Indoleacetic acid enhancement of 14C methionine uptake by tomato cell cultures aids assay of pathogen extracts

Incorporation of one micromolar IAA in the assay system increased the rate of ¹⁴C methionine uptake by tomato cells in suspension and effectively differentiated the rates of uptake such that cells treated with pathogenic and non-pathogenic Fusarium extracts could be easily distinguished in a rapid 3 h assay.     

Author index for volume 219

Proteinase inhibitors I and II were purified to electrophoretic homogeneity from leaves of tomato plants induced by either wounding intact plants or by supplying excised plants with the proteinase inhibitor inducing factor. Affinity chromatography with chymotrypsin-Sepharose was employed as a final purification step for each inhibitor. The tomato leaf inhibitors are very similar to potato tuber inhibitors I and II in subunit molecular weight, composition, and inhibitory activities against chymotrypsin, trypsin, and subtilisin. However, unlike the potato tuber which contains multiple isoinhibitors by isoelectric focusing, the tomato leaf exhibits only two isoinhibitor forms of inhibitor I and a single form of inhibitor II. The molecular weight of native potato inhibitor I was reevaluated by rigorous ultracentrifugal analysis and compared with data from previous analyses. The data confirm that native inhibitor I has a native M_r of about 41,000 and is a pentamer. Inhibitor II has a molecular weight of near 23,000 and is a dimer.     

Renaturation of DNA: a novel reaction of histones.

Histones isolated from several sources, either singly or in combination promote the renaturation of complementary single strands of DNA, as measured by the acquisition of resistance to S1 nuclease. The reaction is rapid (T1/2 less than 1 min), and is stoichiometric rather than catalytic. Renaturation is stimulated by Mg2+, Mn2+, and Ca2+, but is strongly inhibited by Zn2+. Crude extracts of early embryos of Drosophila melanogaster possess renaturation activity which is protease sensitive, heat-stable, and acid-soluble, suggesting that most or all of it can be attributed to histones. This observation thus provides a functional assay for histones that should prove useful in studies of chromatin and histone-DNA interactions, as well as for the identification and isolation of histones and histone-like proteins in crude extracts.     

Comparison of bone marrow extracellular matrices.

We have compared the structure and composition of adult and fetal bovine bone marrow extracellular matrices. In contrast to fetal bone marrow, adult bone marrow has more oval fenestration and accumulation of adipocytes as well as lower protein content. These differences could be due to remodeling of bone marrow tissue as it develops. Zymogram analysis of matrix metalloproteinase (MMP) and tissue inhibitor of MMP (TIMP) activities showed that fetal, but not adult bone marrow extract contained a 96-kDa MMP and TIMP-1 and -2. These activities may contribute to the structural differences between adult and fetal bone marrow tissues.     

Post-Spaceflight (STS-135) Mouse Splenocytes Demonstrate Altered Activation Properties and Surface Molecule Expression

Alterations in immune function have been documented during or post-spaceflight and in ground based models of microgravity. Identification of immune parameters that are dysregulated during spaceflight is an important step in mitigating crew health risks during deep space missions. The in vitro analysis of leukocyte activity post-spaceflight in both human and animal species is primarily focused on lymphocytic function. This report completes a broader spectrum analysis of mouse lymphocyte and monocyte changes post 13 days orbital flight (mission STS-135). Analysis includes an examination in surface markers for cell activation, and antigen presentation and co-stimulatory molecules. Cytokine production was measured after stimulation with T-cell mitogen or TLR-2, TLR-4, or TLR-5 agonists. Splenocyte surface marker analysis immediate post-spaceflight and after in vitro culture demonstrated unique changes in phenotypic populations between the flight mice and matched treatment ground controls. Post-spaceflight splenocytes (flight splenocytes) had lower expression intensity of CD4⁺CD25⁺ and CD8⁺CD25⁺ cells, lower percentage of CD11c⁺MHC II⁺ cells, and higher percentage of CD11c⁺MHC I⁺ populations compared to ground controls. The flight splenocytes demonstrated an increase in phagocytic activity. Stimulation with ConA led to decrease in CD4⁺ population but increased CD4⁺CD25⁺ cells compared to ground controls. Culturing with TLR agonists led to a decrease in CD11c⁺ population in splenocytes isolated from flight mice compared to ground controls. Consequently, flight splenocytes with or without TLR-agonist stimulation showed a decrease in CD11c⁺MHC I⁺, CD11c⁺MHC II⁺, and CD11c⁺CD86⁺ cells compared to ground controls. Production of IFN-γ was decreased and IL-2 was increased from ConA stimulated flight splenocytes. This study demonstrated that expression of surface molecules can be affected by conditions of spaceflight and impaired responsiveness persists under culture conditions in vitro.     

Composition and properties of the extracellular polysaccharide produced by arthrobacter stabilis NRRL B-3225

The extracellular polysaccharide produced by Arthrobacter stabilis NRRL B-3225 contains d-glucose, d-galactose, pyruvic acid, O-succinyl, and O-acetyl in the approximate molar ratio of 6:3:1:1:1.5. Succinyl is linked as its half-ester, making it a readily removable, acidic function that imparts versatility to the polysaccharide both for fundamental research and for commercial use. The viscosity of aqueous, salt-free solutions of both native and deacylated polymer is relatively low, but atypical of anionic polysaccharides, increases rapidly in the presence of salts, acids, or alkali.     

Preliminary analysis of drinking from seawater sources by free-ranging rhesus macaques (Macaca mulatta)

Rhesus macaques on Key Lois Island were observed drinking seawater that flowed into four holes they had excavated in the sand. Data were gathered to determine the salinity (TDS) and pH levels of the water and which animals were using the holes. Average TDS level (4,506.8 ± 1,750.8), but not pH (8.1 ± .30) level, of water from the holes differed from the surrounding seawater (TDS = 29,000, pH = 8.0). There were significant variations in TDS and pH levels of water between holes. A total of 249 drinking and 11 digging bouts were observed. Adult females drank and dug most often (46.9% of total bouts). Of the 260 total drinking and digging bouts, 76.1% (N = 198) were concentrated at one hole. This hole had the lowest average TDS level (3,714.2 ± 1,504.4) and one of the highest average pH levels (8.1 ± .29). Age/sex class differences in drinking bout frequencies may have been due to differential social status. We suggest that the holes were excavated to overcome a temporary shortage of provisioned water. © 1993 Wiley-Liss, Inc.     

Rolling circle DNA replication by extracts of herpes simplex virus type 1-infected human cells.

Whole-cell extracts of herpes simplex virus type 1-infected human cells (293 cells) can promote the rolling circle replication of circular duplex DNA molecules. The products of the reaction are longer than monomer unit length and are the result of semiconservative DNA replication by the following criteria: (i) resistance to DpnI and susceptibility to MboI restriction enzymes, (ii) shift in density on a CsCl gradient of the products synthesized in the presence of bromo-dUTP to a position on the gradient consistent with those of molecules composed mainly of one parental DNA strand and one newly synthesized DNA strand, and (iii) the appearance in the electron microscope of molecules consisting of duplex circles with multiunit linear appendages, a characteristic of a rolling circle mode of DNA replication. The reaction requires ATP and is dependent on herpes simplex virus type 1-encoded DNA polymerase.