Alterations in cholesterol and fatty acid biosynthesis in rat liver homogenates by aryloxy acids (Short Communication)

2,4-Dichlorophenoxyacetic acid and 2,4,5-trichlorophenoxyacetic acid inhibited the incorporation of [2-(14)C]mevalonate into cholesterol and non-saponifiable lipids. Both compounds inhibited the conversion of [1-(14)C]isopentenyl pyrophosphate into cholesterol and the synthesis of cholesterol and fatty acids from [2-(14)C]acetate. There was no inhibition of the conversion of [1-(14)C]mevalonate into CO(2). At low concentrations (0.5mm) of the compounds there was a stimulation of acetate incorporation into fatty acids.     

Effect of CryIC toxin from Bacillus thuringiensis on larval feeding behavior of Spodoptera exigua

The lack of data on the effect of Bacillus thuringiensis (B.t.) toxins on larval feeding behavior of the pest Spodoptera exigua (Hübner) (Noctuidae: Amphypyrini) prompted us to investigate the effect of three delivery systems of CryIC, a commercial formulation, inclusion bodies, and the activated CryIC toxin. The commercial formulation was the least and CryIC toxin the most lethal form to neonates of susceptible colonies. All but two of the treatments in choice tests with neonates and third instars showed significant avoidance of B.t. treated diet, with greater proportion of larvae from susceptible (UCR-S and AUBURN-S) and resistant (AUBURN-R) colonies on untreated diet than on diet treated with any of the CryIC forms and concentrations tested. Furthermore, third instars consumed significantly more control than treated diet for all CryIC forms, colonies and concentrations. The avoidance of CryIC toxin by neonates and third instars strongly suggests that CryIC, which also is present in the commercial formulation and in the inclusion bodies, is responsible for eliciting avoidance behavior by S. exigua larvae. Behavioral observations of third instars in a no-choice test on either treated or control diet indicated that questing behavior in susceptible larvae appears to be positively related with presence of CryIC toxin in the diet. Furthermore, resistant third instars were on the whole more active than susceptible thirds on both treated and control diet. Resistant thirds raised on CryIC treated diet (AUBURN-RC) spent more time eating treated diet than resistant larvae raised on control diet (AUBURN-R), suggesting that diet conditioning plays an important role on feeding behavior of S. exigua. The implications of these results are discussed.     

Effects of plant chemical extracts and physical characteristics of Apium graveolens and Chenopodium murale on host choice by Spodoptera exigua larvae

Choice tests with whole plants and leaf discs indicated that fourth instar Spodoptera exigua (Hübner) (Noctuidae: Amphypyrini) were found more frequently and ate significantly more of the weed Chenopodium murale than the associated crop plant Apium graveolens. In order to explain the preference, plant extracts, plant volatiles, soluble protein concentrations, water contents, and leaf toughness of the two plants were investigated. Bioassays of aqueous methanol (90%) and hexane extracts of leaves on cellulose discs indicated that neither attractants in C. murale nor repellents in A. graveolens could account for the observed preference. No significant difference could be found between the effects of plant volatiles from C. murale, A. graveolens and a control on larval dispersal by S. exigua. Selective feeding for higher levels of proteins also was not a factor, because A. graveolens had nearly twice the soluble protein of C. murale. Water content was approximately 6% higher (by weight) in C. murale than A. graveolens but most polyphagous larvae do not typically show compensatory feeding for water alone. However, the potentially related characteristic of leaf toughness was significantly different, with A. graveolens exhibiting 1.53 times the toughness of C. murale. Studies comparing five types of larval behavior on both plant species showed that the time spent in swallowing behavior was significantly greater on the tougher A. graveolens leaves relative to C. murale. To test the hypothesis that leaf toughness was affecting larval host choice, both plants were finely ground and incorporated into agar blocks. No differences in feeding behavior were detected. The implications of leaf toughness for larval diet and host choice are discussed.     

A permanent prosthesis for converting in situ muscle contractions into hydraulic power for cardiac assist

Trumble, Dennis R., and James A. Magovern. A permanentprosthesis for converting in situ muscle contractions into hydraulic power for cardiac assist. J. Appl.Physiol. 82(5): 1704-1711, 1997.The key toutilizing muscle power for circulatory support lies with thedevelopment of a practical scheme by which contractile energy may becollected and efficiently delivered to the bloodstream. This workdescribes initial in vitro testing of a prototype muscle energyconverter (MEC) designed to transform the power of in situ musclecontractions into hydraulic form. The MEC resembles a simple pistonpump and is designed for implant beneath the humeral insertion of thelatissimus dorsi muscle. Bench tests were conducted to measurecomponent function and to characterize device performance under varioushydraulic loads. Under simulated muscle-pull conditions, MEC energytransfer capacity was found to be 170 mJ/stroke while operating at peakefficiencies (i.e., >98% of input power converted into hydraulicenergy and preload work). Transfer efficiencies dropped from 96 to 38%as mean generated pressures increased from 23 to 36 N/cm² due to metal bellowsflexion. These results demonstrate that a significant amount ofcontractile energy can be efficiently transformed to hydraulic powervia this mechanism.

     

Site-directed mutagenesis of cys148 in the lac carrier protein of Escherichia coli

The lac y gene of Escherichia coli which encodes the lac carrier protein has been modified by oligonucleotide-directed, site-specific mutagenesis such that cys₁₄₈ is converted to a glycine residue. Cells bearing the mutated lac y gene exhibit initial rates of lactose transport that are about 4-fold lower than cells bearing the wild type gene on a recombinant plasmid. Furthermore, transport activity is less sensitive to inactivation by N-ethylmaleimide, and strikingly, galactosyl 1-thio-β-D-galactopyranoside affords no protection against inactivation. The findings suggest that although cys₁₄₈ is essential for substrate protection against sulfhydryl inactivation, it is not obligatory for lactose:proton symport and that another sulfhydryl group elsewhere within the lac carrier protein may be required for full activity.     

Calcium content and distribution as a function of growth and transformation in the mouse 3T3 cell

Total Ca content and that fraction of Ca sensitive to removal by the chelator ethylene glycol-bis(β-aminoethyl ether)N,N,N',N'-tetraacetate (EGTA) have been investigated in the mouse 3T3 cell as a function of growth stage, transformation with SV40 virus, and serum levels of the media. Cells were allowed to grow through several doublings in media containing (45)Ca. The cellular content of (45)Ca was used to access total cell Ca. That fraction of (45)Ca removed by EGTA was presumed to represent primarily surface-localized Ca. The data are expressed on a per cell volume basis to compensate for size differences as a function of growth stage and transformation. During exponential growth phase, the 3T3 cell contains 525pmol Ca/μl cell volume. Of this, approx. 457 pmol/μl is not removable by EGTA and, presumably, is cytoplasmically located. This value is in close agreement with previous studies on the HeLa cell (470 pmol Ca/μl cell water after the removal of the surface Ca). The low level of EGTA- removable Ca present in the 3T3 cell during early exponential growth (68 pmol Ca/μl cell volume) increases progressively with increasing cell density, and upon quiescence it is sevenfold greater. In contrast, SV40- transformed 3T3 cells growing exponentially possess total levels of Ca which are approximately two-thirds the levels of the normal 3T3 cell. However, their EGTA-sensitive Ca is not significantly different from that of exponentially growing, normal 3T3 cells. As the transformed cells continue to grow at high density, their total ca and their sensitivity to EGTA do not change, in contrast to the normal 3T3 cell. Thus, an increase in Ca associated with the cell surface appears to be correlated with growth inhibition. This has been investigated further by regulating growth of the normal and transformed cell with alterations in the serum level of the media. In 4 percent calf serum the normal cell is stopped from continued proliferation. Growth stoppage under these conditions is characterized by a nearly fourfold increase in EGTA-removable Ca, similar to the increase observed upon quiescence in depleted 10 percent serum. Similar treatment of the transformed cell does not reduce its growth rate, nor does it significantly alter Ca distribution. However, at 0.5 percent medium serum levels, the SV40 3T3 growth rate is substantially reduced and, under these conditions, EGTA-removable Ca increases twofold.     

Resistance to Spodoptera exigua in Apium prostratum

Two Apium accessions were compared with the commercial cultivar Tall Utah 52–70R (A. graveolens [L.]) for resistance to Spodoptera exigua (Hübner)(Lepidoptera: Noctuidae). Oviposition rate was not significantly different between the three genotypes. In all accessions, eggs were usually placed on the upper half of the plants. Implications of this oviposition pattern on S. exigua management in celery are discussed. The wild species A. prostratum ssp prostratum var filiform (A230) showed a significantly higher resistance to S. exigua than 52–70R. The levels of carcinogenic and mutagenic linear furanocoumarins in the commercial cultivar 52–70R (1.41 g/g in the petioles; 5.85 g/g in the leaves) and in the plant accession A. nodiflorum (5.40 g/g in the petioles; 2.99 g/g in the leaves) were far below the concentration reported to produce acute contact dermatitis (18.0 g/g). The levels of furanocoumarins in A. prostratum petioles (186.14 g/g) and leaves (326.45 g/g) were 10 and 18 times higher, respectively, than the concentration known to cause contact dermatitis. However, resistance in A. prostratum was primarily due to non-preference and the linear furanocoumarins did not induce non-preference. Therefore, the resistance shown by this plant accession does not appear to be furanocoumarin-based and may be suitable for transfer to commercial celery for use in S. exigua management.     

Recombinant adeno-associated virus type 2-mediated gene delivery into the <Emphasis Type="Italic">Rpe65</Emphasis><Superscript>-/-</Superscript> knockout mouse eye results in limited rescue

Background  

Leber's congenital amaurosis (LCA) is a severe form of retinal dystrophy. Mutations in the RPE65 gene, which is abundantly expressed in retinal pigment epithelial (RPE) cells, account for approximately 10–15% of LCA cases. In this study we used the high turnover, and rapid breeding and maturation time of the Rpe65 ^-/- knockout mice to assess the efficacy of using rAAV-mediated gene therapy to replace the disrupted RPE65 gene. The potential for rAAV-mediated gene treatment of LCA was then analyzed by determining the pattern of RPE65 expression, the physiological and histological effects that it produced, and any improvement in visual function.     

Interspecific and intraspecific differences in two Liriomyzaleafminer species in California

In recent years, the pest status of Liriomyza trifolii (Burgess) and L. huidobrensis (Blanchard) (Diptera: Agromyzidae) has changed in California, as well as other areas of the world. In California, L. huidobrensis has become the predominant Liriomyza species in valleys along the central coast, while L. trifolii remains the predominant species in southern California. To investigate possible reasons for this change in status, differences in host plant use and reproductive success of intraspecific populations were examined for Liriomyza trifolii and L. huidobrensis from both central and southern California. The southern L. trifolii fed, oviposited and reproduced successfully on all five hosts tested, but the central population fed significantly less on all hosts and was restricted to reproducing on pepper only. With the exception of pepper, southern L. trifolii had significantly greater larval survival on all hosts than central L. trifolii. In contrast, the central L. huidobrensis population had greater reproductive success than the southern population of that species on all hosts plants tested. However, pepper was not a suitable host for either L. huidobrensis population. Both species showed positive assortative mating, with homotypic mating occurring more frequently than heterotypic mating; however, the difference between L. trifolii populations was much more pronounced than between L. huidobrensis populations. These data indicate that central and southern California populations of each species are distinct biotypes. Furthermore when coupled with previous genetic data, our results suggest the possible existence of cryptic species within L. trifolii.