An evolutionary relationship between the ColE5-099 and the ColE9-J plasmids revealed by nucleotide sequencing

The nucleotide sequence of a 1124 bp fragment of the ColE5-099 plasmid which encodes colicin E5 immunity, a lys gene involved in colicin release from the host cell, and the 3' end of the colicin E5 structural gene has been determined. Open reading frames corresponding to the three genes have been located by analogy with similar sequences from other E colicin plasmids. The location of these open reading frames corresponds with the position of the genes as determined by subcloning and transposon mutagenesis. The amino acid sequence of the carboxy-terminal 107 amino acid residues of the colicin E5 gene shows no homology with any other E colicin, suggesting a different mode of action in killing sensitive cells. A comparison of the nucleotide sequence of this region of the ColE5-099 plasmid with that of the equivalent region of the ColE9-J plasmid suggests a close evolutionary relationship between these two plasmids.     

Eradication of ear mites from naturally infested conventional research rabbits using ivermectin

Rabbits naturally infested with ear mites were treated with ivermectin injection for cattle, subcutaneously at the rate of 400 mcg/kg; which was repeated in 15 to 17 days. Rabbits treated as described and housed in a conventional vivarium environment were found to be free of mites during a subsequent 33 to 139 day observation period. Side effects were minimal and associated with occasional transient discomfort at the injection site. Ivermectin appears to be safe and effective for treating rabbits with ear mites. The prospects of eradicating mites from infested rabbit colonies using this method of treatment is promising.     

Influence of ammonium and extracellular pH on the amino and organic acid contents of suspension culture cells of Acer pseudoplatanus

The effects of supplied ammonium and nitrate on the amino and organic acid contents and enzyme activities of cell suspension cultures of Acer pseudoplatanus L. were examined. Regardless of nitrogen source the pH of the culture medium strongly affected the malate and citrate contents of the cells; these organic acid pools declined at pH 5, but increased at pH 7 and 8. Over a period of two days, ammonium had little effect on the responses of the organic acid pool sizes to the pH of the medium. In contrast, ammonium had a strong influence on amino acid pool sizes, and this effect was dependent on the pH of the medium. At pH 5 there was no increase in cell ammonium or amino acid contents, but at higher pH values cellular ammonium content rose, accompanied by accumulation of glutamine, glutamate and asparagine. Over several days, supplied ammonium led to an increase in activity of glutamate dehydrogenase irrespective of any changes in internal ammonium and amino acid contents. If the pH of the medium was allowed to fall below pH 4 in the presence of ammonium, phosphoenolpyruvate (PEP) carboxylase activity declined to a very low value over several days; at higher pH, the activity of this enzyme, and that of NAD malic enzyme and NAD malate dehydrogenase, remained substantial irrespective of whether the nitrogen source was NH⁺₄ or NO-₃.     

The fragmentation of 670A MeV neon-20 as a function of depth in water. II. One-generation transport theory

The results of an experiment to study the interaction of a beam of 670A MeV neon ions incident on a water column set to different thicknesses were compared with a "first principles" transport calculation in the straight-ahead approximation. This calculation assumes that the nuclear interactions of the incident particles lead to a secondary particle with the velocity of the incident projectile at the interaction point moving in the direction of the incident projectile. Subsequent nuclear interactions of the fragments were taken into account partially, by calculating the nuclear attenuation of the fragments in the residual material, but were not taken into account as a source of further nuclear interaction products. Fluence spectra were calculated per unit incident neon fluence for 14 absorber thicknesses. The acceptance for each fragment was calculated based on a knowledge of the material in the beam and of the beam extraction energy. The theoretical spectra were multiplied by the calculated acceptance and convoluted with the LET resolution associated with the experiment. The stopping power used in the transport calculation was found to predict a range approximately 1.6% shorter than that given by experiment; this small difference resulted in significant discrepancies between theory and experiment in the stopping region. For particles not stopping in the absorber, the transport calculation was accurate to within 30% for depths less than approximately 15 cm; the effects of tertiary particles become significant at greater depth.     

Extending CD spectra of proteins to 168 nm improves the analysis for secondary structures

The CD spectra for 10 proteins with known secondary structure have been extended from 178 to 168 nm. Combined with the data for 6 other proteins investigated previously, this produces a basis set of 16 proteins, which can be used to analyze CD spectra for secondary structure. Extending the spectra adds another CD band to the data and increases the information content from the equivalent of five to six. Analyzing the CD for each of the 16 proteins in the basis set with the 15 other proteins shows a modest improvement in the prediction of secondary structure with the extended CD spectra.     

Circular dichroism and laser Raman spectroscopic analysis of the secondary structure ofCerebratulus lacteus toxin B-IV

The secondary structure ofCerebratulus lacteus toxin B-IV, a neurotoxic polypeptide containing 55 amino acid residues and four disulfide bonds, was experimentally estimated by computer analyses of toxin circular dichroism (CD) and laser Raman spectra. The CD spectrum of the toxin displayed typical α-helical peaks at 191, 208, and 222 nm. At neutralpH, the α-helix estimates from CD varied between 49 and 55%, when nonrepresentative spectrum analytical methods were used. Analysis of the laser Raman spectrum obtained at a much higher toxin concentration yielded a 78% α-helix estimate. Both CD and Raman spectroscopic methods failed to detect any β-sheet structure. The spectroscopic analyses revealed significantly more α-helix and less β-sheet for toxin B-IV than was predicted from its sequence. To account for the difference between the 49–55% helix estimate from CD spectra and the 78% helix estimate from the Raman spectrum, we postulate that some terminal residues are unfolded at the low toxin concentrations used for CD measurements but form helix at the high toxin concentration used for Raman measurements. Our CD observations showing thatCerebatulus toxin B-IV helix content increases about 15% in trifluoroethanol or at highpH are consistent with this interpretation.     

Dust and ammonia in horse stalls with different ventilation rates and bedding

Airborne particles and ammonia were monitored in horse stalls managed under four conditions. Two ventilation rates, high (27 air changes per h) and low (5 air changes per h) and two bedding types, paper and straw, were employed. At both ventilation rates, the number of airborne particles generated while the stalls were mucked out was higher with straw than with paper. Particles were more efficiently cleared at the higher ventilation rate in both the straw and paper stalls. Ammonia measurements reflected an accumulation over time. In the stalls with low ventilation, ammonia levels were significantly higher than in those stalls with high ventilation regardless of bedding type. Management decisions and their relationships to respiratory disorders are discussed.     

Survey of leaf anatomy,especially secretory structures,of tribeCaesalpinieae (Leguminosae,Caesalpinioideae)

We studied leaflet anatomy, emphasizing secretory structures, from herbarium specimens of 128 species of 44 genera of tribeCaesalpinieae, using clearings, resin sections, and scanning electron microscopy. These observations, combined with those from our three earlier papers, provide a survey of 210 species representing all genera. Seventy-three species had secretory structures: 21 had glands or gland-like trichomes, 40 had living mesophyll idioblasts, and nine had cavities (three species each had two different types). Five additional species, all inCercidium (Caesalpinia group), had paired or clustered large spheroidal, thick-walled, empty cells (veinlet idioblasts) interconnected by perforation plate-like gaps. Secretory structures have systematic significance at various taxonomic levels.     

Molecular basis and haplotyping of the alphaII domain polymorphisms of spectrin: application to the study of hereditary elliptocytosis and pyropoikilocytosis.

Hereditary elliptocytosis (HE) and hereditary pyropoikilocytosis (HPP) are inherited disorders of erythrocyte shape that are frequently associated with abnormalities in alpha-spectrin, one of the principal structural proteins of the erythrocyte membrane skeleton. Five polymorphisms of the alpha-spectrin gene, located in a 6-kb interval of genomic DNA, were identified and analyzed in normal and mutant alpha-spectrin alleles. Three of these polymorphisms are due to single nucleotide substitutions in the alpha-spectrin gene coding region that lead to changes in the amino acid sequence. In combination, these three polymorphisms are responsible for the different peptide phenotypes of the alphaII domain previously observed following limited tryptic digestion of spectrin protein. The most common haplotype, type 1, was found predominantly in Caucasians and was the only haplotype identified in Asians. Haplotypes 2, 3, and 4 were identified predominantly in individuals of African ancestry and were commonly found in patients with HE or HPP. Analysis of coinheritance of alphaII domain polymorphisms with alpha-spectrin gene mutations causing HE or HPP in African-American patients with HE and HPP suggests that, with one exception, a given HE/HPP mutation is present in an alpha-spectrin gene of only one haplotype, indicating a founder effect. The other two polymorphisms located in this region of the alpha-spectrin gene do not change the amino acid sequence of the encoded alpha-spectrin chain and are not in linkage disequilibrium with three of the four alphaII domain haplotypes. A model is proposed for the evolutionary origin of the different haplotypes.     

Spatial Analysis of the Canopy of Kiwifruit Vines as it Relates to the Physical, Chemical and Postharvest Attributes of the Fruit

The position of individual fruit on kiwifruit vines (Actinidiadeliciosa var. deliciosa) grown on a horizontal trellis (pergola)and on a T-bar trellis was determined using a theodolite. Thephysical, chemical, and postharvest attributes of the fruitwere related to their position on the vine during development. Fruit from the pergola vines were more numerous, of lesser weight,with lower concentrations of most mineral nutrients, but greaterconcentrations of soluble solids, and similar flesh firmnessafter 12 weeks of storage at 0 °C, than fruit from the T-barvines. The position on the vine accounted for most of the variationin the attributes of the fruit. Differences between fruit ona single lateral accounted for 43-56% of the variation. Variationbetween vines was relatively small (< 4% of the total variance). The heavier fruit were located at the apical ends of the laterals,while greater concentrations of soluble solids were associatedwith fruit located closer to the cordon. The larger fruit fromthe pergola vines developed from the early opening flowers.A similar relationship existed initially for the T-bar vines,but a reduction in growth of fruit from the early opening flowers8 weeks after anthesis resulted in a more even distributionof fruit size at harvest. The strongest relationship between mineral composition and postharvestattributes of the fruit was with soluble solids concentration(29-46% of the variance). The relationship with flesh firmnesswas weak (r = -0·14 to -0·32). Individual elementscould not be considered in isolation but rather in groups ofelements. Nitrogen was grouped strongly with phosphorus, sulphur,potassium, and copper, while calcium was linked with a secondgroup which included manganese and zinc. These two groups werenegatively related to one another. The greatest proportion of fruit with superior characteristicswas located in the denser parts of the canopy. Fruit with lessdesirable attributes were from the extremities of the canopywhere the leaf area index was low.Copyright 1994, 1999 AcademicPress Actinidia deliciosa, kiwifruit, fruit position, fruit quality, within-vine variation