Effects of Ultraviolet Irradiation on the Cell Cycle

Cultured mouse Cloudman melanoma cells, EMT6 breast carcinoma cells, and 3T3 fibroblasts all accumulated in the G2/M phase of the cell cycle when exposed to UVB radiation. The effects of UVB were maximal at 20–30 mJ/cm² for all three cell lines, and could be observed by flow cytometry as early as 12 hr post irradiation. It has been known since the mid-1970s that MSH receptor binding activity is highest on Cloudman melanoma cells when they are in the G2/M phase of their cycle. Here we show that either UVB irradiation or synchronization of Cloudman cells with colchicine results in a stimulation of MSH binding within 24 hr following treatment, a time when both treatments have resulted in accumulation of cells in the G2/M phase of the cycle. Furthermore, the two treatments performed together on the melanoma cells stimulated MSH receptor activity to the same extent as either treatment performed separately, suggesting that each may be influencing MSH receptor activity solely through a G2/M accumulation of cells. Together, these results raise the possibility that an increase in the number of cells in the G2 phase of the cell cycle is a generalized cellular response to injury, such as UV irradiation. However, in the case of pigment cells this response includes a mechanism for increasing melanin formation, i.e., increased MSH receptor activity. Should this be the case, similar G2/M “injury responses” of other cell types might be expected, consistent with their differentiated phenotypes.     

Review of the genus Hippidion Owen, 1869 (Mammalia: Perissodactyla) from the Pleistocene of South America

This review of Hippidion is based on a multivariate analysis of the foot, and some morphological characteristics of the skull and dentition. We recognize only one genus ( Hippidion ) including all the hippidiform horses, with three different species: H. principale, H. devillei and H. saldiasi. The latter species is stratigraphically and geographically restricted to the period from 13000 to 8000 years BP in the southern part of South America. Hippidion principale and H. devillei have a large geographical distribution (Argentina, Bolivia, Chile, Perú, Uruguay, Brazil) through the Upper Pliocene-Upper Pleistocene. Both species show some morphometric variations across their geographic range; these features may result from the environmental characteristics.     

Chemical and Electron Microscopic Studies of Cattle (Bos taurus) With Four Types of Phenotypic Pigmentation

The biological behavior of the pigmentary phenotypes of four breeds of cattle has been analysed: the black pigmentation of Holstein Friesian; the red pigmentation of Limousin; the dilution in Charolais; and the postnatal disappearance of red pigmentation in Chianina. The analytic techniques included the characterization of melanins by high-performance liquid chromatography, the examination of follicular melanocytes by light microscopy, and the examination of melanosomes by electron microscopy. The black phenotype was very strongly eumelanogenic. The red phenotype in Limousin is polymorphic: individual follicular melanocytes contain both mature eumelanosomes and pheomelanosomes. Charolais and Chianina cattle exhibited a dramatic reduction in melanogenic activity, which was characterized by the almost exclusive presence of prephaoemelanosomes in Charolais and of immature premelanosomes in Chianina. In the dilute Charolais phenotype, the density of distribution of follicular melanocytes also seemed to be reduced. The genes that are responsible for these four phenotypes seem to act on the maturation, differentiation, and density of distribution of the melanosomes.     

Epidemiology of apple proliferation (AP) in northwestern Italy: evaluation of the frequency of AP-positive psyllids in naturally infected populations of Cacopsylla melanoneura (Homoptera: Psyllidae)

Adults of Cacopsylla melanoneura, vector of the apple proliferation (AP) phytoplasma, were collected every 2 weeks from January until May in 2000 and 2001 by the beating tray method in eight apple orchards of the Aosta Valley (northwestern Italy). Total DNA was extracted from batches of five insects and amplified with the universal phytoplasma primers P1/P7 in direct PCR. A nested PCR assay was then performed on P1/P7 amplicons using the primers fO1/rO1, specific for the AP‐ phytoplasma group. The digestion of fO1/rO1 amplicons with Ssp I restriction endonuclease confirmed that C. melanoneura adults harboured the AP phytoplasma. The data obtained with PCR were used to estimate the proportion of AP‐positive insects in over wintered and offspring adults. Percentages of AP‐positive insects of 3.6% and 0.8% were estimated in 2000 among over wintered and offspring psyllids respectively. In 2001 only the over wintered insects were found infected, with an estimated proportion of 2.8%. The seasonal abundance of the vector was measured using yellow sticky traps. C. melanoneura was always present at a low population level, and the highest density was recorded from mid‐February until mid‐March in both years. The results show that the overwintered population is higher and spends a longer period in apple orchards, suggesting the crucial role of the overwintered adults in vectoring AP.     

Plasma Membrane Glycoproteins of Ciona intestinalis Spermatozoa that Interact with the Egg1

In the ascidian Ciona intestinalis the species-specific interaction between the spermatozoon and the egg occurs between the vitelline coat (VC) of the egg and the plasma membrane of the apical part of the head of the spermatozoa. Concanavalin A (Con A)-binding sites are present on this area of the sperm surface. We used Con A to identify and isolate the spermatozoon plasma membrane components that may be involved in the interaction with the VC. These glycoproteins have been identified on SDS-PAGE of a sperm membrane fraction (SMF) enriched with the extermal proteins, after incubation of the gel with ³H-Con A. Affinity chromatography on Con A-agarose has been used for the purification of sperm plasma membrane proteins with and affinity for the lectin. The biological activity of the Con A-retained fraction was determined with binding and fertilization assays.     

Physiological and Biochemical Aspects of Symbiotic Nitrogen Fixation in Cowpea (Vigna unguiculata (L.) Walp. var. Tuy) Plants Infected by Cowpea Mosaic Virus

Three-day-old cowpea seedlings were inoculated with the severestrain of Cowpea Mosaic Virus (CpMV) and 24 h later with Bradyrhizobiumsp. cowpea, strain I-125, when virus translocation to rootsstill had not taken place. Plants were harvested at 22, 30,45 and 59 d after germination. Active virus replication wasassociated with increased protein content, detected in the leavesof 22-d-old plants. CpMV infection reduced the total leaf area,dry weight of shoots and the chlorophyll content of the firsttrifoliolate leaf at all experimental times. Low sugar contentwas recorded in leaves of 22- and 30-d-old CpMV-infected plantsand in nodules and roots of 30- and 45-d-old CpMV-infected plants.Up to 45 d, the nodule mass of CpMV-infected plants was lowerthan in controls, but reached control values at the 4th harvest.In CpMV-infected nodules, massive agglomeration of virus particles,crystalline virus inclusions and the proliferation of the endoplasmicreticulum were observed only in those cells containing bacteroids.In 30- and 45-d-old plants, CpMV infection decreased the contentof ureides in nodules, roots, and petioles. Virus infectiondid not alter the -amino-N content of roots and nodules butinduced a transient 74% reduction in the level of -amino-N inpetioles of 45-d-old plants. At the 1st and 2nd harvests theactivity of uricase (EC 1.7.3.3 [EC] ) in the nodules and of pyruvatekinase (EC 2.7.1.40 [EC] ) in the nodules and leaves were decreasedseverely by virus infection. CpMV did not hinder the allantoinase(EC 3.5.2.5 [EC] ) activity in the leaves but caused a 9% transitorydecrease in the activity of this enzyme in nodules of 45-d-oldplants. Measurements of NAD-malic enzyme (EC 1.1.1.38 [EC] ) in nodulesalso showed the non-effect of CpMV on this enzyme, except fora temporary 16% reduction at the 2nd harvest. As compared tocontrols, the relative abundance of ureides in 30-d-old CpMV-infectedplants indicated a 15%, 10%, and 51% reduction in the nodules,roots, and petioles, respectively. Results indicate that atthe time of the 4th harvest the symbiotic process, measuredin terms of ureide content and enzymatic activities, was functioningat a near normal level despite nodule infection by CpMV. Key words: Cowpea Mosaic Virus, nitrogen fixation, cowpea, enzymes, ultrastructure     

Leafminers help us understand leaf hydraulic design

Leaf hydraulics of Aesculus hippocastanum L. were measured over the growing season and during extensive leaf mining by the larvae of an invasive moth (Cameraria ohridella Deschka et Dimic) that specifically destroy the palisade tissue. Leaves showed seasonal changes in hydraulic resistance (R_lamina) which were related to ontogeny. After leaf expansion was complete, the hydraulic resistance of leaves and the partitioning of resistances between vascular and extra‐vascular compartments remained unchanged despite extensive disruption of the palisade by leafminers (up to 50%). This finding suggests that water flow from the petiole to the evaporation sites might not directly involve the palisade cells. The analysis of the temperature dependence of R_lamina in terms of Q₁₀ revealed that at least one transmembrane step was involved in water transport outside the leaf vasculature. Anatomical analysis suggested that this symplastic step may be located at the bundle sheath where the apoplast is interrupted by hydrophobic thickening of cell walls. Our findings offer some support to the view of a compartmentalization of leaves into well‐organized water pools so that the transpiration stream would involve veins, bundle sheath and spongy parenchyma, while the palisade tissue would be largely by‐passed with the possible advantage of protecting cells from short‐term fluctuations in water status.     

Responses of riparian trees and shrubs to flow regulation along a boreal stream in northern Sweden

1. Flow dynamics is a major determinant of riparian plant communities. Therefore, flow regulation may heavily affect riparian ecosystems. Despite the large number of dams worldwide, little specific information is available on the longitudinal impacts of dams on vegetation, for example how far downstream and at what degree of regulation a dam on a river can influence riparian woodlands. 2. We quantified the long‐term responses of riparian trees and shrubs to flow regulation by identifying their lateral distribution and habitat conditions along a boreal river in northern Sweden that has been regulated by a single dam since 1948. The regulation has reduced annual flow fluctuations, this effect being largest at the dam, downstream from which it progressively decreases following the entrance of free‐flowing tributaries. 3. We related changes in the distribution patterns, composition, abundance and richness of tree and shrub species to the degree of regulation along the river downstream from the dam. Regulation has triggered establishment of trees and shrubs closer to the channel, making it possible to measure ecological impacts of flow regulation as differences in vegetation attributes relative to the positions of tree and shrub communities established before and after regulation. 4. Trees and shrubs had migrated towards the mid‐channel along the entire study reach, but the changes were largest immediately downstream of the dam. Shrubs were most impacted by flow regulation in terms of lateral movement, but the effect on trees extended furthest downstream. 5. The species composition of trees progressively returned to its pre‐regulation state with distance downstream, but entrance of free‐flowing tributaries and variation in channel morphology and substratum caused local deviations. Species richness after regulation increased for trees but decreased for shrubs. The changes in species composition and richness of trees and shrubs showed no clear downstream patterns, suggesting that other factors than the degree of regulation were more important in governing life form.     

Two closely linked tomato HKT coding genes are positional candidates for the major tomato QTL involved in Na+/K+ homeostasis

The location of major quantitative trait loci (QTL) contributing to stem and leaf [Na⁺] and [K⁺] was previously reported in chromosome 7 using two connected populations of recombinant inbred lines (RILs) of tomato. HKT1;1 and HKT1;2, two tomato Na⁺‐selective class I‐HKT transporters, were found to be closely linked, where the maximum logarithm of odds (LOD) score for these QTLs located. When a chromosome 7 linkage map based on 278 single‐nucleotide polymorphisms (SNPs) was used, the maximum LOD score position was only 35 kb from HKT1;1 and HKT1;2. Their expression patterns and phenotypic effects were further investigated in two near‐isogenic lines (NILs): 157‐14 (double homozygote for the cheesmaniae alleles) and 157‐17 (double homozygote for the lycopersicum alleles). The expression pattern for the HKT1;1 and HKT1;2 alleles was complex, possibly because of differences in their promoter sequences. High salinity had very little effect on root dry and fresh weight and consequently on the plant dry weight of NIL 157‐14 in comparison with 157‐17. A significant difference between NILs was also found for [K⁺] and the [Na⁺]/[K⁺] ratio in leaf and stem but not for [Na⁺] arising a disagreement with the corresponding RIL population. Their association with leaf [Na⁺] and salt tolerance in tomato is also discussed.     

Lectin-Binding Sites Involved in Paramecium primaurelia Mating Pair Formation

ABSTRACT. Mating pair formation in Paramecium primaurelia was shown to be inhibited by incubating mating-competent mating type (mt) I and mt U cells with Limulus polyphemus agglutinin (LPA) or wheat germ agglutinin (WGA). Preincubation of LPA and WGA with their specific binding-monosaccharides, N-acetylneuraminic acid (NeuAc) and N-acetylglucosamine (GlcNAc), respectively, prevented the lectin effect on pair formation. Addition either of NeuAc or GlcNAc resulted in a reversal of cell pairing inhibition by LPA or WGA, respectively. Both NeuAc and GlcNAc monosaccharides inhibited pair formation when their concentration exceeded a threshold value. The pattern of the relative distribution of NeuAc and GlcNAc molecules on the cell surface was analyzed using fluorescence resonance energy transfer techniques combined with imaging systems. Mt I1 cells showed a high lectin-binding site density localized just on the surface region engaged in conjugation. The pattern of mt I cells, consisting of a quite homogeneous lectin-binding site density spread on the cell surface, was also common to nonmating-competent cells and to immature cells. These findings suggest that in P. primaurelia pair formation involves both NeuAc and GlcNAc residues and that the development of mating-competence is related to a modification in NeuAc and GlcNAc relative distribution on the cell surface of mt 11 cells.