Quantifying Single Microvessel Permeability in Isolated Blood-perfused Rat Lung Preparation

The isolated blood-perfused lung preparation is widely used to visualize and define signaling in single microvessels. By coupling this preparation with real time imaging, it becomes feasible to determine permeability changes in individual pulmonary microvessels. Herein we describe steps to isolate rat lungs and perfuse them with autologous blood. Then, we outline steps to infuse fluorophores or agents via a microcatheter into a small lung region. Using these procedures described, we determined permeability increases in rat lung microvessels in response to infusions of bacterial lipopolysaccharide. The data revealed that lipopolysaccharide increased fluid leak across both venular and capillary microvessel segments. Thus, this method makes it possible to compare permeability responses among vascular segments and thus, define any heterogeneity in the response. While commonly used methods to define lung permeability require postprocessing of lung tissue samples, the use of real time imaging obviates this requirement as evident from the present method. Thus, the isolated lung preparation combined with real time imaging offers several advantages over traditional methods to determine lung microvascular permeability, yet is a straightforward method to develop and implement.     

Inhibition of oxidative phosphorylation by Ca or Sr: A competition with Mg for the formation of adenine nucleotide complexes

Intramitochondrial Sr²⁺, similar to Ca²⁺, inhibits oxidative phosphorylation in intact rat-liver mitochondria. Both Ca²⁺ and Sr²⁺ also inhibit the hydrolytic activity of the ATPase in submitochondrial particles. Half-maximal inhibition of ATPase activity was attained at a concentration of 2.5 mM Ca²⁺ or 5.0 mM Sr²⁺ when the concentration of Mg²⁺ in the medium was 1.0 mM. The inhibition of ATPase activity by both cations was strongly decreased by increasing the Mg²⁺ concentration in the reaction medium. In addition, kinetical data and the determination of the concentration of MgATP, the substrate of the ATPase, in the presence of different concentrations of Ca²⁺ or Sr²⁺ strongly indicate that these cations inhibit ATP hydrolysis by competing with Mg²⁺ for the formation of MgATP. On the basis of a good agreement between these results with submitochondrial particles and the results of titrations of oxidative phosphorylation with carboxyatractyloside or oligomycin in mitochondria loaded with Sr²⁺ it can be concluded that intramitochondrial Ca²⁺ or Sr²⁺ inhibits oxidative phosphorylation in intact mitochondria by decreasing the availability of adenine nucleotides to both the ADP/ATP carrier and the ATP synthase.     

猪肠毒素源性大肠杆菌K88ac粘附抗原的核苷酸序列分析

 本文对国内流行的猪肠毒素源性大肠杆菌K88ac抗原的结构基因的核苷酸序列进行了测定。该基因由849对核苷酸组成,编码了283个氨基酸的蛋白亚单位及21个氨基酸的信号肽。与国外报道的K88ac序列的不同是我们发现一个碱基的点突变,导致在抗原决定簇内一个氨基酸的改变。从核苷酸序列推导出的氨基酸序列与另两种亚型进行了比较。     

Presence of unique repeated insertion sequences in nodulation genes of Rhizobium ‘hedysari’

Cloning and sequencing of DNA from a symbiotic large plasmid in Rhizobium hedysari strain IS 123 required for its nodulation of the mediterranean legume crop Hedysarum coronarium (sulla) and complementation studies of nod^- mutant derivatives led to the characterization of a 30-kb region containing common and host-specific nod genes. This DNA region also contained at least six copies of a novel insertion sequence-like structure, some of which appeared to have suffered deletions. This 0.8 kb novel element carries two 17-bp flanking inverted repeats and an open reading frame showing homology with a transposase from Staphylococcus aureus. Hybridization studies revealed that several strains of Rhizobium hedysari carry this element in various copy number. The six copies in strain IS 123 appear clustered specifically within the pSym nod region.The significance of this IS element in rhizobia and its possible use as a probe for taxonomic and phylogenetic studies of Rhizobiaceae is addressed.     

Shoot Sr concentrations in relation to shoot Ca concentrations and to soil properties

This work was aimed to investigate whether shoot Sr concentrations of plant species are related to respective Ca concentrations and to soil properties and to compare the Sr-Ca observed ratios (OR), defined as the quotient of the ratios Sr/Ca in shoots and in the soil solution or in the extractable form, among species and soils. Ten pasture plant species were grown in pots (1-L volume) filled with eight soils differing in the various physicochemical characteristics. Each pot received 50 mg Sr except those of the soil with the highest cation exchange capacity (C.E.C.) that received 100 mg Sr per pot. For each soil, shoot Sr concentrations of species were linearly and positively related with the respective Ca concentrations. C.E.C, organic matter content and Ca in the soil solution or in the extractable form were the only soil properties that were related, all negatively, with shoot Sr concentrations. The ratio of extractable Sr and Ca was positively and linearly related with the ratio of Sr and Ca. in the soil solution. OR was affected by both species and soils. Most of OR values of all species in all soils ranged between 0.8 and 1.5, except for the grass Agrostis capillaris which had the highest values for most of soils. This indicates that Agrostis capillaris compared to other species, takes up proportionally more Sr than Ca.     

Interaction of poly(ADP-ribose)polymerase with DNA polymerase α

Homogeneously purified poly(ADP-ribose) polymerase (PARP) specifically stimulated the activity of immunoaffinity-purified calf or human DNA polymerase by about 6 to 60-fold. Apparently, poly(ADP-ribosyl)ation of DNA polymerase was not necessary for the stimulation. The effects of PARP on DNA polymerase were biphasic: at very low concentrations of DNA, it rather inhibited its activity, whereas, at higher DNA concentrations, PARP greatly stimulated it. The autopoly(ADP-ribosyl)ation of PARP suppressed both its stimulatory and inhibitory effects. By immunoprecipitation with an anti-DNA polymerase antibody, it was clearly shown that PARP may be physically associated with DNA polymerase . Stimulation of DNA polymerase may be attributed to the physical association between the two, rather than to the DNA-binding capacity of PARP, since the PARP fragment containing only the DNA binding domain showed little stimulatory activity. The existence of PARP-DNA polymerase complexes were also detected in crude extracts of calf thymus.     

Isolation of glucosinolate degrading microorganisms and their potential for reducing the glucosinolate content of rapemeal

Abstract K88ab fimbriae are filamentous protein structures at the surface of certain enterotoxigenic Escherichia coli strains. Electron microscopy analysis of K88ab fimbriae showed that these structures have different morphological appearances dependent on the medium in which cells expressing these fimbriae or in which purified fimbriae were suspended. Thin and curled structures, thin and flexible fimbriae, a wider and rigid form of the fimbriae, and, in addition, paracrystalline structures were detected. Optical diffraction analysis of the paracrystalline structures indicated a helical conformation of K88ab fimbriae.     

Pod development of groundnut (Arachis hypogaea L.) in solution culture

Normal pods (containing seed) of groundnut (Arachis hypogaea L.) (cv. TMV-2) were successfully raised in darkened, aerated, nutrient solution, but not in the light. The onset of podding was evident 7 to 8 d after gynophores were submerged in the darkened nutrient solution. An examination of pods and submerged portions of gynophore surfaces by scanning electron microscopy showed the presence of two distinctly different protuberances: unicellular root-hair-like structures that first developed from epidermal cells of the gynophores and developing pods; and branched septate hairs that developed later from cells below the epidermal layer. The septate hairs became visible only after the epidermal and associated unicellular structures had been shed by the expanding gynophore and pods. Omission of Mn and Mg from the podding environment increased pod and seed weight, whilst omission of Zn reduced pod and seed weight.     

Evidence for linkage between K88ab, K88ac intestinal receptors to Escherichia coli and transferrin loci in pigs

Segregation at the loci coding for the K88ab and K88ac small intestinal receptors to E. coli adhesins (K88abR, K88acR) and at the transferrin (TF) locus was studied in 38 pig families including 273 piglets. The TF locus showed a segregation deviation towards the B variant while each of the K88 receptors behaved as a single autosomal dominant gene. Recombinants between K88abR and K88acR provide evidence that they are under the control of two different loci. Thirty-two triple backcross families were selected to test linkage and estimate recombination rates (θ). Our results demonstrate that the two K88 receptor loci are closely linked (θ= 0.02) with a maximum lod score value (Z_m) of 46.0. In addition, they are linked to the TF locus, θ= 0.14, Z_m= 19.6 for the K88abR locus and θ= 0.16, Z_m= 17.9 for the K88acR locus. The estimated recombination rates, smaller in males than in females, are consistent with the order TF-K88abR-K88acR. This linkage thus localizes the K88 loci, as the TF locus, on chromosome 13.