Rumen microbes and fermentation of wild sika deer on the Shiretoko peninsula of Hokkaido Island,Japan

A total of 32 wild Hokkaido sika deer (Cervus nippon yesoensis) were shot (13 in summer, nine in autumn and 10 in winter) in the Syari district, Shiretoko Peninsula of Hokkaido Island, Japan. The ingested foods, rumen fermentation parameters and microbes were determined to evaluate digestive strategy and food availability in each season. Ingested foods and ruminal characteristics greatly varied by season. Rumen digesta mainly comprised of graminoids in summer, graminoids and agricultural products in autumn, and bark and twigs in winter. Rumen pH showed seasonal differences (P<0.05) and was lowest in summer, highest in winter, and intermediate in autumn, reflecting the seasonal differences in ruminal concentration of total volatile fatty acids which were significantly lower (P<0.05) in winter than in summer and autumn. Acetate proportions were significantly higher in winter than in other seasons (P<0.05), while the opposite trend was seen in proportions of propionate and butyrate. Rumen ammonia levels showed significant seasonal differences (P<0.05), decreasing from summer to autumn to winter. Rumen protozoa levels in autumn and winter decreased to 28 and 10% of the levels observed in summer, respectively (P<0.05 for both). The rumen bacteria level in winter was lower (P<0.05) than that in autumn, but no difference was seen for the other seasonal comparisons. Gram negative cocci were present in significantly higher proportions in winter than in other seasons (P<0.05), while Gram negative curved rods were less frequently observed in winter (P<0.05). Based on these results, wild sika deer in this area are shown to survive with rumen microbial populations altered with the dietary conditions that vary greatly by season.     

Monitoring the mitochondrial transmembrane potential with the JC-1 fluorochrome in programmed cell death during mesophyll leaf senescence

Summary. Analysis of the mitochondrial transmembrane potential (_m) with the help of the JC-1 fluorochrome (5,5,6,6-tetrachloro-1,1,3,3-tetraethylbenzimidazolcarbocyanine iodide) during mesophyll leaf senescence was performed in order to determine whether a reduction of _m takes place during mesophyll senescence and whether plant mitochondria, like mammalian ones, might be involved in the induction of programmed cell death. Fluorescence analysis of mesophyll protoplasts of Pisum sativum in a confocal microscope, fluorescent spectra analysis and time dependence of fluorescence intensity of monomers and of J-aggregates revealed that JC-1 is incorporated and accumulated specifically in plant mitochondria. Analysis of _m during mesophyll protoplast senescence revealed that two subpopulations of mitochondria which differ in _m exist in all analyzed stages of leaf senescence. The first subpopulation contains mitochondria with red fluorescence of J-aggregates due to an unperturbed high _m. The second subpopulation comprises mitochondria with green fluorescence of monomers due to a low _m, proving total depolarization of mitochondrial membranes. Fluorescence analysis demonstrated that even in the latest analyzed stages of leaf senescence, mitochondria with a high _m still exist. Fluorometric measurements revealed that the fluorescence intensity of J-aggregates decreases with the age of plants, which indicates that a reduction of _m during the mesophyll senescence process takes place; however, it does not take place within the whole population of mitochondria of the same protoplast. The reason of this can be due to a dramatic reorganization of mitochondria in mesophyll cells and the appearance of large mitochondria with local heterogeneity of _m in the oldest analyzed stages. All mitochondria in every stage of senescence maintained their membrane organization even when their size, distribution, and spatial organization in protoplasts changed dramatically. We stated that the reduction of _m does not directly induce programmed cell death in mesophyll cells, as opposed to animal apoptosis.Correspondence and reprints: Department of Plant Anatomy and Cytology, Institute of Experimental Biology of Plants, Warsaw University, Miecznikowa 1, 02-096 Warszawa, Poland.     

Sterol composition of Candida curvata yeast

Summary Six sterols were extracted from lipids produced by Candida curvata yeast. Ergosterol (main component) and five minor compounds: ergosta-7, 24(28)-dien-3 -ol, ergosta-5, 7-dien-3 -ol, ergosta-5, 7, 9, (11), 22-tetraen-3 -ol, ergosta 7, 22-dien-3 -ol, ergost-7-en-3 -ol were identified by gas chromafography/mass spectrometry (GCMS) and quantitated by GLC.     

Enhancement of somatic embryogenesis in orchardgrass leaf cultures by epinephrine

Epinephrine at 10–100 M stimulated somatic embryogenesis from orchardgrass (Dactylis glomerata L.) leaves cultured on SH medium with 30 M of indole-3-acetic acid (IAA). Ethylene emanation was increased at epinephrine concentrations greater than 10 M. Decarboxylation by the leaves of [1-¹⁴C]IAA included in the medium was decreased almost 3-fold by 10 M epinephrine. Epinephrine at 10 M enhanced the number of regenerated plants on SH medium with 30 M dicamba (SH-30). Ethylene emanation was increased by epinephrine concentrations of 500 M and greater included in SH-30 but somatic embryogenesis was decreased. Addition of 8 M CoCl₂, 6H₂O (an ethylene biosynthesis inhibitor) to medium with 500 M epinephrine decreased ethylene emanation to the control level but did not alleviate the decreased embryogenic response.     

Long-term shoot regeneration from pampas grass (Cortaderia selloana Schult.) through manipulation of growth regulators in vitro

A regeneration system for Pumila pampas grass (Cortaderia selloana Schult.) that yields plants over many months and allows control of morphogenesis was developed. Immature inflorescence explants cultured for three 4-week passages on MS basal medium supplemented with 4.5 M 2,4-D and 8.9 M BA yielded a dark green callus that organized into shoot primordia. Rate of shoot development was increased after transfer of shoot primordia to medium supplemented with 9.8 M IBA. Subculture every 4–6 weeks onto medium containing IBA yielded a continuous production of shoots. Control of morphogenesis was achieved by transferring cultures to medium containing 4.5 M 2,4-D and 8.9 M BA for shoot bud proliferation and to medium containing IBA for shoot production.Abbreviations BA 6-benzylaminopurine - 2,4-D 2,4-dichlorophenoxyacetic acid - IBA indole-3-butyricacid - MS Murashige and Skoog (1962)     

Characterization of two different Ca2+ uptake and IP3-sensitive Ca2+ release mechanisms in microsomal Ca2+ pools of rat pancreatic acinar cells

We have examined the effect of the Ca²⁺ (Mg²⁺)-ATPase inhibitors thapsigargin (TG) and vanadate on ATP-dependent ⁴⁵Ca²⁺ uptake into IP₃-sensitive Ca²⁺ pools in isolated microsomes from rat pancreatic acinar cells. The inhibitory effect of TG was biphasic. About 40–50% of total Ca²⁺ uptake was inhibited by TG up to 10 nm (apparent K_i4.2 nm, Ca²⁺ pool I). An additional increase of inhibition up to 85–90% of total Ca²⁺ uptake could be achieved at 15 to 20 nm of TG (apparent K_i12.1 nm, Ca²⁺ pool II). The rest was due to TG-insensitive contaminating plasma membranes and could be inhibited by vanadate (apparent K_i10 m). In the absence of TG, increasing concentrations of vanadate also showed two phases of inhibition of microsomal Ca²⁺ uptake. About 30–40% of total Ca²⁺ uptake was inhibited by 100 m of vanadate (apparent K_i18 m, Ca²⁺ pool II). The remaining 60–70% could be inhibited either by vanadate at concentrations up to 1 mm (apparent K_i300 m) or by TG up to 10 nm (Ca²⁺ pool I). The amount of IP₃-induced Ca²⁺ release was constant at 25% over a wide range of Ca²⁺ filling. About 10–20% remained unreleasable by IP₃. Reduction of IP₃ releasable Ca²⁺ in the presence of inhibitors showed similar dose-response curves as Ca²⁺ uptake (apparent K_i 3.0 nm for IP₃-induced Ca²⁺ release as compared to 4.2 nm for Ca²⁺ uptake at TG up to 10 nm) indicating that the highly TG-sensitive Ca²⁺ pump fills the IP₃-sensitive Ca²⁺ pool I. At TG concentrations >10 nm which blocked Ca²⁺ pool II the apparent K_i values were 11.3 and 12.1 nm, respectively. For inhibition by vanadate up to 100 m the apparent K_i values were 18 m for Ca²⁺ uptake and 7 m for Ca²⁺ release (Ca²⁺ pool II). At vanadate concentrations up to 1 mm the apparent K_i values were 300 and 200 m, respectively (Ca²⁺ pool I). Both Ca²⁺ pools I and II also showed different sensitivities to IP₃. Dose-response curves for IP₃ in the absence of inhibitors (control) showed an apparent K_m value for IP₃ at 0.6 m. In the presence of TG (inhibition of Ca²⁺ pool I) the curve was shifted to the left with an apparent K_m for IP₃ at 0.08 m. In the presence of vanadate (inhibition of Ca²⁺ pool II), the apparent K_m for IP₃ was 2.1 m. These data allow the conclusion that there are at least three different Ca²⁺ uptake mechanisms present in pancreatic acinar cells: TG- and IP₃ insensitive but highly vanadate-sensitive Ca²⁺ uptake occurs into membrane vesicles derived from plasma membranes. Two Ca²⁺ pools with different TG-, vanadate- and IP₃-sensitivities are most likely located in the endoplasmic reticulum at different cell sites, which could have functional implications for hormonal stimulation of pancreatic acinar cells.This work was supported by the Deutsche Forschungsgemeinschaft, Sonderforschungsbereich 246. The authors wish to thank Dr. KlausDieter Preuß for valuable discussions and Mrs. Gabriele Mörschbächer for excellent secretarial help.     

Single mid-chain GlcNAcβ1-6Galβ1-4R sequences of linear oligosaccharides are resistant to endo-β-galactosidase ofBacteroides fragilis

Endo--galactosidase (EC 3.2.1.103) ofBacteroides fragilis, at 250 mU ml^–1, did not cleave the internal galactosidic linkage of the linear radiolabelled trisaccharide GlcNAc1-6Gal1-4GlcNAc, or those of the tetrasaccharides Gal1-4GlcNAc1-6Gal1-4GlcNAc and Gal1-4GlcNAc1-6Gal1-4Glc. The isomeric glycans which contained the GlcNAc1-3Gal1-4GlcNAc/Glc sequence were readily cleaved.Abbreviations GlcNAc 2-acetamido-2-deoxy-d-glucose - Lact lactose - MT maltotriose - MTet maltotetraose - R _MTet chromatographic migration rate in relation to that of maltotetraose     

The influence of ethanol on the functional status of GABA(A) receptors

Gamma aminobutyric acid (GABA) is one of the main inhibitory neurotransmitters in the mammalian brain. Its effects are realized via GABA_A, GABA_B, and GABA_C receptors. GABA_A is the most abundant type of GABA receptors. It consists of six classes of subunits, , , , , , and . Acute and chronic exposures to ethanol are accompanied by changes in structure and function of GABA_A receptors. These changes may be a basis for altered behavior seen in alcoholism.     

Stem distribution and clonal structure of Chamaecyparis pisifera growing in an old-growth beech-conifer forest

To elucidate sapling patch formation and the mode of regeneration occurring in Chamaecyparis pisifera, we investigated stem distribution and clonal structure of this species in an old-growth beech-conifer forest in central Japan. The study was carried out in a 1-ha plot using small and large sapling (d.b.h.<5cm, 0.5mheight<2 m and d.b.h.<5cm, height2m, respectively) and adult (d.b.h.5cm) stems of Chamaecyparis pisifera. The numbers of small and large sapling stems were 677 and 149 in the plot, respectively, and they were strongly aggregated. Adult stems occurred at much lower density (90 stems in the plot) and were weakly clustered. Fifty multilocus genotypes at nine allozyme loci were detected among sampled stems. Small and large sapling stems with identical genotypes showed aggregated distribution, while some neighboring adult stems had the same genotypes. Spatial autocorrelation of alleles in small and large sapling stems revealed a strongly positive association among stems within a distance of 10m; but a negative association as distance increased. In adult stems, a strongly positive value was found only in the shortest distance class. These results indicate that clonal growth by layering occurred at the sapling stage and contributed to patch formation. Our results also suggest that the clonal growth by layering is ecologically significant throughout the life history of Chamaecyparis pisifera, particularly as an important strategy for the sapling establishment in the early stage of regeneration process.     

The cytology of phytophthora infestans

Summary Phytophthora infestans has three kinds of somatic nuclei: an oval shaped nucleus (approx. 3.1×2.7 ) which stains diffusely except for a crescent shaped Feulgen positive cap which stains intensely; a granular nucleus whose contents are organized into a fairly constant number of stained bodies, and, a deeply staining condensed nucleus. The capped nucleus is thought to be metabolic or resting and the granular nucleus is thought to be dividing as it is most commonly found in hyphal tips. Attenuated forms of all three kinds of nuclei are found.Nuclear division is mitotic and intranuclear. Eight—ten chromosomes are seen at metaphase.Sporangia have a mean of 6.3 nuclei which is constant for age and strain of culture. Sporangia become multinucleate as a result of nuclear migration and not by division in the developing sporangium. Zoospores are usually uninucleate.The nuclear cap is persistent throughout nuclear division when it also divides. It is associated with flagella production and nuclear migration and has some of the properties of a blepharoplast.     

Evidence on the nature and origins of endosperm dosage requirements in Solanum and other angiosperm genera

Success of seed development following sexual crosses is primarily dependent on proper endosperm function and development. The failure to produce triploids, or triploid block in 4x×2x crosses served as the impetus for numerous studies of embryo and endosperm to attempt to explain cross failure. Early explanations were based upon a concept of a 232 ploidy balance between maternal tissue, endosperm, and embryo. Subsequent studies done with maize demonstrated that normal endosperm development in intraspecific maize crosses is dependent solely on having a 21 maternal to paternal genome dosage in the endosperm. These results have been modified and extended to solanaceous species in the form of an endosperm dosage system in which empirically determined factors must bear the same 21 relationship for crosses to succeed. Crossing behavior of these species suggest that the system is polygenically controlled and regulates both interspecific and intraspecific crosses. Endosperm dosage systems explain many aspects of species evolution, but the system appears to have originated as an ancient means of ensuring diploid fidelity.     

Histochemical studies on the distribution of β-glucuronidase and succinic dehydrogenase in young and old spinal ganglion cells of the rat

Summary The present study compares the distribution of -glucuronidase and succinic dehydrogenase in young and old spinal ganglion cells of rat. In young cells there are indications of cyclic activity of these enzymes, i.e., in some stages there are perinuclear concentrations of the enzymes, at other times -glucuronidase and succinic dehydrogenase are uniformly distributed throughout the cytoplasm. These stages have been discussed with the identical distribution of mitochondria. However, in old spinal ganglion cells both -glucuronidase and succinic dehydrogenase become mainly concentrated in the pigment areas, suggesting thereby their possible role in the production of pigment, through the medium of the mitochondria.     

Changes in density of chromatin in the meristematic cells ofSinapis alba during transition to flowering

Summary Changes in the density of nuclear chromatin in the shoot apical meristem ofSinapis alba L. during floral transition (floral evocation) are described using Feulgen-stained 2 m thick semi-thin sections and scanning cytophotometric techniques. In both G1 and G2 nuclei the chromatin becomes less heterogeneous and less dense in evoked meristems compared to vegetative meristems. When chromatin is resolved into two fractions the dispersed fraction increases relative to the condensed fraction at evocation. This decondensation process occurs earlier in G1 than in G 2 nuclei. These chromatin changes are presumably closely related to the dramatic stimulation of biosynthetic activity and cell division during floral transition.     

The solution structure of rat Aβ-(1–28) and its interaction with zinc ion: insights into the scarcity of amyloid deposition in aged rat brain

The amyloid -peptide (A) is a major component of insoluble amyloid deposits in Alzheimers disease, and the ability of the -peptide to exist in different conformations is dependent on residues 1–28 [-(1–28)]. However, different from humans, no A amyloid deposition has been found in aged rats brains. Studying the three-dimensional solution structure of rat A-(1–28) and the binding circumstance of Zn²⁺ is beneficial to a clear understanding of the potential role of Zn²⁺ in Alzheimer-associated neuropathogenesis and to suggest why there is no amyloid deposition in aged rats brains. Here we used nuclear magnetic resonance (NMR) spectroscopy to determine the solution structure of rat A-(1–28) and the binding constant of Zn²⁺ to rat A-(1–28). Our results suggest that (1) the three-dimensional solution structure of rat A-(1–28) is more stable than that of human A-(1–28) in DMSO-d₆ and that a helical region from Glu16 to Val24 exists in the rat A-(1–28); (2) the affinity of Zn²⁺ for rat A-(1–28) is lower than that for human A-(1–28) and the NMR data suggest that Arg13, His6, and His14 residues provide the primary binding sites for Zn²⁺; and (3) the proper binding of Zn²⁺ to rat A-(1–28) can induce the peptide to change to a more stable conformation.Electronic Supplementary Material Supplementary material is available in the online version of this article at http://dx.doi.org/10.1007/s00775-004-0556-xAbbreviations A amyloid -peptide - AD Alzheimers disease - hA-(1–28) human A-(1–28) - rA-(1-28) rat A-(1–28) - REM restrained energy minimization     

Effects of high ambient temperatures on the metabolism of West African dwarf goats. I

Thirty-two West African dwarf goats were exposed to temperature treatments: 20, 25, 30, 35, 35, 35, 30, 25, 20°C each lasting three days.Sixteen goats were kept in individual pens (I), the others in two group pens (G). Heat production (HP) and activity were recorded during 48 hours in each temperature treatment.Mean HP and ME intake were similar for G and I animals, but I animals had lower values than G animals at low chamber temperatures and higher values than G animals at high temperatures. Upper critical temperature was between 25°C and 30°C under an increasing T and between 30 and 35°C under a decreasing T. Adaptation of heat production and ME intake to a change in temperature of 5°C required at least six and possibly more than nine days.Diurnal variation in HP was large, up to 44% between extremes. This was largely due to variation in activity.     

Homocysteine stimulates the expression of monocyte chemoattractant protein-1 in endothelial cells leading to enhanced monocyte chemotaxis

The effect of intraperitoneal administration of tocopherol (100 mg/kg wt/24 h) on ascorbate (0.4 mM) induced lipid peroxidation of mitochondria and microsomes isolated from rat liver and testis was studied. Special attention was paid to the changes produced on the highly polyunsaturated fatty acids C20:4 n6 and C22:6 n3 in liver and C20:4 n6 and C22:5 n6 in testis. The lipid peroxidation of liver mitochondria or microsomes produced a significant decrease of C20:4 n6 and C22:6 n3 in the control group, whereas changes in the fatty acid composition of the tocopherol treated group were not observed. The light emission was significantly higher in the control than in the tocopherol treated group. The lipid peroxidation of testis microsomes isolated from the tocopherol group produced a significant decrease of C20:4 n6 , C22:5 n6 and C22:6 n3, these changes were not observed in testis mitochondria. The light emission of both groups was similar. The treatment with tocopherol at the dose and times indicated showed a protector effect on the polyunsaturated fatty acids of liver mitochondria, microsomes and testis mitochondria, whereas those fatty acids situated in testis microsomes were not protected during non enzymatic ascorbateFe²⁺ lipid peroxidation. The protector effect observed by tocopherol treatment in the fatty acid composition of rat testis mitochondria but not in microsomes could be explained if we consider that the sum of C20:4 n6 + C22:5 n6 in testis microsomes is 2-fold than that present in mitochondria.     

Summer forage biomass and the importance of litterfall for a high-density sika deer population

The biomass of summer forage and their contributions were surveyed to show that litterfall supported a high-density population of sika deer (Cervus nippon Temminck) in summer on Nakanoshima Island, Toya Lake, Japan. In July 1974, the grassland had the highest productivity among understory vegetations (228±55kgha^–1: mean±SE). In deciduous forests, palatable plants occupied only 0.1% of the biomass of 0.872±0.366kgha^–1, and deciduous leaves within the reach of deer (=220cm at height) produced 0.208±0.070kgha^–1. However, litterfall during this period had the highest productivity, 28.7± 5.3kgha^–1. The deer consumed litterfall (75.6% in dry weight), short grasses (17.2%), deciduous forest understory (4.1%), deciduous leaves within the reach of deer (3.0%) and conifer plantation under story (0.1%). It is suggested that the high-density deer population would be maintained by litterfall through the year instead of browsing in deciduous forests, which has been overlooked.     

Ultrastructure of four types of striated muscle fibers in the atlantic hagfish (Myxine glutinosa,L.)

Summary Four types of striated muscle fibers with distinctive ultrastructure were defined in the Atlantic hagfish (Myxine glutinosa, L.): white, intermediate, and red fibers of m. parietalis, and red fibers of m. craniovelaris.White fibers are thick, contain very few mitochondria and fat vacuoles, and possess distinct and separate myofibrils with thin Z-disks and distinct M-lines. Intermediate fibers are thinner, possess largely similar myofibrils that often are even better separated due to a higher content of fat vacuoles and especially mitochondria and glycogen granules. Red fibers of m. parietalis contain large amounts of mitochondria, fat vacuoles, and glycogen granules. Their myofibrils possess M-lines, and although branching more, the myofibrils of red fibers conform with a Fibrillenstruktur pattern like those of white and intermediate fibers. Red fibers of m. craniovelaris are very thin, possess many smaller fat vacuoles, and large amounts of mitochondria and glycogen granules. The myofibrils are significantly thinner than in m. parietalis fibers, run as quite independent well separated units, possess thicker Z-disks, and lack M-lines. Large amounts of myosatellite cells are associated with these red fibers.Triads are located near A/I-junctions in all four fiber types and occur irregularly, the density of triads being different in the various fiber types.We are indebted to Dr. Finn Walvig, Biological Station, University of Oslo, Drøbak, for supply of hagfishes, and we also wish to thank Dr. Jan K. S. Jansen, Institute of Physiology, University of Oslo, for valuable suggestions during this study.     

Rhizogenesis in stem discs of Malus pumila rootstock M9 “Jork”: I. Hormonal and environmental effects on root induction and callus formation

To establish a protocol suited for the molecular characterization of root induction the influences of explant position, etiolation state and orientation as well as temperature and light intensity on root and callus formation were investigated. Depending on these factors stem discs of Malus incubated on indole-3-butyric acid containing medium and subsequently on hormone-free medium regenerated roots and callus of filamentous and smooth texture to a varying extent. Concentration and incubation duration of indole-3butyric acid strongly affected rooting and the production of smooth callus. Moreover smooth callus was profuse at the low light levels applied during rooting. Rooting efficiency decreased and filamentous callus increased between 19°C and 25°C. Explants close to the shoot apex displayed reduced rooting efficiency and profuse filamentous callus. There was a strong effect of explant orientation on root and filamentous callus formation.Abbrevations IBA indole-3-butyric acid - IAA indole-3-acetic acid - BA 6-benzylaminopurine - MS Murashigeand Skoog - rts roots - rt rooted - rtd rooted discs     

Further studies on cell adhesion based on Lex-Lex interaction,with new approaches: embryoglycan aggregation of F9 teratocarcinoma cells,and adhesion of various tumour cells based on Lex expression

We previously proposed specific interaction of Le^x (Gal1 4[Fuc1 3]-GlcNAc1 3Gal) with Le^x as a basis of cell adhesion in pre-implantation embryos and in aggregation of F9 teratocarcinoma cells, based on several lines of evidence (Eggenset al., J Biol Chem (1989)264:9476–9484). We now present additional evidence for this concept, based on autoaggregation studies of plastic beads coated with glycosphingolipids (GSLs) bearing Le^x or other epitopes, and affinity chromatography on Le^x-columns of multivalent lactofucopentaose III (Le^x oligosaccharide) conjugated with lysyllysine. Comparative adhesion studies of Le^x-expressing tumour cellsvs their Le^x-non-expressing variants showed that only Le^x-expressing cells adhere to Le^x-coated plates and are involved in tumour cell aggregation, in analogy to F9 cell aggregation. The major carrier of Le^x determinant in F9 cells is not GSL but rather polylactosaminoglycan (embryoglycan), and we demonstrated autoaggregation of purified embryoglycan in the presence of Ca²⁺, and reversible dissociation in the absence of Ca²⁺ (addition of EDTA). Defucosylated embryoglycan did not show autoaggregation under the same conditions. Thus, Le^x-Le^x interaction has been demonstrated on a lactosaminoglycan basis as well as a GSL basis. A molecular model of Le^x-Le^x interaction based on minimum energy conformation with involvement of Ca²⁺ is presented.Abbreviations BSA bovine serum albumin - CHO carbohydrate - DMEM Dulbecco's modified Eagle's medium - EDTA ethylenediaminetetraacetic acid - GP glycopeptide - GSL glycosphingolipid - LAG lactosaminoglycan - Le^x Gal1 4[Fuc-1 3]GlcNAc1 R - LFP lacto-N-fucopentaose - LysLys-OH lysyllysinol - Mr relative molecular weight - PBS phosphate-buffered saline - PG paragloboside (Gal1 4GlcNAc1 3Gal1 4Glc1 1Cer) - TBS Tris-buffered saline (10mM Tris-HCl, pH 7.4, containing 0.15M NaCl) - TC tumour cell