Toxicity of pendimethalin to nontarget soil organisms.
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Biomedical subjects
Publications and source records attributed to T A Phillips.
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After application, organophosphorus insecticides (OPs) are often strongly adsorbed to soil constituents. Because of their relatively low water solubility, OPs may be transferred from field to stream adsorbed on suspended solids. However, we are not aware of research done to evaluate the bioavailability (i.e., toxicity) of OPs transported on suspended solids to fish. We conducted 48-h static toxicity tests to determine the toxicity of chlorpyrifos in aqueous solution and adsorbed on calcium-saturated humic acid (HA) to three larval stages of walleye (Stizostedion vitreum). Three concentrations of chlorpyrifos adsorbed on HA, a HA control, and a chlorpyrifos-only treatment were tested. Fish that survived the 48-h static toxicity tests were analyzed to determine total cholinesterase (ChE) activity. In general, survival of all larval stages of walleye exposed to chlorpyrifos-HA complexes was less than that of walleye exposed to HA controls and the chlorpyrifos-only treatment, which were not toxic to walleye. Cholinesterase inhibition of larval walleye exposed to chlorpyrifos-HA complexes was similar to the ChE inhibition observed in larval walleye exposed to chlorpyrifos in the aqueous phase. These laboratory experiments indicate potential toxicity of chlorpyrifos-soil complexes to larval fish.
Organophosphorus (OP) insecticides have high acute toxicity toward many nontarget vertebrate and invertebrate organisms, but direct measurement of OPs in environmental samples is difficult because their concentrations may fall below detection limits within hours to days after entering aquatic ecosystems. Because OPs exert toxicity through cholinesterase (ChE) inhibition, which may persist for up to several weeks, ChE inhibition has been widely used in aquatic ecosystems as a biomarker for OP exposure in aquatic organisms. However, the biological, environmental, and methodological factors affecting ChE activity have not been well documented and must be considered and understood before ChE activity can be used as a dependable indicator of OP exposure to aquatic organisms. This study examined the influence of water temperature, size of larval and juvenile walleye (Stizostedion vitreum), stress, long-term storage, postmortem changes, and methods of euthanasia on ChE activity. Water temperature (17.2, 20.9, and 24.6 degrees C), stress, long-term storage (up to 180 days), postmortem changes, and method of euthanasia had no effect on ChE activity of walleye. There was a strong positive correlation (r = 0.87) between whole body ChE activity and total length (7.2-17.9 mm) for larval walleye, but a negative correlation between brain ChE activity and total length (59-164 mm) for juvenile walleye (r = 0.75). Because size, age, and development may affect ChE activity, fish of similar size should be used when evaluating the effects of ChE inhibitors. If fish of similar size are not available, it is recommended that relations between size, age, and development be understood so estimates of variation in ChE activity can be made.
The placenta utilizes both active and passive mechanisms to evade rejection by the maternal immune system. Recently, the mRNA for two newly cloned members of the B7 family of immunomodulatory cell-associated proteins have been identified in the human term placenta. In this article, we review the current knowledge of the B7 family member B7-H1, and discuss how it may participate in modulation of the maternal immune system at the maternal-fetal interface. B7-H1 has been found to possess immunostimulatory or immunoinhibitory properties, and immunohistological examination of first trimester and term placenta has revealed that this protein is abundant in the placenta. B7-H1 is highly expressed by both the syncytiotrophoblast and extravillous cytotrophoblast, both of which lie in direct contact with maternal blood and tissue. Further, treatment of the choriocarcinoma cell line, JEG-3, with recombinant human interferon (IFN)-gamma resulted in a dose-dependent increase in the abundance of the message for B7-H1, suggesting that IFN-gamma could regulate expression of B7-H1 by the trophoblast. These studies document that the positioning of B7-H1 at the maternal-fetal interface is such that it could participate in suppression of activated maternal leukocytes.
Mechanisms accounting for protection of the fetal semiallograft from maternal immune cells remain incompletely understood. In other contexts, interactions between TRAIL (TNF-related apoptosis-inducing ligand/Apo-2L) and its receptors kill activated lymphocytes. The purpose of this study was therefore to investigate the potential of the TRAIL/TRAIL-R system to protect the placenta against immune cell attack. Analysis by Northern blotting demonstrated mRNAs encoding TRAIL as well as the four TRAIL receptors (DR4, DR5, DcR1/TRID, DcR2/TRUNDD) in human placentas. Immunohistochemical experiments demonstrated that TRAIL protein is prominent in syncytiotrophoblast, an uninterrupted placental cell layer that is continuously exposed to maternal blood, as well as in macrophage-like placental mesenchymal cells (Hofbauer cells). Studies on cell lines representing trophoblasts (Jar, JEG-3 cells) and macrophages (U937, THP-1 cells) showed that both lineages contained TRAIL mRNA and that steady state levels of transcripts were increased 2- to 11-fold by IFN-gamma. By contrast, cell lineage-specific differences were observed in expression of the TRAIL-R genes. Although all four lines contained mRNA encoding the apoptosis-inducing DR5 receptor, only trophoblast cells contained mRNA encoding the DcR1 decoy receptor and only macrophages contained DcR2 decoy receptor transcripts. DR4 mRNA was present only in THP-1 cells and was the only TRAIL-R transcript increased by IFN-gamma. Cytotoxicity assays revealed that the two trophoblast cell lines were resistant, whereas the two macrophage lines were partially susceptible to killing by rTRAIL. Collectively, the results are consistent with a role for the TRAIL/TRAIL-R system in the establishment of placental immune privilege.
The inducible protein p71/73 marks the response of mouse macrophages to one of several stimuli (e.g., bacterial lipopolysaccharide or poly I:C) that trigger the expression of cytolytic activity when these cells have previously been primed for tumor cell killing by interferon-gamma (IFN-gamma). The results reported here identify this marker protein as the inducible prostaglandin endoperoxide synthase (PES), TIS10/PES-2. Identification was based on four findings: (1) p71/73, like the TIS10/PES-2 protein, was associated with cellular membranes; (2) the sequence of amino acids in the NH2 terminus of both p71 and p73 was 96% identical to the predicted NH2-terminal sequence of the TIS10/PES-2 protein; (3) a polyclonal antiserum raised against the COOH-terminal region of the TIS10/PES-2 gene product recognized p71/73 in immunoblots; and (4) dexamethasone, which blocks induction of TIS10/PES-2 expression, inhibited the induction of both p71/73 synthesis and tumoricidal activity in macrophage. Several regulatory roles for this protein in the activation process are possible.
The activation-associated markers, p47b and p71/73, have been recognized as minor proteins in peritoneal and bone marrow culture-derived macrophages activated for tumor cell killing. Proteins with identical characteristics of migration on 2-dimensional gels and comparable Cleveland peptide maps are described here in macrophage cell lines that could be activated for tumor cell killing (J774A.1, RAW 264, UNC-2). Macrophage cell lines that could not be activated (P388D1 and PU5-1.8) did not express the markers. The expression of these markers in activatable macrophage cell lines strengthens their association with the activation process and provides a bulk source of the proteins for purification studies.
During depletion of 4.5S RNA, cells of Escherichia coli displayed a heat shock response that was simultaneous with the first detectable effect on ribosome function and before major effects on cell growth. Either 4.5S RNA is involved directly in regulating the heat shock response, or this particular impairment of protein synthesis uniquely induces the heat shock response. Several hours later, lambda prophage was induced and the cells lysed.
In its most useful form a cellular protein database should be genomically based, because it is the genome which determines both the total number of proteins a cell can make and the particular ones that will be made under any given condition. Such a database should trace each protein back to its structural gene, and should account for every structural gene of a cell. Recent advances in molecular biology greatly facilitate the construction of such gene-protein databases. The mapping of genes of unidentified proteins resolved from total cell extracts on two-dimensional gels can now be accomplished by largely biochemical methods, without the necessity of isolating mutants or performing genetic crosses. Other techniques permit one to search gels for the product of any newly discovered gene (or open reading frame) suspected of encoding a protein. Consequently, gene-protein indices can be built independently and simultaneously from either direction--deducing the genetic map from the protein pattern, or finding the protein pattern from information encoded in the genome. A database of this sort is being constructed for the bacterium, Escherichia coli. Given the current pace of DNA nucleotide sequencing, the development of total gene-protein indices for a variety of cells can be anticipated in the near future.
The Escherichia coli rho-15 mutant, which is highly defective in transcription termination, was examined to see whether its reduced DNA superhelicity could be explained by altered expression of proteins that may affect DNA structure. Levels of DNA gyrase and topoisomerase I were normal; levels of single-stranded-DNA-binding protein, DNA polymerase I, and a protein tentatively identified as Lon were significantly altered.
The product of the pleiotropic gene lon is a protein with protease activity and has been tentatively identified as protein H94.0 on the reference two-dimensional gel of Escherichia coli proteins. Purified Lon protease migrated with the prominent cellular protein H94.0 in E. coli K-12 strains. Peptide map patterns of Lon protease and H94.0 were identical. A mutant form of the protease had altered mobility during gel electrophoresis. An E. coli B/r strain that is known to be defective in Lon function contained no detectable H94.0 protein under normal growth conditions. Upon a shift to 42 degrees C, however, the Lon protease was induced to high levels in K-12 strains and a small amount of protein became detectable at the H94.0 location in strain B/r. Heat induction of Lon protease was dependent on the normal allele of the regulatory gene, htpR, establishing lon as a member of the high-temperature-production regulon of E. coli.
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High-pressure liquid chromatographic procedures were developed for the determination of chlorothiazide and hydrochlorothiazide in plasma and urine. The plasma assay incorporates a preextraction procedure that eliminates interference by endogenous substances. Chromatography is carried out on an octadecyl reversed-phase column. Mobile phases are 15% methanol in 0.01 M acetic acid for plasma and 4% acetonitrile in 0.01 M sodium perchlorate, adjusted to pH 4.6, for urine. At a flow rate of 2.5 ml/min, the retention times for chlorothiazide and hydrochlorothiazide are 3.5 and 4.6 min for plasma and 10.5 and 13.5 min for urine, respectively. Preliminary results of a clinical study in fasting male volunteers showed that the plasma levels and urinary excretion rate of chlorothiazide peaked at 1-2 hr following a 500-mg oral dose and subsequently declined irregularly. On the other hand, the plasma levels and urinary excretion rate of hydrochlorothiazide peaked at 2-3 hr following a 50-mg oral dose and subsequently declined in biphasic fashion. Urinary excretion rates of both chlorothiazide and hydrochlorothiazide closely resemble their concentration profiles in plasma.
Protein B56.5 is a major Escherichia coli protein, originally identified on two-dimensional gels as an abundant cellular protein with unique regulation. The groE gene product is a bacterial protein essential for the assembly of many diverse bacteriophages. The ribosomal A-protein is a large, acidic protein of unknown function associated with isolated, washed ribosomes. On the basis of comigration in two-dimensional gels, oligopeptide map patterns, amino acid composition, immunological specificity, physical properties, and genetic analysis, protein B56.5 has now been shown to be the groE gene product and to be identical with the A-protein.
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The pharmacokinetics of fluorouracil were examined after single 250-mg iv doses and 500-mg oral doses to female patients with breast cancer. In five patients who received intravenous fluorouracil, the mean peak plasma level of unchanged drug was 13.4 microgram/ml, the elimination half-life was 6.3 min, and the plasma clearance was 1410 ml/min. The last value is similar to the hepatic blood flow. In six patients who received oral fluorouracil, the mean peak value of unchanged drug in plasma, which occurred within 20 min of dosing, was 8.3 microgram/ml, and the fluorouracil elimination half-life was 7.2 min. The overall bioavailability of oral fluorouracil as unchanged drug was 28%, and the variation in plasma drug levels between individuals was similar following oral and intravenous doses. The data provide additional evidence of saturable hepatic metabolism of fluorouracil during the first pass.
The two-dimensional gel electrophoresis method of P. H. O'Farrell readily resolves approximately 1,000 proteins from whole-cell homogenates. We have found that the location of most individual proteins is sufficiently reproducible and precise to permit different laboratories to exchange information about them. We present the location of 81 Escherichia coli structural proteins, binding proteins, enzymes, and factors, identified with the aid of purified proteins supplied to us by many investigators.