Polymorphonuclear leukocyte migration through human amnion membrane

A new in vitro model has been developed for studying migration of human polymorphonuclear leukocytes (PMN) through living native cellular and matrix barriers. Human amnion membrane consists of a single layer of epithelium bound to a continuous basement membrane interfacing an avascular collagenous stroma. Living amnion was placed in plastic chambers with separate compartments on each side of the membrane. PMN were introduced on the epithelial side of the amnion, and a Millipore filter (Millipore Corp., Bedford, Mass.) was placed against the stromal side. In response to N-formylmethionyl-leucyl- phenylanlanine (FMLP) chemoattractant, PMN penetrated the full thickness of the amnion and were collected and counted on the filter. The rate of PMN traversal of the amnion was dependent on the concentration of FMLP (optimal at 10(-8)M) as well as the slope of the FMLP gradient across the amnion. The route of PMN migration was studied by transmission electron microscopy. PMN first attached to the epithelial surface, then infiltrated between intercellular junctions. PMN migrated around or through tight junction and hemidesmosome attachments. The PMN then penetrated the basement membrane and migrated through the dense collagenous stroma. The present amnion migration system has characteristics of the in vivo inflammatory state not described in any previous method for monitoring PMN migration in vitro. Prior methods have not used native epithelium, whole basement membrane, or collagenous stroma. PMN penetration of these barriers occurs in the normal inflammatory response and probably involves biochemical mechanisms not required for simple migration through the pores of an artificial filter. The amnion system can be useful for future biochemical and morphological studies of PMN penetration of these barriers and possible repair processes that may follow.     

Stomatal Responses to Sulphur Dioxide and Vapour Pressure Deficit

Stomatal conductances (g_s) of plants of Vicia faba, Raphanussativus, Phaseolus vulgaris, Heilanthus annuus, and Nicotianatabacum were measured in chambers containing either clean airor air containing between 18 and 1000 parts 10^–9 SO₂ atwater vapour pressure deficits (vpd) ranging from 1·0to 1·8 kPa. When vpd was low (<1·3 kPa at 22 °C) and stomatawere open, exposure to SO₂ induced rapid and irreversible increasesin g_s in V. faba. This response persisted throughout the exposure(3 d). The increase in g_s, 20–30% compared with cleanair, was independent of SO₂ concentration up to 350 parts 10^–9Stomatal conductances of polluted plants at night were greaterthan controls. When stomata were closed before exposure to SO₂,there was no effect on g_s. When vpd was varied, g_s of unpolluted plants of P. vulgarisshowed no response, but that of R. sativus increased slightlywith increasing vpd. In both species exposure to SO₂ causedan increase in g_s at all vpd values. g_s of unpolluted plantsof V. faba, H. annuus, and N. tabacum decreased with increasingvpd. At low vpd values exposure to SO₂ in these species causedan increase in g_s, but, above a certain value of vpd, dependingon species, g_s decreased with exposure to SO₂. It is postulated that SO₂, once in the substomatal cavity, entersthe stomatal complex via adjacent epidermal cells and at lowconcentrations leads to a reduction in turgor in these cellsand consequently to stomatal opening. In vpd-sensitive species,increased transpiration from guard cells or epidermal cellsadjacent to the stomata induced by SO₂ may lead to stomatalclosure at large vpd levels. Stomatal sensitivity to vpd insuch cases may be enhanced because adjacent epidermal cell turgoris lowered by SO₂. At high SO₂ concentrations direct disruptionof guard cell structure may lead to a loss of turgor and stomatalclosure.     

Photoperiodic time signals during twilight

Abstract. Although daylength has a major effect on flowering and several other aspects of plant development, the actual environmental time signals for the beginning and the end of day are obscure. An intensive spectroradiometric study was carried out in three contrasting environments: namely, unshaded sites, a mature oak woodland and a sugar beet crop. Spectral photon distributions were obtained describing numerous twilight phases and intervening photoperiods throughout the year. From each, absolute photon fluence rates, photon fluence rate ratios and phytochrome photoequilibria were calculated. Although substantial changes in spectral composition occurred during twilight, they were less capable of providing reliable and accurate time signals than the absolute fluence rate; this was especially apparent beneath the canopies. Thus, spectral changes are unlikely to be valuable in photoperiodic perception. The results are discussed in relation to the possible involvement of the known plant photoreceptors in photoperiodism.