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  • 1990-1994  (17,892)
  • 1955-1959  (1,475)
  • 1890-1899  (195)
  • Life and Medical Sciences  (11,936)
  • Analytical Chemistry and Spectroscopy  (7,463)
  • Agrobacterium
  • genetic engineering
  • protoplasts
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Year
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  • 1
    Electronic Resource
    Electronic Resource
    A reporter gene under the control of tms or aux promoters is differentially expressed in tobacco and barley protoplasts (1994)
    Springer
    Plant cell reports 13 (1994), S. 155-158 
    Details
    ISSN: 1432-203X
    Keywords: Agrobacterium ; auxin biosynthesis genes ; barley and tobacco protoplasts ; transient expression
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Agrobacterium tumefaciens and some Agrobacterium rhizogenes strains possess auxin biosynthesis genes (tms and aux genes respectively), responsible for a de novo auxin biosynthetic pathway in transformed plant cells. A comparison is presented of the potential expression of these genes in a monocotyledonous (barley) and a dicotyledonous plant (tobacco). The promoters of the genes were translationally fused to the β-glucuronidase reporter gene and analysed in transient expression experiments. The tms and aux fusions were highly expressed in tobacco, but not in barley. However, the aux enhancer active in tobacco, conferred low β-glucuronidase expression in barley when fused to a truncated cauliflower mosaic virus 35S promoter. The results are discussed in relation to the differential responses to Agrobacterium infection in monocots and dicots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00239883
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  • 2
    Electronic Resource
    Electronic Resource
    Stable expression of the GUS reporter gene in chrysanthemum depends on binary plasmid T-DNA (1994)
    Springer
    Plant cell reports 14 (1994), S. 59-64 
    Details
    ISSN: 1432-203X
    Keywords: Agrobacterium ; transformation ; T-DNA
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three chrysanthemum (Dendranthema grandiflora) cultivars were cocultivated with 2 Agrobacterium tumefaciens strains in combination with 4 pBIN19 derived binary plasmids, all carrying the Nosnptll selection gene and 35Sgus(intron) reporter gene. All binary plasmids transferred DNA to chrysanthemum explants but only pMOG410 gave good stable expression of GUS. This plasmid differs from the other plasmids in 2 aspects: 1) It carries a restored nptll gene and 2) the selection gene is positioned at the left border side of the reporter gene. Cocultivation with AGLO(pMOG410) yielded up to 13 GUS positive shoots per 100 explants. The presence of the gus and nptll gene in recovered shoots was confirmed by PCR and Southern blot analysis.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233300
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  • 3
    Electronic Resource
    Electronic Resource
    Agrobacterium-mediated transformation of commercial melon (Cucumis melo L., cv. Amarillo Oro) (1994)
    Springer
    Plant cell reports 13 (1994), S. 145-148 
    Details
    ISSN: 1432-203X
    Keywords: muskmelon ; gene transfer ; Agrobacterium ; regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Cotyledon explants of muskmelon (Cucumis melo L., cv. Amarillo Oro) seedlings were co-cultivated with disarmed Agrobacterium tumefaciens strain LBA4404 that contained the binary vector plasmid pBI121.1. The T-DNA region of this binary vector contains the Nopaline synthase/neomycin phosphotransferase II (NPTII) chimeric gene for kanamycin resistance and the Cauliflower Mosaic Virus 35S/β-glucuronidase (GUS) chimeric gene. After infection, the cotyledon pieces were placed in induction medium containing 100 mg/l kanamycin. Putative transformed shoots were obtained, followed by the development of morphologically normal plantlets. The transgenic nature of regenerants was demonstrated by polymerase chain reaction, Southern blot analysis, plant growth on medium selective for the transgene (NPTII) and expression of the co-transformed GUS gene. Factors affecting the transformation procedure are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00239881
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  • 4
    Electronic Resource
    Electronic Resource
    Isolation and transformation of rice aleurone protoplasts (1994)
    Springer
    Plant cell reports 13 (1994), S. 394-396 
    Details
    ISSN: 1432-203X
    Keywords: Rice ; α-amylase ; protoplasts ; aleurone ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Protoplasts isolated from the aleurone have been used extensively in molecular studies focusing on hormone-mediated regulation of gene expression in barley seed. To extend the use of aleurone protoplasts to other species, we have determined the conditions necessary for the isolation of protoplasts from rice aleurone layers of germinated seed. Many of the common cell wall degrading enzymes used in making protoplasts were tested for their ability to release protoplasts from rice aleurone layers. Cellulysin was found to be the most effective. Transformation of these aleurone protoplasts was accomplished using polyethylene glycol and DNA constructs containing the firefly luciferase reporter gene under the control of two different promoters were tested. Luciferase expression was 24-fold greater when the reporter gene was under the control of the CaMV 35S promoter than when the promoter from the alcohol dehydrogenase 1 gene was used. With the isolation and transformation of aleurone protoplasts from rice, it is now possible to investigate molecular events occurring in this tissue during germination.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234145
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  • 5
    Electronic Resource
    Electronic Resource
    A reproducible procedure for plant regeneration from seedling hypocotyl protoplasts of Vigna sublobata L. (1994)
    Springer
    Plant cell reports 14 (1994), S. 175-179 
    Details
    ISSN: 1432-203X
    Keywords: Vigna sublobata ; protoplasts ; microcalli ; shoot bud formation ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Viable protoplasts of Vigna sublobata L. were isolated enzymatically from hypocotyls of axenic seedlings. Protoplast yields were dependent upon seedling age, with maximum yields (2.25 ± 0.35 × 106 g fwt−1) from seedlings aged 6 d. Protoplasts regenerated cell walls and underwent sustained divisions when cultured in either agarose-solidified or liquid K8P medium. The plating density affected the division frequency and plating efficiency; the division frequency (68 ±0 6.0%) was maximum at 4.0 × 104 ml−1 while plating efficiency was maximum (1.3 ± 0.1%) at 5.0 × 104 ml−1. Dividing protoplasts developed into microcalli, which produced glossy green compact nodular calli on transfer to 8.0 gl−1 w/v agar-solidified medium containing MS salts, B5 organic components, 30 g l−1 sucrose, NAA (0.2–0.5 mg l−1), zeatin riboside (0.5–2.0 mg l−1) and GA3 (0.5–1.0 mg l−1). These calli, after sub-culture on the same medium, produced shoot buds which underwent elongation following transfer of tissues to 6.0 g l−1 agar-solidified B5 medium containing 30g l−1 sucrose, IBA (0.01 mg l−1) and BAP (1.0 mg l−1). Elongated shoots developed roots after transfer to 8.0g l−1 agar-solidified, hormone-free MS medium with 30 g l−1 sucrose.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233785
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  • 6
    Electronic Resource
    Electronic Resource
    Synergistic enhancement of protoplast growth by oxygenated perfluorocarbon and Pluronic F-68 (1994)
    Springer
    Plant cell reports 13 (1994), S. 251-255 
    Details
    ISSN: 1432-203X
    Keywords: Petunia hybrida ; protoplasts ; oxygen delivery ; perfluorochemicals ; Pluronic F-68 ; surfactant ; cell division
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary Cell suspension-derived protoplasts of albino Petunia hybrida were grown for 10 d at the interface between aqueous culture medium (KM8P) and an oxygenated (10 mbar for 15 min) perfluorocarbon liquid, perfluorodecalin. Protoplasts synthesised new cell walls and divided normally at the perfluorodecalin/culture medium interface, with a mean viability after 10 d of 〉 92.0%. The mean plating efficiency of protoplasts was elevated by 37% (P〈0.05) following culture at the perfluorodecalin/medium interface, but was unaltered by perfluorodecalin or oxygen separately. The mean plating efficiency of protoplasts cultured at the interface was further increased to a maximium of 52% above control, in the presence of oxygenated perfluorodecalin and KM8P medium supplemented with the non-ionic, co-polymer surfactant, Pluronic F-68 at 0.01% (w/v). These findings demonstrate the effectiveness of oxygenated perfluorodecalin for promoting protoplast growth, by facilitating oxygen delivery. The finding that Pluronic F-68 further increased the plating efficiency of protoplasts cultured at the perfluorocarbon/aqueous interface suggests that these agents improve growth through separate, but cumulative, mechanisms.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00233314
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  • 7
    Electronic Resource
    Electronic Resource
    Effect of plant genotype on the transformation of cultivated alfalfa (Medicago sativa) by Agrobacterium tumefaciens (1994)
    Springer
    Plant cell reports 13 (1994), S. 330-334 
    Details
    ISSN: 1432-203X
    Keywords: Medicago sativa ; Alfalfa ; Transformation ; Agrobacterium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The trait for somatic embryogenesis is being introduced sexually into alfalfa (Medicago sativa) breeding populations to facilitate genetic transformation of this crop. Cocultivation experiments were conducted with an agronomically-improved embryogenic clone from one such population as well as with two other embryogenic clones, one of which was the source of the embryogenic trait in the breeding populations. Transgenic plants were produced from the agronomically-improved clone whereas none were produced from the other two clones. Among the 16 transgenic plants analyzed there was a range in both copy number and number of integration sites for the NPT-II gene; those plants regenerated after a prolonged selection phase in vitro generally had the highest numbers in both respects. There was no evidence of sectoral chimerism of the transgene in a subsample of transgenic plants analyzed by PCR.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00232631
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  • 8
    Electronic Resource
    Electronic Resource
    Identification of Agrobacterium strains by PCR-RFLP analysis of pTi and chromosomal regions (1994)
    Springer
    Archives of microbiology 161 (1994), S. 300-309 
    Details
    ISSN: 1432-072X
    Keywords: Agrobacterium ; 16S rDNA ; 16S rDNA-23S rDNA Intergenic spacer ; tmr Gene ; nos Gene ; virA Gene ; virB2 Gene ; Polymerase chain reaction ; Restriction fragment length polymorphism ; Crown gall
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Chromosomes and Ti plasmids of 41 Agrobacterium strains, belonging to biovars 1, 2, 3, and Agrobacterium rubi species were characterized by the restriction fragment length polymorphism of PCR-amplified DNAs. Profiles that were obtained by the analysis of the amplified 16S rDNA confirmed the grouping of the strains according to their species. Higher polymorphism was detected in the intergenic spacer between the 16S rDNA and 23S rDNA genes, allowing efficient discrimination of strains. Identification of most strains was possible, and the genetic relatednesses of Agrobacterium strains could be estimated. The analysis of the plasmid Ti encoded regions between the tmr and nos genes, and the virA and virB2 genes, allowed fingerprinting of Ti plasmids. Genomic typing by the rapid PCR-RFLP method is thus shown to be useful for an independant identification of strains and of the conjugative Ti plasmids.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00303584
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  • 9
    Electronic Resource
    Electronic Resource
    Several phenotypic changes in the cell envelope of Agrobacterium tumefaciens chvB mutants are prevented by calcium limitation (1994)
    Springer
    Archives of microbiology 161 (1994), S. 310-315 
    Details
    ISSN: 1432-072X
    Keywords: Agrobacterium ; β-1,2-Glucan ; chvB Mutants—Ca2+
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The chvB gene of Agrobacterium tumefaciens encodes a 235 kDa proteinaceous intermediate involved in the synthesis of β-1,2-glucan. chvB mutants show a pleiotropic phenotype. Besides not to produce cyclic β-1,2-glucan, chvB mutants have been reported to be avirulent, attachment-deficient, and nonmotile. In this study we report additional differences from the parent strain, probably all linked to changes in the cell envelope. This pleiotropic phenotype — except for attachment and virulence — could largely be prevented by growing chvB cells with low levels of calcium. Although a role for β-1,2-glucan in osmoadaptation has been proposed, the mode of action of β-1,2-glucan is not known. We speculate that in A. tumefaciens β-1,2-glucan stabilizes membranes, which would be important especially in hypotonic media containing calcium.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00303585
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  • 10
    Electronic Resource
    Electronic Resource
    Expression of biologically active hordothionins in tobacco. Effects of pre- and pro-sequences at the amino and carboxyl termini of the hordothionin precursor on mature protein expression and sorting (1994)
    Springer
    Plant molecular biology 24 (1994), S. 83-96 
    Details
    ISSN: 1573-5028
    Keywords: anti-bacterial protein ; genetic engineering ; precursor processing ; synthetic gene ; thionin ; transgenic
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hordothionins (HTHs) are small anti-bacterial proteins present in barley endosperm which are processed from larger precursor proteins, consisting of an amino-terminal signal peptide (SP), the mature highly basic HTH and a carboxy-terminal acidic peptide (AP). Different HTH precursor proteins were expressed in tobacco to study the effects of the pre-sequences (SP) and pro-sequences (AP) on expression, processing, sorting and biological activity and hence the feasibility of engineering bacterial disease resistance into crops which lack these proteins. Maximum HTH expression levels of approximately 0.7% (11 μmol/kg) of total soluble protein in young tobacco leaves were obtained using a semi-synthetic gene construct encoding a complete chimaeric HTH precursor protein. Tenfold lower HTH expression levels (maximum 1.3 μmol/kg) were obtained using synthetic gene constructs without the AP-coding sequence and no expression was found in plants containing synthetic HTH gene constructs without SP-and AP-coding sequences. In both cases where expression was found, the precursors were apparently correctly processed, although the HTH produced in plants containing a construct without AP sequence appeared to be slightly modified. No effect on plant phenotype was observed. Localization studies indicated that the HTH was in identical fractions of plants expressing the two different precursors, albeit at a different ratio, and was not secreted into the intercellular spaces of leaves or culture medium by protoplasts. Our results indicated that the AP is not involved in sorting and suggested that it might facilitate transport through membranes. The in vitro toxicity of HTH isolated from transgenic tobacco plants expressing the two different precursor proteins for the bacterial plant pathogen Clavibacter michiganensis subsp. michiganensis appeared similar to that of the HTH purified from barley endosperm.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040576
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  • 11
    Electronic Resource
    Electronic Resource
    Localization of the VirA domain involved in acetosyringone-mediatedvir gene induction inAgrobacterium tumefaciens (1994)
    Springer
    Plant molecular biology 25 (1994), S. 899-907 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; gene regulation ; sensor protein ; signal transduction ; VirA protein ; acetosyringone
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The VirA protein ofAgrobacterium tumefaciens is thought to be a receptor for plant phenolic compounds such as acetosyringone. Although it is not known whether the interaction between VirA and the phenolics is direct or requires other phenolic-binding proteins, it is shown in this study that the first 280 amino acids of the VirA protein are not essential for the acetosyringone mediatedvir gene induction response. Considering the fact that the cytoplasmic region between the amino acids 283 and 304 is highly conserved between the different VirA proteins, and that deletion of this region abolishes VirA activity, we suggest that the acetosyringone receptor domain is located in this cytoplasmic domain of the VirA protein.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00028884
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  • 12
    Electronic Resource
    Electronic Resource
    The small, versatilepPZP family ofAgrobacterium binary vectors for plant transformation (1994)
    Springer
    Plant molecular biology 25 (1994), S. 989-994 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; binary vectors ; gentamycin resistance ; kanamycin resistance ; tobacco
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The newpPZP Agrobacterium binary vectors are versatile, relatively small, stable and are fully sequenced. The vectors utilize the pTiT37 T-DNA border regions, the pBR322bom site for mobilization fromEscherichia coli toAgrobacterium, and the ColE1 and pVS1 plasmid origins for replication inE. coli and inAgrobacterium, respectively. Bacterial marker genes in the vectors confer resistance to chloramphenicol (pPZP100 series) or spectinomycin (pPZP200 series), allowing their use inAgrobacterium strains with different drug resistance markers. Plant marker genes in the binary vectors confer resistance to kanamycin or to gentamycin, and are adjacent to the left border (LB) of the transferred region. A lacZ α-peptide, with the pUC18 multiple cloning site (MCS), lies between the plant marker gene and the right border (RB). Since the RB is transferred first, drug resistance is obtained only if the passenger gene is present in the transgenic plants.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00014672
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  • 13
    Electronic Resource
    Electronic Resource
    Purification and partial characterization of a glycoprotein from pea (Pisum sativum) with receptor activity for rhicadhesin, an attachment protein of Rhizobiaceae (1994)
    Springer
    Plant molecular biology 24 (1994), S. 171-183 
    Details
    ISSN: 1573-5028
    Keywords: Agrobacterium ; attachment ; plant receptor ; rhicadhesin ; Rhizobium
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Attachment of Rhizobium and Agrobacterium bacteria to cells of their host plants is a two-step process. The first step, direct attachment of bacteria to the plant cell wall, is mediated by the bacterial protein rhicadhesin. A putative plant receptor molecule for rhicadhesin was purified from cell walls of pea roots using a bioassay based on suppression of rhicadhesin activity. This molecule appeared to be sensitive to treatments with pronase or glycosidase. Its isoelectric point is 6.4, and its apparent molecular mass was estimated to be 32 kDa before and 29 kDa after glycosidase treatment, as determined by sodium dodecyl sulphate-polyacrylamide gel electrophoresis and ultrafiltration. The sequence of the first 29 N-terminal amino acids was determined: A-D-A-D-A-L-Q-D-L-C(?)-V-A-D-Y-A-S-V-I-L-V-N-G-F-A-S-K(Q)-(P/Q)-(L)-(I). No homology with known proteins was found. In the course of this research project the extracellular matrix protein vitronectin was reported to inhibit attachment of A. tumefaciens to carrot cells [29]. A variety of adhesive proteins, including vitronectin, contain a common cell attachment determinant with the sequence R-G-D. Since we could not detect other cell wall components able to suppress rhicadhesin activity, and since an R-G-D containing hexapeptide was also active as a receptor, we speculate that the plant receptor for rhicadhesin is a glycoprotein containing an R-G-D attachment site.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00040583
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  • 14
    Electronic Resource
    Electronic Resource
    Coordinate expression of antibody subunit genes yields high levels of functional antibodies in roots of transgenic tobacco (1994)
    Springer
    Plant molecular biology 26 (1994), S. 1701-1710 
    Details
    ISSN: 1573-5028
    Keywords: Antibody ; coordinated expression ; genetic engineering ; protein assembly ; root ; secretion
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract To explore the feasibility of employing antibodies to obtain disease resistance against plant root pathogens, we have studied the expression of genes encoding antibodies in roots of transgenic plants. A model monoclonal antibody was used that binds to a fungal cutinase. Heavy and light chain cDNAs were amplified by PCR, fused to a signal sequence for secretion and cloned behind CaMV 35S and TR2′ promoters in a single T-DNA. The chimeric genes were cloned both in tandem and in a divergent orientation. The roots of tobacco plants transformed with these constructs produced antibodies that were able to bind antigen in an ELISA. Immunoblotting showed assembly to a full-size antibody. In addition, a F(ab′)2-like fragment was observed, which is probably formed by proteolytic processing. Both antibody species were properly targeted to the apoplast, but the full-size antibody was partially retained by the wall of suspension cells. The construct with divergent promoters showed a better performance than the construct with promoters in tandem. It directed the accumulation of functional antibodies to a maximum of 1.1% of total soluble protein, with half of the plants having levels higher than 0.35%. The high efficiency of this construct probably results from coordinated and balanced expression of light and heavy chain genes, as evidenced by RNA blot hybridization.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00019485
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  • 15
    Electronic Resource
    Electronic Resource
    A simple and efficient gene transfer system of trifoliate orange (Poncirus trifoliata Raf.) (1994)
    Springer
    Plant cell reports 13 (1994), S. 541-545 
    Details
    ISSN: 1432-203X
    Keywords: Trifoliate orange ; Epicotyl segment ; Agrobacterium ; Transformation ; rolC promoter ; β-glucuronidase
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Summary A simple and efficient gene transfer system of trifoliate orange (Poncirus trifoliata Raf.) was developed using epicotyl segments. The segments were infected with Agrobacterium harboring the binary vector pBI121 or pBI101-O12-p1. Both vectors contained the neomycin phosphotransferase II (NPTII) and the β-glucuronidase (GUS) genes. In the plasmid pBI101-O12-p1, the GUS gene was directed to the promoter region of ORF12 (rolC) of the Ri plasmid. On a selection medium containing 100 or 200 μg/ml kanamycin, adventitious shoots were formed from 21.7–44.6% of the segments. Histochemical GUS assay showed that 55.4–87.7% of the shoots expressed the GUS gene. The stable integration of this gene was also confirmed by polymerase chain reaction (PCR) analysis and by Southern blot analysis. When the pBI101-O12-p1 plasmid was used, the GUS activity was found to be located in phloem cells of leaf, stem and root. More than 100 transformed plants were obtained using this method within 2–3 months.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00234507
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  • 16
    Electronic Resource
    Electronic Resource
    Expression and inheritance pattern of two foreign genes in petunia (1994)
    Springer
    Theoretical and applied genetics 88 (1994), S. 433-440 
    Details
    ISSN: 1432-2242
    Keywords: Agrobacterium ; Transformation ; Gene expression ; Petunia
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transgenic petunia (Petunia hybrida Vilm.) plants were obtained from Agrobacterium-mediated shoot apex transformation. Studies at the phenotypic as well as molecular level established both the presence of the NPT II (neomycin phosphotransferase II) and GUS (β-glucuronidase) genes and their level of activity. Twenty-nine primary transformed plants showed varying patterns of phenotype expression of both genes. NPT II and GUS expression in 7 primary plants over a 4-month interval showed varying levels of gene expression within and among individual plants. All primary transgenic plants were self-pollinated and backcrossed to establish the inheritance patterns of both genes. Mendelian and non-Mendelian inheritance patterns for both genes were observed. Analysis of the progeny showed poor transmission of the foreign genes through the pollen especially when two or more bands were present in the Southern hybridization. Most plants whose progeny segregated in Mendelian ratios for either the NPT II or GUS gene had just one copy of the gene. In this study where both foreign genes were examined in both self and test crosses, no transgenic plant showed Mendelian patterns of inheritance for both foreign traits.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00223657
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  • 17
    Electronic Resource
    Electronic Resource
    Cybridization in Nicotiana tabacum L. using double inactivation of parental protoplasts and post-fusion selection based on nuclear-encoded and chloroplast-encoded marker genes (1994)
    Springer
    Theoretical and applied genetics 88 (1994), S. 1017-1022 
    Details
    ISSN: 1432-2242
    Keywords: Agrobacterium ; Iodoacetamide Nicotiana protoplast fusion ; Nitrosomethyl urea Rhodamine 6G ; X-irradiation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract An effective selection system preceded by double inactivation of parental protoplasts was used to transfer Nicotiana suaveolens Leh. cytoplasmic male sterility into a commercial tobacco (N. tabacum L.) breeding line. Mesophyll protoplasts from transformed plants of N. tabacum cultivar WZ2-3-1-1 possessing a neomycin phosphotransferase II gene were used as the nuclear donors, while those isolated from N. suaveolens plants carrying a chloroplast mutation for resistance to spectinomycin, induced using nitrosomethyl urea, were the cytoplasm donors in somatic cybridizations. Prior to fusion, nuclear donor protoplasts were inactivated with iodoacetamide or rhodamine 6G, while those of the cytoplasm donor were inactivated by X-irradiation. The resultant microcalli were cultured on a shoot regeneration medium containing both kanamycin and spectinomycin to select cybrids. Only regenerants that had typical characteristics of the N. tabacum cultivar were selected for transfer to the glasshouse. Four putative cytoplasmic male-sterile (CMS) plants, out of a total of 44 regenerated plants transferred to the glasshouse, were obtained. Intraspecific somatic transfers of the CMS trait between N. tabacum cultivars with distinctlydifferent morphologies using single inactivation and nonselective shoot regeneration medium were demonstrated. The implications of the results for practical tobacco breeding as a means of circumventing lengthy backcrossing procedures are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00220810
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  • 18
    Electronic Resource
    Electronic Resource
    Transformation of protoplasts and intact cells from slowly growing embryogenic callus of wheat (Triticum aestivum L.) (1994)
    Springer
    Theoretical and applied genetics 89 (1994), S. 577-582 
    Details
    ISSN: 1432-2242
    Keywords: Wheat ; Transformation ; Agrobacterium ; Electroporation ; Polyethylene glycol
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract A procedure for culturing protoplasts from slowly growing embryogenic calli of wheat was developed. The procedure was dependent on the ability to isolate large numbers of culturable protoplasts from slowly growing embryogenic callus. Approximately 68% of the isolated protoplasts divided, and 22% formed colonies; of the latter, 67% continued to proliferate. Plating efficiency was reduced when protoplasts were transformed by polythylene glycol, electroporation, and/or Agrobacterium. Intact cells were also directly transformed by electroporation. Direct electroporation of the Agrobacterium binary vector into intact cells resulted in a significant increase of GUS activity over the control.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00222451
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  • 19
    Electronic Resource
    Electronic Resource
    T-DNA-insert-independent mutations induced in transformed plant cells duringAgrobacterium co-cultivation (1994)
    Springer
    Transgenic research 3 (1994), S. 317-325 
    Details
    ISSN: 1573-9368
    Keywords: Agrobacterium ; mutagenesis ; Nicotiana plumbaginifolia ; nitrate reductase ; ploidy ; transformation
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Transformation frequencies were determined for 1n, 2n, and 4n Nicotiana plumbaginifolia protoplast cultures inAgrobacterium-mediated gene transfer experiments. An unexpected large drop (50%) in plating efficiencies was observed in the non-selected (control) 1n populations after transformation treatment with virulent strains. This effect was not observed in the 2n or 4n cultures or in the 1n cultures when treated with avirulent bacteria. The mortality was disproportionally high and could not be explained by the low (0.1–0.5%) transformation efficiency in the 1n population, indicating mutagenesis of the cell populations independently from the T-DNA insertions. Mutagenesis was also indicated in gene tagging experiments where nitrate reductase-deficient (NR−) mutants were selected from haploidNicotiana plumbaginifolia protoplasts, as well as from leaf disc cultures or protoplasts of diploid plants that were heterozygotic for a mutation either in the NR apoenzyme gene (nia/wt) or one of the molybdenum-containing cofactor genes (cnxA/wt), afterAgrobacterium co-cultivation. The chlorate-resistant isolates were tested for the T-DNA-specific kanamycin resistance trait only after NR-deficiency had been established. Thirty-nine independent NR-deficient mutants were analysed further by Southern blot hybridization. There was no indication of integrated T-DNA sequences in the mutated NR genes, despite the fact that NR-deficient cells were found more frequently in cell populations which became transformed during the treatment than in the populations which did not. These observations suggest that transformation-competent cells undergo mutagenesis during theAgrobacterium gene transfer process not only as a result of stable integration events, but also through accompanying events that do not result in major changes in the mutated loci. The nature of these changes at the molecular level remains to be elucidated.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01973592
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    Electronic Resource
    Electronic Resource
    Spurious localizations of diX-indigo microcrystals generated by the histochemical GUS assay (1994)
    Springer
    Transgenic research 3 (1994), S. 176-181 
    Details
    ISSN: 1573-9368
    Keywords: Agrobacterium ; β-glucuronidase ; cytoenzymology ; reporter gene ; transgenic plants
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract For several models expressing theuidA orgus reporter gene with or without a presequence for mitochondrial targeting, we have demonstrated that the compartmentation of β-glucuronidase (E.C. 3.2.1.31) activity was not in agreement within situ localization of the diX-indigo microcrystals generated by the cytoenzymological GUS assay. These crystals were generally associated with the various cytomembranes and lipid inclusions. Experiments with purified β-glucuronidase or withgus-expressing bacteria incubated with 5-bromo-4-chloro-3-indolyl-β-d-glucuronide and maize oil-phosphate buffer emulsion indicated that the intermediate products resulting from the GUS assay actively diffused and crystallized preferentially in association with lipids, sometimes far from the site of enzyme activity. This phenomenon could not be suppressed by the addition of potassium ferricyanide in the incubation medium. These findings are discussed with regard to previously reported biochemical and histochemical data on animal tissues, and focus on the necessity for caution in studies of tissue-specific gene expression using the GUS assay, particularly for lipid-rich plant models.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF01973985
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  • 21
    Electronic Resource
    Electronic Resource
    The NocR repressor-activator protein regulates expression of the nocB and nocR genes of Agrobacterium tumefaciens (1994)
    Springer
    Molecular genetics and genomics 244 (1994), S. 367-373 
    Details
    ISSN: 1617-4623
    Keywords: Agrobacterium ; Nopaline ; Repressor Activator ; LysR family
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The NocR protein of Agrobacterium tumefaciens was found to regulate expression of the divergently transcribed nocB and nocR genes of the pTiT37 nopaline catabolism (noc) region. Experiments using the firefly luciferase (luc) gene as reporter demonstrated that NocR represses and activates transcription from the nocB promoter in the absence and presence of nopaline, respectively. NocR also negatively autoregulates its own synthesis irrespective of the presence of nopaline. Regulation of expression of both nocB and nocR is mediated by binding of the NocR protein to the nocR promoter. A 12 by symmetrical sequence, which lies 3 by downstream of the −10 hexamer of the nocR promoter, was confirmed to be essential for binding of the NocR protein. Functional localization of the nocB and nocR promoters verified that they do not overlap at all, and that the interrupted dyad, at which NocR binds, is 137 by upstream of the regulated nocB promoter. The in vivo and in vitro results described here and those published previously suggest that a novel type of regulatory mechanism, which may involve changes in DNA topology, controls gene expression in the noc operon of pTiT37.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00286688
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  • 22
    Electronic Resource
    Electronic Resource
    Agrobacterium vitis nopaline Ti plasmid pTiAB4: relationship to other Ti plasmids and T-DNA structure (1994)
    Springer
    Molecular genetics and genomics 245 (1994), S. 493-505 
    Details
    ISSN: 1617-4623
    Keywords: Agrobacterium ; Bacterial evolution ; Nopaline strains ; Ti plasmids ; Grapevine
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The Ti plasmid of the Agrobacterium vitis nopaline-type strain AB4 was subcloned and mapped. Several regions of the 157 kb Ti plasmid are similar or identical to parts of the A. vitis octopine/cucumopine (o/c)-type Ti plasmids, and other regions are homologous to the nopaline-type Ti plasmid pTiC58. The T-DNA of pTiAB4 is a chimaeric structure of recent origin: the left part is 99.2% homologous to the left part of the TA-DNA of the o/c-type Ti plasmids, while the right part is 97.1 % homologous to the right part of an unusual nopaline T-DNA recently identified in strain 82.139, a biotype Il strain from wild cherry. The 3′ non-coding regions of the ipt genes from pTiAB4 and pTi82.139 are different from those of other ipt genes and contain a 62 by fragment derived from the coding sequence of an ipt gene of unknown origin. A comparison of different ipt gene sequences indicates that the corresponding 62 by sequence within the coding region of the AB4 ipt gene has been modified during the course of its evolution, apparently by sequence transfer from the 62 by sequence in the 3′ non-coding region. In pTi82.139 the original coding region of the ipt gene has remained largely unmodified. The pTiAB4 6b gene differs from its pTi82.139 counterpart by the lack of a 12 by repeat in the 3′ part of the coding sequence. This leads to the loss of four glutamic acid residues from a series of ten. In spite of these differences, the ipt and 6b genes of pTiAB4 are functional. Our results provide new insight into the evolution of Agrobacterium Ti plasmids and confirm the remarkable plasticity of these genetic elements. Possible implications for the study of bacterial phylogeny are discussed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00302262
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  • 23
    Electronic Resource
    Electronic Resource
    Genetic manipulation in vesicular-arbuscular mycorrhizal fungi (1994)
    Springer
    Plant and soil 159 (1994), S. 171-178 
    Details
    ISSN: 1573-5036
    Keywords: Agrobacterium ; Gigaspora margarita ; Glomus intraradix ; PCR ; mycorrhizae ; rDNA gene ; root organ culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Agriculture, Forestry, Horticulture, Fishery, Domestic Science, Nutrition
    Notes: Abstract The symbiosis between vesicular-arbuscular mycorrhizal (VAM) fungi and host plants develops after successful interactions between both partners. These interactions probably involve signal molecules produced by the host plant, by the fungi, or by both. So far the biotrophic status of VAM fungi has hampered the understanding of the processes regulating their physiology. However, among different methods for co-cultivating VAM fungi, root organ cultures (ROC) appear to be a useful technique for studying VAM development. This system has been useful in defining the nutritional requirements of VAM fungi in the precolonization stage and in obtaining axenic fungal material in various developmental stages. The work discussed here focuses on the application of Polymerase Chain Reaction (PCR) technology and the potential of promoting hyphal growth in the absence of the plant. These techniques are being used to study VAM fungi in two main areas. The first concerns the determination of the DNA sequences coding for the SSU ribosomal RNA of two VAM fungi. This approach has allowed the design of specific primers for the rapid identification and quantification of VAM fungi. The second area of research concerns the potential use of PCR technology to study selective expression of specific genes during fungal spore development in defined in vitro conditions. The achievement of this future prospect depends on the ability to prepare PCR-based cDNA libraries from small amounts of fungal material after stimulation of hyphal growth with CO2 and plant flavonols.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00000106
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  • 24
    Electronic Resource
    Electronic Resource
    The biosynthetic pathway of the S-alk(en)yl-L-cysteine sulphoxides (flavour precursors) in species of Allium (1994)
    Springer
    Plant cell, tissue and organ culture 38 (1994), S. 181-188 
    Details
    ISSN: 1573-5044
    Keywords: Allium cepa ; biosynthesis ; compartmentation ; γ-glutamyl peptides ; onion ; protoplasts ; S-alkenyl-L-cysteine sulphoxide
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Pulse labelling experiments with 35SO4 2- fed for 24h to intact plants (shooted onion sets)of Allium cepa (onion) showed that 〉70% of the label appeared in the S-alkenyl-L-cysteine sulphoxides within 18h, reached a maximum at 48h and thereafter decreased. The amount of label detected in the γ-glutamyl peptide fractions was below 20% of the total label at any time. It is concluded that in intact plants (at the growth stage used) the γ-glutamyl peptides are not the immediate precursors of the S-alkenyl-L-cysteine sulphoxides. The major S-alkenyl-L-cysteine sulphoxide in onion was found to be compartmentalized mainly within the endoplasmatic reticulum.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033876
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  • 25
    Electronic Resource
    Electronic Resource
    Traits of transgenic Atropa belladonna doubly transformed with different Agrobacterium rhizogenes strains (1994)
    Springer
    Plant cell, tissue and organ culture 38 (1994), S. 257-262 
    Details
    ISSN: 1573-5044
    Keywords: Agrobacterium ; Atropa belladonna ; double transformation ; phytohormone content ; tropane alkaloids ; viviparous leaves
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Hairy root cultures of Atropa belladonna L. were established by infection either with Agrobacterium rhizogenes ATCC 15834 or MAFF 03-01724, and transgenic plants were obtained from both hairy root cultures. Doubly transformed roots were induced by re-infection of the leaf segments of transgenic Atropa belladonna plants (A. rhizogenes 15834) with MAFF 03-01724. Shoots and viviparous leaves were regenerated from the doubly transformed roots. The genetic transformation was determined by the opine assay (agropine, mannopine and/or mikimopine) and polymerase chain reaction. Physiological changes and tropane alkaloid biosynthesis in the hairy roots (singly and doubly transformed) were investigated. The alkaloid content in the doubly transformed root strain was intermediate as compared to the root strains which were singly transformed. On the other hand endogenous IAA levels in doubly transformed roots were significantly decreased compared to both singly transformed roots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00033885
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  • 26
    Electronic Resource
    Electronic Resource
    Production and characterization of intergeneric somatic hybrids through protoplast electrofusion between potato (Solanum tuberosum) and Lycopersicon pennellii (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 137-144 
    Details
    ISSN: 1573-5044
    Keywords: electrofusion ; L. pennellii ; protoplasts ; S. tuberosum ; salt tolerance ; somatic hybrids
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Mesophyll protoplasts of Lycopersicon pennelli Corr., a wild relative of tomato, were electrofused with those from a dihaploid potato clone, cv Nicola, with the objectives of transferring saline tolerance from L. pennellii to cultivated potato. 150 calli were selected from the fusion experiments, finally giving 2 hybrid shoots. Their hybrid nature was verified by examining isoenzyme patterns for esterases (EST), peroxidase (PRX), phosphogluconate dehydrogenase (6-PGD), and glutamate oxaloacetate transaminase (GOT). The hybrid plants had an intermediate morphology, and grew vigorously in vitro. When transplanted to soil, they were less vigorous, due to difficulties in rooting, but were still capable of flowering, and forming short stolons and mishaped tubers, probably resulting from the effects of gene dosage due to the novel association of two genomes from a tuberizing (potato) and a non tuberizing species (L. pennellii). The characteristics of such mishaped tubers provided strong evidence of a hybrid nature for the selected plants. The hybrid plants were highly sterile, producing only 3–7% viable pollen. Tests for salt tolerance showed that the growth of the somatic hybrid plants was reduced by 50% as for L. pennellii, whilst potato did not grow at all under saline conditions.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00043607
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  • 27
    Electronic Resource
    Electronic Resource
    Isolation of cells and protoplasts from leaves of in vitro propagated peach (Prunus persica) plants (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 99-105 
    Details
    ISSN: 1573-5044
    Keywords: cell division ; peach ; protoplasts ; Prunus persica ; tissue culture
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Yields of 106–108 peach mesophyll cells and protoplasts · gfw-1 were obtained depending on factors such as digesting enzymes, and leaf size. Onozuka R-10 (2%) in combination with Macerase (0.5%) was found best for protoplast isolation and mediocre for cell isolation among several enzyme combinations tested. Viability was 90% for protoplasts and 60% for cells. Pectolyase Y23 was found to be ineffective in our investigation. Small leaves, 4–10 mm in length, were a superior source for protoplast isolation than medium or big expanded leaves, 22–30 mm in length. The high yields of protoplasts could be obtained only when keeping the ratio of leaf biomass to volume of digesting enzyme solution under 20 mg ml-1. Purification of protoplasts on a sucrose gradient yielded about 107 protoplasts · gfw-1, however, the preparation was still contaminated by intact cells. Protoplasts were cultured under different growth regulators and physical conditions. Limited growth and division of protoplasts embedded in agarose drops were observed.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048320
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  • 28
    Electronic Resource
    Electronic Resource
    Highly efficlent plant regeneration from mesophyll protoplasts of Indian field cultivars of tomato (Lycopersicon esculentum) (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 255-258 
    Details
    ISSN: 1573-5044
    Keywords: tomato ; protoplasts ; plant regeneration
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Three Indian cultivars ofL. esculentum were assessed for shoot regeneration from protoplast-derived calli. Consistent yields of viable protoplasts (〉9.0×106 g f.wt.-1) were obtained from leaflets of 14 days old cultured shoots. Protoplast viability (88–94%) and planting efficiency (55–70%) were recorded for the three cultivars. Up to 71% of the protoplast-derived tissues regenerated shoots.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037728
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  • 29
    Electronic Resource
    Electronic Resource
    Expression of human α-interferon cDNA in transgenic rice plants (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 197-204 
    Details
    ISSN: 1573-5044
    Keywords: interferon ; Lipofectin ; protoplasts ; rice ; transgenic plant
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract The plasmid pIG3031 containing human α-interferon cDNA and the neomycin phosphotransferase II coding sequence was successfully transferred into rice protoplasts (Indica type rice) by Lipofectinmediated transformation. Assays for NPT II enzyme activity indicated that the transformation frequency was 10%. Transgenic plants were regenerated from transformed calli. Southern blots showed that the human α-interferon cDNA sequence was present in rice DNA. RNA slot blots indicated apparent transcription of human α-interferon cDNA under the control of the plant-active promoter PI'. Extracts of transgenic cell cultures and plants contained apparent interferon activity as measured by resistance of a human amniotic cell line to viral infection in the presence of plant extracts. These studies demonstrate that human α-interferon cDNA may be correctly expressed in rice cells.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037720
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  • 30
    Electronic Resource
    Electronic Resource
    Isolation and culture of protoplasts from young sporophytes ofSalvinia natans aseptically obtained by co-culture of female and male gametophytes (1994)
    Springer
    Plant cell, tissue and organ culture 36 (1994), S. 237-242 
    Details
    ISSN: 1573-5044
    Keywords: aseptic culture ; female gametophyte ; heterospory ; male gametophyte ; protoplasts ; Salvinia natans
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Sporophytes were aseptically obtained by co-culture of female and male gametophytes derived from two types of spores (megaspores and microspores) of the heterosporous fernSalvinia natans All. Protoplasts isolated enzymatically from juvenile leaflets of sporophytes were cultured in a 1/10 Murashige and Skoog's medium containing 2.2 μM naphthalene acetic acid, 2.2 μM 6-benzyl-aminopurine, 0.35 M mannitol, and 0.05 M sucrose. Cell division took place within 6 days of culture, and cell-clusters composed of 9–10 cells were observed after 30 days of culture.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00037725
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  • 31
    Electronic Resource
    Electronic Resource
    Single-cell cloning of a crown gall from protoplasts regenerated in hormone-free medium. Establishment of pure transformed cell-lines of Catharanthus roseus (1994)
    Springer
    Plant cell, tissue and organ culture 37 (1994), S. 25-30 
    Details
    ISSN: 1573-5044
    Keywords: Catharanthus roseus ; crown gall tumor ; heterokaryons ; protoplasts
    Source: Springer Online Journal Archives 1860-2000
    Topics: Biology
    Notes: Abstract Protoplasts enzymatically isolated from cell line of Catharanthus roseus G. Don crown gall, were cultured at high density (105 P ml-1) in modified B5 liquid medium (Gamborg et al. 1976). In the absence of growth regulators C. roseus protoplasts were able to regenerate a cell-wall, divide and, subsequently, yield very numerous clones in the absence of growth regulators. After two weeks, the cultures were greatly diluted in order to obtain clones of single-cell origin. Most of the clones individually transferred onto solid medium can proliferate indefinitely, without growth regulators. Among analyzed clones, 90% were nopaline positive. Their ajmalicine and serpentine content was compared with that of the parental crown gall line, and was found to be low. The CR10 protoplasts were very easy to grow, they were an interesting model for the development of pure tumorous lines. Moreover, we found that the tumorous protoplasts were useful for cell fusion experiments or for the delicate culture of tree protoplasts.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1007/BF00048113
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  • 32
    Electronic Resource
    Electronic Resource
    A note on chromatographic analysis of enantiomers by indirect method (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 52-52 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080114
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  • 33
    Electronic Resource
    Electronic Resource
    Relations between blood pressure and plasma norepinephrine concentrations after administration of diltiazem to rats: HPLC-peroxyoxalate chemiluminescence determination on an individual basis (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 19-21 
    Details
    ISSN: 0269-3879
    Keywords: Rat plasma norepinephrine ; HPLC ; ethylenediamine condensation ; peroxyoxalate chemiluminescence ; blood pressure reduction ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A calcium antagonist, diltiazem, was infused continuously into Sprague-Dawley rats through the left femoral vein at four different flow rates. The mean arterial blood pressure and concentrations of plasma norepinephrine (NE) were measured in each single rat (n = 5) and the correlations between them were studied. Blood (150 μL) was collected 13 times during the infusion. Plasma NE was determined by HPLC-ethylenediamine condensation reaction-peroxyoxalate chemiluminescence detection system (HPLC-ED-PO-CL).In four cases from 5 rats, the blood pressure reduction caused by diltiazem was inversely correlated to logarithm of plasma NE concentration. The relation was expressed as Y = -α logX + m. The coefficients of correlation were -0.9506, -0.9293, -0.9341 and -0.8675, respectively. The correlation for the last rat was worse (r = -0.0799). The good correlation would imply that the sympathetic nervous system released NE to maintain blood pressure up to the normal level, responding to the blood pressure reduction caused by diltiazem.The present experiment proved the feasibility of the determination method of NE utilizing HPLC-ED-PO-CL detection in applying to the individual rats.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080105
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  • 34
    Electronic Resource
    Electronic Resource
    Experimental models for optimization of insulin separation on reversed phase columns (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 32-36 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Experimentally defined relationships have been found to describe adequately the retention and separation of bovine and porcine insulins as well as their desamido products on reversed phases C4 30 nm and C18 10 nm pore diameter. The equations are valid for a region of initial acetonitrile concentrations from 16 to 31% and gradient rates from 0.04 to 0.60%/min. The peak heights showed an exact non-linear relationship with the time interval between the first and last peak of interest, independent of their retention time.The number of experiments required for obtaining the parametric estimates of the models depends on the particular task, but in all cases is less than eight.The relationships found permit the correct choice to be made in advance both for analysis conditions for a particular column and for different requirements of insulin analysis.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080109
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  • 35
    Electronic Resource
    Electronic Resource
    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994) 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080201
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  • 36
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    Electronic Resource
    Quantitative analysis of neuroactive amino acids in brain tissue by liquid chromatography using fluorescent pre-column labelling with o-phthalaldehyde and N-acetyl-L-cysteine (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 114-118 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A high-performance liquid chromatographic method for the determination of aspartic acid, glutamic acid, glutamine, glycine, taurine, and γ-aminobutyric acid in brain tissue is described. Amino acids were derivatized with o-phthalaldehyde/N-acetyl-L-cysteine, a fluorescent labelling mixture, in the presence of 0.1 M borate buffer pH 9.5. The derivatization reaction was sensitive to the pH and concentration of borate buffer. A drift in the fluorescent response less than 4% was obtained with the reported conditions after 4 h of reaction. The resolution of the amino acid derivatives was accomplished in a reversed-phase column with a methanol gradient in 50 mM acetate buffer pH 5.5. These conditions also allowed the separation of the major tissue free physiological amino acids. L-Norvaline was used as an internal standard for both peak identification and quantification. Within-day and between-day precision were less than 6.2%, and the accuracy ranged from 99.1 to 104%. The applicability of the method was demonstrated in a study in rats in which the levels of the assayed amino acids in discrete areas of brain were examined.
    Additional Material: 4 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080304
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  • 37
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    Electronic Resource
    Glutathione and cysteine measurement in biological samples by HPLC with a glassy carbon working detector (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 134-136 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An HPLC method using a glassy carbon working detector for measuring cysteine and reduced glutathione (GSH) in biological samples was developed. Oxidized glutathione (GSSG) was also measured after reduction with glutathione reductase. This study was conducted in human plasma. Cysteine and GSH standard curves were linear in the physiological range, presenting detection limits of 22.0 pmol and 6.0 pmol respectively. Plasmatic results found were 43.92 ± 4.15, 4.50 ± 0.65, and 0.19 ± 0.12 mM (means ± SE), for cysteine, GSH and GSSG, respectively. Peak specificity for cysteine and reduced glutathione was certified by their disappearance after treatment with N-ethylmaleimide.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080308
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  • 38
    Electronic Resource
    Electronic Resource
    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994) 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080401
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  • 39
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    Simultaneous determination of antidepressant drugs and metabolites by HPLC. Design and validation of a simple and reliable analytical procedure (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 158-164 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An analytical procedure suitable for routine use has been developed and validated for the simultaneous separation and quantification of most of the antidepressant drugs (bicyclic, tricyclic and tetracyclic) and their metabolites in human serum by reversed-phase high performance liquid chromatography with ultraviolet detection. The method has good sensitivity and specificity, without the need for long and complex extraction procedures. The results obtained with three different analytical columns were compared and an improvement in specificity by selection of column and detection wavelength used was achieved.
    Additional Material: 1 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080403
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    Gas chromatography-mass spectrometry determination of etodolac in human plasma following single epicutaneous administration (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 180-183 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A highly sensitive gas chromatographic-mass spectrometric method for the determination of etodolic acid, as methyl ester, in plasma was developed. The feasibility and specificity of the method was ascertained monitoring the concentration levels in plasma samples collected from 12 male healthy volunteers given epicutaneously 5 g of 10% etodolac gel formulation.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080407
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    TLC - A rapid method for liposome characterization (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 193-195 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: One of the most important problems for the use of liposomes as a drug delivery system is the modification of the vesicle induced by the liquid medium in which they are introduced (blood plasma for in vivo studies and the saline buffer solution for in vitro studies).Using thin-layer chromatography (TLC) we compared the behaviour of phosphatidylcholine (used for liposomes preparation) to that of the following unfilled liposomes: multilamellar liposomes (MLV); small unilamellar vesicles (SUV); and reverse phase evaporation vesicles (REV), before and after storage for 15 min in Krebs-Henseleit solution (37°C, pH 7.4, aereated continuously with 95% O2 + 5% CO2). All variants contained the same amount of phosphatidylcholine.Thin-layer chromatography was performed on silica gel 60 as adsorbant. Two types of solvents were tested: one based on chloroform/alcohol (n-butanol or n-propanol or methanol)/water mixture (in different ratios) and another based on alcohol/alcohol/water mixture (n-butanol/n-propanol/water in 4/3/3 volume ratio). In all variants of chloroform containing solvents no differences were found between phosphatidylcholine and all types of liposomes.When using as solvent n-butanol/n-propanol/water significant differences were found between all types of liposomes before and after storage in Krebs-Henseleit solution. Their presence, after TLC treatment, was shown in electron microscopy studies.
    Additional Material: 1 Ill.
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    Improved resolution of tryptic digest fragments from haemoglobin variants using phytic acid in free zone capillary electrophoresis (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 202-204 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Capillary electophoresis can be applied to the rapid characterization of tryptic digests of proteins. The addition of phytic acid to the separation buffer was found to improve resolution considerably when the technique was applied to differentiate between tryptic digests derived from variant haemoglobins. Moreover, analysis time was of the order of 15 min, which is considerably shorter than that obtained using gradient reversed-phase high-performance liquid chromatography or two-dimensional paper chromatography-electrophoresis.
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    URL: http://dx.doi.org/10.1002/bmc.1130080413
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  • 43
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    A non-radioactive iothalamate and p-aminohippuric acid high-performance liquid chromatographic method for simultaneously measuring glomerular filtration rate and renal blood flow in the rat (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 224-229 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A high-performance liquid chromatographic (HPLC) assay method has been developed for the quantitative determination of iothalamate and p-aminohippuric acid (PAH) concentrations in serum and urine samples in the male rat. Glomerular filtration rate (GFR) was measured as clearance of iothalamate, while effective renal blood flow (ERBF) was measured as clearance of PAH. The method is simple, rapid and sensitive and detects iothalamate and PAH in rat serum and urine following administration of bolus doses and continuous infusions of iothalamate and PAH. Samples of serum and urine were deproteinized with two volumes of acetonitrile containing the internal standard, and an aliquot chromatographed on a C18 reversed-phase column. The mobile phase was comprised of 0.1 M sodium phosphate with 1.2 mM tetrabutylammonium phosphate: methanol, 85:15 (v/v), at a flow rate of 1.0 mL/min. The analytical column eluate was monitored with a UV detector at 254 nm with quantitation achieved using peak-height ratios. The precision of the method was 6.6 and 3.6% for iothalamate in serum and urine, and 5.6 and 4.9% for PAH in serum and urine, respectively. The lower limit of quantitation was 0.63 μg/mL for iothalamate and 1.25 μg/mL for PAH in serum, and 3.1 μg/mL for iothalamate and 1.5 μg/mL for PAH in urine. Recovery of iothalamate from serum and urine was 99.9 and 93.5%, respectively. Recovery of PAH from serum and urine was 99.8 and 92.6%, respectively.The present study demonstrated that non-radioactive iothalamate and PAH can be measured simultaneously using a HPLC assay to measure GFR and ERBF in the male rat.
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    URL: http://dx.doi.org/10.1002/bmc.1130080506
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    Use of capillary electrophoresis for thalassaemia screening (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 175-179 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This paper describes the use of high-performance capillary electrophoresis (HPCE) for screening of thalassaemias. Preparation of globin was performed by adding 1mL of haemolysate to 20 volumes of cold acidified acetone. Separation of globins was accomplished in a 25 mM disodium phosphate buffer at pH 11.8 using an uncoated capillary column of 50 cm × 75 μm (i.d.). Distinct peaks of alpha, beta and gamma were resolved within 6min. The coefficient of variations for within-day and between-day runs were 4.7% and 6.37% respectively. Using this simple and rapid procedure up to 30 specimens could be analysed in a single day. This method appears to be reliable and can be used for mass screening of various haemoglobinopathies.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080406
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    Measurement of theophylline metabolites produced by reaction with hepatic microsome by high performance liquid chromatography following solid phase extraction (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 189-192 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An analytical method has been developed with which to measure the microsomal enzyme activities responsible for oxidative theophylline metabolism. Three metabolites: 3-Methylxanthine (3-MX); 1-methylxanthine (1-MX); 1,3-dimethyluric acid (1,3-DMU), with acetaminophen as an internal standard (IS), were separated by solid phase extraction using a Sep-Pak C18 cartridge, followed by high performance liquid chromatography on a reversed-phase column with isocratic elution using 25 mM acetate buffer containing 4% acetonitrile and 2.5 mM tetra-n-butylammonium hydrogen sulphate (pH 5.25) as the mobile phase. The analytes were clearly resolved and no interference with foreign peaks was observed. A linear relationship was obtained for the metabolites over the concentration range of 0.5-5.0 μg/mL, and their analytical recovery was almost 100%. This method can be used to assess drug interactions involving alterations in the biotransformation of theophylline.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080409
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    Editorial (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 205-205 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080502
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  • 47
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    A comparison of fluorescence versus chemiluminescence detection for analysis of the fluorescamine derivative of histamine by HPLC (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 207-211 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Fluorescence and chemiluminescence detection were compared for HPLC analysis of the fluorescamine derivative of histamine. The kinetic behaviour of the chemiluminescent response for the derivative was characterized in a static system. An HPLC method was optimized for the derivative using fluorescence detection. Fluorescence detection was linear over the range of 166-1666 pg on column for the fluorescamine-histamine derivative with a limit of detection of 13 pg on column. Using a detector designed for optimal use with chemiluminescence, the chemiluminescence response of the fluorescamine derivative was linear over a range of 1.66-16.6 ng on column with a limit of detection of 1.0 ng on column. These results exemplify a case in which superior detectibility is provided by fluorescence over chemiluminescence, and contradicts many reports comparing fluorescence to chemiluminescence. The authors conclude that chemiluminescence should be considered when indicated by conditions established for separation that are favourable for the observation of chemiluminescence. These conditions include sufficiently low excitation energies corresponding to an excitation maximum greater than 400 nm, favourable dipole character of analytes, mobile phases of high organic content, and an appropriate pH of the mobile phase.
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  • 48
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    HPLC separation of pentazocine enantiomers in serum using an ovomucoid chiral stationary phase (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 255-257 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A stereospecific HPLC method was developed for the analysis of (-) and (+) pentazocine in human serum. Each enantiomer and the internal standard nalophine were isolated from serum using a liquid-liquid extraction procedure. Recoveries of 99.05 ± 5.37 and 97.42 ± 2.78% were obtained for (-) and (+) pentazocine, respectively. Resolution of the enantiomers was obtained by using an ovomucoid chiral stationary phase with a mobile phase of methanol: acetonitrile: 10 mM phosphate buffer, pH 5.8 (20:5.3:74.7 v/v/v). A resolution (Rs) value of 1.80 was obtained for the pentazocine enantiomers. Linear calibration curves were obtained in the 10-100 ng/mL range for each enantiomer in serum. The detection limit based on a signal-to-noise ratio of 3 was 5 ng/mL for each enantiomer in serum using fluorescence detection with excitation at 275 nm and emission set at 335 nm. The lowest quantifiable level was found to be 10 ng for each enantiomer. Precision and accuracy of the method were in the 3.8-4.8% and 1.3-4.2% ranges, respectively.
    Additional Material: 2 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080512
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  • 49
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    Determination of fluvoxamine concentration in plasma by reversed-phase liquid chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 278-282 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Fluvoxamine, a serotonin re-uptake inhibitor, was quantified in plasma by modifying a previously published procedure for monitoring plasma concentrations of tricyclic antidepressants. Alkalinized plasma samples were extracted with n-hexane/isoamyl alcohol, followed by back-extraction with diluted phosphoric acid, The extracts were analysed by reversed-phase liquid chromatography using a C-18 column, with phosphate/acetonitrile as the mobile phase. The assay was linear from 10 to 800 μg/L. Precision studies showed within-run and day-to-day coefficients of variation to be 4.5 and 6.8%, respectively. Desipramine interfered with the detection of fluvoxamine. The assay was used to measure a total of 8 plasma samples from 4 alcohol-dependent patients medicated with fluvoxamine as an adjunct to relapse prevention psychotherapy. In these patients, the plasma concentrations ranged from 54 to 241 μg/L. Dosage of fluvoxamine, duration of treatment, interval between last dosage and blood collection were associated with effects on plasma concentrations that were consistent with the pharmacokinetic profile of fluvoxamine.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080605
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  • 50
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    Simultaneous determination of p-aminobenzoic acid and its metabolites in urine by high performance liquid chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 294-296 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: We developed a new HPLC method for the determination of p-aminobenzoic acid (PABA) and its metabolites (p-aminohippuric acid, N-acetyl-p-aminohippuric acid, N-acetyl-p-aminobenzoic acid) in urine. As the internal standard m-hydroxybenzoic acid was used. In the isocratic elution the mobile phase consisted of methanol and 0.02 M. ammonium acetate (20:80 v/v, pH 4.0). The separation was carried out on the C18, reversed-phase column, particle size 5 μm. The separated components were detected at 280 nm. The method can be used in the assessment of the response of pancrease (secretion of digestive enzymes) to soya feeding as well as in the diagnosis of the exocrine pancreatic diseases of animals.
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    URL: http://dx.doi.org/10.1002/bmc.1130080609
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    Solid phase extraction and high performance liquid chromatographic determination of dobutamine in plasma of dialysed patients (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 309-312 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An isocratic reversed-phase high performance liquid chromatographic method has been developed for the determination of dobutamine in the plasma of dialysed patients. A solid phase extraction method with a Sep-Pak C18 cartridge was used to isolate the drug and isoxsuprine (internal standard) from plasma. The separation was carried out on an ODS-Hypersil column with 0.1 M phosphate buffer: acetonitrile: methanol (72 : 20 : 8 v/v/v) as the mobile phase. The recovery of dobutamine added to plasma by the extraction procedure was 87 ± 2.3% (mean ± SD). The accuracy and reproducibility of the method were within acceptable limits over the concentration range 0-1000 ng/ mL. Quantification was by fluorescence detection at 275 nm excitation and 310 nm emission wavelengths with a detection limit of 5 ng/ mL for dobutamine. This procedure was applied to ascertain the pharamacokinetics of dobutamine infusion in nine patients with cardiogenic shock and end-stage renal disease undergoing haemodialysis.
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    URL: http://dx.doi.org/10.1002/bmc.1130080613
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    Simple determination of formaldehyde in dimedone adduct form in biological samples by high performance liquid chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 313-314 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A high performance liquid chromatographic method for the determination of endogenous formaldehyde in dimedone adduct form in biological samples is described. The simple procedure involves extraction of the formaldehyde of different binding force in biological samples with methanol containing dimedone as the capture molecule and separation on a C18 reversed phase column with methanol as eluent.
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    URL: http://dx.doi.org/10.1002/bmc.1130080614
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    Regional and age variations in growing tendon (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 221 (1994), S. 309-320 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Gastrocnemius tendons of 10 White Leghorn chickens at 6, 8, and 12 weeks of age were divided into proximal, middle, and distal portions to assess regional variability in composition and growth. Body weight increases ∼ 150% during the period examined, whereas the lateral gastrocnemius muscle and tendon increase ∼ 193% and 227%, respectively. No significant changes in cellularity (DNA concentration) or hydroxypyridinium (OHP) crosslinks occur with increasing age. Hydroxyproline (HYP) concentration increases by 12 weeks of age, as hexuronate, glucosamine, and galactosamine decrease. Composition shows some regional variation: the distal region of the tendon has a lower HYP concentration, and increased GAGs and OHP crosslinks compared to either the proximal or middle regions, which do not differ from each other. The mean collagen fibril diameter increases with age, but the oldest tendons also contain more small diameter fibrils (〈40 nm). There is a unimodal fibril distribution at all three ages, although this has broadened by 12 weeks. The data from this study suggest that rapid tendon growth occurs throughout the time period examined and that changes characteristic of mature tendon, such as increased OHP crosslink concentration, have not yet developed in hatchlings because of the large amount of new tissue being produced. Whereas all three regions of the tendon are similar in size, composition of the distal region differs from that of the proximal and middle regions, suggesting that this portion of the tendon should be avoided when sampling a tendon. © 1994 Wiley-Liss, Inc.
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    URL: http://dx.doi.org/10.1002/jmor.1052210306
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    Microscopic anatomy of pycnogonida: I. Cuticle, epidermis, and muscle (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 33-48 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The integument of Pycnogonida (Arthropoda) consists of an epicuticle decorated with tubercles and a filamentous coat, an exocuticle with a small number of ill-defined layers, and an endocuticle whose numerous layers are composed of conspicuously cross-banded fibrils. This cuticular periodicity, attributable to cross-linked chitin, has been observed previously in uncalcified and untanned cuticle of many lower crustaceans, especially branchiopods and copepods, and in scattered examples of thin respiratory or excretory cuticles of other arthropods. It is uniformly present in all representatives of all nine pycnogonid families examined to date. Stomodeal, proctodeal, and arthrodial cuticles are devoid of the endocuticular periodicity. The cuticle is decorated with sensory filaments and setae, but is more noteworthy for a dense coverage by glands, up to 1,400/mm2. Myocuticular junctions have desmosomal fine structure previously found only in chelicerates. Muscle fine structure is that of slow fibers with long sarcomeres and a high actin to myosin filament ratio, except for cardiac muscle, which has short sarcomeres. Among the arthropods, only merostomates resemble the pycnogonids in the lack of fast somatic muscle fibers. Pycnogonids display a hybrid array of fine structural features that variously serve to relate them to some arthropod subphyla and distance them from others. © 1994 Wiley-Liss, Inc.
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    Erratum (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 111-111 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052220111
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  • 56
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    Development and growth of compound tooth plates in Callorhinchus milii (chondrichthyes, holocephali) (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 73-89 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The chimaeroid holocephalian fishes are distinguished among extant chondrichthyans by the possession of three pairs of tooth plates, evergrowing and partially hypermineralized, that are not shed and replaced like the teeth of living elasmobranchs. Although derivation of the chimaeroid tooth plate from the fusion of members of a plesiomorphic chondrichthyan tooth family has been proposed, evidence for this hypothesis has been lacking. A new analysis of the development and structure of the tooth plates in Callorhinchus milii (Holocephali, Chimaeriformes) reveals the compound nature of the tooth plates in a chimaeroid fish. Each tooth plate consists of an oral and aboral territory that form independently in the embryo and maintain separate growth surfaces through life. The descending lamina on the aboral surface of the tooth plate demarcates the growth surface of the aboral territory. Comparison with the tooth plates of Chimaera monstrosa indicates that compound tooth plates may be a feature of all chimaeroids in which a descending lamina is present. The tooth plates in these fishes represent the fusion of two members of a reduced tooth family. The condition of the tooth plates in C. milii is plesiomorphic for chimaeroids and is of evolutionary significance in that it provides further evidence to support a lyodont dentition in chimaeroid fishes similar to that found in other chondrichthyans. © 1994 Wiley-Liss, Inc.
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    URL: http://dx.doi.org/10.1002/jmor.1052220108
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    Morphological and functional body reconstruction in a teleost of the genus Oreochromis (Cichlidae) (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 1-6 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The process of morphological and functional regeneration was followed on a tilapid fish, a cross of Oreochromis aureus × Oreochromis niloticus, by observations on movements and the use of X-rays. A four-year-old adult fish that lost its tail as post larva, including ten vertebrae, was able to reconstruct a novel and shorter central skeleton, including a specially modified urostyle. The enlarged and strengthened pterygiophores and their junctions with the dorsal and anal spine formed a fast-holding base for the fins, the posterior part of which largely performed the functions of the missing caudal fin. Although the fish was much shorter than usual, this male behaved and functioned normally. © 1994 Wiley-Liss, Inc.
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    URL: http://dx.doi.org/10.1002/jmor.1052190102
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    Ultrastructure of the paraphysis in Bufo bufo larvae (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 7-13 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: A study of the ultrastructure and function of the paraphysis in Bufo bufo larvae was carried out. The structure is a tubular-ramified gland made up of numerous tubules with monolayered epithelial walls surrounded by connective tissue and sinusoids. The epithelial cells secrete glycoprotein to contribute to production of the cephalorachidian fluid. The role of the paraphysis in the transport of fluids and electrolytes from the blood to the cephalorachidian fluid in regulation of ionic and osmotic homeostasis is discussed. © 1994 Wiley-Liss, Inc.
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    URL: http://dx.doi.org/10.1002/jmor.1052190103
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  • 59
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    Characteristics of the flagellar axoneme in Neuroptera, Coleoptera, and Strepsiptera (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 15-20 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Spermatozoa from representatives of the five insect orders in superorder Neuropteroidea were examined by electron microscopy following a new fixation method that includes tannic acid in the primary fixative but has uranyl acetate rather than osmium tetroxide as the secondary fixative. The sperm axoneme was found to be similar in the four orders Megaloptera, Raphidioptera, Neuroptera, and Coleoptera, and is characterized above all by its so-called intertubular material being divided into two portions, one located outside, but in contact with the doublet, and the other projecting from the accessory tubule and having a beak-like shape. These features have not been seen in insects from other orders and may be a synapomorphy for these neuropteroid orders. The accessory tubules in these four orders have 16 protofilaments. The shape of the accessory bodies adjacent to the mitochondrial derivatives is nearly the same in insects from the more primitive neuropteroid orders and in Coleoptera. The sperm tail of the examined strepsipteran deviates in several respects from that of other neuropteroids: the particle row in the wall of accessory tubules is incomplete, an intertubular material is missing, and the mitochondria contain no crystal. © 1994 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052190104
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    Spiral cleavage and cell position contribute to the specification of the dorsoventral axis in the embryo of the archaeogastropod Haliotis tuberculata (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 21-33 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: In the embryo of Haliotis tuberculata spiral cleavage induces size differences between the quadrants in the 4-cell embryo. These size differences, together with the formation of compact cell configurations, induce asymmetrical positions of equivalent cells in the 8- and 16-cell embryo. The asymmetries in size and position influence the final specification of the dorsoventral asymmetry in the 32-cell embryo, as well as formation of the mesentoblast. © 1994 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052190105
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    Eversible abdominal vesicles and some observations of the male reproductive system of the spoon wing lacewing Palmipenna (Neuroptera: Nemopteridae) (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 47-58 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The anatomy and histology of the abdominal eversible vesicles and the male reproductive tract of the spoonwing lacewing Palmipenna (Neuroptera: Nemopteridae) have been examined. The eversible vesicles open as a pair of large bulbous sacs between tergites five and six, each folding into halves during retraction. They consist of highly pleated cuticle, beneath which are typical gland cells, each having a circular or oval end apparatus surrounded by closely packed microvilli. These communicate to the surface via cuticularized channels. In spite of considerable behavioral observations, male Palmipenna were never noted with everted vesicles. Even during mating trials, where females were presented to males in the field, the vesicles were never everted during the attempted copulation that ensued. Our observations indicate that mate attraction is mediated by the release of a female pheromone. The function of the eversible vesicles and their associated gland cells remains unknown, and their structure appears to be unique to the Nemopteridae. The reproductive tract is similar to that of other Neuroptera, consisting of a pair of five-lobed testes, a medium-to-large pair of seminal vesicles, and three pairs of accessory glands. The major accessory glands are surrounded by circular and longitudinal muscle, and are lined by an epithelium, the cells of which presumably secrete the amorphous rods of material always present in this pair of glands. The sperm in the seminal vesicles are elongate, with a pointed head and a 9 + 9 + 2 configuration in the flagellum. A single spermatophore, similar in shape to that described for other Neuroptera, was found occluding the bursa copulatrix of a teneral female. © 1994 Wiley-Liss, Inc.
    Additional Material: 24 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052190107
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    In vivo fluorescence imaging of the functional organization of trophotaenial placental cells of goodeid fishes (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 35-46 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Embryos of viviparous goodeid fishes undergo a 10 to 150 × increase in dry weight during gestation. Maternal nutrients are transferred across a trophotaenial placenta comprised of the ovarian lumenal epithelium and the trophotaeniae of the embryo. Trophotaeniae are externalized projections of the embryonic hindgut. Epithelial cells of the ribbon trophotaenia (Ameca splendens) resemble intestinal absorptive cells of suckling mammals and endocytose macromolecules. They possess an apical brush border, endocytotic complex, endosomal-lysosomal system, and apical and basal clusters of mitochondria. Cells of the rosette trophotaenia (Goodea atripinnis) lack an endocytotic apparatus, have small lysosomes, two mitochondrial clusters, and transport small molecules. Organelle-specific fluorescent probes were employed to characterize the functional organization of the two types of trophotaenial cells. In A. splendens, Lucifer Yellow, a membrane-impermeable tracer of vesicular transport, first appears in peripheral vesicles (15-45 sec), then passes into elongated tubular endosomes (1-3 min) and later appears in large central vacuoles (10-15 min). These vacuoles accumulate Acridine Orange, a classical probe for lysosomes, and have been shown to contain lysosomal enzymes. Endosomelysosome fusion was observed. In both A. splendens and G. atripinnis, Rhodamine 123 fluorescence was localized in two clusters of fine spots that corresponded to mitochondria. 4′,6-diaminido-2-phenyl-indole (DAPI) staining of nuclei established the positional relationships of cell organelles with respect to the nuclei. 3,3′-dihexyloxacarbo-cyanine iodide (DiOC6) revealed the perinuclear distribution of the endoplasmic reticulum. In order to compare in vivo fluorescence of Lucifer Yellow with previous ultrastructural observations, we employed fluorescence photoconversion and electron microscopy. © 1994 Wiley-Liss, Inc.
    Additional Material: 15 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052190106
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    Ontogenesis and ultrastructure of seminal vesicles of the catfish, Clarias gariepinus (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994), S. 59-71 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The ontogenesis and structural characteristics of the seminal vesicles in Clarias gariepinus (sharptooth catfish) were studied by light and electron microscopy and are described in detail. The seminal vesicles, beginning as simple protrusions from the vas efferentia, becomes more complex with age. Their distal ends become fingerlike and the bases form palm-like extensions. Juvenile male organs do not reveal any signs of seminal vesicles although spermatogenic tissue is already well delineated. The developing gonads contain clusters of large cells, close to the sperm duct and cysts of the testis, from which seminal vesicles are formed. Secretory epithelium lines the tubules of the seminal vesicles and becomes columnar as the tissue matures. Electron micro-graphs of these epithelial cells reveal two types of cells: opaque cells and cells with very vacuolized cytoplasm. Dense pinocytotic vesicles are present between the membranes of neighbouring seminal tubules and apical cell membranes facing the lumen. Maturation and onset of secretion by the secretory cells is accompanied by morphological changes. Protruding cylindrical cells become shortened, modified to cuboidal, rounded cells that send tubular extensions into the lumen. In the final stage of differentiation, only connective tissue membranes supporting the tubule walls remain intact. At the points of contact between the testis, seminal vesicles, and sperm duct, the epithelia of these organs often become confluent. The distal parts of the seminal vesicles, rarely contain sperm; during spawning sperm accumulated in the proximal tubules of the vesicles. © 1994 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052190108
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    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 219 (1994) 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052190201
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    Identification and verification of the anabolic steroid boldenone in equine blood and urine by HPLC/ELIS (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 63-68 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An enzyme linked immunosorbent assay (ELISA) was developed to detect the anabolic steroid boldenone in equine blood and urine. The polyclonal antiserum was raised in rabbits, employing boldenone-17-hemisuccinate-bovine serum albumin as antigen. Boldenone-17-hemisuccinate-horseradish peroxidase served as enzyme conjugate. Sensitivity of the assay was 26.0 ± 3.0 pg/well. Among the endogenous steroids tested only progesterone and testosterone exhibited moderate cross-reactivities, 3.4 and 2.5%, respectively. These crossreactivities are of no importance for the boldenone assay. For the reduction of background levels, srceening for boldenone of equine serum was performed after extraction. Urine samples were determined directly after dilution, omitting hydrolysis of boldenone conjugates. Positive screening results were confirmed by means of two independent HPLC systems combined with off-line detection, employing the boldenone ELISA. Methandienone served as internal standard to ascertain retention factors. In horses treated with boldenone-17-undecylenate the presence of boldenone in serum was confirmed up to 28 days and in unhydrolysed urine up to 56 days post applicationem.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080204
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    Automated HPLC analyses of drugs of abuse via direct injection of biological fluids followed by simultaneous solid-phase extraction and derivatization with fluorescence detection (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 53-62 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: An automated system is described for the simultaneous extraction and derivatization of nucleophilic compounds from various biological media. The method includes the use of a solid-phase reagent containing a 9-fluorenylacetate activated ester. The reagent is based on a controlled pore, polystyrene divinylbenzene support prepared through a silica template procedure. An X-Y-Z robotic arm equipped with a needle is used in conjunction with a syringe pump for aspirating and dispensing samples and standards into the HPLC system. A precolumn cartridge containing the solid-phase reagent is put on-line in place of the fixed-volume injection loop. Injections of biological fluids such as urine or plasma with minimal sample treatment and handling are made directly into this reactor. The analytes are dervatized as they are extracted, allowing virtually unlimited sample volumes to be injected. The polymeric cartridge can be used for up to 100 injections without accruing unacceptable reductions in sensitivity. A detection limit of 500 p.p.t. (parts per trillion) of amphetamine in urine was achieved with this system.
    Additional Material: 6 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080203
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    Reversed-phase HPLC separation and chromatographic-spectral characterization of 17β-(2′-thiazolyl)androst-5-en-3β-ols and their acetates (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 95-98 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Simple isocratic HPLC separation of a series of thiazolyl steroids with the 17β-2′ linkage is described. The chromatographic and spectral characterization utilizes both on-the-fly UV spectral maxima and absorbances changes.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080210
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    Electrophoresis in practice: A guide to theory and practice. R. Westermeier VCH, Weinheim, 1993, ISBN 3-527-30012-0, £28.00 (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 105-105 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080213
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    A Novel electrophilic reagent, 4-(N-chloroformylmethyl-N-methyl)amino-7-N,N-dimethylaminosulphonyl-2, 1,3-benzoxadiazole (DBD-COCI) for fluorometric detection of alcohols, phenols, amines and thiols (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 107-113 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A novel electrophilic reagent for alcohols, phenols, amines and thiols, 4-(N-chloroformylmethyl-N-methyl)amino-7-N,N-dimethylaminosulphonyl-2, 1, 3-benzoxadiazole (DBD-COCI) was synthesized. The reactivity of the reagent to these nucleophiles was studied using high-performance liquid chromatography (HPLC) with precolumn derivatization techniques.DBD-COCI reacted in benzene solution at the room temperature or 60°C with androsterone (a representative of hydroxyls), (-)-mandelic and D,L-lactic acid (hydroxy acid), estrone (phenol), benzylamine (primary amine), phenetidine (aromatic amine) and α-mercapto-N,2-naphthylacetamide (thiol) to give fluorescent products bearing fluorescence wavelengths at between 543 nm and 555 nm (excitation at between 437 nm and 445 nm). The base catalyst, quinuclidine was required to complete the reaction with the nucleophiles except aromatic amines. The reaction solution was subjected to a reversed phase HPLC and the detection limits of the derivatives were at the femto mol range on column.
    Additional Material: 8 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080303
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    Proteinergic profiles in cerebrospinal fluid from alcoholic subjects (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 137-141 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Minute amounts of cerebrospinal fluid samples from alcoholics were subjected to separation by HPLC-molecular sieving, combined with multispectral UV analysis of the acquired data. A significant difference in the protein/polypeptide pattern within the molecular weight range of 7-10 kDa has been observed between samples, taken directly after detoxification and 2 weeks later. Spectral analysis of the results suggests that the components are of peptidergic nature. On the other hand, albumin content did not differ significantly, suggesting that the blood-brain barrier was not affected. An enzyme marker, dynorphin converting enzyme, remained unchanged in both groups.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080309
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    A need for a uniform approach for indexing data of the stereoisomers (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 151-151 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 1 Tab.
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    URL: http://dx.doi.org/10.1002/bmc.1130080313
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    Separation of vitamin B complex and folic acid using TLC plates impregnated with some transition metal ions (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 196-198 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The separation of Vitamin B complex and folic acid was achieved on silica gel layers impregnated with Mn++, Fe++, Co++, Ni++, Cu++, Cd++, Zn++ or Mg++ (0.1%, 0.2%, 0.3%, 0.4% of each ion). Three new solvent systems n-propanol: n-butanol: water: ammonia (7:5:1:2, by vol), n-propanol: n-butanol: water: ammonia (7:5:1:1.5, by vol), n-propanol: n-butanol: water: ammonia (7:5:0.75:2, by vol) were developed. The spots of vitamins were located by exposing the chromatoplate to iodine vapours.
    Additional Material: 4 Tab.
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    URL: http://dx.doi.org/10.1002/bmc.1130080411
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    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994) 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080501
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    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994) 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080601
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    Interaction of carboxymethyl-γ-cyclodextrin with anticancer drugs studied by charge-transfer thin-layer chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 267-272 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: The interaction between 22 anticancer drugs and carboxymethyl-γ-cyclodextrin (CM-γ-CD) was studied by reversed-phase charge-transfer thin-layer chromatography and the relative strenth of interaction was calculated. CM-γCD formed inclusion complexes with 11 compounds, the complex always being more hydrophilic than the uncomplexed drug. The inclusion forming capacity of drugs differed considerably according to their chemical structure. Both principal component analysis and cluster analysis found a relationship between the inclusion complex forming capacity and hydrophobicity parameters of anticancer drugs. This result suggests that the preponderant role of hydrophobic interactions is in inclusion complex formation.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080603
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    Comparison of standard chromatographic procedures for the optimal purification of soluble human brain acetylcholinesterase (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 259-266 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: With the view of purifying soluble human brain acetylcholinesterse (AChE) into its separate isoforms, various preparative chromatographic procedures were compared. Chromatofocusing of cerebrospinal fluid (CSF) AChE revealed two major activity peaks, whilst that of caudate nucleus AChE showed one major peak, Both CSF and caudate nucleus AChE eluted at isoelectric points (pI) of between 5.5 and 5.2 Chromatofocusing failed to separate AChE into its individual isoforms, based on qualitative isoelectric focusing. Preparative purification by affinity chromatography showed a better AChE yield with the use of procainamide as a ligand, vis-à-vis acridinium. Maximum recovery for CSF and caudate nucleus AChE was 10 and 43% using acridinium and procainamide, respectively. Qualitative analysis by SDS-PAGE of affinity-purified AChE revealed four major bands between 50 and 62kDa, corresponding to the catalytic subunits of AChE as verified by an anti-AChE polyclonal antibody. A size-exclusion column also allowed brain AChE purification, with the latter eluting at a putative molecular mass of 310 kDa. Unfortunately, cation-exchange using the state-of-the-art SMART system failed to separate AChE into its isoforms. AChE aggregation is given as one major obstacle precluding good resolution of isoforms.
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    URL: http://dx.doi.org/10.1002/bmc.1130080602
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    Direct determination of free phenylacetic acid in human plasma and urine by column-switching high performance liquid chromatography with flurescence detection (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 283-287 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A simple and highly sensitive column-switching high performance liquid chromatographyic method with fluorescence detection for the determination of free phenylacetic acid (PAA) in human plasma and urine is described. The method is based on the direct derivatization of plasma and urine PAA with 6,7-dimethoxy-1-methyl-2(1H)-quinoxalinone-3-propionylcarboxylic acid hydrazide (DMEQ-hydrazide). The derivatization reaction proceeds in aqueous solution in the presence of pyridine and 1-ethyl-3-(3-dimethylaminopropyl)-carbodiimide at 37°C. The resulting DMEQ derivative of PAA is separated from endogenous interfering substances by a column-switching chromatographic system consisting of a precolumn (YMC-Pack C4) for sample clean-up and an analytical column (L-Column ODS) for the complete separation of the derivative. The derivative is detected fluorimetrically at 445 nm wih excitation at 367 nm. The detection limits (signal to noise ratio = 3) for PAA is 10 fmol in a 10 μL injection volume. The recoveries from plasma and urine are 75 and 96%, respectively. The present method is highly sensitive and simple compared to conventional liquid-liquid extraction procedures. The sensivity allows the direct determination of free PAA in an extremely small amount (5 μL) of plasma and urine.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080606
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    Optical purity determination of thyroxine enantiomers in bulk materials by chiral thin layer chromatographyAccepted in part for presentation at the 5th International Symposium on Chiral Discrimination, 26-28 September 1994, Stockholm, Sweden. (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 317-318 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: L-thyroxine and D-thyroxine were separated on ligand exchange chiral thin layer chromatographic plates, using a solvent system consisting of acetonitrile:methanol:water 60:15:15 v/v, at a wavelength of 254 nm. The methodology, chiral recognition mechanism (s) involved and its application are discussed.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080616
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    Fluidized-bed receptor-affinity chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 236-241 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A multipurpose fluidized-bed receptor-affinity purification system based upon the biological recognition between an immobilized receptor and its soluble protein ligands is described. The fluidized affinity sorbent consists of a soluble form of interleukin-2 receptor chemically bonded to an aldehyde derivative of controlled pore glass beads, which have a pore diameter of 1000 Å and a particle density of 1.2-1.3 g/mL. The fluidized-bed separation device used in this study consists of a specially designed column fitted at the inlet end with a perforated distributor plate covered with a screen and the top outlet with an adjustable piston. The fluidized-bed consisting of a loose gel matrix permits the unimpeded passage of cell debris and particulate matter, while the target protein is captured by the affinity beads. Purification of the humanized-anti-Tac monoclonal antibody is used as a model system to determine the operational parameters. Also, fluidized-bed receptor-affinity chromatography has been successfully employed in the purification of recombinant interleukin-2 and single chain anti-Tac(Fv)-Pseudomonas exotoxin immunotoxin from unclarified inclusion body extracts. Overall, fluidized-bed receptor-affinity chromatography is found to be a productive affinity method suitable for the purification of recombinant human interleukin-2 and related molecules.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080508
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    High performance liquid chromatographic analysis of polypeptide hormones in transplanted rat islets (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 301-305 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: This report describes high performance liquid chromatographic analysis of transplanted pancreatic islets. A reversed phase ODS column made it possible to measure rat insulin I, II, rat C-peptide I, II and glucagon simultaneously in isolated rat islets without using radioisotopes. Freshly isolated islets contained 118.0 ± 9.7 ng (mean ± SE, n = 6) insulin and 3.01 ± 0.60 ng glucagon per islet. The insulin I/II ratio was 1.22 ± 0.03. Isolated islets were then cultured in vitro or transplanted into mice under the renal capsule. Transplantation induced mild hypoglycemia in the recipients. The graft mean survival time was 7.2 ± 0.4 days (n = 5). Both cultured (n = 7) and transplanted (n = 6) islets showed similar alterations of polypepide hormones on day 4. Insulin decreased to one third and glucagon remained unchanged. The insulin I/II ratio increased twofold. In conclusion, it was suggested that the general fate of isolated islets was caused by ischemia and denervation. Relatively, ischemia may not damage α cells but may damage β cells because α cells are peripherally located. Denervation may release β cells from a resting state under neural tonic inhibition. Mild hypoglycemia and an increased insulin I/II ratio were related to the accelerated insulin synthesis in the isolated islets.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080611
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    External morphology and abundance of antennal sensilla in australian gryllacrididae (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 11-18 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: The long (49-93 mm) antennae of two species of Australian gryllacridids have high total numbers of sensilla consisting of five sensillar types. Ametrus sp. 7 has 22,300 (♀) and 26,250 (♂) sensilla; although the antennae of males are 33% longer than those of females, their sensillar density was 11% less. Bothriogryllacris pinguipes has 26,700 (♂) and 31,900 (♀) sensilla; antennae of females are 55% longer than those of males but sensillar density is 23% less. Aporous sensilla chaetica form 94.5 to 99.5% of all sensilla; they are presumably mechanoreceptors. Uniporous trichoid contact chemoreceptors range from 75-900 in number. Olfactory, multiporous, basiconic sensilla range from 22-440 and olfactory, coeloconic sensilla from 16-235. Two to five multiporous lenticular organs occur on all but female A. sp. 7. Differences in sensillar abundance between males and females are discussed as well as are the relationships between sensillar diversity on gryllacridid mouthparts and antennae. © 1994 Wiley-Liss, Inc.
    Additional Material: 7 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052220103
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    Immunofluorescent studies on titin and myosin in developing hearts of normal and cardiac mutant axolotls (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 19-32 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Homozygous recessive cardiac mutant gene c in the axolotl, Ambystoma mexicanum, results in a failure of the embryonic heart to initiate beating. Previous studies show that mutant axolotl hearts fail to form sarcomeric myofibrils even though hearts from their normal siblings exhibit organized myofibrils beginning at stage 34-35. In the present study, the proteins titin and myosin are studied using normal (+/+) axolotl embryonic hearts at stages 26-35. Additionally, titin is examined in normal (+/c) and cardiac mutant (c/c) embryonic axolotl hearts using immunofluorescent microscopy at stages 35-42. At tailbud stage-26, the ventromedially migrating sheets of precardiac mesoderm appear as two-cell-layers. Myosin shows periodic staining at the cell peripheries of the presumptive heart cells at this stage, whereas titin is not yet detectable by immunofluorescent microscopy. At preheartbeat stages 32-33, a myocardial tube begins to form around the endocardial tube. In some areas, periodic myosin staining is found to be separated from the titin staining; other areas in the heart at this stage show a co-localization of the two proteins. Both titin and myosin begin to incorporate into myofibrils at stage 35, when normal hearts initiate beating. Additionally, areas with amorphous staining for both proteins are observed at this stage. These observations indicate that titin and myosin accumulate independently at very early premyofibril stages; the two proteins then appear to associate closely just before assembly into myofibrils. Staining for titin in freshly frozen and paraffin-embedded tissues of normal embryonic hearts at stages 35, 39, and 41 reveals an increased organization of the protein into sarcomeres as development progresses. The mutant siblings, however, first show titin staining only limited to the peripheries of yolk platelets. Although substantial quantities of titin accumulate in mutant hearts at later stages of development (39 and 41), it does not become organized into myofibrils as in normal cells at these stages. © 1994 Wiley-Liss, Inc.
    Additional Material: 21 Ill.
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    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994) 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052220201
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  • 84
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    Quantitative design of the skeleton in bird hatchlings: Does tissue compartmentalization limit posthatching growth rates? (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 113-131 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Based on a detailed description of hatchling skeletons of the precocial buttonquail (Turnix suscitator) and the altricial budgerigar (Melopsittacus undulatus), this report presents the hypothesis that the rate of avian posthatching growth is limited by the quantitative design (i.e., relative volumes of cartilage, bone, and marrow) of the hatchling skeletons. A Jarge portion of bone in the skeletal elements and fast growth are hypothesized to be mutually exclusive. This hypothesis is tested by morphometric techniques and by statistical comparison of morphometric and growth data. All predictions are met by the data, and the design of hatchling skeletons is described as determined by a tradeoff between tissue composition of skeletal elements and maximum rates of posthatching growth. The precocial design shows large bony areas that supposedly resist mechanical stress of locomotion; however, the relatively small cartilaginous areas exclude high growth rates. The altricial design shows the reverse relationship with small bony areas and a lack of locomotion on the one side but large cartilaginous areas and fast posthatching growth on the other side. © 1994 Wiley-Liss, Inc.
    Additional Material: 8 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052220202
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  • 85
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    Kinematic analysis of jaw protrusion in orectolobiform sharks: A new mechanism for jaw protrusion in elasmobranchs (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 175-190 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Jaw protrusion is an important component of prey capture in fishes, although the mechanics of protrusion have thus far been studied largely in teleosts. Elasmobranchs are also able to protrude their jaws (Tricas and McCosker [1984] Proc. Cal. Acad. Sci. 43: 221-238; Tricas [1985] Mem. S. Calif. Acad. Sci. 8:81-91.; Frazzetta and Prange [1987] Copeia 4:979-993). Several related features of the feeding apparatus contribute to jaw protrusion in sharks. Labial cartilages form an extendible series attached dorsally to the anterolateral face of the palatoquadrate and ventrally to the anteroventral surface of Meckel's cartilage. The labial cartilage chain swings anterolaterally as the lower jaw is depressed, thrusting the labial margins forward to form a circular oral opening and displacing the jaw apparatus towards the food; this pattern is analogous to halecomorph and primitive actinopterygian fishes in which the maxilla swings forward (Lauder [1979] J. Zool. Lond. 187:543-578). The palatoquadrate and Meckel's cartilage also project anteriorly and represent the major contribution to protrusion. These movements occur simultaneously with enlargement of the oral cavity to generate suction. The wobbegong sharks (Orectolobidae) are specialized for jaw protrusion. The spotted wobbegong protrudes its jaw by 33% of its chondrocranial length using two different mechanical systems. In the first mechanism of jaw protrusion, the intermandibularis and interhyoideus muscles medially compress the lower jaw and hyomandibulae. Compression of the lower jaw results in a more acute symphyseal angle so that the anteroposterior alignment of the lower jaw increases due to the rotation of each lower jaw towards a saggital orientation. Distal compression of the hyomandibulae at their attachments to the jaws swings the jaws forward. The second mechanism involves rotation of the ceratohyal around a posterior process of the lower jaw, pushing the hyomandibulae anteroventrally, thereby pushing the jaw articulation ventrally and anteriorly to protrude the jaws. © 1994 Wiley-Liss, Inc.
    Additional Material: 13 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052220205
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    Localization and distribution of gap junctions in normal and cardiomyopathic hamster heart (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 203-213 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: Gap junctions in mammalian heart function to provide low-resistance channels between adjacent cells for passage of ions and small molecules. It is clear that the almost unrestricted passage of ions between cells, ionic coupling, is required for coordinate and synchronous contraction. This knowledge of gap junction function has made it important to study their properties in normal and abnormal tissues. In the present study, we analyzed gap junction distribution in normal and cardiomyopathic heart tissue utilizing immunofluorescent and electron microscopy techniques. Frozen, unfixed sections of age-matched normal and cardiomyopathic cardiac tissues were immunofiuorescently stained using an antibody directed against a specific peptide sequence of the connexin-43 gap junction protein. These studies revealed a characteristic punctate staining pattern for the intercalated discs in normal tissues. Some of the intercalated discs in cardiomyopathic hearts appeared to stain normally; however, others stained diffusely. The pixel intensity distribution of the confocal images demonstrated a marked difference of up to 90% increase in the number of pixels in cardiomyopathic myocardium (CM), yet the pixel intensity of gap junctions had a decrease of approximately 60%. This suggests the possibility that connexin-43 is present in CM cells in significant quantity; however, it does not become localized on the membranes as in normal cells. Electron-microscopic findings corroborate these observations on CM cells by showing an irregular distribution of intercalated discs relatively smaller in size with abnormal orientation and distribution. © 1994 Wiley-Liss, Inc.
    Additional Material: 11 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052220207
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  • 87
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    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994) 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/jmor.1052220301
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  • 88
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    Phylogenetic implications of structure of adult ovary and oogenesis in the penicillate diplopod, Eudigraphis nigricians (miyosi) (diplopoda: Myriapoda) (1994)
    New York, NY : Wiley-Blackwell
    Journal of Morphology 222 (1994), S. 223-230 
    Details
    ISSN: 0362-2525
    Keywords: Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Medicine
    Notes: We describe some significant structures of the adult ovary in a Japanese penicillate diplopod, Eudigraphis nigricans, with respect to phylogenetic implications. The ovary is a long, saclike organ lying between the alimentary canal and the ventral nerve cord from the fourth through the ninth body segment. The ovarian wall consists of a thin ovarian epithelium and a sparse muscle covering. There are two types of oogenetic sites: a single, mound-shaped germarium sitting on the center of the ventral ovarian epithelium, and ∼ 10 pairs of patchlike vitellarial areas metamerically arranged anterior and posterior to the germarium. The germarium consists of oogonia, early previtellogenic oocytes, and some somatic interstitial cells. In contrast, the vitellarial areas are composed of more advanced oocytes, follicle cells surrounding the oocytes, and some interstitial cells, but no oogonia. A few larger previtellogenic oocytes rise up from each vitellarial area into the ovarian lumen. Each of these oocytes is still connected with its own vitellarial area by a partial extension of its follicle. Vitellogenesis takes place in these oocytes rising in the ovarian lumen. The ripe primary oocytes leave their follicles to be transported forward into the oviducts. Some phylogenetic implications of the basic characteristics in ovarian structure and oogenesis of E. nigricans are discussed. © 1994 Wiley-Liss, Inc.
    Additional Material: 10 Ill.
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    URL: http://dx.doi.org/10.1002/jmor.1052220302
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  • 89
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    Determination of nitrite by high-performance liquid chromatography system with electrochemical detector: Measurement of nitric oxide synthase activity in rat cerebellum cytosol (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 14-18 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A simple and sensitive assay method for NO synthase activity is described. Using glassy carbon as electrode and 30% methanol solution with 10 mM NH4Cl as mobile phase, NO2- can be measured without disturbing ECD-detectable substance in NO synthase assay mixture. The NO2- production in the assay mixture of rat cerebellum NO synthase increased with protein and in a time-dependent manner. The Km value for the substrate, L-arginine, was 1.25 μM. The enzyme activity was inhibited in a concentration-dependent manner by a NO synthase inhibitor, NNA. The Ki value for NNA was 0.166 ± 0.060 μM. This ECD-HPLC method for determining NO synthase activity has advantages compared with the diazo-coupling method of the Greiss reagent and the isotopic method in which the conversion of the substrate, [14C]L-arginine, to the product, [14C]L-citrulline is measured; it is simple, sensitive and is convenient for studying the NO synthase activity with various compounds as the substrate.
    Additional Material: 5 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080104
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  • 90
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    A liquid chromatographic method for the determination of fenoprofen in equine plasma and urine (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 29-31 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A high performance liquid chromatographic method to measure plasma and urine fenoprofen levels in equine biofluids is described. Liquid-liquid extraction with diethylether was used to isolate the drug from plasma and urine. The accuracy and reproducibility of the method were within acceptable limits over the concentration range 0-10 μg/mL and 0-20 μg/mL respectively from plasma and urine. Detection limits were 0.05 μg/mL (2 mL plasma) and 0.2 μg/mL (0.5 mL urine). This procedure was applied to ascertain the pharmacokinetics of a 3 g dose of fenoprofen calcium in a horse.
    Additional Material: 3 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080108
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  • 91
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    Masthead (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994) 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080101
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  • 92
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    A fully automated HPLC method for the determination of catecholamines in biological samples utilizing ethylenediamine condensation and peroxyoxalate chemiluminescence detection (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 1-8 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A fully automated in-line extraction reversed-phase high-performance liquid chromatography (HPLC) method with chemiluminescence detection was developed for the analysis of human and rat plasma catecholamines (CAs), norepinephrine (NE), epinephrine (E) and dopamine (DA). N-Methyldopamine (N-MeDA) was used as an internal standard. The method involves collection of plasma samples, which are first diluted with a sample dilution buffer containing N-MeDA, and in-line extraction of CAs using a carboxylic acid small resin precolumn (SERUMOUT-CEX). This pre-extraction process was coupled with an HPLC system including reversed-phase mode separation on an analytical column (TSK gel ODS-80Ts), fluorogenic derivatization with ethylenediamine (ED) and finally postcolumn peroxyoxalate chemiluminescence reaction detection using bis [4-nitro-2-(3,6,9-trioxadecyloxycarbonyl)phenyl]oxalate (TDPO) and hydrogen peroxide. The optimized mobile phase compositions, flow rates, operation timing for the adsorption and desorption of CAs in the precolumn, the separation in the analytical column and the optimum fluorogenic and chemiluminogenic reaction conditions were investigated.The detection limit for all the CAs was about 1 fmol (signal-to-noise ratio is 2). Excellent linearity of the calibration curves for CAs was observed in the range from 5 to 500 fmol for each CA using the internal standard. The relative standard deviations of the method for determining NE (183 fmol), E (23.6 fmol) and DA (6.1 fmol) in 50 μL of human plasma (n = 3) were 2.8, 2.7 and 3.1%, respectively, for the within-day assay and 5.0, 3.8 and 4.0%, respectively, for the between-day assay. The method was applicable to the determination of CAs in 25-50 μL of human or rat plasma.
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  • 93
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    Rapid assay of iopanoic acid in dog plasma using a Hisep column (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 26-28 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A rapid method for the determination of iopanoic acid (IOP) in dog plasma utilizing a Hisep column was developed. A mobile phase of 12% methanol, 88% 0.05 M phosphate buffer pH 3.4 yielded a k′ of 8.5 with no interference from proteins present in plasma. Recoveries of IOP from spiked plasma ranged from 97% to 103% at 270 μmol/L and 1.75 mmol/L respectively. Replication was ±2.8% at 1.75 mmol/L and ±6% at 21 μmol/L. A method utilizing 2,4,6-triiodobenzoic acid as internal standard was also developed for comparison.
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    URL: http://dx.doi.org/10.1002/bmc.1130080107
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  • 94
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    Erratum (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. ii 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080202
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  • 95
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    A new matrix for affinity chromatography and its application in the separation of a human monoclonal antibody (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 142-144 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A method for the preparation of a p-aminobenzene sulphonyl ethyl containing crosslinked Sepharose 4B (ABSE-Sepharose 4B-CL) is described. trypsin, bovine serum albumin (BSA) and concanavalin A (Con A) were immobilized onto this matrix by diazotization. Conditions for the coupling reaction were investigated. The activity of immobilized trypsin reached 1.4 × 104U/g, and 25 mg BSA can be coupled onto 1 g ABSE-Sepharose 4B-CL under the optimal conditions. An affinity medium with immobilized Con A as ligand was prepared by this method, and was used in the separation of a human monoclonal antibody.
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    URL: http://dx.doi.org/10.1002/bmc.1130080310
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  • 96
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    Separation and identification of proteins obtained from Agkistrodon acutus snake venom by capillary zone electrophoresis and laser desorption/ionization mass monitoring (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 148-150 
    Details
    ISSN: 0269-3879
    Keywords: Capillary zone electrophoresis ; laser desorption/ionization mass monitoring ; Agkistrodon Acutus snake venom ; Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: Fractions of seven protein principles with fibrinolytic or thrombin-like activities obtained from Agkistrodon acutus snake venom purified by two steps of normal pressure chromatography were separated further by capillary zone electrophoresis (CZE). Mass determination for these fractions were achieved by performing laser desorption/ionization mass monitoring (LDIM). The comparative study between CZE and LDIM on the separation of these fractions was made.
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    URL: http://dx.doi.org/10.1002/bmc.1130080312
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  • 97
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    Enantiospecific determination of ibuprofen in rat plasma using chiral fluorescence derivatization reagent, (-)-2-[4-(1-aminoethyl)-phenyl]-6-methoxybenzoxazole (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 170-174 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A highly sensitive and reversed-phase high-performance liquid chromatographic method for the determination of the enantiomeric composition of ibuprofen in rat plasma is described. The method is based on the resolution of the diastereomeric amides formed on reaction of the ibuprofen enantiomers with (-)-2-[4-(1-aminoethyl)phenyl]-6-methoxybenzoxazole ((-)-APMB) in the presence of 2,2′-dipyridyl disulphide (DPDS) and triphenylphosphine (TPP) in dichloromethane. The reaction mixture was allowed to stand at room temperature for 5 min, and the reaction was completed by evaporation with a stream of nitrogen at 40°C. The minimum quantifiable concentrations were 0.2 μg/mL and 0.4 μg/mL for S-ibuprofen and R-ibuprofen, respectively, in a 10 μL injection volume. The method was applied to the determination of enantiomeric ibuprofen in plasma after oral administration to rats.
    Additional Material: 4 Ill.
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    URL: http://dx.doi.org/10.1002/bmc.1130080405
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  • 98
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    Simultaneous determination of a mixture of organophosphorus and carbamate pesticides by high performance liquid chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 153-157 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A simple HPLC procedure has been developed for the simultaneous determination of a mixture of standard carbaryl, carbofuran, carbendazim, dimethoate, malathion and methyl parathion from a spiked agricultural soil sample. The pesticide mixture has been extracted using methanol-water. A concave gradient elution with acetonitrile-phosphate buffer for 15 min followed by linear elution for 5 min, using a wavelength of 224 nm for detection, has been found to resolve the mixture efficiently.
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    URL: http://dx.doi.org/10.1002/bmc.1130080402
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  • 99
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    Measurement of amiloride in airway surface liquid utilizing HPLC and fluorescence detection (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 219-223 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Notes: A high-performance liquid chromatographic method for the determination of amiloride in airway surface liquid is described. It involves extraction of the drug with methanol from filter paper on which the sample is absorbed and chromatography on a Zorbax Rx column; the mobile phase is 25% acetonitrile in 0.05 M phosphate buffer; detection by fluorescence at 360/420 nm. Triamterene is used as an internal standard. The range of the assay is 2.0-2033.4 ng/sample, with adequate precision and accuracy. A power curve y = axb best describes the relationship between the peak-height ratio and concentration. Recovery of amiloride is non-linear, probably due to an adsorption process. Accuracy of the assay at lower amounts may be affected by the choice of filter paper a well as by presence of endogenous plasma components. The assay was used to measure amiloride amount in airway surface liquid after administration of 2.5 and 4.5 mg of nebulized amiloride.
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    URL: http://dx.doi.org/10.1002/bmc.1130080505
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  • 100
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    Determination of flutamide and hydroxyflutamide in dog plasma by a sensitive high performance liquid chromatography method utilizing mid-bore chromatography (1994)
    New York, NY : Wiley-Blackwell
    Biomedical Chromatography 8 (1994), S. 251-254 
    Details
    ISSN: 0269-3879
    Keywords: Chemistry ; Analytical Chemistry and Spectroscopy
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Medicine
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/bmc.1130080511
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