DMBA induced DNA damage and repair in mammary epithelial cells in vitro measured by a nick translation assay

A new E. coli DNA polymerase I directed nick translation assay was used for measuring 7,12-dimethylbenz[a]anthracene-induced in situ DNA damage and repair in mouse mammary epithelial cells in monolayer culture. The nick translation assay was capable of detecting a DMBA-dose dependent significant increase of DNA damage, and the same assay also allowed monitoring of the DNA repair activity provoked by DMBA treatment of the epithelial cells. This relatively simple method thus provides a rapid assay for carcinogen-induced in situ DNA damage and repair in an epithelial cell tumorigenic system.     

Effect of biological toxins on gap-junctionai intercellular communication in Chinese hamster V79 cells

Since chemical modulation of gap junctional intercellular communication has been implicated in several toxicological endpoints, a study to examine the ability of several biological toxins to inhibit this process was undertaken. Eight biological toxins were tested for their ability to inhibit metabolic cooperation, a measure of gap-junctional intercellular communication, in the Chinese V79 cell system. Aplysiatoxin, anhydrodebromoaplysiatoxin and debromoaplysiatoxin showed the strongest ability to inhibit metabolic cooperation while T2-toxin and vomitoxin inhibited metabolic cooperation to a lesser degree. Afatoxin B₁, afatoxin B₂ and palytoxin were inactive in the Chinese V79 system. Palytoxin, which was extremely cytotoxic, might act as a tumor promoter if it induces compensatory hyperplasia in vivo.Abbreviations 6-TG 6-thioguanine - TPA 12-0-tetradecanoylphorbol-13-acetate     

Effect of castration on plasma luteinizing hormone in postpubertal boars

Two experiments were conducted to test the working hypothesis that mean plasma concentrations of luteinizing hormone (LH) increase as a result of an increase in the frequency and amplitude of the pulsatile releases of LH in postpubertal boars after removal of gonadal steroid hormones by castration. It was further hypothesized that these changes in secretion of LH would be the result of changes in sensitivity of the pituitary to gonadotropin releasing hormone (GnRH). In Experiment 1, plasma LH was monitored in 10 postpubertal crossbred boars (13 to 14 mo old and weighing 159 +/- 6.0 kg) at 12-min intervals for 6 h before and 1 h after GnRH (375 ng/kg of body weight) on Days -1, 7, 14, 21 and 29 relative to castration. In Experiment 2, plasma LH was monitored in four castrated and five intact postpubertal boars (11 to 12 mo old and weighing 150 +/- 5.1 kg) after each of three doses of GnRH (94, 188 and 375 ng/kg) were administered to each animal. Sample collection occurred 5 wk after castration. Mean LH and frequency of pulsatile releases of LH increased as a result of castration (P<0.0001), with changes evident by Day 7 after castration. However, the amplitude of the LH pulses increased minimally after castration (P<0.10). The response to exogenous GnRH increased throughout Experiment 1 (P<0.0001), even though the amplitude of the pulsatile releases of LH (response to endogenous GnRH) did not change. Castrated animals in Experiment 2 had a greater response of LH to GnRH stimulation than intact boars (P<0.05). The dose-response curve of castrated animals was not parallel (P<0.001) to that of intact boars, and indicated that sensitivity of the pituitary to GnRH had increased in the absence of gonadal steroids. Thus, the hypotheses stated above can be accepted with the exception that castration may have a minimal effect on LH pulse amplitude. Based on the results of these experiments, we suggest that gonadal steroid hormones modulate both the size of releasable stores of LH and pituitary sensitivity to GnRH in boars.     

Expression of a Clostridium thermocellum endoglucanase gene in Lactobacillus plantarum

Recombinant plasmid pM25 containing the celE gene of Clostridium thermocellum, which codes for an enzymatically active endoglucanase, was transformed into Lactobacillus plantarum by electroporation. Strains harboring pM25 expressed thermostable endoglucanase, which was found predominantly in the culture medium. Two other plasmids, pGK12 and pSA3, were transformed into L. plantarum, and the stability of each plasmid was evaluated.     

Effect of estradiol on secretion of luteinizing hormone during the follicular phase of the bovine estrous cycle

Mean concentrations of luteinizing hormone (LH) increase during the follicular phase of the estrous cycle in cows. The working hypotheses in the present study were (1) that increasing concentrations of 17 beta-estradiol (E2) during the follicular phase of the estrous cycle cause an increase in mean concentration of LH by increasing amplitude of pulses of LH, and (2) that increasing E2 concentrations during this stage of the estrous cycle decrease frequency of pulses of LH in bovine females. Day of estrus was synchronized in seventeen mature cows. Treatments were initiated on Day 16 of the experimental estrous cycle (Day 0 = estrus). At Hour 0 (on Day 16), 4 cows were lutectomized. Lutectomy of these cows (EE; n = 4) allowed for endogenous secretion of E2. The remaining cows were ovariectomized at Hour 0 and were assigned to one of three E2 treatments: luteal phase E2 (LE, n = 5), increasing then decreasing E2 (DE, n = 5), and no E2 (NE, n = 3). Cows in the group that received LE were administered one E2 implant at Hour 0, which provided low circulating concentrations of E2 similar to those observed during the luteal phase of the estrous cycle. Cows in the group that received DE were administered one E2 implant at Hour 0, and additional implants were administered at 8-h intervals through Hour 40; then, two implants were removed at Hours 48 and 56, and one implant was removed at Hour 64.(ABSTRACT TRUNCATED AT 250 WORDS)     

Chemokinetic Motility Responses of the Cyanobacterium Oscillatoria terebriformis

Oscillatoria terebriformis, a gliding, filamentous, thermophilic cyanobacterium, exhibited an inhibition of gliding motility upon exposure to fructose. The observed response was transient, and the duration of nonmotility was directly proportional to the concentration of fructose. Upon resumption of motility, the rate of motility was also inversely proportional to the concentration of fructose. Sulfide caused a similar response. The effect of sulfide was specific and not due to either anoxia or negative redox potential. Exposure to glucose, acetate, lactate, or mat interstitial water did not elicit any motility response.     

TGF-Beta inhibits the in vitro induction of lymphokine-activated killing activity

Summary Employing serum-free media, human peripheral blood mononuclear cells, and purified recombinant interleukin-2 (IL-2), conditions were observed in which the development of IL-2-driven cytotoxic activity was suppressed. The cytotoxic activity of such IL-2-generated lymphokine activated killing (LAK) was tested against natural killer-resistant cultured tumor cells (Daudi, Raji, and a glioma). LAK generation was inhibited by addition of some normal sera, normal platelets, or some tumor cells. Because recent reports have indicated that transforming growth factor-beta (TGF-beta)-like factors are often secreted by tumors and the acidic alpha granules of platelets and can be present in sera, we tested the effect of purified human TGF-beta on the activation of LAK. Our results indicated that TGF-beta is very suppressive for LAK induction, and can completely prevent both the IL-2-driven proliferation and cytotoxicity at concentrations as low as 5 ng/ml. Titrations of IL-2 and of TGF-beta indicated that the suppression is dose-dependent and can be avoided by employing higher levels of IL-2. It was also found that the suppressive effect of TGF-beta can be overcome by washing suppressed cell populations and further culture in low levels of IL-2. Collectively, these data indicate that TGF-beta can be a potent inhibitor of LAK generation under standard activation conditions, but that this effect is regulated by the relative level of IL-2 and may be overcome and/or reversed in vitro.     

The production of haploid wheat plants from wheat x maize crosses

Summary Hybrid embryos from hexaploid wheat x maize crosses rapidly lose the maize chromosomes to produce haploid wheat embryos. Such embryos almost always aborted when left to develop on the plant, and only 1 was recovered from 2440 florets (0.17% of the expected number). Embryos had greater viability in spikelet culture, 47 (26.5% of the expected number) being recovered from 706 ovaries. Thirty-two of these embryos germinated to give green plants, 31 of which were haploid (21 wheat chromosomes) and 1 of which was euploid (42 wheat chromosomes). Spikelet culture enabled 17.1% of the expected number of embryos to be recovered as haploid plants, a 100-fold improvement on allowing embryos to develop in vivo. Ten haploid plants of Chinese Spring (kr1, kr2), 13 plants of Chinese Spring (Hope 5A) (kr1, Kr2), and 8 of Hope (Kr1, Kr2) were recovered. The potential of wheat x maize crosses for wheat haploid production and for gene transfer from maize to wheat is discussed.