Cytokine production in a model of wound healing: the appearance of MIP-1, MIP-2, cachectin/TNF and IL-1

Macrophages are essential for normal wound repair and many of their effects on healing wounds are likely to be mediated by the secretion of cytokines. This study examines the appearance of messenger RNA (mRNA) for cachectin/tumor necrosis factor (TNF), IL 1, and macrophage inflammatory proteins 1 and 2 (MIP-1 and MIP-2), as well as the mature peptides, in a model of wound healing using wound chambers. RNA for all four cytokines can be detected in wound inflammatory cells by polymerase chain reaction amplification throughout the first 7 days. Cachectin/TNF and IL 1 protein levels peaked on the first day after wound chamber implantation, and MIP-1 and MIP-2 were detected only on day 3. The data suggest that these cytokines participate in the early inflammatory response to wounding.     

A growth inhibitory protein secreted by human diploid fibroblasts. Partial purification and characterization

We have identified and partially purified a growth inhibitor protein secreted by human diploid fibroblast cells. This protein is not secreted constitutively but only after induction with the double stranded hetero duplex polyriboinosinic:polyribocytidylic acid. The growth inhibitory activity has been purified 3,800-fold and has an estimated molecular mass of 12,000 daltons. The protein will inhibit the growth in culture of human diploid fibroblast cells, human cells derived from tumors, and mouse L cells. Although interferon-beta is secreted with the growth inhibitory protein, the partially purified growth inhibitory protein has no antiviral activity, and its activity is not neutralized by antibodies to interferon-alpha, interferon-beta, and interferon-gamma. We believe this growth inhibitory activity to reside in a newly defined protein and have named it fibroblast-derived growth inhibitor.     

Somatic embryogenesis from three commercially important inbreds of Zea mays

Zea mays (maize) genotypes B73, Mo17 and LH38 were evaluated for their capacity to undergo somatic embryogenesis. Over 1500 immature embryos (ie's) of B73, 2900 ie's of LH38 and 400 ie's of Mo17 were excised 10–17 days after pollination and plated on six different media. Overall response, reported as a percentage of the ie's plated that developed embryogenic callus, was 2.1%, 1.6% and 26% for LH38, B73 and Mo17, respectively. Best response on a given medium for each of these genotypes was 9.2% (LH38), 4.4% (B73) and 100% (Mo17). Other parameters examined for their effects on production of embryogenic callus included self vs. sib pollination, ear ranking (1st, 2nd or 3rd ear), and temperature shock, all of which had no significant effect. Plantlets regenerated from selected treatments of B73 have been grown to maturity, selfed or sibbed and seed collected for field evaluation.Abbreviations i.e. immature embryo - 2,4-D 2,4-di-chlorophenoxyacetic acid     

Mutational analysis of antigen receptor regulation of B lymphocyte growth. Evidence for involvement of the phosphoinositide signaling pathway.

Stimulation of the antigen receptor of WEHI-231 B lymphoma cells with anti-receptor antibodies (anti-IgM) induces irreversible growth arrest. Anti-IgM stimulates two kinds of transmembrane signaling events, phosphorylation of proteins on tyrosyl residues and breakdown of inositol phospholipids, which results in increases of inositol phosphates, diacylglycerol, and calcium. The roles of these reactions in mediating the growth arrest of the B lymphoma cells have not been established. To examine this issue, we took a genetic approach. Mutants of WEHI-231 cells were isolated that were resistant to anti-IgM-induced growth arrest. Five out of seven independent mutants analyzed had normal cell-surface expression of antigen receptors. Although each of these five mutants had tyrosine protein phosphorylation patterns comparable to wild-type cells, they exhibited alterations in the phosphoinositide signaling pathway. Four of the mutants had decreased phosphoinositide breakdown, probably due to an alteration in phospholipase C. Decreased second messenger production may be responsible for the growth-resistant phenotype. Full growth arrest was restored upon addition of the calcium ionophore ionomycin, suggesting that the limiting second messenger was intracellular free calcium. The final mutant appeared to be altered in a component(s) that responds to diacylglycerol and calcium. Taken together, these results provide further evidence that the phosphoinositide pathway is at least partly responsible for mediating antigen receptor regulation of B lymphoma cell growth.     

Effects of energy level and monensin on reproductive performance and lactation of beef cows

This experiment consisted of a 2 year drylot study involving 80 multiparous, suckled beef cows (40 Angus and 40 Herefords). Experimental treatments (breed, monensin and year) were arranged in 2(3) factorial to evaluate the effect of breed and monensin on reproductive performance and lactation. Cows received 85% of the NRC total digestible nutrient (TDN) requirement for the first 56 days of the 140 day trial. Cows were synchronized with Synchro-Mate B and artificially inseminated 30 days into each trial with blood samples collected for luteinizing hormone (LH) analysis from one-half of each treatment group from 24 to 36 hours after implant removal. Progesterone determinations were made on plasma samples collected at day 9 and 16 after implant removal. On day 56, milk yield estimates were obtained by the weigh-suckle-weight technique. Following these collections, energy levels were increased by allowing ad lib consumption of forage. Calves, which were removed from the cows during feeding, were given access to a 75% TDN creep ration after day 56. Milk estimates were again evaluated at 140 days. Monensin supplementation did not result in a difference in cow weight change through the restricted energy period (first 56 days) or throughout the entire 140 day period. Milk yield estimates at 56 and 140 days and calf gains throught the trial, were unaffected by monensin supplementation. Monensin resulted in no effect on conception rate or services per conception. The time of the LH peak was shifted slightly forward by monensin although not significantly. Progesterone levels were not consistently affected by monensin supplementation.     

Snow depth manipulation and its influence on soil frost and water dynamics in a northern hardwood forest

Climate change will likelyresult in warmer winter temperatures leading toless snowfall in temperate forests. Thesechanges may lead to increases in soil freezingbecause of lack of an insulating snow cover andchanges in soil water dynamics during theimportant snowmelt period. In this study, wemanipulated snow depth by removing snow for twowinters, simulating the late development of thesnowpack as may occur with global warming, toexplore the relationships between snow depth,soil freezing, soil moisture, and infiltration.We established four sites, each with two pairedplots, at the Hubbard Brook Experimental Forest(HBEF) in New Hampshire, U.S.A. and instrumentedall eight plots with soil and snow thermistors,frost tubes, soil moisture probes, and soillysimeters. For two winters, we removed snowfrom the designated treatment plots untilFebruary. Snow in the reference plots wasundisturbed. The treatment winters (1997/1998 and1998/1999) were relatively mild, withtemperatures above the seasonal norm and snowdepths below average. Results show the treatedplots accumulated significantly less snow andhad more extensive soil frost than referenceplots. Snow depth was a strong regulator ofsoil temperature and frost depth at all sites.Soil moisture measured by time domainreflectometry probes and leaching volumescollected in lysimeters were lower in thetreatment plots in March and April compared tothe rest of the year. The ratio of leachatevolumes collected in the treatment plots tothat in the reference plots decreased as thesnow ablation seasons progressed. Our data showthat even mild winters with low snowfall,simulated by snow removal, will result inincreased soil freezing in the forests at theHBEF. Our results suggest that a climate shifttoward less snowfall or a shorter duration ofsnow on the ground will produce increases insoil freezing in northern hardwood forests.Increases in soil freezing will haveimplications for changes in soil biogeochemicalprocesses.