Induction of autocrine epidermal growth factor receptor ligands in human keratinocytes by insulin/insulin-like growth factor-1

Autocrine activation of the epidermal growth factor (EGF) receptor on keratinocytes has been recognized as an important growth regulatory mechanism involved in epithelial homeostasis, and, possibly, hyperproliferative diseases. Insulin-like growth factor (IGF)-1 and insulin have been shown to be paracrine keratinocyte mitogens that bind to the type I IGF receptor which is expressed on actively proliferating keratinocytes in situ. In this report, we demonstrate that IGF-1/insulin induced production of keratinocyte-derived autocrine growth factors that bind to the EGF receptor. Increased steady-state mRNA levels for transforming growth factor alpha (TGF-α) and for amphiregulin (AR) were observed upon incubation of keratinocytes with mitogenic concentrations of IGF-1. IGF-1 also induced production and secretion of TGF-α and AR proteins as detected by immunoassays. An EGF receptor antagonistic monoclonal antibody abolished the mitogenic effect of IGF-1 on cultured keratinocytes. These results suggest that stimulation of keratinocyte growth by IGF-1 requires activation of an EGF receptor-mediated autocrine loop. © 1995 Wiley-Liss, Inc.     

An Arabidopsis lipid map reveals differences between tissues and dynamic changes throughout development

Mass spectrometry is the predominant analytical tool used in the field of plant lipidomics. However, there are many challenges associated with the mass spectrometric detection and identification of lipids because of the highly complex nature of plant lipids. Studies into lipid biosynthetic pathways, gene functions in lipid metabolism, lipid changes during plant growth and development, and the holistic examination of the role of plant lipids in environmental stress responses are often hindered. Here, we leveraged a robust pipeline that we previously established to extract and analyze lipid profiles of different tissues and developmental stages from the model plant Arabidopsis thaliana. We analyzed seven tissues at several different developmental stages and identified more than 200 lipids from each tissue analyzed. The data were used to create a web-accessible in silico lipid map that has been integrated into an electronic Fluorescent Pictograph (eFP) browser. This in silico library of Arabidopsis lipids allows the visualization and exploration of the distribution and changes of lipid levels across selected developmental stages. Furthermore, it provides information on the characteristic fragments of lipids and adducts observed in the mass spectrometer and their retention times, which can be used for lipid identification. The Arabidopsis tissue lipid map can be accessed at http://bar.utoronto.ca/efp_arabidopsis_lipid/cgi-bin/efpWeb.cgi .     

Characterization of chlorobenzoate degraders isolated from polychlorinated biphenyl-contaminated soil and sediment in the Czech Republic

Two polychlorinated biphenyl-contaminated sites in the Czech Republic, a soil at Zamberk and a sediment sludge at Milevsko, were screened for the presence of chlorobenzoate degraders. Sixteen different chlorobenzoate degraders were isolated from the soil compared with only three strains isolated from the sediment. From these strains, only four soil degraders and one strain isolated from the sediment, respectively, were shown to possess a complete chlorobenzoate (CB) pathway. Bacteria isolated from the soil have expressed more flexibility for CB degradation, namely in the case of ortho-chlorinated benzoates. They all possessed large plasmids, the restriction patterns of which were compared. Plasmids in Pseudomonas sp. A7, A8, A18 and A19, respectively, were cured and found to encode at least part of the metabolic pathway involved in the growth on ortho-chlorinated benzoates.     

Effect of gibberellic acid on pod set in soybean

Field-grown soybean plants (Glycine max (L.) Merr. cv. Evans) were treated with gibberellic acid (GA₃; 10gl^–1) and/or (2-chloroethyl)-trimethylammonium chloride (CCC; 0.8gl^–1) in 1983 and 1984, and subsequent anthesis, pod set, seed size, seed number, and seed yield were determined at one node. The treatments were applied to five leaves in the center of each plant (typically leaves 7–11) and reproductive development at the node in the center of those leaves was monitored. Gibberellin A₃ applied Early (about 3d before anthesis of the first flower at the monitored node) had no effect on the number of flowers produced, but decreased the fraction of flowers that set pods in both experimental years (by 32% in 1983 and 76% in 1984). Seed size was slightly decreased by the GA₃ treatment in 1983 but not in 1984. The Middle GA₃ treatment (applied about 3 days after the Early treatment) slightly decreased the number of pods set; and Late treatments (9 days after) had no effect. None of the monitored parameters were affected by CCC.The Early experiments were repeated with two additional genotypes, Lincoln and T210. Genotype T210 is a single-gene, dwarf mutant of Lincoln whose stem elongation and leaf expansion are insensitive to GA₃. Gibberellin A₃ affected the reproductive parameters in Lincoln very similarly to Evans but those in T210 were unaffected. This indicates that GA₃ exerts its effect by increasing the mass of vegetative tissue and thus diverting assimilates away from the pods. However, since the mutation in T210 might affect a receptor that is in flowers as well as shoots, it is possible that GA₃ exerted its effect on the normal genotypes directly on the developing pods, rather than indirectly by diverting photoassimilates.     

Annual cycle of liveweight and reproductive changes of farmed male fallow deer (Dama dama) and the effect of daily oral administration of melatonin in summer on the attainment of seasonal fertility

Entire bucks (N = 7) exhibited pronounced liveweight gains over spring and summer months (October-February), to reach a peak mean weight of 59.8 kg, and rapid liveweight losses over the rutting period (April-May) with a minimum mean liveweight of 54.2 kg. Mean neck girth and serum testosterone levels increased during late summer (January-March) and peaked at 387 mm and 12 ng/ml respectively immediately before the onset of the rut (April). Thereafter both measures declined during winter and spring months (June-December). Bucks castrated prepubertally (N = 11) exhibited similar but less pronounced changes in mean liveweight and neck girth, in the absence of any change in testosterone secretion. Peak mean testicular diameter of entire bucks (39 mm) occurred immediately before the rut and was followed by testicular regression over winter and spring months (June-November), such that the testes attained their minimum mean size of 18 mm diameter in early summer (November). Motile spermatozoa were absent from ejaculates collected in summer (November 1983, 1984; January 1984). However, ejaculates collected pre-rut (late March), immediately post-rut (June) and in early spring (September) contained successively increasing numbers of motile spermatozoa. A further 14 polled, entire bucks were given orally 5 mg (N = 7; Group A) or 20 mg (N = 7; Group B) melatonin at 15:30 h daily from 1 December 1983 to 14 January 1984 (45 days). Seven control bucks (Group C) received vehicle ration only. The measurements taken for bucks in Groups A and B were not significantly different (P greater than 0.1) on any sampling date and the data for these 2 groups were pooled. Mean serum testosterone concentrations and mean ejaculate volume were not significantly different between melatonin-treated and control bucks on any sampling date, although other measures exhibited significant differences (P less than 0.05) at various treatment or post-treatment dates: melatonin-treated bucks showed a transiently greater increase in neck muscle development during and immediately after treatment, a slight retardation of liveweight gain between 45 and 75 days after treatment, an earlier peak in maximum mean testicular diameter and an earlier onset of sperm presence in ejaculates.