Jasmonic acid spraying does not induce tuberisation in short-day-requiring potato species kept in non-inducing conditions

The potato species Solanum andigena (Juz. and Buk.) and Solanum demissum (Lindl.) that both require short days for tuberisation were kept in either long days (16 h light), or short days (8 h light) with a 30-min night break mid-way through the dark period. Tuberisation of these species was inhibited under both conditions. Repeated spraying of these plants with up to 100 μM jasmonic acid did not induce them to tuberise even though jasmonic acid was shown to be taken up and transported within the plant. This result argues against jasmonic acid itself being the transported tuber-inducing signal, although it does not exclude a role for jasmonic acid later in tuber formation and development once induction has taken place.     

Differential recovery of polymorphonuclear neutrophils,B and T cell subpopulations in the thymus,bone marrow,spleen and blood of mice following split-dose polychemotherapy

In these studies, we examined the effect of a maximum-tolerated, split-dose chemotherapy protocol of cyclophosphamide, cisplatin, and 1,3-bis(2-chloroethyl)-1-nitrosourea carmustine on neutrophil and lymphocyte subpopulations in the peripheral blood (PBL), thymus, bone marrow and spleen. It was found that this protocol of polychemotherapy, modeled after the induction protocol used with autologous bone marrow transplantation for breast cancer, suppressed both B and T cell populations and T cell function at times when the absolute neutrophil count had returned to normal or supernormal numbers. In the peripheral blood, 7 days following initiation of chemotherapy, there was a twofold increase in the percentage of granulocytes as compared to the level in control animals on the basis of a differential count. The polymorphonuclear neutrophil (PMN) frequency in the bone marrow was increased on day 14 and statistically identical to that in control mice on all other days analyzed. In contrast to the bone marrow cells and PBL on day 7, the frequency of PMN in the spleen and thymus was depressed. B cells (B220+) were depressed in the PBL, spleen and bone marrow and took 18–32 days to return to their normal frequency, while the frequency of B cells in the thymus was increased owing to a loss of immature T cells. The percentage of CD3+ cells in the thymus, spleen and bone marrow was significantly increased and required 10–18 days to return to normal levels, while the absolute number of CD3+ cells in the blood varied around the normal value. The ratio of CD4+ to CD8+ cells in all the organs studied varied only slightly owing to a similar reconstitution of CD4+ and CD8+ cells. In contrast to the phenotypic recovery of the CD3+, CD4+ and CD8+ cells, the ability of the splenic lymphocytes to respond to concanavalin-A was depressed and remained depressed, despite the phenotypic reconstitution of the T cell subsets, on the basis of both percentage and absolute cell number. These results show a selective T and B cell depression following multi-drug, split-dose chemotherapy in tissue and blood leukocyte populations and a chronic depression in T cell function.     

Identification of the Shiga toxin A-subunit residues required for holotoxin assembly.

Recent X-ray crystallographic analyses have demonstrated that the receptor-binding (B) subunits of Shiga toxin (STX) are arranged as a doughnut-shaped pentamer. The C terminus of the enzymatic (A) subunit presumably penetrates the nonpolar pore of the STX B pentamer, and the holotoxin is stabilized by noncovalent interactions between the polypeptides. We identified a stretch of nine nonpolar amino acids near the C terminus of StxA which were required for subunit association by using site-directed mutagenesis to introduce progressive C-terminal deletions in the polypeptide and assessing holotoxin formation by a receptor analog enzyme-linked immunosorbent assay, immunoprecipitation, and a cytotoxicity assay. Tryptophan and aspartic acid residues which form the N-terminal boundary, as well as two arginine residues which form the C-terminal boundary of the nine-amino-acid sequence, were implicated as the stabilizers of subunit association. Our model proposes that residues 279 to 287 of the 293-amino-acid STX A subunit penetrate the pore while the tryptophan, aspartic acid, and 2 arginine residues interact with other charged or aromatic amino acids outside the pore on the planar surfaces of the STX B pentamer.     

Occurrence ofBacillus popilliae and two nematode pathogens in populations ofAmphimallon solstitialis (Col.: Scarabaeidae) near darmstadt,Germany

Populations of the June beetle,Amphimallon solstitialis (Coleoptera: Scarabaeidae), were sampled from grasslands near Darmstadt, Germany, and found to be infected by a number of diseases and parasitic nematodes. Several milky diseased larvac were found, andBacillus popilliae type A 1 isolated as the causative agent. The bacterium readily infected healthy larvae ofA. solstitialis when administeredper os in the laboratory, but was not infective to larvae ofMelolontha hippocastani by this route. A large mermithid nematode was found parasitising 10% of theA. solstitialis larvae at one site andHeterorhabditis bacteriophora was found infecting larvae at the other.

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Résumé Des populations deAmphimallon solstitialis (Col.: Scarabaeidae) ont été échantillonnées dans deux prairies distinctes près de Darmstadt (Allemagne) et se sont révélées infestées par des maladies et des nématodes parasites. Plusieurs larves dans chaque site étaient infectées par la maladie laiteuse etBacillus popillae, type A1 a été isolé comme étant l'agent responsable. La bactérie infeste facilement des larves saines deA. solstitialis quand elle est administrée par voie orale au laboratoire mais n'a pas d'effet infectieux pour les larves deMelolontha hippocastani par cette même voie. Un grand nématode mermithide a été trouvé parasite de 10% des larves deA. solstitialis sur un seul site, mais comme seuls les jeunes ont été trouvés, la détermination à l'espèce n'a pu être faite.Heterorhabditis bacteriophora était responsable de l'infection de 1,5% des larves sur seulement un site. Plusieurs larves deA. solstitialis présentaient une couleur ambrée, identique à celle trouvée sur le scarabéeCostelytra zealandica infecté parSerratia spp., et ne se nourrissaient pas de carotte au laboratoire. Les bactéries isolées à partir de ces larves ne provoquent aucun effet visible quand elles sont données en nourriture à des larves saines.

     

Anti-predator defences of a web-building spider, Holocnemus pluchei (Araneae, Pholcidae)

Anti-predator defence behaviour of Holocnemus pluchei (Araneae, Pholcidae) was studied in the laboratory and the field. Bouncing, in which this web-building spider moves its body rapidly up and down while keeping its legs on the silk, was the most frequent response to disturbance. Less often, H. pluchei left the web when disturbed. Experiments were carried out to determine the types of stimuli that elicited bouncing and leaving the web: tactile stimulation (striking the spider with a brush) and air movement (blowing on the spider) elicited bouncing and leaving the web, but there was no evidence that chemical stimuli from potential predators were important. Tactile stimulation elicited stronger responses than air movement. There were no evident differences in how H. pluchei responded to tactile stimulation in the laboratory and the field.     

Structurally defined synthetic cancer vaccines: analysis of structure, glycosylation and recognition of cancer associated mucin, MUC-1 derived peptides

Translation of an immune response into therapy is probably the toughest task in designing vaccines for cancer due to the heterogeneity of the cell surface antigens which display tremendous variations in glycoforms. Consequently, a small segment (antigen) of the cancer-associated mucin, in spite of generating antigen-specific immune responses, may be limited in therapeutic value. It is important that the synthetic segment resembles the native cancer-associated mucin in both structure and conformation. Synthetic cancer associated mucin derived 16 amino acid peptide GVTSAPDTRAPAPGSTA and its partially glycosylated forms have demonstrated specific binding to two monoclonal antibodies, B27.29 and BCP8, raised against the native cancer associated mucin, MUC-1 and a MUC-1 derived synthetic peptide, respectively. In spite of the structural similarities at the core peptide level of both glycosylated and unglycosylated peptides, it appears that partial glycosylation does not inhibit and even slightly enhances binding to the MAb B27.29 indicating that the glycosylated synthetic peptide more closely resembles the native mucin epitope recognized by MAb B27.29. From molecular dynamic simulations using NMR derived distance constraints, both glycosylated and unglycosylated peptides have shown a type I turn involving the same amino acids in both glycosylated and unglycosylated peptides. The GalNAc attached to the threonine (T³) and serine (S⁴) in the 16 amino acid sequence has not imposed any conformational changes to the peptide backbone nor has offered severe steric resistance to the binding of either antibody to the glycopeptides as indicated by hapten inhibition studies. Nevertheless, all peptides have displayed glycosylation dependent specificities in binding to these antibodies, i.e. the glycosylated peptides demonstrated relative higher affinities to the native mucin antibody B27.29 while the unglycosylated peptide is more specific to the MAb BCP8. Immune responses generated by these synthetic glycopeptides are highly specific in recognizing the native cancer associated mucin.     

Incidence of acute symptomatic toxoplasma retinochoroiditis in south London according to country of birth.

OBJECTIVE--To determine the incidence of acute symptomatic toxoplasma retinochoroiditis presenting to ophthalmologists for patients born in Britain and elsewhere. DESIGN--Population based, cross sectional study. SETTING--11 districts in south Greater London. SUBJECTS--All patients presenting to NHS ophthalmologists with symptoms due to acute toxoplasma retinochoroiditis in 1992-3. MAIN OUTCOME MEASURE--Intraocular inflammation in association with a retinochoroidal scar, active adjoining retinitis, and IgG serum antibodies to toxoplasma. RESULTS--The estimated incidence of acute symptomatic retinochoroiditis for all people born in Britain was 0.4/100,000/year. If a mean of two symptomatic episodes per lifetime is assumed, 100 people born in Britain may be affected each year, about a fifth of the estimated 500-600 congenitally infected people born each year. CONCLUSIONS--A substantial proportion of people with acute symptomatic toxoplasma retinochoroiditis were born outside the country, and the number born in Britain was smaller than the number previously estimated to develop retinochoroidal lesions due to congenital toxoplasmosis. These findings suggest that prenatal screening for toxoplasmosis in Britain may be of limited benefit.     

Inhibition of lipid peroxidation in muscle homogenates by phospholipase A2 inhibitors

Chlorpromazine, mepacrine, tetracaine, dibucaine, chloroquine, and procaine have been shown to inhibit the iron- and ascorbate-induced lipid peroxidation of skeletal-muscle hornogenates in vitro. These compounds are known to be inhibitors of phospholipase activity, but they were also found to be effective in blocking free-radical-mediated damage to lipids in denatured homogenates, to linoleate suspensions, and to glutamic acid solutions where phospholipase activity was not a relevant factor. The inhibitory action did not appear to be related to any iron-binding activity of the compounds.