Stimulation of corneal differentiation by interaction between cell surface and extracellular matrix: II. Further studies on the nature and site of transfilter “induction”

Corneal epithelial differentiation (primary stroma production) is dependent on the underlying extracellular matrix (ECM), for if the developing epithelium is enzymatically removed from the embryo, it fails to produce stroma in vitro unless it is cultured on collagenous ECM. We have previously shown that the stimulatory effect is mediated across Nucleopore filters in direct proportion to the surface area created by epithelial cell processes traversing the filter to contact ECM. Since collagenous ECM is insoluble under physiological conditions, transfilter stimulation of stroma production is probably due to an interaction of the epithelial cell surface with “inducer” ECM (killed lens capsule or purified collagen). We grew 5-day-old corneal epithelia on Nucleopore filters atop [³H]proline-labeled lens capsules and used both autoradiography and scintillation counting to show that radioactive collagen does not enter the epithelial cells in detectable amounts. We also show here that the stimulatory effect of collagen on collagen synthesis is not dependent on trapping of serum or binding of conditioned medium factors by ECM. Finally, we demonstrate that the stimulatory effect is reduced by removal of transfilter ECM after 6–12 hr in vitro. By 18–24 hr, however, cultured epithelium is less dependent on the substratum, probably because it has produced its own ECM. We conclude that: (1) the contact mediated collagen-cell surface interaction under study here requires the continuous presence of collagen in vivo and in vitro for maintenance of “stimulated” epithelial stroma synthesis; (2) the collagenous “inducer” interacts directly with epithelium rather than indirectly via trapped intermediates; (3) collagen acts at the epithelial cell surface without entering the cells.     

Leaching loss of NO3-N and dissolved P from manure and fertilizer during turfgrass establishment

Cycling of manure nutrients through turfgrass sod could affect groundwater quality. The fate of nutrients in transplanted fertilizer- or manure-grown sod of Tifway bermudagrass (Cynodon dactylon L. Pers. X C. transvaalensis Burtt-Davey) was compared with that in composted dairy manure (CDM) applied to a sprigged treatment. Leaching loss of NO₃-N and dissolved P (DP) in filtrate (<0.45 μm) of leachate was compared among sodded and sprigged treatments during periods 0–50, 60–110, and 330–380 d after planting in lysimeters. In addition, recovery of N and P in turfgrass clippings and a sand medium was quantified. Maintenance applications of CDM or fertilizer P were top-dressed starting 60 d after planting. Leachate was collected and sampled over three simulated rain events during each of the three sampling periods. From 0 d to 50 d after planting, leaching loss of NO₃-N from sprigged Tifway totaled 2.0 g m⁻² and was 10 times greater than loss from CDM-grown or fertilizer-grown sod. In contrast, DP loss in leachate was ≤0.02 g m⁻² and similar among treatments. Surface applications of CDM and fertilizer P and N increased concentration and mass of total Kjeldahl N (TKN) and soil-test P (STP) in surface or subsurface layers of the sand medium. Yet, NO₃-N mass in leachate collected over three simulated rain events ranged from 0.0 to only 1.0% of applied N from 60–110 d and 330–380 d after planting. Leaching loss of NO₃-N did not differ between the sodded and sprigged treatments after two topdressings of CDM. Similarly, the DP mass recovered in leachate was small (≤0.013 g m⁻¹) and did not differ among treatments during the latter two sampling periods. The mass loss of DP in leachate was typically less than the DP mass applied through irrigation or simulated rain. Importing CDM in sod reduces NO₃-N leaching loss compared to sprigged turfgrass amended with CDM, but NO₃-N and DP leaching losses are similar during maintenance of CDM-grown and fertilizer-grown sod from 60–110 d to 330–380 d after transplanting. Responsible Editor: Bernard Nicolardot.     

Geographic Variation in Camouflage Specialization by a Decorator Crab

In North Carolina, the decorator crab Libinia dubia camouflages almost exclusively with the chemically noxious alga Dictyota menstrualis. By placing this alga on its carapace, the crab behaviorally sequesters the defensive chemicals of the plant and gains protection from omnivorous consumers. However, Dictyota is absent north of North Carolina, whereas Libinia occurs as far north as New England. Crabs from three northern locations where Dictyota is absent (Rhode Island, Connecticut, and New Jersey) camouflaged to match their environment, rather than selectively accumulating any one species. When D. menstrualis was offered to crabs from northern sites, they did not distinguish between it and other seaweeds for camouflage, whereas crabs from Alabama and two locations in North Carolina used D. menstrualis almost exclusively. In addition, in winter and spring, when Dictyota was seasonally absent in North Carolina, Libinia selectively camouflaged with the sun sponge Hymeniacidon heliophila, which was chemically unpalatable to local fishes. Thus, southern crabs were consistent specialists on chemically defended species for camouflage, while northern crabs were more generalized. The geographic shift in crab behavior away from specialization coincides with a reported decrease in both total predation pressure and the frequency of omnivorous consumers. These shifts in the nature and intensity of predation pressure may favor different camouflage strategies (generalist vs. specialist), contributing to the observed geographic differences in camouflage behavior.     

Market-resource Links and Fish Vendor Livelihoods in the Upper Zambezi River Floodplains

This paper examines small-scale fish vending in a southern African floodplain from two perspectives: as a link between natural resource use and consumption, and as a livelihood in itself. We used a combination of observation, surveys and semistructured interviews in a market in Katima Mulilo, Namibia, to determine sources of fish, preferences and constraints to vending, average investment and profit, as well different routes into fish vending and perceptions regarding vending. Most vendors come from fishing households, but their stock is often an accumulation of purchases from other fishers. There is little evidence of formal arrangements between fishers and vendors, yet most adapt to the highly variable natural and social environments of the region. Although all vendors ranked selling fish as their most important livelihood activity, a wide range of investment and profit exists among individuals. Our findings indicate that fisheries management proposed for the area must be developed with a careful understanding of how changes in access and use will affect vending livelihoods.

The action of the beta-agonist clenbuterol on protein metabolism in innervated and denervated phasic muscles.

1. Clenbuterol treatment in innervated and denervated phasic extensor digitorum longus, plantaris and gastrocnemius muscles from rats caused a significant increase in RNA and protein contents in all muscles except denervated extensor digitorum longus. 2. All muscles showed an increase in the fractional rate of protein synthesis (Ks) with clenbuterol, but the temporal response varied. 3. The data suggest that the effect of clenbuterol on protein metabolism in innervated muscles is muscle-type specific, and demonstrate the homology of response for denervated muscles.     

The immunocytochemical demonstration of Toxoplasma antigen in the brains of congenitally infected mice

The peroxidase anti-peroxidase immunocytochemical staining method was used to identify Toxoplasma antigen in paraffin embedded sections of the brains of 22 mice congenitally infected with the parasite. Intact Toxoplasma tissue cysts were readily demonstrated in the brain in all cases. In 4 of the 22 infected mice there was evidence of rupture of the cyst wall and/or presence of extra-cystic Toxoplasma antigen. Further support for the extra-cystic location of Toxoplasma antigen was obtained by electron microscopy of reprocessed tissue which revealed endozoites in the area immediately surrounding a ruptured cyst. The possible implications of these findings in relation to the pathogenesis of congenital toxoplasmic meningo-encephalitis are discussed.     

Inhibition of Chk1 by activated PKB/Akt

We have shown recently that DNA damage effector kinase Chk1 is phosphorylated in vitro by protein kinase B/Akt (PKB/Akt) on serine 280. Activation of Chk1 by DNA damage in vivo is suppressed in presence of activated PKB. In this study we show that Chk1 is phosphorylated by PKB in vivo, and that increased phosphorylation by PKB on serine 280 correlates with impairment of Chk1 activation by DNA damage. Our results indicate a likely mechanism for the negative effects that phosphorylation of serine 280 has on activation of Chk1. The Chk1 protein phosphorylated by PKB on serine 280 does not enter into protein complexes after replication arrest. Moreover, Chk1 phosphorylated by PKB fails to undergo activating phosphorylation on serine 345 by ATM/ATR. Phosphorylation by ATM/ATR and association with other checkpoint proteins are essential steps in activation of Chk1. Inhibition of these steps provides a plausible explanation for the observed attenuation of Chk1 activation by activated PKB after DNA damage.