bluuebunny/crossref_metadata_2025_split huggingface dataset:
Screening of umbilical cord blood hemoglobins by isoelectric focusing in capillaries
Authors: Conti Monica , Gelfi Cecilia , Righetti Pier Giorgio 
Published: May 2005

AbstractSeparation and quantitation of the three main hemoglobin components of umbilical cord blood (fetal, acetylated fetal and adult hemoglobins; Hb F, Fac, A) by capillary isoelectric focusing (IEF) in a pH 6–8 gradient is reported. Even in coated capillaries (with covalently bound chains of linear acrylamido derivatives, notably N‐acryloylaminoethoxyethanol), no base line separation is obtained between Hb F and A, although this is routinely achieved in gel slab IEF. However, when the carrier ampholyte buffers were added to 3% short‐chain liquid linear polyacrylamide, base line resolution and stabilization of peak transit times were obtained. This suggests that even in the best coating procedures, patches of the inner capillary surface could still be naked, so that the static coating is complemented by a dynamic coating on the unoccupied sites. An additional improvement in separation occurs if the above mixture, comprising 5% carrier ampholytes in the pH 6–8 range and 3% soluble polyacrylamide, is made to contain 50 mM β‐alanine, a “separator” known to flatten the pH gradient around pH 7. In the normal newborns analyzed (n = 30), the following average values were obtained: Hb F, 70.1% (range 65–75%); Hb A, 20.2% (range 15–25%); and HbFac, 9.5% (range 7–11%).
Coelectrophoresis of cardiac tissue from human, dog, rat and mouse: Towards the establishment of an integrated two‐dimensional protein database
Authors: Corbett Joseph M. , Wheeler Colin H. , Dunn Michael J. 
Published: May 2005

AbstractWe have investigated the feasibility of identifying homologous proteins in whole tissue protein extracts of dog, mouse and rat hearts by comparison with our human heart two‐dimensional (2‐D) database. Samples of ventricular myocardial tissue from each of these species were coelectrophoresed with a human tissue sample. Gels were silver stained and patterns were analysed using PDQUEST. The number of proteins comigrating with human proteins was 301, 201 and 356 for the dog, mouse and rat, respectively. In the dog pattern, 33 of these comigrating proteins were tentatively identified from the similarity between their migration properties and those of known human proteins. Twentynine such proteins were identified in the mouse pattern while 30 comigrating rat proteins were identified. While these tentative identifications require confirmation, we feel that this technique offers a useful shortcut in the characterisation of proteins present in similar tissue samples from different species and avoids the necessity for duplicating laborious procedures, such as protein microsequencing, otherwise used in the identification of these proteins in each species.
Increased microsatellite variability in Macaca mulatta compared to humans due to a large scale deletion/insertion event during primate evolution
Authors: Kayser Manfred , Nürnberg Peter , Bercovitch Fred , Nagy Marion , Roewer Lutz 
Published: May 2005

AbstractHuman (GATA)n microsatellites D12S66 and D12S67 could be successfully amplified by polymerase chain reaction (PCR) in various species of apes and monkeys. In 86 unrelated animals of the most intensively studied species Macaca mulatta we demonstrated five alleles at “D12S66” differing in size in increments of 4 bp (159–175 bp), whereas 17 alleles were observed at locus “D12S67”. The alleles of the latter locus are distributed in two separate groups with no alleles of intermediate size. Six alleles were found between 108–128 bp and 11 alleles between 181–249 bp. Mendelian inheritance of the codom‐inant alleles was proven by family studies. Sequencing of the “D12S67” locus revealed that the shorter alleles are characterized by a single perfect (GATA)n stretch whereas the longer alleles consist of two blocks of (GATA)n repeats separated by an intervening sequence of 9 bp. The composite structure of the longer alleles closely resembles that of the 12 human D12S67 alleles (229–273 bp). The enormous species variation in the fragment size range, with the smallest allele found in Macaca mulatta (108 bp) and the largest (364 bp) in Gorilla gorilla gorilla strongly indicates that D12S67 has been subjected to recurrent mutations over the course of primate evolution including a large deletion and/ or insertion event.
Arbitrarily amplified DNA in systematics and phylogenetics
Authors: Schierwater Bernd 
Published: May 2005

AbstractDifferent polymerase chain reaction (PCR) based techniques which use single primers of arbitrary sequence have become prominent and powerful tools for taxonomic and genealogical studies. While the use of arbitrarily amplified DNA (AAD) markers for the identification of lower taxonomic units (e.g. at the species level) has been proven to be more or less straightforward, their potential usefulness in phylogenetic studies has not been sufficiently addressed yet. The main advantages of AAD markers include that they require only very small amounts of DNA, no sequence information of the taxa under investigation and the relative ease of generating high numbers of AAD markers in short time. On the other hand, the generation of AAD markers is susceptible to certain random and systematic errors, and for phylogenetic studies the taxonomic level at which informative markers can be generated is severely limited.
Genetic self‐management in a captive colony of mandrills (Mandrillus sphinx) as revealed by DNA minisatellite fingerprints
Authors: Wickings E. Jean 
Published: May 2005

AbstractDNA fingerprints using three oligonucleotide probes, a (GTG)5 short tandem repeat sequence, and 2 corresponding to the core sequences of minisatellites 33.15 and 33.6, have been used to generate DNA fingerprints of a captive colony of mandrills (Mandrillus sphinx) housed at the Centre International de Recherches Médicales de Franceville (CIRMF), in order to monitor paternity. A colony of 6 males and 7 females was established in a 6 ha. rain‐forested enclosure in 1983. Band sharing scores indicated that none of the founders were related. By April 1994 the colony had increased to 84 individuals. Forty‐nine of the 77 births were first‐generation infants, a further 27 were second‐generation offspring, and one was a third‐generation mandrill. Five animals died during the study, and one was removed for husbandry reasons. Overall band sharing between individuals increased markedly and the power to discriminate between first‐ and second‐generation infants was reduced; the probability of two individuals sharing the same pattern increased 1000‐fold from 10−6 to 10−3. The dominant male in the group effectively guards access to all receptive females during the annual breeding season. Paternity analysis confirmed that the dominant male is the father of the great majority of offspring born during his term of dominance. Daughters born before 1991 to either of the two dominant males reached sexual maturity during his tenure. Incestual inbreeding occurred with both males, but only 25% of all possible cases resulted in the birth of a viable infant; one male fathered infants on 3 of 13 possible occasions and a second male on 2 of 7 occasions. Female avoidance strategies and shifts in male dominance in this closed mandrill colony appear to be timed in a way that reduces incest.
Immunoprinting reveals different genetic bases for (auto)immune diseases
Authors: Epplen Cornelia , Buitkamp Johannes , Rumpf Heike , D'Souza Markus , Epplen Jörg T. 
Published: May 2005

AbstractThe genetic basis of complex (auto)immune diseases has been studied for an ovine nematode infection, human rheumatoid arthritis (RA), early onset pauciarticular arthritis (EOPA) and multiple sclerosis (MS). Immunoprinting combines the powerful simplicity of polymerase chain reaction (PCR)‐based amplification of discrete, highly informative microsatellite loci with the principle of genetic associations. This approach has allowed us to define novel genetic risk factors in adult RA patient categories whereas EOPA forms in juveniles display other prominent genetic contributions. Differentially regulated tumor necrosis factor (TNF) expression may lead to a better understanding of the causal pathogenesis of EOPA while T cell receptor (TCR) gene polymorphisms appear crucial for RA manifestation in certain patient groups. Statistically significant marker associations have still to be defined for MS in larger panels of patient and control cohorts. The clinical course of the disease will probably have to be taken into account when associations with lymphokine levels are evaluated. In essence a convoluted myriad of negative and a few positive disease association data have been generated efficiently by immunoprinting. As expected, the interrelationships are truly complicated between the polymorphic genetic instances predisposing to autoimmune disease. Nevertheless, risk factors may be defined on an individualized basis by indirect gene diagnosis revealing predispositions and providing a more solid basis for differential diagnosis and treatment.
Parallel genome analysis by one‐ and two‐dimensional DNA fingerprinting in human gliomas
Authors: Nürnberg Peter , Marczinek Karola , Thiel Gundula , Hampe Jochen 
Published: May 2005

AbstractThe detection of DNA variation in cancers is an important step in elucidating the mechanism of tumorigenesis. Using the strategy of multipoint genome analysis we detected many differences between glioma‐derived and constitutional DNA by customary DNA fingerprinting with simple repetitive oligonucleotide probes. Amplification of the epidermal growth factor receptor (EGFR) gene has been found to be easily detectable as new or highly intensified bands in one‐dimensional (1‐D) DNA fingerprints of glioblastoma DNA generated with probes (GTG)5 or (GT)8. However, in most low‐grade astrocytomas, 1‐D DNA fingerprinting has failed to reveal any genomic abnormalities. In these cases a two‐dimensional (2‐D) technique was successfully employed that is based on size separation in neutral gels followed by sequence‐dependent separation in denaturing gradient gels and hybridization with several mini‐ and microsatellite core probes. The hundreds of spots visualized with this technique were used to detect subtle changes probably occurring as the initial steps of tumorigenesis in human gliomas. On average, five of the approximately 580 sports generated by probes CAC and 33.6 were found to be altered in tumor DNA; 80% of the alterations were spot losses, the rest being spot gains or amplifications. Computer‐based image analysis using an external lambda marker provided a stringent way to compare spot patterns generated by 2‐D DNA finger‐printing. In comparisons performed between typing patterns generated on the same gel, 99% of truly identical spots were confirmed by the sofware. In intergel comparisons 84% of identical spots were matched on the basis of the marker information alone.
DNA fingerprinting of Brassica juncea cultivars using microsatellite probes
Authors: Bhatia Sabhyata , Das Sandip , Jain Ajay , Lakshmikumaran Malathi 
Published: May 2005

AbstractThe genetic variability in the Brassica juncea cultivars was detected by employing in‐gel hybridization of restricted DNA to simple repetitive sequences such as (GATA)4, (GACA)4 and (CAC)5. The most informative probe/enzyme combination was (GATA)4/EcoRI, yielding highly polymorphic fingerprint patterns for the B. juncea cultivars. This technique was found to be dependable for establishing the variety specific patterns for most of the cultivars studied, a prerequisite for germplasm preservation. The results of the present study were compared with those reported in our earlier study in which random amplification of polymorphic DNA (RAPD) was used for assessing the genetic variability in the B. juncea cultivars.
Highly repetitive DNA sequence elements from Orseolia oryzae (Wood‐Mason) discriminate between the Indian isolates of the Asian rice gall midge and the paspalum midge
Authors: Ehtesham Nasreen Z. , Bentur Jagdish S. , Bennett John 
Published: May 2005

AbstractWe describe multicopy DNA clones isolated from a partial genomic library of Orseolia oryzae, based on reverse genomic hybridization, suitable for studying genetic variation in the Asian rice gall midge and other isomorphic species. Three clones produced monomorphic DNA band patterns between biotypes of O. oryzae but polymorphic patterns were produced between O. oryzae and O. fluvialis, the paspalum midge. These probes detect changes in the repetitive sequence structure between species and constitute the first genetic markers for distinguishing between field isolates of rice gall midge and related species of Orseolia. These will be useful in identifying and perhaps eradicating alternative hosts for this pest, and detecting early‐season outbreaks of O. oryzae from light trap collections.
Hypervariable genomic variation to reconstruct the natural history of populations: Lessons from the big cats
Authors: Menotti‐Raymond Marilyn , O'Brien Stephen J. 
Published: May 2005

AbstractThe extent and nature of variation in hypervariable regions of DNA have been used in the past as a means to infer the natural histories of populations. We review the interpretation of the extent of genetic diversity for minisatellite DNA in the cheetah to estimate the timing of a population bottleneck in the species and the potential application of a second class of hypervariable DNA, microsatellite DNA, as a molecular tool to examine the natural histories of felid populations. A calibration curve relating the degree of allele fragment sharing in individuals to relatedness in a captive pedigree of cheetahs is presented. This measurement has important applications for management of potential matings in captive management situations.
Social and related implications of DNA fingerprinting and other new DNA technologies
Authors: Bhargava Pushpa M. 
Published: May 2005

AbstractThe discovery of the structure of DNA at the beginning of this half of the century, and the recognition that it is virtually the universal genetic material, made it imperative for man, sooner or later, to apply this knowledge for unexpected ends. Genetic engineering, DNA fingerprinting, sequencing of whole genomes (be they of man, plants or microorganisms), or exploitation of the differences between the DNA of the male and the female (for example, of the X and the Y sperm) have thus all been historical imperatives. Today, many such predicted technologies are a reality or have come close to being so. Such DNA‐based technologies have enormous social implications, discussed in this article. The impact of these technologies on different societies is likely to be different, in view of the different social conditions that prevail, and the different historical background and cultural traditions. In India, for example, the use of DNA fingerprinting could prevent the large‐scale exploitation of women, and several kinds of litigation. On a global scale, the availability of this technique and its refinement to a stage where it would become virtually foolproof could act as a deterrent to certain kinds of crime. There will also be unprecedented benefits to agriculture but also new problems that will need to be faced and resolved. Wisdom demands that we prepare ourselves for a largescale application of the new DNA technologies that are a historical imperative, understand the problems that are likely to arise, and plan to cope with them before they descend on us in full force.This is the text of a popular public lecture given by the author during the Third International Conference on DNA Fingerprinting held in Hyderabad, India, in December 1994.
Fluorescence‐based classification of microsatellites using a single‐wavelength semiautomatic sequencer: Genotype assignment and identity tests by analysis of comigrating peak profiles
Authors: Moscetti Alessandra , Boschi Ilaria , Dobosz Marina , Destro‐Bisol Giovanni , Pescarmona Marina , d'Aloja Ernesto , Pascali Vincenzo L. 
Published: May 2005

AbstractMicrosatellite analysis, based on fluorescein labeling and reading through a semiautomatic single wavelength sequencer, is described. Paris of labeled polymerase chain reaction (PCR) samples, mixed in equimolar proportion, were electrophoresed and the specific peaks read in a single gel lane. Identity was asserted when peaks overlapped in a unique fluorescent signal which, compared with individual sample profiles, had a twofold intensity. Classification was achieved by blending individual PCR products to ‘locus specific allelic ladders’ (composite samples containing a repertory of fragments allelic to a given locus) and by noticing the specific peak enhancement. The resulting protocol of analysis assigned no size and classified allelic forms by tandem repeat number. Applied to a large repertory of PCR products and compared with manual electrophoresis, this protocol proved to be reliable and reduced times and costs of genotype analysis. Analysis of comigrating peak profiles is highly objective and provides convincing evidence for diagnostics and identity tests.
Flanking region sequences and internal repeat structure of the pYNH24 (D2S44) 2 kbp insert analyzed by polymerase chain reaction and partial digestion with RsaI
Authors: Holmlund Gunilla , Lindblom Bertil 
Published: May 2005

AbstractThe 2 kbp YNH24 pUC18 insert was partially sequenced, amplified with fluorescence‐labeled primers, and characterized by incomplete digestion with the restriction enzyme RsaI. A characteristic RsaI profile with one restriction site in each repeat unit was obtained when the fragments were analyzed on a DNA sequencer. We developed this procedure with the specific aim of analyzing the internal repeat structure in recombinant alleles often observed in the YNH24 variable number of tandem repeat system (D2S44). Nonetheless, it should also be possible to apply the method when analyzing other tandem repeats, even when little information regarding their sequences is available. In addition, when the repeat array is amplified, either a 5′ or a 3′ fluorescence‐labeled primer can be used to enable analysis from both directions of fairly long (> 3.6 kbp) alleles. The RsaI fragments obtained by partial digestion of the amplified pYNH24 insert corresponded well to the RsaI sites found in the sequenced regions. The 5′ flanking region contained eight 30–34 bp sequences similar to the repeat unit and also several RsaI sites, whereas only a few RsaI sites and no repeat units were found in the 3′ flanking region. Twenty‐five consecutive RsaI sites were revealed, in the center of the pYNH24 insert, indicating a region with strictly repeated core sequences; this repetitive block comprised less than half of the total insert. Two approximately 2.24 kbp HinfI alleles from two different heterozygous individuals were found to have identical internal structure, which was very similar to that of the insert although it exhibited only 15 RsaI repeats.
Theoretical basis of one‐dimensional genome scanning: A direct method to identify the site of a mutation
Authors: Gondo Yoichi , Brilliant Murray H. 
Published: May 2005

AbstractGenome scanning is a technique designed to uncover a net genetic difference between otherwise identical DNA samples. As such, it can be used to directly identify the site of a gene mutation, facilitating the cloning of DNA fragments from that site. Unlike other conventional positional cloning methods, one‐dimensional genome scanning does not require prior knowledge of the location of the gene or mutation nor does it require closely linked markers. Rather, this method can directly identify the site of a net genomic change, such as a deletion or duplication caused by a mutation. Thus, the genome scanning method can be used in place of classic positional cloning strategies because prior positioning or mapping of the objective gene is unnecessary. By using this approach, we have identified and cloned a DNA fragment duplicated in the pun mutation of the mouse pink‐eyed dilution locus (Brilliant et al., Science 1991, 252, 566–569). However, no other similar attempt using one‐dimensional genome scanning has been reported so far, in spite of the simplicity of the procedure and its success in identifying and ultimately characterizing the pink‐eyed dilution gene of the mouse. The lack of other reports of its success are perhaps not because of the practical difficulties of this method, but may be due to the false presumption that the probability for directly identifying the mutation site using genome scanning is extremely low. The theoretical probability was calculated and is presented here. We calculate that there is about a 50% chance in identifying a 200 kbp net genomic change (due to a mutation) by scanning 10 000 random sites in the mammalian genome. The practical key elements for calculating the probability of success are also discussed.
Capillary electrophoresis for screening of adenylosuccinate lyase deficiency
Authors: Gross Manfred , Gathof Birgit S. , Kölle Pirkko , Gresser Ursula 
Published: May 2005

AbstractWe report a new screening method for adenylosuccinate lyase (ASase) deficiency using capillary electrophoresis (CE). This enzyme defect causes secondary autism and psychomotor retardation in early childhood. In all body fluids of these patients, two succinylpurine metabolites can be found that are normally not detectable: succinyladenosine and succinylaminoimidazole carboxamide (SAICA) riboside. A Beckman P/ACE 2050 capillary electrophoresis system was used with a 47.1 cm capillary, 75 μm ID, and the P/ACE Beckman UV absorbance detector. Untreated urine, injected for 1 s, was separated in a pH 8.63 borate buffer at 20 kV. The two succinylpurines (migration times 13.36 and 13.60 min) were detected at 254 nm only in urine of patients with ASase deficiency but not in control samples.
Impact of electroosmosis on isotachophoresis in open‐tubular fused‐silica capillaries: Analysis of the evolution of a stationary steady‐state zone structure by computer simulation and experimental validation
Authors: Thormann Wolfgang , Caslavska Jitka , Mosher Richard A. 
Published: May 2005

AbstractA dynamic computer model for simulation of open‐tubular capillary electrophoresis that includes in situ calculation of electroosmosis along the fusedsilica capillary column has been applied to the characterization of an anionic isotachophoretic system in presence of a cathodic electroosmotic flow. For each column segment, electroosmosis is calculated with the use of a wall mobility, the voltage gradient and the degree of dissociation of the silanol surface groups of the capillary wall. Then, the bulk capillary flow is taken to be the average of all of the segment flows and considered to represent a plug flow. This simple approach enables the combined simulation of the temporal behavior of an isotachophoretic zone structure in presence of electroosmosis. For a model anionic isotachophoretic configuration at pH 6, simulation data reveal the asymptotic formation of a stationary steady‐state zone configuration in which electrophoretic and electroosmotic zone displacements are opposite and of equal magnitude. The position of the stationary boundaries are predicted to be dependent on the selected wall pK and mobility values. For two different instruments, qualitative agreement between experimental data and simulation results obtained with a wall pK between 5 and 6 is demonstrated. However, for the two experimental setups, significant differences in electroosmotic pumping (i.e. wall mobility values) are noted.