High D-glucose does not affect binding of α-tocopherol to human erythrocytes

The role of -tocopherol uptake system in human erythrocyte in the uptake of plasma -tocopherol has been suggested. However no information is available on -tocopherol uptake activity of human erythrocytes in the presence of high levels of D-glucose which is known to lead to pathological alterations in different cells including human erythrocytes. Therefore, in order to examine the effect of D-glucose on the binding of -tocopherol to human erythrocytes, the binding characteristics of -tocopherol to these cells were established first. Binding of [3H]-tocopherol to human erythrocytes was both saturable and specific. Scatchard analysis of -tocopherol binding to these cells showed the presence of two independent classes of binding sites with widely different affinities. The high affinity binding sites had a dissociation constant (Kd1) of 90 nM with a binding capacity (n1) of 900 sites per cell, whereas the low affinity binding sites had a dissociation constant (Kd2) of 5.2 M and a binding capacity (n2) of 105,400 sites per cell. Trypsin treatment abolished all the -tocopherol binding activity. Competition for the binding of -tocopherol to human erythrocytes was effective with other homologues of -tocopherol (-tocopherol, -tocopherol and -tocopherol) and their potency was almost equal to -tocopherol itself. The order of preference was -tocopherol > -tocopherol -tocopherol -tocopherol. Incubation of human erythrocytes with various concentrations of D-glucose did not affect -tocopherol uptake activity. Our data demonstrate the presence of an -tocopherol uptake system in human erythrocytes and that the -tocopherol uptake activity is not modulated by the presence of D-glucose.     

Genetic polymorphism at the κ chain locus in mice: Comparisons of restriction enzyme hybridization fragments of variable and constant region genes

Variable (V) and constant (C) region genes of the mouse kappa light chain have been compared in inbred strains and in geographically isolated or genetically separated populations of mice by Southern blot analysis of endonuclease-restricted germline DNA. In most cases, the C gene is found on a single restriction fragment while the V genes of the V19 and V21 groups are each found on several (6–18) fragments. The restriction fragment (RF) patterns of V19 and V21 groups are both polymorphic when compared among inbred mouse strains. Southern blot patterns of V21 and V19 of inbred strains are also found among some geographically isolated populations of mice, suggesting that inbred strains acquired kappa loci from different subspecies. Some populations of geographical isolates show V21, V19, and C contexts similar to inbred mice while more distantly related species within the genus Mus and laboratory rats show no apparent similarity in context to inbred strains. Variable region genes determining the RF patterns of V19 and V21 appear to be linked to each other and to the C and Lyt-3 loci.     

Estuarine and mangrove systems and the nursery concept: which is which? The case of the Sine Saloum system (Senegal)

The mangrove system of Sine Saloum in Senegal is characterized by the lack of permanent river flow, in the context of the Sahelian drought which began in the 70s. The main environmental consequence is that Sine Saloum has become a so called reversed estuary with salinity increasing upstream and reaching 100 and more, with a mean salinity between 45–50. A threeyear survey of the juvenile fish community was undertaken with the aim of verifying whether or not this environment is still suitable as a nursery area for exploited fish populations. The main sampling gear used were small fykenets in addition to gill nets and a limited rotenone sampling. One of the six mangrove stations included in the survey exhibited a relatively high species diversity. This station is the only one where salinity may reach levels as low as 25 at the end of the rainy season, although salinity is much higher in the upstream region near this station. Such a low salinity is likely due to an underground freshwater connection or underwater springs. These observations highlight the relative importance of estuaries and mangroves in the nursery function.     

Dry miombo – ecology of its major plant species and their potential use as bio-indicators

Results of an extensive vegetation survey of 214 miombo woodland sitesare reported, encompassing the full spectrum of site types in the northernregion of Zimbabwe. At each sample site the vascular species (altogether640) were recorded, as well as 38 environmental variables (climatic,topographic, edaphic factors; geology, vegetation features, exploitation andland tenure). The sampling design was orientated along the environmentalgradients: altitude, slope, precipitation and geology. A reduced species set(286 species; frequency > 3%)was analysed by Canonical Correspondence Analysis (CCA), complementedby the Monte Carlo Permutation Test. The first axis represents acombination of the major environmental variables, altitude, precipitation andsoil texture, and the second – being of marginal importance – thePiriwiri and Lomagundi sediments. The matrix–consisting of the species and sites arranged according to the firstordination-axis – shows a diagonal feature embodying the moistureavailability gradient of the study area, subsequently divided into sevenordinalmoisture classes. The weighted averages of the species scores of therelevés – the scores being fixed by the membership of the singlespecies to one of the respective moisture classes – permits ranking thesites due to moisture availability, thereby assessing their land use capacity.     

Invertebrate Invasions on Pacific Islands and the Replacement of Unique Native Faunas: A Synthesis of the Land and Freshwater Snails*Contribution no. 2001-001 of Bishop Museum's Pacific Biological Survey.

The once immense diversity of native Pacific island land snail species, with high single island or archipelago endemism, is declining dramatically. The native/endemic species are being replaced by a much smaller number of widespread tropical tramps, that is, those species that are most readily transported by humans. The 82 introduced (including 14 cryptogenic) land snail species recorded include some that were distributed accidentally by Pacific islanders before European exploration of the Pacific and that are now widespread. However, the majority are modern introductions, with many recent accidental introductions often associated with the horticultural trade. Native freshwater faunas were less diverse than the terrestrial faunas and exhibited much lower endemism. Among the 59 alien freshwater species recorded (including 38 cryptogenic species), the most diverse and widespread are the thiarids. Predation by and competition with these aliens (as well as habitat loss) are probably important mechanisms underlying the loss of native taxa, but almost no quantitative or experimental work has been done to demonstrate such ecological interactions. Prevention of further spread and of new introductions should be the main approach. Increased public education and development of public trust is essential to the success of these efforts.     

Freshwater protozoa: biodiversity and ecological function

The purpose of this article is to pull together various elements from current knowledge regarding the natural history of free-living protozoa in fresh waters. We define their functional role, set the likely limits of biodiversity, and explore how the two may be related. Protozoa are unicellular, phagotrophic organisms, and 16 phyla of protists contain free-living freshwater protozoan species. They are the most important grazers of microbes in aquatic environments and the only grazers of any importance in anoxic habitats. In sediments, ciliates are usually the dominant protozoans. Benthic ciliate biomass accounts for slightly less than 10% of total benthic invertebrate biomass, but ciliate production may equal or even exceed invertebrate production. Freshwater protozoan species are probably ubiquitous, although many may persist locally for long periods in a cryptic state – as potential rather than active biodiversity. As protozoa are among the largest and most complex of micro-organisms, it follows that bacteria and all other smaller, more numerous microbes are also ubiquitous. The number of protozoan species recorded in local surveys (232) is about 10% of the estimated global species richness (2390). The 'seedbank of protozoan (and microbial) species ensures that local microbial diversity is never so impoverished that it cannot play its full part in ecosystem functions such as carbon fixation and nutrient cycling.     

Specialized and generalized dispersal systems: where does ‘the paradigm’ stand?

I explore the specialization versus generalization paradigm in frugivory and seed dispersal. This view predicts that some tropical trees produce nutritious fruits adapted for use by a small coterie of specialized frugivores that provide reliable seed dissemination. Other tree species are expected to offer superabundant fruits of lower nutritional reward, relying instead on common opportunistic frugivores that are individually less reliable, but collectively disperse seeds effectively. Though widely referenced, many aspects of the paradigm are untested with tropical trees and avian frugivores, primarily because plant ecologists rarely determine whether specialist or generalist foragers are responsible for different patterns of seed distribution, while students of foraging behavior rarely determine the effects seed dispersal by different animals for survival of seeds or seedlings of specialist or generalist trees.Ecological paradigms provide alternative hypotheses, without evolutionary arguments. Keystone species have ecological effects disproportionate to their abundance; it is important for management considerations to know whether fruiting trees or frugivores serve as keystone mutualists in tropical forests. Alternatively, the extent to which vertebrate seed dispersers influence density-dependent seed, seedling, sapling, or adult mortality may have important consequences for spatial dispersion and population dynamics of tree species in tropical forests.     

Two novel isoneolacto-undecaglycosylceramides carrying Galα1→3Lewisx on the 6-linked antenna and N-acetylneuraminic acidα2→3 or Galactose α1→3 on the 3-linked antenna, expressed in porcine kidney

Three sialosylated and three neutral glycosphingolipids sharing a common iso-neolacto core were isolated from porcine kidney cortex. They were purified by preparative HPTLC, and were characterized by partial exoglycosidase hydrolysis followed by thin layer chromatography and immunostaining with anti-Gal13Gal, anti-type 2 lactosamine and anti-Lewisx antibodies, methylation analysis, MALDI-TOF mass spectrometry and 1H-NMR spectroscopy. Among neutral glycolipids, one was a known structure, VI3VI3(Gal)2-iso-nLc8Cer, and two were novel structures differing by the number of Gal3Lewisx determinants: VI3VI3(Gal)2V3Fuc-iso-nLc8, and VI3VI3(Gal)2 V3V3(Fuc)2-iso-nLc8. The single Gal3Lewis x determinant was found on the 6-linked antenna. Among sialosylated glycolipids, two had been previously found in other species and tissues, VI3VI3(NeuAc)2-iso-nLc8, and VI3NeuAcVI3Gal-iso-nLc8. A novel structure was discovered presenting a Gal3Lewisx determinant on the 6-linked antenna and a N-acetylneuraminic acid on the 3-linked antenna, VI3NeuAcVI3GalV3Fuc-iso-nLc8. These results indicate that, in vivo, the porcine kidney 3fucosyltransferase synthesizes the Gal3Lewisx determinant, acting on the 6-linked before the 3-linked Gal3neolactosamine, and appears unable to synthesize the sialosylated Lewisx determinant on neolactoseries glycolipids.     

Variation in the karyotype of three cultivars of Narcissus tazetta L. (Amaryllidaceae)

Three cultivated varieties of Narcissus tazetta, Grand Soleil d'Or, Chinese Sacred Lily and Cypri, are triploid (2n = 3x+30) with the basic number 10. Grand Soleil d'Or has three homomorphic sets, each comprising 2 long submetacentrics, 4 long acrocentrics and 4 short acrocentrics. Karyotypes of the other two varieties are heteromorphic. Both possess one telocentric satellited chromosome. In addition, Cypri shows translocation between two chromosomes belonging to the seventh and eighth triplets. The number of secondary constrictions varies between 3 (Chinese Sacred Lily) and 4 (Grand Soleil d'Or and Cypri) which is also the number of nucleoli observed in the respective varieties.     

Nuclear Overhauser effect investigation on GM1 ganglioside containingN-glycolyl-neuraminic acid (II3Neu5GcGgOse4Cer)

The conformational properties of the oligosaccharide chain of GM1 ganglioside containingN-glycolyl-neuraminic acid, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer, were studied through NMR nuclear Overhauser effect investigations on the monomeric ganglioside in dimethylsulfoxide, and on mixed micelles of ganglioside and dodecylphosphocholine in water. Several interresidual contacts for the trisaccharide core--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-were found to fix the relative orientitation of the three saccharides, while the glycosidic linkage of the terminal -Gal-was found to be quite mobile as the -Gal-(1-3)--GalNAc-disaccharide exists in different conformations. These results are similar to those found for two GM1 gangliosides containingN-acetyl-neuraminic acid and neuraminic acid [1].Abbreviations Ganglioside nomenclature is in accordance with Svennerholm [23] and the IUPAC-IUB Recommendations [24] GM3(Neu5Ac) II³Neu5AcLacCer, -Neu5Ac-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM3(Neu5Gc) II³Neu5GcLacCer, -Neu5Gc-(2-3)--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Ac) II³Neu5AcGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu5Gc) II³Neu5GcGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu5Gc-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GM1(Neu) II³NeuGgOse₄Cer, -Gal-(1-3)--GalNAc-(1-4)-[-Neu-(2-3)]--Glc-(1-1)-Cer - GD1a IV³Neu5AcII³Neu5AcGgOse₄Cer, -Neu5Ac-(2-3)--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - GalNAc-GD1a IV⁴GalNAcIV³Neu5AcII³Neu5AcGgOse₄Cer, -GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-3)--GalNAc-(1-4)-[-Neu5Ac-(2-3)]--Gal-(1-4)--Glc-(1-1)-Cer - Neu neuraminic acid - Neu5Ac N-acetyl-neuraminic acid - Neu5Gc N-glycolyl-neuraminic acid - Cer ceramide     

Enzymatic regioselective synthesis of sucrose acrylate esters

Using a protease (at 100 g l^–1) from Bacillus licheniformis, enzymatic acryloylation of sucrose (1 M) with vinyl acrylate (4 M) was carried out in anhydrous pyridine and yielded sucrose acrylate esters with more than 90% of sucrose converted in 24 h. After 5 days of reaction, the ester products consisted of 70% sucrose monoacrylate and 30% sucrose diacrylate. The monoester product was a sucrose 1-acrylate and the diester products consisted of sucrose 6,1-diacrylate and sucrose 6,1-diacrylate in the ratio of 3:2.     

Molecular mapping of a new gene Rrs4 CI 11549 for resistance to barley scald (Rhynchosporium secalis)

Doubled haploid (DH) progeny from a cross between the scald susceptible barley (Hordeum vulgare L.) cultivar Ingrid and the resistant accession CI 11549 (Nigrinudum) was evaluated for resistance in the pathogen Rhynchosporium secalis (Oudem) J.J. Davis. Two linked and incompletely dominant loci confer resistance CI 11549 against isolate 4004. One is an allele at the complex Rrs1 locus on chromosome 3H close to the centromere; the other is located 22 cM distally on the long arm. The latter locus is designated Rrs4. In BC₃-lines into Ingrid from CI 2222 (another Nigrinudum) resistance seems governed by one locus close to the telomeric region of chromosome 7H, probably allelic to Rrs2. In neither case did we find any trace of the recessive gene rh8 reported to be present in Nigrinudum. Various resistance donors of Ethiopian origin designated as Nigrinudum, Jet or Abyssinian were identical to a great extent with respect to markers, but differed in resistance to different isolates of scald or in barley yellow dwarf virus (BYDV) resistance. The implications for their use as differentials in scald tests and screening of germplasm collections are discussed.     

A survey of the response of Rhododendron to in vitro culture

The responses of 7 genotypes of Rhodendron to culture conditions and their establishment as shoot cultures are described. The genotypes represent a broad genetic diversity in the genus. After sterilization and an acclimation period of 3 to 12 months, all the selections were established as shoot cultures on Woody Plant Medium (WPM) supplemented with N⁶(-²-isopenteny) adenine (2iP). Plants with strong episodic growth cycles required the longer acclimation periods. Utilizing shoots from these cultures, the response to a cytokinin series of 0 to 32 M 2iP or BAP (6-Benzylaminopurine) was analyzed. BAP proved toxic to all but the elipidote and lepidote rhododendrons (R. mucronulatum, R. x Boule de Neige, and R. x PJM); however, even with these selections, 2iP stimulated greater shoot multiplication rates. The optimum 2iP level for shoot multiplication varied little with the genotype and levels of 4 to 16 M generally proved optimal, depending on the specific selection. Adventitious shoot production was observed in 3 selections (R. canadense, R. x Boule de Neige and R. x PJM), but only at 2iP levels above 8 M. Shoot multiplication rates of 7 to 21 times were observed, depending on the selection. Using an average utilizable shoot production rate of 40 shoots per culture per 6 week subculture period, some 75,000 shoots can be generated per square meter of culture space per year. The harvested shoots (microcuttings) rooted readily out-of-culture and the resultant plants grew like seedlings.     

Identification and characterization of the functional alpha origin of DNA replication of the R6K plasmid and its relatedness to the R6K beta and gamma origins

Summary The functional R6K origin is composed of two DNA elements, one of 580 bp carrying the origin sequences and the other of 277 bp containing the seven 22 bp direct repeats previosly identified as also required for and origin activity. These two genetic elements are separated by approximately 3,000 bp of R6K sequences which are dispensable for origin activity. The function of the origin depends on the presence in cis of the 580 bp and the 277 bp fragments and requires that they be oriented as in the intact R6K. Activation of the origin depends on the R6K replication initiation protein .Within the 580 bp of the origin, there is a sequence of 98 bp which appears as an inverted repeat of 96 bp in the replicon. Deletion of the 96 bp or 98 bp results in inactivation of the and the origins respectively. These long repeats are palindromic and it is suggested that these may serve as the recognition signals for initiation of DNA replication in the and the origins of R6K. DNA homology analysis performed on , and origin sequences, also reveals 10–23 bp sequences in the and the origins that are related to the family of 22 bp direct repeats in the origin which were shown previously to be binding sites for the protein.     

Postmitotic ‘isodiametric’ cell growth in the maize root apex

The onset of rapid cell elongation occurred at different distances from the apex in various tissues of the primary root of maize (Zea mays L.). Furthermore, the comparison of these distances with those determined for the cessation of mitotic divisions revealed a considerable discrepancy. The onset of rapid cell elongation was realized much farther from the root apex than the cessation of cell divisions and therefore a distinct region could be distinguished in every examined maize root tissue. This region was denoted the region of postmitotic isodiametric cell growth. Cells in this region grew in width as well as in length and obtained approximately a square-isodiametric shape. They were also characterized, as are cells in the meristem, by intense nucleic-acid metabolism. This prominent postmitotic isodiametric cell growth was observed in both polyploid and diploid tissues, and indicates that postmitotic isodiametric cell growth, like mitotic division and cell elongation growth, represents an important developmental stage in plant cell ontogeny.The authors dedicate this paper to Dr. M. Luxová on the occasion of her 65th birthday     

Expression of a chimeric stilbene synthase gene in transgenic wheat lines

A chimeric stilbene synthase (sts)gene was transferred into wheat. Stilbene synthases play a role in the defence against fungal diseases in some plant species (e.g. groundnut or grapevine) by producing stilbenetype phytoalexins like resveratrol. Resveratrol is also claimed to have positive effects to human health. Embryogenic scutellar calli derived from immature embryos of the two commercial German spring wheat cultivars Combi and Hanno were used as target tissue for cotransformation by microprojectile delivery. The selectable marker/reporter gene constructs contained the bargene either driven by the ubiquitinpromoter from maize (pAHC 25, also containing the uidAgene driven by the ubiquitinpromoter), or by the actinpromoter (pDM 302) from rice. The cotransferred plasmid pStil 2 consisted of a grapevine stscoding region driven by the ubiquitin promoter. Eight transgenic Combi and one Hanno T_Oplant were obtained and, except one Combi T_Oplant, found to be cotransformants due to the integration of both the stsgene and the selectable marker or reporter genes. Expression of the stsgene was proven by RTPCR, and, for the first time, by detection of the stilbene synthase product resveratrol by HPLC and mass spectrometry. The stsgene was expressed in four of the seven transgenic Combi T_{_o}plants. Two of the respective T₁progenies segregated in a Mendelian manner were still expressing the gene. Investigations into methylation of the stsgene showed that in three nonexpressing progenies inactivation was paralleled by methylation.     

Average phase difference theory and 1∶1 phase entrainment in interlimb coordination

The dynamics of coupled biological oscillators can be modeled by averaging the effects of coupling over each oscillatory cycle so that the coupling depends on the phase difference between the two oscillators and not on their specific states. Average phase difference theory claims that mode locking phenomena can be predicted by the average effects of the coupling influences. As a starting point for both empirical and theoretical investigations, Rand et al. (1988) have proposed d/dt= — K sin ), with phase-locked solutions =arcsin( /K), where is the difference between the uncoupled frequencies and K is the coupling strength. Phase-locking was evaluated in three experiments using an interlimb coordination paradigm in which a person oscillates hand-held pendulums. was controlled through length differences in the left and right pendulums. The coupled frequency _c was varied by a metronome, and scaled to the eigenfrequency _v of the coupled system K was assumed to vary inversely with _c. The results indicate that: (1) and K contribute multiplicatively to (2) =0 or = regardless of K when =0; (3) 0 or regardless of when K is large (relative to ); (4) results (1) to (3) hold identically for both in phase and antiphase coordination. The results also indicate that the relevant frequency is _c/_v rather than _c. Discussion high-lighted the significance of confirming =arcsin(/K) for more general treatments of phase-locking, such as circle map dynamics, and for the 11 phase-entrainment which characterizes biological movement systems.     

Molecular characterization of β-thalassemia in Czechoslovakia

Summary We have identified different -thalassemia mutations in 93 members of 34 families of Czech or Slovakian descent using gene amplification, hybridization with specific ³²P-labeled oligonucleotide probes, sequencing of amplified DNA, and gene mapping. The GA mutation at IVS-I-1 was found in 18 families; other Mediterranean mutations were IVS-II-1 (GA), IVS-II-745 (CG), IVS-I-110 (GA), and codon 39 (CT); these were present in 9 additional families. The GT mutation at codon 121, known to cause Heinzbody -thalassemia, was present in 3 families, and the frameshift at codons 82/83 (-G), first described in the Azerbaijanian population, in 2 families. A newly discovered allele was a frameshift at codons 38/39 (-C). One -thalassemia allele was incompletely characterized. We observed in 2 families a TC mutation at position +96 UTR (untranslated region) relative to the termination codon; this mutation likely is a rare polymorphism, -Thalassemia was rare; only one person carried the -^3.7 heterozygosity, and one other had a yet to be identified -thalassemia-1, while seven had the ^{anti 3.7} triplication.     

Capacity of dispersal of a colonizing species: Drosophila subobscura

The European species Drosophila subobscura has colonized, in a very short time, large areas in South America (Chile and Argentina) and North America (The Pacific Coast, from Vancouver to Southern California), where it has been established as a very common species. Its capacity of dispersal was analyzed and results compared with the few data available from other authors. The estimates of the mean distance travelled during one activity period (159.20 meters) are considerably higher than those obtained by other authors. The Brownian motion model seems most probable because the values of DF and BM are very much alike. The effects on the estimates of some environmental factors, such as wind, are also discussed.     

Phosphoglucoisomerase variation in Trifolium repens

An electrophoretic survey carried out at the PGI/2 locus of white clover revealed the existence of five alleles. These alleles were shown to be controlled by single co-dominant genes, behaving in a normal Mendelian manner. The results also indicate that white clover behaves as an amphidiploid, in which one of its parental species is normally homozygous for the c allele at the PGI/2 locus, whilst the other parent has a multi-allelic series at this locus.