Roles of glycosaminoglycans and glycanmimetics in tumor progression and metastasis

Various tumor cells exhibit structural alterations in the sulfated modifications to glycosaminoglycans (GAGs). The altered expression of chondroitin sulfate (CS) and heparan sulfate (HS) on the surfaces of tumor cells is known to play a key role in malignant transformation and tumor metastasis. The receptor molecule for the CS chains containing E-disaccharide units (CS-E) expressed on Lewis lung carcinoma (LLC) cells was recently revealed to be Receptor for Advanced Glycation End-products (RAGE). RAGE is also involved in the development of various pathological conditions including aging, infection, pulmonary fibrosis, diabetes, and Alzheimer’s disease, by binding to a wide range of ligands. RAGE binds strongly not only to CS-E, but also to HS-expressing LLC cells. Recombinant RAGE bound CS-E and HS with high affinity. Furthermore, in a mouse model, the colonization of the lungs by LLC cells was inhibited by intravenously injected CS-E, an anti-CS-E antibody, or an anti-RAGE antibody. These findings demonstrated that RAGE is at least one of the critical receptors for CS and HS chains expressed on the tumor cell surface and is involved in experimental lung metastasis, and also that CS/HS and RAGE are potential molecular targets for the treatment of pulmonary metastasis. We, hence, reviewed these findings and also several chemically synthesized small GAGmimetics that exhibit potent anti-metastatic and/or anti-tumor activities.     

Agarose plating and a bead type culture technique enable and stimulate development of protoplast-derived colonies in a number of plant species

Two novel techniques improve division and colony formation from protoplasts:

1. Plating in agarose stimulates colony formation of protoplasts from a wide range of species. Protoplasts from Nicotiana tabacum developed to colonies from lower initial population densities in agarose than in agar or liquid. Protoplasts from Hyoscyamus muticus which do not divide in agar divided and formed colonies in agarose at higher efficiencies than in liquid medium.
2. Culture of gel embedded protoplasts in large volumes of liquid medium on a gyrotatory shaker (‘bead culture’) further improved plating efficiencies in some species (e.g. Lycopersicon esculentum and Crepis capillaris) and enabled sustained proliferation of protoplasts which had not previously developed beyond the few cell colony stage (Brassica rapa and a mutator gene variety of Petunia hybrida).

The combination of ‘agarose plating’ and ‘bead culture’ dramatically improved plating efficiencies of protoplasts in all species tested.     

Somatic embryogenesis in cell cultures of Glycine species

This report describes the development of procedures for the production of somatic embryos in cell cultures of Glycine species including soybean. The conditions for callus induction and initiation of rapidly growing cell suspension cultures were defined. Methods for inducing embryogenesis were tested on 16 lines of several Glycine species and cultivars of soybean. The SB-26 Culture of a G. soja gave the best results and was used in the experiments. Embryogenesis required the presence of picloram or 2,4-D. AMO 1618, CCC, PP-333 and Ancymidol enhanced the embryogenesis frequency. Plants of the G. soja (SB-26) were grown to maturity from seed-derived shoot tips. Characteristics of the plants are discussed.     

High production of caffeine and related enzyme activities in callus cultures of Coffea arabica L.

Callus tissue culture of Coffea arabica L. cv Hybrido de Timor prepared from apical portions of orthotropic branches produced 49 to 92 times as much caffeine per unit weight of tissue as did the original explant. Cell-free extracts made from 42 to 54-day-old callus cultures in which active biosynthesis was occurring exhibited N-methyl-N ⁹-nucleoside hydrolase and N-methyltransferase enzyme activities. Similar cell-free extracts exhibited selective biodegradative activity in forming urea from xanthine. Biosynthetic substrate specificities are similar to those of the enzyme obtained from green coffee fruit and tea leaves, suggesting that callus cultures of C. arabica form caffeine in the same way as the coffee fruit and tea leaves.     

Inactivation of aphidicolin by plant cells

Plant cells are endowed with an aphidicolin inactivating activity. Data on cultured cells show that the rate of inactivation depends on the cell type, Daucus carota cells being the most effective among the other tested materials (Oryza sativa and Nicotiana plumbaginifolia). Also germinating seedling of Haplopappus gracilis and of Citrullus vulgaris inactivate aphidicolin. Inactivation, which may lead to unexpected results when a prolonged incubation with the drug is required, as in the case of the induction of synchrony of the cell cycle by aphidicolin, can be controlled by appropriately choosing the experimental conditions.     

Low Energy Conformations for Gonadotropin-Releasing Hormone with d- and l-Amino Acid Substitutions for Gly 6: Possible Receptor-Bound Conformations

In the preceding paper, using ECEPP, including the effects of water, and the chain build-up procedure, we computed the low energy structures for GnRH and found that there were no distinct low energy structures or structures with high statistical weights. To attempt to deduce possible structures of GnRH that may bind to the GnRH receptor, we computed the low energy structures for GnRH peptides that have l- and d-amino acids substituting for Gly 6. The l-amino acid-substituted peptides (l-Ala and l-Val) have very low or no affinity for the receptor and on activity (release of FSH and LH) while the d-Ala-, d-Leu-, d-Trp- and d-Phe-substituted peptides have significantly higher relative affinities and activities than those for native GnRH; the d-Val-substituted peptide has about one-third of the affinity and activity as native GnRH. Unlike native GnRH, our computations suggest that both sets of peptides form well-defined structures in water: the l-amino acid-substituted peptides are predominantly α-helical while the d-amino acid-substituted peptides adopted E*A A A E D*(C*) A E C A(C*) and minor variants of these structures. By eliminating structures that lay in common to the d-Ala and l-Val peptides and further eliminating structures that differed between the d-Ala and d-Leu peptides, we reduced the number of possible distinct binding conformations to 254. Searching for structures among these 254 conformations that had relative statistical weights that paralleled their relative affinities, we found two candidate structures: D*E A A E C*A E C A and D*G A A E D*A E C G*, both of which have conformations for residues 3–9 that are similar to the computed most probable structures for the d-amino acid-substituted GnRH peptides in water.     

ATPase activities in partially purified membranes of Acetabularia

Partly purified membranes (with plasmalemma material) of Acetabularia mediterranea were studied with respect to ATPase activity in alkali- and Ca⁺⁺-free media and its sensitivity to pH (5 – 9), oligomycin (200 ?g/mg protein), 100 ?M N-N′-dicyclohexylcarbodiimide (DCCD), and 50 ?M vanadate. Besides activities which may originate from mitochondrial H⁺ ATPase (oligomycin-sensitive, alkaline pH optimum) and tonoplast H⁺ ATPase (DCCD-sensitive, pH optimum 7.5), there is ATPase activity with a pH optimum around pH 6.5, sensitive to vanadate and insensitive to DCCD. These results strongly suggest that the electrogenic Cl^? pump in the plasmalemma of Acetabularia is an ATPase. Effects of Mg⁺⁺, Mg-ATP, ADP, GTP, UTP, CTP and HCO₃ ^? versus Cl^? on this ATPase activity are described.     

Growth and infectivity of callus cultures of tomato plants infected with a mycoplasma disease — Potato witches' broom

Callus tissues were derived from the stem of healthy tomato plants (Lycopersicum esculentum Mill. ev. Pr?honické) and of plants infected with potato witches' broom—a disease caused by mycoplasma. Callus cultures were established on modified fully synthetic media described byMorel (1948) and byMurashige andSkoog (1962). Callus cultures obtained from diseased plants were grown and subcultured on both media, growth in primary isolates from healthy plants took place on the Murashige and Skoog medium only. Growth of callus tissue derived from diseased plants was more vigorous even after several subcultivations in comparison with callus tissues isolated from healthy plants. Variations in the morphology in these callus cultures were not noted. Callus cells of diseased plants varied in size; they were about 50% larger than those from healthy ones. Implantation of primary and subcultivated callus tissues into tomato stems of healthy plants did not show any symptoms of infection on test plants.     

Effect of sugars,Na+-, and K+-ions on biopterin transport in Crithidia fasciculata

Biopterin uptake by Crithidia fasciculata is pH dependent with optimum at pH 6 and is strongly inhibited by 0.5 mM NAA and DNP,respectively. Both inhibitors also reduce respiration by 40% (NAA) and 97% (DNP). K⁺-ions (1.1%) and K⁺/Na⁺ (0.5% each) stimulate biopterin uptake to the same high extent, but ouabain has no effect, thereby ruling out involvement of Na⁺/K⁺ pump. In absence of these ions, even in 5% glucose solution biopterin uptake is reduced to minimum. Proton excretion seems to be linked to sugar uptake. Both these sugars seem to have the same site of entry, demonstrated by competitive uptake, though D-glucose is taken up much faster by Crithidia than D-galactose. DNP (0.5 mM) causes greater proton excretion in glucose than in galactose medium. With NAA (0.5 mM) proton excretion is inhibited in both glucose and galactose media. D-glucose promotes greater biopterin uptake than D-galactose.     

siMBa—a simple graphical user interface for the Bayesian phylogenetic inference program MrBayes

MrBayes is a program that uses a Bayesian framework for inferring phylogenetic relationships. As MrBayes is a command-line-driven program, users acquainted to programs with graphical user interfaces will not find it easy to operate, especially as it requires a complex input format for the data to be analysed. We thus developed siMBa (simple MrBayes), a simple graphical user interface for MrBayes. This tool gives the user interactive control over most of the parameters and also facilitates the input of a multiple sequence alignment, as any widely used format can be used. siMBa is coded in Perl using the Tk module. Executables are provided for Windows, Linux, and Macintosh. The Perl codes, along with executables for different operating system, are freely available to download from [http://www.thines-lab.senckenberg.de/simba].     

Backbone resonance assignments of the α sub-domain of Brevibacillus thermoruber Lon protease

Lon is an ATPases associated with diverse cellular activities protease and belongs to a unique group that binds DNA. The α sub-domain of Lon protease is responsible for DNA-binding, but the structural information for its DNA-recognition mode is still limited. Here, we report ¹H, ¹⁵N and ¹³C backbone assignment for the α sub-domain from Brevibacillus thermoruber Lon protease as the basis for the elucidation of its structure and interactions with DNA, necessary for understanding the allosteric regulatory mechanism of the enzymatic function.     

Physiological activities of ergoline alkaloids in submerged cultures ofClaviceps paspali andClaviceps purpurea

Ergoline alkaloids do not belong to a group of physio ogically inert secondary metabolites. Ergometrine as well as ergotamine increased the acetyl-CoA carboxylase activity and inhibited the activities of citrate synthase and isocitrate lyase. The slightly inhibitory effect of ergotamine and stimulatory effect of ergometrine on malate synthase was established. Supplied ergometrine brought about a change in the culture growth rate,i.e. inhibition (10^?4 m) or stimulation (10^?3 m). Addition of 10^?4 m ergometrine to the culture ofClaviceps paspali MG-6 increased the total alkaloid yield by 120%. The stimulatory influence was inversely proportional to the concentration of added alkaloid. It is assumed that accumulating alkaloids suppress attainment of the yield that would be achieved by continuous control of the alkaloid level. A positive relationship is assumed between the process of alkaloid synthesis and the viability of the culture.     

Effects of plant extracts on the rates of parasitization ofAnagasta kuehniella [Lep.: Pyralidae] eggs byTrichogramma pretiosum [Hym.: Trichogrammatidae] under greenhouse conditions

In the greenhouse, rates of parasitization ofAnagasta kuehniella (Zeller) eggs byTrichogramma pretiosum (Riley) were significantly increased when treated with water extracts of the weed,Amaranthus retroflexus L.: 1) Parasitization rates appeared to be dose dependent with greatest percentage parasitization obtained fromVicia faba L. plants sprayed with 2.5 ml of the extract, 2) Parasitization rates increased with an exposure time of 24 h, and 3) Plants sprayed withA. retroflexus in mosaic patterns interspersed with 2 additional plant extracts (Chenopodium album L. andPortulaca oleracea L.) did not show a total increase of parasitization over control plants treated with water.     

Changing status of the parasites of potato tuber mothPhthorimaea operculella [Lepidoptera: Gelechiidae] in Australia

Three introduced parasites ofPhthorimaea operculella (Zeller) are now widespread, and a fourth established in a limited area, in Australia.Apanteles subandinus Blanchard, which occurs from Tasmania to Queensland, is the most effective species and the outstanding parasite in Victoria. The extent of parasitism and the status of the various parasites were monitored from the time of their release until 1971. In many areas whereCopidosoma koehleri Blanchard was the only parasite found in 1966, a substantial increase in parasitism was recorded in subsequent years, following the liberation of other species. In the 1969–70 season there was an explosive increase inOrgilus lepidus Muesebeck, which became the dominant species in some coastal areas of New South Wales, displacingA. subandinus. Its change of status to a dominant position was associated with a change of sex ratio from 1.6 ♀: 1♂ to 1.0 ♀: 1 ♂.     

Cell cycle arrest induced by theophylline in root meristem of Haplopappus gracilis

Theophylline, an inhibitor of cyclic nucleotide phosphodiesterase, induced a block of the cell cycle in G₁, a temporary arrest in G₂ and 70% decrease in the uptake of labelled thymidine in roots of Haplopappus. These effects are compared to those previously found with aminophylline and discussed in view of the possible involvement of cAMP in the regulation of the cell cycle in plants.     

Observation of algal colonization on Acropora aspera killed by Acanthaster planci

Algae started colonizing branches of the coral Acropora aspera (Dana) killed by the sea star Acanthaster planci (Linnaeus) within less than 24 hours. Two blue-greens ((Microcoleus lyngbyaceus (Crouan) Ag. and Hormothamnion solutum B. & F.)) dominated the early community but became less abundant than a brown ((Giffordia indica (Sonder) Papenfuss & Chihara)) after 26 days.