Inheritance of a mutant histocompatibility gene and a new mammary tumor virus genome in the B6.KH-84 mouse strain

The potential association between integration or deletion of mouse mammary tumor virus (MMTV) retroviral sequences and the appearance of non-H-2 histocompatibility (H) antigen mutations was investigated. Genomic blots from inbred strains carrying 22 loss, gain-loss, and gain mutations on the BALB/c and C57BL/6 backgrounds were hybridized with probes homologous to the long terminal repeat (LTR) and envelope (env) regions of MMTV. Twenty-one mutants were identical in restriction patterns to the respective background strains with all tested restriction enzymes and both probes. However, genomic blots of one gain mutant, B6.C-KH-84, exhibited restriction fragments which were not exhibited by either of the parental strains, C57BL/6 or BALB/c. An additional 5.5 kb Eco RI fragment was observed with the env probe and additional 9.2 kb and 5.5 kb fragments were observed with the LTR probe. These observations were substantiated by hybridization of these two probes with genomic blots generated with additional restriction enzymes. Assuming that the new provirus contains a single, internal Eco RI site as has been observed for other MMTV proviral sequences, it is presumed that the new provirus includes both 5 and 3 LTRs in addition to the env region. Based on the unique sizes of the observed restriction fragments relative to other identified MMTV proviral sequences, this provirus has been designated Mtv-22. The potential role of Mtv-22 in the genesis of the gained histocompatibility antigen in B6.C-KH-84 is discussed.On leave of absence from Istituto Nazionale per to Studio e la Cura dei Tumori, Milano, Italy     

Doxorubicin and epirubicin iron-induced generation of free radicalsIn vitro. A comparative study

To ascertain any differences in myocardial injury exerted by the anthracyclines doxorubicin and epirubicin, their ability to generate oxygen free radicals when mixed with Fe(II) was examined in vitro using an oxygen electrode. 5–250 g/ml doxorubicin or epirubicin consumed oxygen when mixed with 50 or 100 mol/1 Fe(II). Addition of 75 mol/1 cytochrome C showed that of the consumed oxygen, approximately 80% entered the monovalent pathway of oxygen reduction. The strong inhibitory effect of 250 mg/1 catalase indicates that most of the superoxide radicals generated are further reduced to hydrogen peroxide by both anthracyclines. Addition of metal chelators DTPA (100/mol/1), or DDTC (50 mol/1) did not affect oxygen consumption, whereas EDTA (100/mol/1) or desferrioxamine (100 mol/1) with anthracyclines and Fe(II) rather stimulated oxygen consumption. It is concluded that there are no significant differences in the amount or proportion of generated oxygen free radicals between doxorubicin and epirubicin when mixed with Fe(II) in a cell-free system in vitro. Thus, the ability of the anthracyclines, in conjunction with iron alone, to generate radicals does not explain the differences of the drugs in causing myocardial injury.     

Aluminum,Fe, Ca,Mg, K,Mn, Cu,Zn and P in above- and belowground biomass. I.Abies amabilis andTsuga mertensiana

In a mature mixed subalpine stand ofTsuga mertensiana andAbies amabilis, significantly higher Al levels were found in foliage, branch and root tissues ofT. mertensiana.Tsuga mertensiana had significant increases in Al, Ca and Mn levels with increasing foliage age. In current foliage,T. mertensiana had lower levels of Ca, similar levels of Mg and P, and higher levels of Mn thanA. amabilis. Both tree species had Cu and Fe present at higher levels in branch than foliage tissues. Fine roots had the highest concentrations of Al, Fe and Cu but the lowest Ca and Mn concentrations of all tissues analyzed. In the roots of both species, phloem tissues always had significantly higher Al levels than xylem. Fine roots (< 1 and 1–2 mm) ofT. mertensiana had higher Al levels than were found inA. amabilis. Roots greater than 2 mm in diameter exhibited no significant differences in Al levels in phloem or xylem tissue betweenA. amabilis andT. mertensiana. The two species show a clear difference in their ability to accumulate specific elements from the soil.     

Proteolytic processing of foot-and-mouth disease virus polyproteins expressed in a cell-free system from clone-derived transcripts

All picornaviral genes are expressed as a single, large polyprotein, which is proteolytically processed into the system produces functional proteins, including viral protease 3C, which plays a major role in processing the precursor proteins. To study the function of the two putative proteases 3C and leader (L) in processing, we constructed several cDNA plasmids encoding various regions of the FMDV type A12 genome. These plasmids, containing FMDV cDNA segments under the control of the T7 promoter, were transcribed in vitro by using T7 RNA polymerase and then translated in rabbit reticulocyte lysates. The expressed FMDV gene products were identified by immunoprecipitation with specific antisera and analyzed by gel electrophoresis. The results demonstrate the following: (i) the leader protein, L, is processed from the structural protein precursor, P1, in the absence of any P2 or P3 region proteins; (ii) protein 2A remains associated with the structural protein precursor, P1, rather than the precursor, P2; (iii) the processing of the P1-2A/P2 junction is not catalyzed by 3C or L; (iv) the proteolytic processing of polyproteins from the structural P1 region (except VP4/VP2) and the nonstructural P2 and P3 region is catalyzed by 3C.     

Stopover and fat deposition by North American wood-warblers (Parulinae) following spring migration over the Gulf of Mexico

Summary Length of stopover and rate of weight gain (fat deposition) were studied in several species of passerine birds that stopped in southwestern Louisiana along the northern coast of the Gulf of Mexico after a trans-Gulf flight. Fatdepleted birds were more common among the birds that arrived at our study site in southwest Louisiana, though variability characterized our samples. Migrants that landed after encountering opposing winds or rain over the northern Gulf of Mexico were, on average, fatter than migrants that landed when weather was favorable for continued migration. Some of the variation in the energetic condition of arrivals may be explained by the location where migrants initiated crossings. Our simulation of flight over the Gulf of Mexico showed that with following winds a warbler can cross the Gulf of Mexico from Yucatan with fat reserves to spare, and stronger tailwinds make flights from as far south as Honduras energetically permissible. The length of stay after a trans-Gulf flight was related to the extent of fat-depletion upon arrival: lean birds stayed longer than fat migrants. Migrants stopped over for 1–7 days and replenished energy reserves at rates that varied from 0.19 g/d for Hooded Warblers (Wilsonia citrina) to 0.87 g/d for Ovenbirds (Seiurus aurocapillus). Within each species, most individuals gained weight at a rapid rate, though a few individuals lost or maintained weight during their stay.     

Expression of the β-subunit of β-conglycinin in seeds of transgenic plants

Soybean (Glycine max (L.) Merr.) seeds contain the storage protein -conglycinin, encoded by a multigene family. -Conglycinin consists of three subunits; , , and . A genomic clone for a -subunit of -conglycinin has been characterized by restriction-enzyme mapping and hybrid selected in-vitro translation followed by immunoprecipitation. In order to determine the developmental regulation of this -subunit gene, its expression was studied in seeds of transgenic petunia (Petunia hybrida) and tobacco (Nicotiana tabacum L.) plants. The -subunit expressed in seeds of petunia and tobacco was recognized by anti--conglycinin serum at a relative molecular mass of 53 000, equivalent to that of the native protein. Separation of the petunia-seed proteins by isoelectric focusing followed by sodium dodecyl sulfate-polyacrylamide gel electrophoresis and immunoblot analysis showed that multiple isoelectric forms of the -subunit were produced. There was approximately a twofold variation in the accumulation of the -subunit protein in the mature seeds of transgenic petunia plants, each containing a single -subunit gene. However, the level of protein accumulation in mature seeds and the amount of -subunit mRNA in developing seeds was not correlated. Accumulation of the -subunit protein in transgenic seeds was less than the -subunit protein that accumulated in transgenic petunia seeds containing a single -subunit gene and less than the amount of the -subunit in mature soybean seeds which contain 8–13 -subunit genes. In transgenic tobacco plants, the accumulation of the -subunit protein in seeds was generally well correlated with the number of genes that were incorporated in the different transformants.Abbreviations kb kilobase - kDa kilodalton - M_r relative molecular mass - SDS-PAGE sodium dodecyl sulfate polyacrylamide gel electrophoresis     

Calcium regulates inositol 1,3,4,5-tetrakisphosphate production in lysed thymocytes and in intact cells stimulated with concanavalin A.

Lysed mouse thymocytes release [3H]inositol 1,4,5 trisphosphate from [3H]inositol-labelled phosphatidyl inositol 4,5-bisphosphate in response to GTP gamma S, and rapidly phosphorylate [3H]inositol 1,4,5-trisphosphate to [3H]inositol 1,3,4,5-tetrakisphosphate. The rate of phosphorylation is increased approximately 7-fold when the free [Ca2+] in the lysate is increased from 0.1 to 1 microM, the range in which the cytosolic free [Ca2+] increases in intact thymocytes in response to the mitogen concanavalin A. Stimulation of the intact cells with concanavalin A also results in a rapid and sustained increase in the amount of inositol 1,3,4,5-tetrakisphosphate, and a much smaller transient increase in 1,4,5-trisphosphate. Lowering [Ca2+] in the medium from 0.4 mM to 0.1 microM before addition of concanavalin A reduces accumulation of inositol 1,3,4,5-tetrakisphosphate by at least 3-fold whereas the increase in inositol 1,4,5-trisphosphate is sustained rather than transient. The data imply that in normal medium the activity of the inositol 1,4,5-trisphosphate kinase increases substantially in response to the rise in cytosolic free [Ca2+] generated by concanavalin A, accounting for both the transient accumulation of inositol 1,4,5-trisphosphate and the sustained high levels of inositol 1,3,4,5-tetrakisphosphate. Inositol 1,3,4,5-tetrakisphosphate is a strong candidate for the second messenger for Ca2+ entry across the plasma membrane. This would imply that the inositol polyphosphates regulate both Ca2+ entry and intracellular Ca2+ release, with feedback control of the inositol polyphosphate levels by Ca2+.     

Circulating steroid hormones during rapid aggressive responses of territorial male mountain spiny lizards, Sceloporus jarrovi

Levels of aggression and circulating steroid hormones were monitored simultaneously in free-living male lizards following a staged territorial aggressive encounter with another male. In the first 15 min following the aggressive encounter, the frequency of territorial patroling and the frequency of agonistic and advertisement displays increased four- to fivefold in resident males. In most cases these increases persisted for at least 90 min after withdrawal of the intruder male and probably persisted for the entire day of the encounter. Blood samples collected at 15-min intervals revealed no changes in circulating levels of testosterone or corticosterone while this behavioral change was occurring. Thus, the increase in aggressive behavior that follows a male-male territorial encounter in this species does not appear to be mediated by simultaneous changes in circulating levels of these hormones. Interspecific comparisons suggest that interspecific variation in steroid hormone involvement in rapid aggressive responses may depend on the mating system and the extent of male parental care.