Characterization of the inhibitory effect of glucocorticoids on the DNA replication of adult rat hepatocytes growing at various cell densities

Dexamethasone inhibited the basal and EGF-stimulated DNA synthesis of adult rat hepatocytes in primary culture. The inhibition was glucocorticoid-specific: It was shown by dexamethasone and hydrocortisone, but not by progesterone, testosterone, or estradiol; and was counteracted by the glucocorticoid antagonist RU-38486 in a concentration-dependent manner. Dexamethasone acted by decreasing the rate of entry into S-phase (kG1/S), while cell cycle parameters were unaffected. The steroid was able to decrease the kG1/S severalfold even when added more than 20 hr after EGF, half-maximal effect occurring 11 hr after the addition of dexamethasone. Densely populated areas were much more sensitive to the inhibition by dexamethasone than sparsely populated areas within the same culture dish: A moderate (10 nM) concentration of dexamethasone nearly abolished the DNA synthesis in densely populated areas of hepatocyte cultures with only marginal effect on sparsely populated cells.     

Microclimates and water budget of algae,lichens and a moss on some nunataks in Queen Maud Land

During the Norwegian Antarctic Expedition of 1984–1985, land parties worked in the area of Mühlig-Hofmannfjella and Gjelsvikfjella in Queen Maud Land (5° 20E, 1° 37W, 1000–1600 m a.s.l.). The nunataks in this area, which represents one of the climatic limits for terrestrial life on earth, is among those areas absorbing the highest quantity of shortwave radiation during a period of 24 h in summer. In spite of this the air temperature never, or very seldom, exceeds 0° C. The limiting factor for photosynthesis over most of the summer was water availability. Melt-water plays an important role in spring. During rest of the growth season, water from condensation is probably the most important water source for plants. On calm nights the rate of condensation during 6 h may reach 0.5 mm, which constitutes only 10%–30% of daily potential evapotranspiration. Plants situated in narrow clefts or shielded by stone blocks have the highest rate of potential photosynthesis. These locations are shielded from direct solar radiation most of the time, but the radiation from surrounding stone surfaces is higher than from the atmosphere and heat loss by turbulence is smaller than for exposed locations. These locations also probably have the highest rate of actual photosynthesis.     

The influence of fixation on the relative amount of cytoplasmic ribosomes in mouse epidermal basal keratinocytes

The nature and significance of so-called dark keratinocytes in the epidermis during chemical carcinogenesis is still a matter of concern and debate. Based on ultrastructural observations it has been suggested that dark cells most often are shrunken cells. Reports on skin carcinogenesis, however, claim that dark cells are a sign of ongoing tumor promotion and represent those stem cells in the epidermis from which the tumors originate. It is therefore important to find out whether these cells are simply injured and shrunken cells, or vital cells of great importance for carcinogenesis. Dark cells are assumed to be rich in ribosomes. There is evidence, however, that the observed number of dark cells is highly dependent on tissue fixation. In the present ultrastructural study, morphometric methods were used to compare the effects of two different fixation procedures on the amount of cytoplasmic ribosomes in dark cells from both untreated and carcinogen-treated hairless mouse epidermis. The results show that the ultrastructural features of both dark and clear cells vary considerably with different fixation procedures. In acetone-treated controls typical dark cells are only observed when the fixative has a lower osmotic activity than the plasma. With iso-osmolal fixation typical dark cells are not observed. After an abortive two-stage carcinogenesis treatment, in which a single application of 9,10-dimethyl-l,2-benzanthracene (DMBA) in acetone was followed by a single application of 12-O-tetradecanoyl-13-acetate (TPA) in acetone, signs of cell injury could be found after both fixation procedures. With DMBA/TPA and hypo-osmolal fixation the number of dark cells seemed to increase, whereas only signs of cell injury with occurrence of some heavily altered “clear cells” dominated the picture with iso-osmolal fixation. Morphometry showed that both the numerical and the volumetric densities of cytoplasmic ribosomes in basal keratinocytes varied most significantly with the fixation procedure used. The cytoplasmic volumes did not vary in a way that could explain these differences. One might therefore assume that the number of ribosomes depends on the fixative. Large swelling artifacts occurred when a fixative with low osmotic activity was used, leading to compression of neighboring cells. Hence, an increased ribosomal density reported previously in dark cells is probably related to such cell volume artifacts and does not reflect an actually increased quantity of ribosomes. With both fixation procedures, a single application of DMBA followed by one of TPA appeared to produce an increased number of ribosomes in basal keratinocytes. When hypo-osmolal fixation was used, however, treatment with DMBA/TPA did not influence the cytoplasmic volume or the numerical density of ribosomes, in dark cells. This might indicate that so-called dark keratinocytes following DMBA/TPA treatment are functionally inactive cells that appear more vulnerable than active cells to compression during hypo-osmolal fixation.     

Variation in numbers, territoriality and flock size of a Goldcrest Regulus regulus population in winter

A Goldcrest Regulus regulus population living in a spruce-dominated forest near Oslo (c.60°N), in southern Norway, was studied during the winters (November-March) from 1966–67 to 1971–72. A significant reduction in population size occurred between November and March, the recorded values varying from 76 to 96%, with a mean reduction for all six winters studied of 86%. This winter decline in numbers was significantly correlated with both precipitation (i.e., snowfall) and ambient temperature, but not with bird density. Spiders were the predominant prey items of the Goldcrests, and it is suggested that the birds experience food shortages that affect their survival rate. Although not statistically significant, a mean change in the ratio of males:females from 2:1 in October-November to 10:1 in December—February was observed, suggesting that a greater mortality of females than males occurs during the winter.
Partly based on data for colour-banded birds, it is suggested that the Goldcrests form social flocks, which move around as individual units within territories of about 9 ha (7.5 12 ha, n = 5) in extent. The mean flock size decreased significantly from 4.57 ( n = 157) birds in November-December to 2.82 ( n = 74) birds in January-February, probably because of the reductions in the size of the local population during the winter. The mean flock size in November-December varied inversely with the ambient temperature ( r = -0.92).     

THE FINE STRUCTURE OF CHONDROCOCCUS COLUMNARIS : I. Structure and Formation of Mesosomes

Cells of Chondrococcus columnaris were sectioned and examined in the electron microscope after fixation by two different methods. After fixation with osmium tetroxide alone, the surface layers of the cells consisted of a plasma membrane, a dense layer (mucopeptide layer), and an outer unit membrane. The outer membrane appeared distorted and was widely separated from the rest of the cell. The intracytoplasmic membranes (mesosomes) appeared as convoluted tubules packaged up within the cytoplasm by a unit membrane. The unit membrane surrounding the tubules was continuous with the plasma membrane. When the cells were fixed with glutaraldehyde prior to fixation with osmium tetroxide, the outer membrane was not distorted and separated from the rest of the cell, structural elements (peripheral fibrils) were seen situated between the outer membrane and dense layer, and the mesosomes appeared as highly organized structures produced by the invagination and proliferation of the plasma membrane. The mesosomes were made up of a series of compound membranes bounded by unit membranes. The compound membranes were formed by the union of two unit membranes along their cytoplasmic surfaces.     

THE FINE STRUCTURE OF CHONDROCOCCUS COLUMNARIS : II. Structure and Formation of Rhapidosomes

When cells of C. columnaris were broken open, treated with PTA, and examined in the electron microscope, tubular structures (rhapidosomes) were present in the preparations. The rhapidosomes are approximately 300 A in diameter. Their length varies from about 500 to about 15,000 A. An axial hole which runs the length of the rhapidosomes appears to widen and narrow with a regular periodicity. End-on views of short segments of rhapidosomes revealed the presence of subunits around their outside peripheries. The results of studies of lysed cells and of sectioned cells indicate that the rhapidosomes are produced during the disintegration of cells. It seems likely that the compound membranes of the mesosomes break down to give rise to the tubular structures. The mesosomal origin of rhapidosomes is postulated only for the rhapidosomes of C. columnaris, since the origin of rhapidosomes from other organisms was not investigated during this study. The rhapidosomes of C. columnaris may be unrelated to those of S. grandis, S. myxococcoides, A. violaceum, and Sorangium 495, since there was a difference in the details of fine structure between rhapidosomes from C. columnaris and those found in the other four organisms.     

THE FINE STRUCTURE OF TWO UNUSUAL STALKED BACTERIA

Two strains of bacteria that produce slender appendages (pseudostalks) from their lateral surfaces were studied using the electron microscope. The pseudostalks were shown to be extensions of the cytoplasm and peripheral membranes of the cell proper. Both strains of bacteria produce holdfasts at the poles of the cells by the means of which attachment can take place. The pseudostalks are not involved in the attachment of cells. No specialized intracytoplasmic structures are present at the point of juncture of pseudostalk and cell. A discussion of the possible functions of the pseudostalks, based on the electron microscope findings, is presented.     

Estimation of the effect of photoinhibition on the carbon gain in leaves of a willow canopy

The occurrence of photoinhibition of photosynthesis in leaves of a willow canopy was examined by measuring the chlorophyll-a fluorescence ratio of F _V/F _M (F_M is the maximum fluorescence level of the induction curve, and F_V is the variable fluorescence, F _V=F _M–F ₀, where F₀ is the minimal fluorescence). The majority of the leaves situated on the upper parts of peripheral shoots showed an afternoon inhibition of this ratio on clear days. This was the consequence of both a decrease in F _M and a rise in F _O. In the same leaves the diurnal variation in intercepted photosynthetic photon flux density (PPFD) was monitored using leaf-mounted sensors. Using the multivariate method, partial least squares in latent variables, it is shown that the dose of PPFD, integrated and linearly weighted over the last 6-h period, best predicts photoinhibition. Photoinhibition occurred even among leaves that did not intercept PPFDs above 1000 mol·m^–2·s^–1. Exposure of leaves to a standard photoinhibitory treatment demonstrated that the depression in the F _V/F _M ratio was paralleled by an equal depression in the maximal quantum yield of CO₂ uptake and a nearly equal depression in the rate of bending (convexity) of the light-response curve of CO₂ uptake. As a result, the rate of net photosynthesis is depressed over the whole natural range of PPFD. By simulating the daily course in the rate of net photosynthesis, it is estimated that in the order of one-tenth of the potential carbon gain of peripheral willow shoots is lost on clear days as a result of photoinhibition. This applies to conditions of optimal temperatures. Photoinhibition is even more pronounced at air temperatures below 23° C, as judged from measurements of the F_V/F_M ratio on clear days: the afternoon inhibition of this ratio increased in a curvilinear manner from 15% to 25% with a temperature decrease from 23° to 14° C.Abbreviations and Symbols F_O minimum fluorescence - F_V variable fluorescence - F_M maximum fluorescence - PLS partial least squares in latent variables - PPFD photosynthetic photon flux density - VPD water vapour-pressure deficit This study was supported by the Swedish Natural Science Research Council. We are indebted to Dr. Jerry Leverenz (Department of Plant Physiology, University of Umeå, Sweden) for guidance with the modelling of the photosynthesis data.     

A new approach for estimating the crosslink density of covalently crosslinked ionic polysaccharide gels

For an ideal polysaccharide gel with a known total polymer chain contour length, crosslinks all of the same functionality and elastic chains all with the same contour length and stiffness, the gel crosslink density can readily be determined from measurements of the maximum volume of the swollen gel (Moe et al., (1991) Food Hydrocolloids, 5, (1/2), 119–123. In the case of randomly crosslinked polysaccharide gels, where the chain contour length between two adjacent crosslinks may vary greatly, it is often much more difficult to determine the crosslink density. This paper reports on an attempt to extend the use of maximum gel volume measurements to estimate crosslink density for the latter type of gel. This is done by calculating the maximum swelling volume for polymer networks with four-functional crosslinks, known elastic chain mean contour length and standard deviation. The numerical analysis involves the calculation of the equilibrium force at each crosslink as the network expands. This allows a detailed study of how the distribution of individual polymer chain contour lengths affects the maximum swelling volume. The computer simulation results are compared with the results from experimental measurements of the maximum volume of swollen covalently crosslinked sodium alginate gels.