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  • 1
    ISSN: 1432-1211
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract  Our analysis of IgG rheumatoid factors (RFs) from three patients with rheumatoid arthritis (RA) revealed that most contained significant numbers of skewed mutations per V region, suggesting that these RFs arose from antigen-driven responses. To further study IgG RFs in RA, we used pComb3 vector to construct an IgG1,λ combinatorial antibody library from a synovial fluid sample. After panning against human IgG, Fab fragments from 71/96 phage clones bound to Fc-coated wells. Sequence analysis of 20 randomly chosen Fc-binders showed that 17 (85%) clones had identical heavy (H) and light (L) chain V regions, represented by Humha311 and Humla211, respectively. Of the remaining three clones, two had the same Humla211 L chain, but each with a different H chain V region. All the putative germline V genes for these RFs also encode RF in RA patients. However, none of these RF V regions are similar to those of the two IgG RFs derived by the hybridoma technique from the same synovial fluid. The Humha311 H chain has two frameshifts: a one-base insertion upstream of the JH region and a four-base deletion near the end of the CH1 region, resulting in a mainly unconventional amino acid sequence in the CH1 region. In the future, it will be important to study the presence of IgG molecules with such unconventional CH1 amino acid sequences, and the effects of these amino acid sequences on the structures and immunological properties of the IgG molecules.
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  • 2
    ISSN: 1432-072X
    Keywords: 3,5-Dichlorocatechol ; 2,4-Dichloromuconate ; Dichloromuconate cycloisomerase ; 2-Chloro-4-carboxymethylenebut-2-en-4-olide ; Alcaligenes eutrophus JMP 134 ; Pseudomonas sp. B 13
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract 2,4-Dichloro-cis,cis-muconate is established as ringcleavage product in the degradation of 3,5-dichlorocatechol by Alcaligenes eutrophus JMP 134. The formerly described isomerization of 2-chloro-trans- to 2-chlorocis-4-carboxymethylenebut-2-en-4-olide as an essential catabolic step could not be certified.
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  • 3
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract To more closely define the risk of infection by the agent of Lyme disease in Europe, we determined whether spirochetal prevalence increases throughout the development of theIxodes ricinus vector tick. Of all ticks that could be flagged from vegetation,I. ricinus were by far the most abundant. Spirochetal infection rates in the adult stage of this tick (15%) are no higher than those in nymphs (18%) but greatly exceed those in larvae (0.7%). This tick therefore appears to attain infection mainly from the host of its larval stage, generally feeds on hosts that are noncompetent as reservoirs in its nymphal stage, and rarely inherits infection. Risk of human infection mainly derives from contact with the nymphal stage of the vector tick because the larva is rarely infected and the adult is large enough to be noticed and promptly removed.
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  • 4
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract For a comparison of the times of day at which the subadult stages ofIxodes ricinus detach from nocturnal vs diurnal hosts, these ticks were placed on a variety of indigenous and experimental animals. The time of detachment appears to depend more on properties of the host than on the periodicity of tick behavior. Ticks on rodents, regardless of host periodicity, tended to detach late in the afternoon; those on a hedgehog detached around midnight, and those on lizards and birds, during the morning. Ticks on carnivores (dog, cat) detached throughout the daylight hours. The ability of these parasites to survive to the next developmental stage and, ultimately, to come into contact with another suitable host may be influenced by the identity of the host and, hence, by the circumstances of detachment. SubadultI. ricinus probably concentrate in the host's nest when feeding on mammals but are scattered over the ground when feeding on lizard or avian hosts an aspect of engorgement behavior that may profoundly affect the capacity of this tick as a vector of agents of Lyme disease and other infections.
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  • 5
    ISSN: 1618-2650
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The hydrolytic stability of organophosphonic acids, comprising 〉NCH2PO3H2 moieties, is studied exemplarily on aqueous solutions of ethylenediaminetetramethylenephosphonic acid (EDTMP). Metabolization of EDTMP increases severely at lower pH values, and is enhanced at higher temperature. The formation of orthophosphate in the course of the hydrolytical degradation can be established unequivocally by several, methodologically independent analytical procedures. By ion chromatography, the presence of (inorganic) phosphonate (phosphite) is affirmed in alkaline, and of hydroxymethylphosphonate in neutral to acid solutions. Formation of all three metabolites is in accord with a general degradation mechanism proposed here. The results of these stability investigations have far-reaching consequences for the analytical practice, as well as for the industrial application of organophosphonic acids.
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  • 6
    ISSN: 1618-2650
    Source: Springer Online Journal Archives 1860-2000
    Topics: Chemistry and Pharmacology
    Notes: Summary The ion-chromatographic separation of organophosphonic acids is investigated systematically in a comparative study of two anion-exchange resins (Dionex HPIC AS7, Biotronik BT I AN). For thirteen structurally diverse phosphonic acids, chromatographic properties are determined with HNO3 eluents (5–30 mmol/l), and compared with those of classical complexones and inorganic anions, e.g. phosphonate (phosphite) and phosphate. While all anions tested may be determined, in principle, both qualitatively and quantitatively (down to a 10 ppm scale) on the two materials, the Dionex column exhibits the far higher separation potential and can be applied to almost any analytical problem in the phosphonic acid field. A systematic optimization of the separation conditions for the Dionex HPIC AS7 (a detailed prescription is given) makes qualitative and quantitative analyses possible which hitherto could not be performed with this material.
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  • 7
    ISSN: 1573-5028
    Keywords: acid phosphatase ; deoxyinosine-containing primers ; polymerase chain reaction ; restriction fragment length polymorphism analysis ; root-knot nematode resistance ; tomato
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract With a view to cloning the root-knot nematode resistance gene Mi in tomato by chromosome walking, we have developed a molecular probe for the tightly linked acid phosphatase-1 (Aps-1) locus. The acid phosphatase-1 allozyme (APS-11), encoded by the Aps-1 1 allele originating from Lycopersicon peruvianum, was purified to apparent homogeneity from tomato roots and suspension cells. Microsequencing of CNBr and tryptic peptides generated from APS-11 provided a partial amino acid sequence, which accounted for approximately 23% of the protein and revealed two stretches of homology with soybean proteins KSH3 and VSP27, comprising 22 matches within 26 amino acid residues. The partial amino acid sequence information enabled us to isolate a 2.4 kb genomic Aps-1 1 sequence by means of the polymerase chain reaction (PCR), primed by degenerate pools of oligodeoxyribonucleotides, synthesized on the basis of the amino acid sequences. Synthesis of the 2.4 kb PCR product was specific for genomic templates carrying the L. peruvianum Aps-1 1 allele. Crucial to the priming specificity and the synthesis of the 2.4 kb genomic sequence was the use of degenerate primer pools in which the number of different primer species was limited by incorporating deoxyinosine phosphate residues at three and four base ambiguities. In using cDNA as a template, a 490 bp sequence was obtained, indicating a high proportion of intron sequences in the 2.4 kb genomic Aps-1 1 sequence. The Aps-1 1 origin of the PCR product was confirmed by RFLP (restriction fragment length polymorphism) analysis, using both a chromosome 6 substitution line and a pair of nearly isogenic lines, differing for a small chromosomal region around the Aps-1/Mi loci.
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  • 8
    ISSN: 1573-0603
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
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  • 9
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract A quantitative, competitive polymerase chain reaction (QC-PCR) assay for the sensitive detection of Wuchereria bancrofti DNA was developed. A competitor sequence was constructed by an exchange of nucleotides in the Wuchereria-specific Ssp I repeat. The PCR products were hybridized to specific DNA probes and their amounts, determined by an enzyme-linked immunosorbent assay (ELISA). In laboratory-prepared samples the QC-PCR-ELISA assay was capable of detecting the amount of DNA equivalent to 0.1 microfilaria (mf) added to 200 μl of blood lysate. The assay was also tested on 78 blood samples collected in endemic areas in Egypt. All 28 samples that were positive both for mf and for circulating antigen were also QC-PCR-ELISA-positive. In addition, one mf-negative but antigen-positive sample was also positive as determined by QC-PCR-ELISA. A positive correlation of mf density with the QC-PCR-ELISA was observed. Samples containing 10 or fewer mf/ml had a mean relative amount of Ssp I PCR product of 19.7 units, whereas samples with 11–100 mf/ml had a mean of 36.3 units and those with more than 100 mf/ml had a mean of 84.6 units. Because of the high standard deviation within each group, estimates of worm burdens in infected individuals using the QC-PCR-ELISA are not recommended. However, we present data indicating that the W. bancrofti QC-PCR-ELISA is a powerful new tool for evaluation of parasitic loads for community-based diagnosis of bancroftian filariasis.
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  • 10
    ISSN: 1432-1955
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology , Medicine
    Notes: Abstract In recent years, bovine Onchocerca species have been used as models for human onchocerciasis in drug screens. They have been suggested for immunology studies and evaluation of vaccine candidates. Therefore, mast cells and their association with other inflammatory cells were studied in five onchocercal species of cattle and deer using immunohistology. Intact mast cells occurred in large numbers in the capsule and septae of nodules, in fibrous tissue adjacent to nonnodular worms, and perivascularly. Inactive and, more frequently, activated and degranulating mast cells were observed within infiltrates in the nodule center or around nonnodular filariae. They were not detected in direct contact with the cuticle of adult worms or of microfilariae or among the macrophages, giant cells, and neutrophils forming the innermost layer around the worms. Eosinophils, but not mast cells, were obviously associated with microfilariae-producing females. The distribution, frequency, and activity of mast cells were similar for all five species and O. volvulus.
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