tryphine. These results demonstrate that tryphine is critical for pollen-stigma interactions and suggest that tryphine lipids are required for fertilization, either by directly signaling the stigma or by stabilizing other tryphine components.
This article reports a series of 31 patients with true phantom pain. Mexiletine was used in an open-label study to evaluate its effectiveness in the treatment of this disorder. Eighteen of these patients had good to excellent results with mexiletine alone. Eleven other patients responded favorably with a combination of mexiletine and clonidine. Two patients did not respond to either drug alone or in combination. Three patients elected to discontinue use of the mexiletine because of persistent nausea, even at low doses.
Phantom sensation, phantom pain, and stump pain have been known since antiquity. For millenia, sensations in the missing body part were thought to be of psychic origin. During this century the psychic explanations have gradually given way to physiological explanations. However, these have been tenuous hypotheses that have poorly explained the reported symptoms. This history is reviewed here. It is now believed that phantom pain probably originated in the pain transmitting neurons of the dorsal horn. This has led to significant advances in the treatment of this disorder. It is now thought that phantom sensation is best explained as a sensory emgram or a part of the neuomatrix as purposed in 1989 by Melzack. This hypothesis is in agreement with newer theories of brain function. The differential diagnosis and treatment of stump pain are also reviewed. There is no current treatment for phantom sensation. The treatment of phantom pain and stump pain must be part of a more comprehensive rehabilitation program aimed at restoration of function in all spheres of the individual's life. Discussion of the role of "phantom" sensation and pain in the spinal cord injured is also presented.
A sympathetic general practitioners who is aware of the problems encountered by these families has a vital role in their health care. Families who face a lifetime of dealing with a myriad of problems appreciate having an accessible counsellor they are comfortable with. Being aware of the process of adaptation undergone by the family enables practitioners to do their job more effectively. Disability and its effect on the family is a worthwhile challenge for holistic family practitioners.
Anthocyanin pathway-specific transcriptional activators R and C1 from the monocot maize were expressed in two dicots, Arabidopsis thaliana and Nicotiana tabacum. Expression of R caused augmented anthocyanin pigmentation in both plant species and augmented trichome (hair) production in Arabidopsis. Alone, C1 had no effect. Hybrid transgenic Arabidopsis expressing both C1 and R produced anthocyanins in root, petal, and stamen tissues that normally never express anthrocyanins. When R was expressed in the transparent testa glabrous (without anthocyanins and trichomes) mutant of Arabidopsis, the deficiency was complemented and both anthocyanins and trichomes were restored.
In Saccharomyces cerevisiae, numerous genes have been identified by selection from high-copy-number libraries based on "multicopy suppression" or other phenotypic consequences of overexpression. Although fruitful, this approach suffers from two major drawbacks. First, high copy number alone may not permit high-level expression of tightly regulated genes. Conversely, other genes expressed in proportion to dosage cannot be identified if their products are toxic at elevated levels. This work reports construction of a genomic DNA expression library for S. cerevisiae that circumvents both limitations by fusing randomly sheared genomic DNA to the strong, inducible yeast GAL1 promoter, which can be regulated by carbon source. The library obtained contains 5 x 10(7) independent recombinants, representing a breakpoint at every base in the yeast genome. This library was used to examine aberrant gene expression in S. cerevisiae. A screen for dominant activators of yeast mating response identified eight genes that activate the pathway in the absence of exogenous mating pheromone, including one previously unidentified gene. One activator was a truncated STE11 gene lacking approximately 1000 base pairs of amino-terminal coding sequence. In two different clones, the same GAL1 promoter-proximal ATG is in-frame with the coding sequence of STE11, suggesting that internal initiation of translation there results in production of a biologically active, truncated STE11 protein. Thus this library allows isolation based on dominant phenotypes of genes that might have been difficult or impossible to isolate from high-copy-number libraries.
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When the recA protein (RecA) of Escherichia coli promotes strand exchange between single-stranded DNA (ssDNA) circles and linear double-stranded DNAs (dsDNA) with complementary 5' or 3' ends a polarity is observed. This property of RecA depends on ATP hydrolysis and the ssDNA that is displaced in the reaction since no polarity is observed in the presence of the non-hydrolyzable ATP analog, ATP gamma S, or in the presence of single-strand specific exonucleases. Based on these results a model is presented in which both the 5' and 3' complementary ends of the linear dsDNA initiate pairing with the ssDNA circle but only one end remains stably paired. According to this model, the association/dissociation of RecA in the 5' to 3' direction on the displaced strand determines the polarity of strand exchange by favoring or blocking its reinvasion into the newly formed dsDNA. Reinvasion is favored when the displaced strand is coated with RecA whereas it is blocked when it lacks RecA, remains covered by single-stranded DNA binding protein or is removed by a single-strand specific exonuclease. The requirement for ATP hydrolysis is explained if the binding of RecA to the displaced strand occurs via the dissociation and/or transfer of RecA, two functions that depend on ATP hydrolysis. The energy for strand exchange derives from the higher binding constant of RecA for the newly formed dsDNA as compared with that for ssDNA and not from ATP hydrolysis.
Homeobox genes encode a large family of homeodomain proteins in animal systems. To test whether such genes are also abundant in higher plants, degenerate oligonucleotides complementary to sequences encoding the recognition helix (helix three) of the homeodomain were used to screen genomic and cDNA libraries from the plant Arabidopsis thaliana. Analysis of 8 of the 41 cDNAs isolated revealed that each encodes a presumptive homeodomain; interestingly, most of these clones also contain a leucine zipper motif tightly linked to the homeodomain. It is concluded that Arabidopsis encodes a large family of homeodomain proteins, including members that contain a homeodomain/leucine-zipper (HD-Zip) motif.
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The PstI K fragment of Autographa californica nuclear polyhedrosis virus (AcMNPV) encodes a protein with a molecular weight of 31,000. To define the role of this protein (pp31) in virus infection further, it was overexpressed in bacteria and used to produce polyclonal antiserum. Radioimmunoprecipitation analysis indicated that pp31 was synthesized during both the early and late phases of virus infection, consistent with previous analyses indicating that the gene was regulated by tandem early and late promoters. Metabolic labeling of cells with carrier-free phosphate indicated that pp31 was phosphorylated. Biochemical fractionation experiments showed that pp31 was localized in the nucleus and that it was more stably associated with the nucleus at later times of infection. Immunoblot analysis of subnuclear fractions indicated that pp31 was associated predominantly with the chromatin and nuclear matrix fractions. Immunofluorescence experiments confirmed that the pp31 protein was localized in the nucleus. Nuclear staining was relatively uniform early but was more centrally nuclear later in infection. Immunoelectron microscopy indicated that the pp31 protein was a component of virogenic stroma. Southwestern (DNA-protein) blot analysis demonstrated that pp31 is a DNA-binding protein. These findings suggest a possible role for pp31 in the virus life cycle.
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Promoters that allow the selective induction of gene expression in vivo constitute an important methodology in eukaryotic organisms such as yeast and the fruit fly, but to date no such system has been described for higher plants. Given the fact that mammalian steroid receptors can function as hormone-dependent inducers of gene expression in heterologous systems, the feasibility of using mammalian steroid hormones as selective inducers of plant gene expression was investigated. Here it is shown that the glucocorticoid receptor expressed in plant cells is capable of activating a test gene linked to glucocorticoid response elements, providing the transfected plant cells are treated with glucocorticoid hormone. Nanomolar concentrations of glucocorticoids are sufficient to induce gene expression more than 150-fold, without causing detectable alterations in the physiology of the cultured plant cells. These findings indicate that glucocorticoid induction of steroid-responsive promoters should provide a general method for regulating gene expression in plant cells and imply that such a system might ultimately function in whole plants such as Arabidopsis thaliana.
In general, synthetic RNA transcripts corresponding to the 3' ends of Saccharomyces cerevisiae genes appear to be accurately cleaved and polyadenylated in vitro under appropriate conditions in yeast cell extracts. Initially, however, the endpoints observed in vitro for the GAL7 gene failed to correlate adequately with those reported in vivo as derived from traditional S1 nuclease protection analyses. This led us to apply an independent method for analyzing mRNA 3' ends, using the polymerase chain reaction, with a first strand primer that incorporated a BamHI restriction site sequence near its 5' end, followed by (dT)17. This proved to be a sensitive and accurate means for determining precisely the major and minor polyadenylation sites of the GAL7 mRNA. Moreover, there was complete agreement between the sites identified with this technique when applied to cellular RNA and those generated in vitro by our 3' end mRNA processing reaction. This provides further support for the likelihood that processing in vitro faithfully reflects the endonucleolytic cleavage and polyadenylation events that occur within the living cell.