Estrogen-dependent trypsin-like activity in the rat uterus. Localization of activity in the 12,000g pellet and nucleus.

Direct evidence was obtained for the presence of hormone-stimulated trypsin-like protease activity in the rat uterus. Ovariectomized rats were either untreated (U), treated with estradiol (E), or estradiol plus progesterone (EP). The uteri were excised and subcellular fractions were prepared. Each fraction was assayed for protease activity using protamine as substrate, the cleavage products being quantitated fluorometrically following reaction with 4-phenylspiro[furan-2(3H),1′-phthalan]-3,3′dione (Fluram). Fractions from U rats yielded negative results, whereas the 12,000g pellets and nuclei from the uteri of E and EP rats exhibited appreciable activities. No significant increase in protease activity was observed in thymus and diaphragm following hormone treatment, indicating organ specificity. The enzyme (or enzymes) from the 12,000g pellet was solubilized and some characteristics were determined. The apparent K_m is about 1.0 × 10^?6m, the temperature optimum is about 44 °C and maximum velocity is achieved in the alkaline range (pH ~ 8.5). The protease is a plasminogen activator and is inhibited by diisopropyl fluorophosphate, Antipain, and Leupeptin. These properties resemble those of trypsin.     

Fluorine nuclear magnetic resonance studies of trifluoroacetylinsulin derivatives. Effects of salts and denaturants.

19F nuclear magnetic resonance spectroscopy has been used to study the effects of salts and denaturants on the structure and aggregation properties of several trifluoroacetyl derivatives of insulin. This technique has been shown to be a powerful tool in the study of specific sites on the protein molecule. Circular dichroic and sedimentation velocity studies were also carried out to aid in the interpretation of the magnetic resonance data. At pH 6.8 Zn2+ had no effect on the 19F magnetic resonance spectrum, however, citrate and acetate ions significantly sharpened the signal from the trifluoroacetyl probe at the N-terminal end (glycine A-1) of the insulin A chain. No alterations were observed in the S20,W value of circular dichroic spectra, suggesting that the probe had gained a considerable degree of motional freedom without changes in aggregation or conformational properties. In the absence of perturbants the trifluoroacetyl group on glycine A-1 showed considerably more motional freedom than on phenylalanine B-1. Guanidine hydrochloride and sodium dodecyl sulfate were used to study the unfolding of several trifluoroacetylinsulin derivatives. The results suggested differential alterations in the environments of the probes located at glycine A-1, phenylalanine B-1, and lysine B-29 in the insulin molecule as the concentration of perturbant was increased.     

Studies on the electrophoretic separability of B and T human lymphocytes.

Cell electrophoresis experiments were performed at physiological, ionic strength on Hypaque-Ficoll separated normal human peripheral blood lymphocytes. The initial lymphocyte preparations averaged 72% T cells and 22% B cells as estimated by resetting techniques. A bimodal electrophoretic distribution of fast T cells and slow B cells such as has been repeatedly shown to exist for experimental animals could not be demonstrated. Neither B- nor T-enriched populations showed any correlation between mobility and degree of enrichment. A strong positive correlation was found between the non-rosetting cells and a very slow electrophoretic subpopulation produced during the B enrichment procedure. It was also found that considerable variability in lymphocyte mobilities existed from person to person and even in the same individual sampled over several months; these observations imply that the pooling of electrophoretic data can be misleading. The importance of these findings lies in the growing interest in relating lymphocyte electrophoresis to disease processes and immunological rejection phenomena.     

Altered Glycoflavone expression in induced Autotetraploids of Phlox drummondii

The effects of colchicine induced autotetraploidy on non-anthocyanin flavonoid expression were determine for 15 cultivars of Phlox drummondii and for the naturally occuring P. drummondii ssp. mcallisteri. Collectively, the taxa express a total of nine glycoflavonoid derivatives (C, O-diglycosides or di-C-glycosides) of either apigenin or luteolin. The autotetraploids of 14 cultivars and those of the natural subspecies exhibit altered glycoflavone profiles relative to their respective diploid sources. The qualitative alternations in the cultivars may be grouped into three phenotypic categories: (1) the expression of novel glycoflavones, (2) the absence of diploid glycoflavones, and (3) the deregulation of tissue-specific glycoflavone production. Alterations in mcallisteri autotetraploids include only the latter two categories. Each of the novel compounds is otherwise expressed among other diploid cultivars or in other wild P. drummondii subspecies. Quantitatively, the phenolic content of most autotetraploid flowers is significantly greater than in respective diploid flowers. However, on a dry weight basis, phenolic titre in comparable 4n and 2n floral or leaf tissues is not significantly different. Floral tissues express from 5 to 10 times the phenolic titre of leaves. The results are discussed in terms of the possible origins of novel flavonoids in natural polyploid Phlox species.     

Balanced familial translocation t(5;19)(q12;p or q11) with phenotypical abnormalities in a girl.

Human diploid fibroblasts cultured in Dulbecco's Modified Eagle's medium (DME) were exposed to different concentrations of 15 antibiotics to determine the limiting toxic concentration. The number of cells surviving after antibiotic treatment was given as the index of toxicity. No visible chromosomal damage could be detected when half the maximal toxic concentration was applied. The maximum limiting concentration was found to be the same for both the preconfluent and postconfluent phases.     

Impaired macrophage function in Friend virus leukemia: restoration by statolon.

Phagocytic and migratory functions of peritoneal macrophages from Friend virus (FV) leukemic mice are significantly depressed as compared with normal controls. Leukemic macrophages exposed in vivo and in vitro to statolon, an extract of the mold Penicillium stoloniferum, shown previously to suppress FV erythroleukemia, regain normal function and release reduced amounts of FV. Statolon's in vivo restoration of leukemic macrophage function is paralleled by restoration of humoral immune competence. Statolon induces interferon in vitro but its effects on leukemic macrophages are probably direct, since restoration of macrophage function occurs at dosage levels far below those that induce interferon. These studies suggest that macrophages play an integral role in both the pathogenesis and the statolon-induced suppression of FV disease.     

Relation of tegumentary phosphohydrolase to purine and pyrimidine transport in Schistosoma mansoni.

Inhibition of the saturable influx of 0.05 mM 14C-labeled adenine or adenosine by AMP in adult Schistosoma mansoni in vitro suggested hydrolysis of this nucleotide at the tegumental surface of the parasite. Adenosine liberated as a result of AMP hydrolysis was the inhibitor of uptake of labeled adenine or adenosine. Inhibition of adenosine uptake by AMP or ATP was relieved by paranitrophenyl phosphate or ammonium molybdate supporting the hypothesis of nucleotide hydrolysis at the tengumental surface. Addition of glucose-1-phosphate, glucose-6-phosphate, NaF, or cysteine did not relieve AMP inhibition of adenosine uptake indicating substrate and inhibitor specificity for the surface enzyme(s). AMP, ATP, UMP, and p-nitrophenyl are hydrolyzed, at least in part, by the same enzyme(s). Apparent absorption of labeled AMP was preceded by hydrolysis, with labeled adenosine as the actual compound absorbed, although there was a small diffusion component for absorption of intact AMP. The site of nucleotide hydrolysis in close proximity to absorption sites provides a kinetic advantage for uptake of products of adenine nucleotide hydrolysis but not for products of uracil nucleotide hydrolysis.     

Bioassay of three strains of Bacillus sphaericus on field-collected mosquito larvae.

Bacillus sphaericus strains 1593, 1404, and SSII-1 were assayed for infectivity against field-collected larvae of Psorophora columbiae, Culex nigripalpus, and Aedes taeniorhynchus in southwest Florida. Results indicate that all three strains are highly active against the Psorophora and Culex species. A. taeniorhynchus is also susceptible but requires higher dosages to achieve lethal responses. Tests were also conducted on the rate of infection and the differences in susceptibility of different instars to B. sphaericus. These tests indicate that nearly 75% of the mortality that occurs in the course of exposure to B. sphaericus occurs within 48 hr post-incubation with the bacteria. Furthermore, our tests indicate P. columbiae larvae decrease in susceptibility to the Bacillus with increase in larval age (instar). This investigation shows B. sphaericus to be a feasible biological control agent that warrants further study.     

The purification and properties of chicken liver RNase: An enzyme which is useful in distinguishing between cytidylic and uridylic acid residues

A heat-stable endoribonuclease isolated from chicken liver has been purified to homogeneity as evidenced by the presence of a single protein band upon polyacrylamide gel electrophoresis. The enzyme can, in limit digests of 5 S rRNA and 5.8 S rRNA, dinstinguish between cytidylic and uridylic acids bonds at a ratio of 61:1 and, therefore, may be useful in RNA sequence analysis. The means by which the enzyme hydrolyzes substrate is unusual in that kinetic data do not support a simple formation and breakdown of an enzyme . substrate complex. Rather, the existence of a second complex, consisting of 2 mol of substrate and one of enzyme, derived from the initial enzyme . substrate complex, is postulated. In common with the other endonucleases, enzyme activity is inhibited by free poly(A) or tracts of the polypurine present at the 3'-terminus of RNA. Reversal of inhibition and restoration of activity may be achieved by the addition of low concentrations of spermidine to reaction mixtures.     

Genetic control of the immune response to ferredoxin: linkage and mapping of T cell proliferation and antibody production genes to the MHC of mice

The genetics of the immune response in the mouse were studied by using the antigenically simple, stable, naturally occurring protein ferredoxin (Fd) from Clostridium pasteurianum. The immune status of mice primed and boosted with Fd was assessed by using two parameters of immunity: T cell proliferation and serum antibody production with the ELISA method. In both assay systems, the response has been shown to be H-2 linked: k, b, and s haplotypes respond to Fd, and H-2d mice are nonresponders. It is apparent that different immunoregulatory events modulate the response in the responder strains; these factors become evident in the recombinant analysis of the response and to date an immunoregulatory gene(s) has been mapped to at least the K/I-A subregions. F1 analysis demonstrated a gene dose-dependent response of the strains studied.