Isoenzymatic characterization of potentially useful forage shrubs belonging to the genus Cytisus and allies (Genistea: Leguminosae)

Twenty-five accessions belonging to the genus Cytisus and allies (Genisteae: Leguminosae), potentially useful for agrosylvopastoral systems in the Mediterranean basin, were analyzed using isoenzymes. The aim was to assess the taxonomic usefulness of these characters at the specific and generic levels, and its value as an aid for the taxonomy. A high level of infrageneric and infraspecific variability was detected. Numerical analysis showed that isoenzymes are useful for the characterization both at the specific and generic levels. The obtained dendrogram supports the generic arrangement proposed by Bisby [(1981) Advances in Legume Systematics, Vol. 1, pp. 409–425], therefore helping to elucidate the difficult taxonomy of the studied species.     

Linkage analysis and allelic imbalance in human breast cancer kindreds using microsatellite markers from the short arm of chromosome 3

Eight Icelandic breast cancer kindreds were subjected to linkage analyses with respect to 28 microsatellite loci dispersed along the short arm of chromosome 3. Breast tumors derived from these kindreds were concurrently scored for allelic imbalance with ten of the markers. Linkage to most markers could be excluded on the basis of negative LOD scores and haplotype analyses, although some moderately positive LOD scores resulted. A high frequency of imbalance in the familial tumors was seen with two of the markers in comparison with results obtained from sporadic material. The highest frequency (68%) of imbalance was detected with the marker D3S1217, which is located on 3p14.2-p14.1. Imbalance at the D3S1211 locus, which is more telomeric (3p24.2-p22), was not significantly elevated in the familial tumors. We suggest that the genetic defect responsible for breast cancer susceptibility in these families either promotes instability in the 3p14.2-p14.1 region or enhances the selective advantage of such changes.     

Meningococcal pilin: a glycoprotein substituted with digalactosyl 2,4-diacetamido-2,4,6-trideoxyhexose

Neisseria meningitidis pili are filamentous protein structures that are essential adhesins in capsulate bacteria. Pili of adhesion variants of meningococcal strain C311 contain glycosyl residues on pilin (PilE), their major structural subunit. Despite the presence of three potential N -linked glycosylation sites, none appears to be occupied in these pilins. Instead, a novel O -linked trisaccharide substituent, not previously found as a constituent of glycoproteins, is present within a peptide spanning amino acid residues 45 to 73 of the PilE molecule. This structure contains a terminal 1-4-linked digalactose moiety covalently linked to a 2,4-diacetamido-2,4,6-trideoxyhexose sugar which is directly attached to pilin. Pilins derived from galactose epimerase ( galE ) mutants lack the digalactosyl moiety, but retain the diacetamidotrideoxyhexose substitution. Both parental (#3) pilins and those derived from a hyper-adherent variant (#16) contained identical sugar substitutions in this region of pilin, and galE mutants of #3 were similar to the parental phenotype in their adherence to host cells. These studies have confirmed our previous observations that meningococcal pili are glycosylated and provided the first structural evidence for the presence of covalently linked carbohydrate on pili. In addition, they have revealed a completely novel protein/saccharide linkage.     

Seasonal and area-related variations in the diet of otters Lutra lutra on Mull

The diet of otters Lurra lutrn living in a sea loch on the Isle of Mull was described by means of spraint analysis. Seasonal and spatial differences in the spraint composition were explored in the context of fluctuations in prey availability, as revealed by trapping. Butterfish Pholis gunnellus and the cottids Myoxocephalus scorpius and Taurulus bubalis were the most commonly occurring prey in spraints. In the winter, the proportion of rocklings ( Ciliata spp. and Gaidropsarus spp.) and non-rockling gadoids (Gadidae) in the diet increased. Their relatively large size makes these latter species an important constituent of the winter diet. Seasonal and spatial variation in fish consumption largely reflected fluctuations in availability. The mean size of principal prey fishes in spraints was similar to that observed in the traps. These observations suggest that otters were not strongly selective with regard to fish species and size. Shore crabs, in contrast, were not taken in proportion to availability. The findings are discussed in the light of other studies of the diet of coastal otters. It is suggested that the lack of selectivity demonstrated in this study, compared with previous studies, may be explained by area-related variations in prey availability.     

A kinetic study of the interaction of vanadate with the Ca2+ + Mg2+-dependent ATPase from sarcoplasmic reticulum.

Ca2+ + Mg2+-dependent ATPase from sarcoplasmic reticulum was inhibited by preincubation with vanadate. When the inhibited enzyme was preincubated in the presence of vanadate and assayed in its absence, a slow reactivation process was observed. This slow, hysteretic, process was exploited to study the influence of Ca2+ and ATP on the dissociation of vanadate. Ca2+ alone slowly displaced vanadate from the inhibited enzyme, and a rate constant of 0.1 min-1, at 25 degrees C, was calculated for this re-activation process. However, ATP re-activated with an apparent constant that hyperbolically depended on ATP concentration, and from it a rate constant for vanadate dissociation induced by ATP of 0.5 min-1 was calculated. It is deduced from the kinetic studies that ATP binds to the enzyme-vanadate complex, forming a ternary complex, with a dissociation constant of 4 microM, and that this binding accelerates vanadate dissociation. Binding experiments with [14C]ATP showed that ATP binds to the enzyme-vanadate complex with a dissociation constant of 12 microM, i.e. the affinities calculated with the isotope technique and the kinetic procedure are of the same order of magnitude.     

Histidine residues in elongation factor EF-tu from Escherichia coli protected by aminoacyl-tRNA against photo-oxidation

Complexes of Escherichia coli elongation factor EF-Tu with GTP or GTP and aminoacyl-tRNA were photo-oxidized by irradiation with visible light in the presence of rose bengal dye. EF-Tu was isolated, digested with trypsin, the resulting tryptic peptides were separated by high-performance liquid chromatography (HPLC), and the position of most of the peptides on the chromatogram was determined. Irradiation of complexes resulted in the inactivation of the factor (as tested by its capacity to interact with aminoacyl-tRNA) and was accompanied by the loss of its histidine residues (as revealed by amino acid analysis) and by the decrease in the amount of some tryptic peptides (as detected by HPLC). Aminoacyl-tRNA, bound to EF-Tu during the irradiation, protected the protein from inactivation, from the loss of histidine residues and some of its peptides from photo-oxidative degradation. Comparison of quantities of individual tryptic peptides recovered from the irradiated EF-Tu X GTP X aminoacyl-tRNA complex with those from the irradiated EF-Tu X GTP complex revealed that histidine-containing peptides T12 and T15 as well as methionine-containing peptide T14 were in the ternary complex markedly protected against the photo-oxidative degradation. This finding suggests that their histidines, i.e. His-66 and His-118 respectively and at least one of the methionines (Met-91, 98 or 112) present in peptide T14 are located near to or at the binding site of EF-Tu for aminoacyl-tRNA and could be involved in the interaction between aminoacyl-tRNA and the factor.     

Injected mitotic extracts induce condensation of interphase chromatin

Although extracts from mitotic cells have been shown to induce chromosome condensation when injected into amphibian oocytes, they have not as yet been shown to induce this response in somatic interphase cells. In the experiments reported here, when mitotic extracts were injected into syncytial frog embryos, whose somatic nuclei were arrested in interphase, chromosome condensation was observed. The inability of interphase extracts, injected at similar concentrations, to induce this event demonstrates the cell cycle-specific accumulation of the factors responsible.     

Naltrexone effects on pituitary neurointermediate lobe and median eminence

The long-acting opiate antagonist naltrexone hydrochloride was administered by intraperitoneal injection, in a dose response protocol, to adult rats. The drug was used to observe effects of opiate receptor blockade on cells of the pituitary gland and adjacent hypothalamus. At higher drug doses (5mg/kg or 10mg/kg), neurites directly innervating pars intermedia cells contained swollen vesicles and disrupted membranous elements. Fibers within the median eminence of the hypothalamus appeared swollen, and contained myelin figures. Despite the consistent degenerative changes appearing in neurites, measurements of levels of dopamine, norepinephrine, and epinephrine in striatum, and hypothalamus did not differ significantly between naltrexone-treated or control animals, although there was a significant elevation of norepinephrine in the pituitary after drug treatment. At all drug dose levels administered, supraependymal neuron-like cells appeared atop the ependyma of the third ventricle above the median eminence. These observations suggest that naltrexone produces specific neurotoxic effects on neurites of the tuberoinfundibular system, and may induce changes in the ventricular environment which stimulate the appearance of supraependymal neurons.