A Fine Structure Map of the Salmonella Histidine Operator-Promoter

Over 100 regulatory mutations linked to the histidine (his) operon of S. typhimurium have been isolated. They all map in a region estimated to be several hundred base pairs in length located at one end of the his operon ("the hisO region"). The mutations are located at sixteen recombinationally separable sites or are deletions encompassing several sites. Data obtained from pairs of reciprocal three-point tests show that "constitutive" (high enzyme levels) and "promoter-like" (low enzyme levels) hisO mutations are interspersed on the genetic map. In a few crosses, recombination was not observed to occur between markers shown to occupy different sites based on behavior in other recombination tests.     

Morphologic Evaluation of Acute Endometritis in Mares with Differing Resistance to Uterine Infections

The study was designed to determine differences between normal mares and mares with endometrial pathology in the inflammatory response after bacterial challenge. Six normal mares (biopsy category I) and 4 mares with pathological endometrial changes (biopsy category II) were given an intrauterine infusion of β-hemolytic streptococci on the second day of estrus. All mares had a similar kind of inflammatory response after the bacterial inoculation as assessed by rectal and vaginal examinations. There were no significant differences in the amount of discharge, uterine tone, uterine size and cervical relaxation between the groups. Leukocytic response, as determined by endometrial smears and biopsies, was of the same magnitude in both groups. Two mares from the pathological group were not able to eliminate the infection, but had vaginal discharge and bacteriologically positive uterine swabs until the end of the experiment. It is concluded that the inability of some mares to clear uterine infections cannot be explained by a deficient inflammatory response.     

Uterine infections in the postpartum cow: II. Possible synergistic effect of Fusobacteriumnecrophorum and Corynebacteriumpyogenes

Clinical conditions, which were observed in primiparous Angus and Hereford heifers with postpartum uterine infections are reported. Forty-three of sixty-four cows (67%) had uterine infections. $\text{Corynebacterium}$$\text{pyogenes}$ and $\text{Fusobacterium}$$\text{necrophorum}$ were the most frequently isolated aerobe and anaerobe, respectively. Twelve of the sixty-four cows (18.8%) had infections that involved these species. Three of these twelve cows were infected only with $\text{C.}$$\text{pyogenes}$, two were infected only with $\text{F.}$$\text{necrophorum}$, and seven were infected with both organisms. All five of the cows which were infected with either $\text{C.}$$\text{pyogenes}$ or $\text{F.}$$\text{necrophorum}$ showed signs of estrus and four of the five cows conceived by 110 days postpartum. The single cow that did not conceive was infected with $\text{C.}$$\text{pyogenes}$. Three of the seven cows which were infected with both organisms showed signs of estrus and none of the seven cows conceived by 110 days postpartum. In addition, when only $\text{C.}$$\text{pyogenes}$ or $\text{F.}$$\text{necrophorum}$ was isolated from the uterus, cows had either mild or no clinical signs of infection. In contrast, the seven cows which were infected with both organisms had severe clinical signs of infection that included excessive vulvar discharge, uterine abscesses and pelvic adhesions. These observations suggested that a pathogenic synergism between $\text{C.}$$\text{pyogenes}$ and $\text{F.}$$\text{necrophorum}$ might have caused the increased severity of postpartum uterine infections, and the subsequent detrimental effect on return to estrus and conception.     

Antigen-reactive cloned helper T cells. II. Exposure of murine cloned helper T cells to IL 2-containing supernatant induces unresponsiveness to antigenic restimulation and inhibits lymphokine production after antigenic stimulation

Cloned murine helper T lymphocytes (HTL) reactive to alloantigen or to ovalbumin (OVA) become unresponsive to antigenic restimulation after exposure to antigen or to culture supernatant fluids (SF) containing multiple lymphokine activities. Unresponsiveness is manifest by a failure of antigen-stimulated cells to incorporate thymidine or to produce lymphokines after antigenic challenge. Antigen-unresponsive HTL, however, will incorporate thymidine when exposed to an exogenous source of interleukin 2 (IL 2). The duration of unresponsiveness to antigen is correlated with the concentration of IL 2 in SF to which the cloned HTL had been exposed. Chromatographic fractionation of IL 2-containing supernatant from EL-4 thymoma cells (EL-4 SF) yielded a pool of SF that was enriched for IL 2 activity. Exposure of HTL to lymphokines contained in this pool induced unresponsiveness to antigen that was comparable to that observed when HTL were exposed to unfractionated EL-4 SF. Unresponsiveness to antigen also developed after cloned HTL were stimulated with concanavalin A (Con A) or with OVA and syngeneic splenic filler cells. We have used monoclonal antibody (mAb) GK1.5 (anti-L3T4) to investigate the role of lymphokine production in the induction of unresponsiveness. This antibody did not inhibit IL 2-induced thymidine incorporation by cloned HTL, and did not inhibit the induction of unresponsiveness after exposure of cloned HTL to EL-4 SF. In the presence of mAb GK1.5, however, HTL that were stimulated with Con A or OVA did not become unresponsive to antigenic restimulation, an effect that was overcome by the addition of EL-4 SF. These results suggest that HTL become unresponsive to antigen after exposure to IL 2-containing SF, and that stimulation by antigen or Con A can induce the unresponsive state by virtue of stimulating lymphokine production.     

Modulation of agonist-induced inositol phosphate metabolism by cyclic adenosine 3',5'-monophosphate in adrenal glomerulosa cells

Activation of the cAMP messenger system was found to cause specific changes in angiotensin-II (All)-induced inositol phosphate production and metabolism in bovine adrenal glomerulosa cells. Pretreatment of [3H]inositol-labeled glomerulosa cells with 8-bromo-cAMP (8Br-cAMP) caused both short and long term changes in the inositol phosphate response to stimulation by All. Exposure to 8Br-cAMP initially caused dose-dependent enhancement (ED50 = 0.7 microM) of the stimulatory action of All (50 nM; 10 min) on the formation of D-myo-inositol 1,4,5-trisphosphate [Ins(1,4,5)P3] and its immediate metabolites. This effect of 8Br-cAMP was also observed in permeabilized [3H]inositol-labeled glomerulosa cells in which degradation of Ins(1,4,5)P3 was inhibited, consistent with increased activity of phospholipase-C. Continued exposure to 8Br-cAMP for 5-16 h caused selective enhancement of the All-induced increases in D-myo-inositol 1,3,4,6-tetrakisphosphate [Ins(1,3,4,6)P4] and myo-inositol 1,4,5,6-tetrakisphosphate. The long term effect of 8Br-cAMP on the 6-phosphorylated InsP4 isomers, but not the initial enhancement of Ins(1,4,5)P3 formation, was inhibited by cycloheximide. The characteristic biphasic kinetics of All-induced Ins(1,4,5)P3 formation were also changed by prolonged treatment with 8Br-cAMP to a monophasic response in which Ins(1,4,5)P3 increased rapidly and remained elevated during All stimulation. In permeabilized glomerulosa cells treated with 8Br-cAMP for 16 h, the conversion of D-myo-inositol 1,3,4-trisphosphate [Ins(1,3,4)P3] to Ins(1,3,4,6)P4 was consistently increased, whereas dephosphorylation of Ins(1,4,5)P3 to D-myo-inositol 1,4-bisphosphate and of D-myo-inositol 1,3,4,5-tetrakisphosphate to Ins(1,3,4)P3, was reduced.(ABSTRACT TRUNCATED AT 250 WORDS)     

Monoclonal antibodies to Rous sarcoma virus pp60src react with enzymatically active cellular pp60src of avian and mammalian origin

The derivation and characterization of 22 hybridoma clones producing monoclonal antibodies (Mabs) specific for the transforming protein of Rous sarcoma virus, pp60src, are described. All Mabs reacted with pp60v-src encoded by Prague, Schmidt-Ruppin, and Bratislava 77 strains of Rous sarcoma virus. Of these Mabs, 10 efficiently immunoprecipitated pp60c-src from chicken embryo cells. Of these 10 Mabs, 2 (GD11 and EB8) readily detected pp60c-src from a variety of rodent and human cultured cells and from rat brain tissue in an in vitro immune complex kinase assay. Mapping experiments have tentatively localized the determinant(s) recognized by GD11 and EB8 to a region of the src protein bounded by amino acid residues 82 to 169, whereas the remaining Mabs appeared to recognize determinants residing within residues 1 to 82 or 169 to 173. Most of the Mabs complexed denatured pp60v-src in a Western immunoblot, and several were used to localize pp60v-src in Rous sarcoma virus-transformed chicken embryo cells by indirect immunofluorescence microscopy.     

Codon Usage Methods for Horizontal Gene Transfer Detection Generate an Abundance of False Positive and False Negative Results

Bacteria acquire new DNA in a process known as horizontal gene transfer (HGT). To investigate the evolutionary impact of this transfer of DNA, various methods have been developed to detect past HGT events. For example, codon usage-based methods detect the presence of transferred genes by identifying atypical patterns of codon usage. However, some inherited genes exhibit atypical codon usage and some transferred genes have codon usage patterns similar to those of the inherited genes. In this study, we used a comparative phylogenetic approach with Methylobacterium and Caulobacter species to demonstrate that even well-designed codon usage methods fail to detect many HGT events and generate a high rate of false positives (60–75 %) and false negatives (23–61 %). Therefore, we recommend caution when employing codon usage methods to identify transferred genes and suggest that the rapidly increasing availability of bacterial genome sequences makes the phylogenetic approach the method of choice.