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  • 1
    Electronic Resource
    Electronic Resource
    Characterization of genetic variants of human serum transferrin by isoelectric focusing: Comparison between conventional and immobilized pH gradients, and application to a protocol for paternity testing (1988)
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 411-417 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The polymorphism of transferrin (Tf) is currently being studied by isoelectric focusing in carrier ampholyte-generated pH gradients, carrier ampholyte-separator pH gradients or in immobilized pH gradients. Details for obtaining reproducible results with each of the three procedures are outlined. The effectiveness of pretreatment of serum samples with ferrous/ferric salts is discussed, and incubation times optimized after spectrophotometric measurement of the monoferric Tf conversion. Most of the presently available commercial batches of carrier ampholytes do not reliably discriminate the six common TfC subtypes. Resolution of C1, C3 and C2 was achieved by adding 20 to 90 mM HEPES slab gels prepared with various carrier ampholytes. Isoelectric focusing in carrier ampholyte-separator pH gradients cannot be recommended as a standard typing procedure because the results strongly depend on the batch of carrier ampholytes. Tf subtype resolution was only achieved by using isoelectric focusing in immobilized pH gradients with pH slopes reliably reproducible from one experiment to another. Two major shortcomings of immobilized pH gradients are a marked tendency to protein precipitation at the application site and an interaction of proteins with the charged matrix. A protocol for Tf subtyping in immobilized pH gradients is described, based on prior desialylation of samples instead of pretreatment with iron. Sample entry into the matrix was optimized by addition of 5 mM Tris to the gels, and initially running them at low voltage. Recommendations are provided for the application of Tf typing for paternity testing.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150090811
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  • 2
    Electronic Resource
    Electronic Resource
    Simplified molding procedure for ultrathin-layer polyacrylamide slab gels using methacrylate supports and unsilanized glass plates: Hybrid isoelectric focusing on ultrathin immobilized pH gradients (1987)
    Weinheim : Wiley-Blackwell
    Electrophoresis 8 (1987), S. 371-373 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A simplified procedure for the preparation of ultrathin polyacrylamide slab gels is reported. The molding cells consist of an unsilanized glass plate and methacrylate plate or acryl-coated glass plate. Owing to the repelling properties of acrylate the ultrathin gels spontaneously detach from the acrylate surface on unfastening the molds, firmly adhering to the unsilanized glass. Gels as thin as 0.20-0.25 mm could be polymerized without resorting to silanized glass plates or polyester films. The procedure is especially suitable for sticky gels, such as highly cross-linked polyacrylamide gels and gels with immobilized pH gradients.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150080808
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  • 3
    Electronic Resource
    Electronic Resource
    Errors in sizing bands of hypervariable DNA profiles on autoradiograms: Are they Gaussian? (1992)
    Weinheim : Wiley-Blackwell
    Electrophoresis 13 (1992), S. 341-345 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: The accuracy of procedures for sizing hypervariable restriction fragments by Southern blot analysis (SBA) has been tested under three different experimental conditions: (i) intrablot serial analyses: three heterozygous DNA profiles were tested 14 times each in the same gel electrophoresis; (ii) intralaboratory analyses: we replicated three profiles (six autoradiographic bands) in over 100 SBA experiments; (iii) interlaboratory analyses: 15 serial measurements produced in a recent collaborative study (Forensic Sci. Int. 1991, 49, 1-15) were taken into account. In these three cases, a typical U-shaped correlation curve between molecular size and coefficient of variation was found. We explain decrease of accuracy at both extremities of the gels in terms of: (i) enlarged shapes of bands and mean electrophoretic resolution at the cathode; (ii) diffusion and blurring of bands at the anodal edge. The three populations of data were subjected to a χ2 test and to the Kolmogorov-Smirnov test in-order to verify their compliance with normal distribution. Twenty-three out of 27 tests indicated no significant deviation from the assumption of Gaussian distribution. We recommend the adoption of tests for normality to validate the use of symmetric confidence intervals for calculating gene frequencies and asserting a match between adjacent bands.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150130171
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  • 4
    Electronic Resource
    Electronic Resource
    Immobilized pH gradients in ultrathin-layer (100 μm) gels: Avoiding capillarity by a ‘squeezing-sealing mold’ technique (1988)
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 514-519 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Several procedures for casting 100 μm ultrathin immobilized pH gradients are described. When acrylate/glass molding cassettes (Pascali et al., Electrophoresis 1987, 8, 371-373) are used two main problems are encountered: (i) a tendency of polymerization solutions, at the beginning of delivery, to spread across the glass surfaces with troublesome effects on the gradient stratification, and (ii) the raising of steep menisci at both extremities of the pH intervals, originating from capillarity phenomena and resulting in nonuniform gradients with bowed electrophoretic patterns. The first shortcoming was acceptably solved by increasing the density of sucrose gradients, and pouring them into prewarmed molding cassettes. The detrimental effect of menisci could be overcome by using a ‘squeezing-sealing mold’ technique. A molding cassette was endowed with a continuous, squared spacer frame, the upper side being open by inserting a wedged clip. A slight excess of polymerization solution was first dispensed into the cassette and squeezed away on removal of the clip. By completely excluding air from the molding cassette, uniform and well reproducible ultrathin gels could be cast. A major advantage of ultrathin immobilized pH gradient gels is the drastically shorter focusing time.
    Additional Material: 9 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150090911
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  • 5
    Electronic Resource
    Electronic Resource
    Efficient, small scale electroelution of high molecular weight DNA from agarose gels by a miniature vertical electrophoresis cell (1991)
    Weinheim : Wiley-Blackwell
    Electrophoresis 12 (1991), S. 317-320 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A miniature, vertical electrophoretic system is described that allows quick elution and efficient harvesting of double-stranded, high molecular weight DNA fragments from agarose gels. A gel slice containing the band of interest is excised from the agarose gel after submarine electrophoresis and placed into an electrophoresis cell, assembled by inserting a truncated micropipette tip into a 2.0 mL polypropylene tube. The tip supports the agarose slice and is connected to dialysis tubing. Platinum wire electrodes are placed over the tip and at the bottom of the test tube. DNA molecules are quickly eluted into the dialysis bag (60 s to 10 min for bands ranging from 2 to 23 kb) and then easily recovered. The method is highly efficient and allows considerable time saving. Of special interest is its applicability to small starting amounts of DNA (〈0.5 μg per band). Purity of the DNA is comparable, if higher, to that obtainable by conventional electroelution on dialysis bags and DEAE-cellulose membrane electrophoresis. The system is simple enough to be used as a routine method for eluting multiple bands in short times.
    Additional Material: 3 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150120417
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  • 6
    Electronic Resource
    Electronic Resource
    Fluorescence-based classification of microsatellites using a single-wavelength semiautomatic sequencer: Genotype assignment and identity tests by analysis of comigrating peak profiles (1995)
    Weinheim : Wiley-Blackwell
    Electrophoresis 16 (1995), S. 1875-1880 
    Details
    ISSN: 0173-0835
    Keywords: Microsatellites ; Semiautomatic sequencers ; Identity tests ; Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Microsatellite 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.
    Additional Material: 5 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.11501601307
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  • 7
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    PLS2 in Metabolomics (2019)
    Molecular Diversity Preservation International
    Details
    Publication Date: 2019-03-15
    Description: Metabolomics is the systematic study of the small-molecule profiles of biological samples produced by specific cellular processes. The high-throughput technologies used in metabolomic investigations generate datasets where variables are strongly correlated and redundancy is present in the data. Discovering the hidden information is a challenge, and suitable approaches for data analysis must be employed. Projection to latent structures regression (PLS) has successfully solved a large number of problems, from multivariate calibration to classification, becoming a basic tool of metabolomics. PLS2 is the most used implementation of PLS. Despite its success, PLS2 showed some limitations when the so called ‘structured noise’ affects the data. Suitable methods have been recently introduced to patch up these limitations. In this study, a comprehensive and up-to-date presentation of PLS2 focused on metabolomics is provided. After a brief discussion of the mathematical framework of PLS2, the post-transformation procedure is introduced as a basic tool for model interpretation. Orthogonally-constrained PLS2 is presented as strategy to include constraints in the model according to the experimental design. Two experimental datasets are investigated to show how PLS2 and its improvements work in practice.
    Electronic ISSN: 2218-1989
    Topics: Biology , Medicine
    Published by Molecular Diversity Preservation International
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  • 8
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    Occupational Exposure to Fine Particles and Ultrafine Particles in a Steelmaking Foundry (2019)
    Molecular Diversity Preservation International
    Details
    Publication Date: 2019-02-01
    Description: Several studies have shown an increased mortality rate for different types of tumors, respiratory disease and cardiovascular morbidity associated with foundry work. Airborne particles were investigated in a steelmaking foundry using an electric low-pressure impactor (ELPI+™), a Philips Aerasense Nanotracer and traditional sampling equipment. Determination of metallic elements in the collected particles was carried out by inductively coupled plasma mass spectrometry. The median of ultrafine particle (UFP) concentration was between 4.91 × 103 and 2.33 × 105 part/cm3 (max. 9.48 × 106 part/cm3). Background levels ranged from 1.97 × 104 to 3.83 × 104 part/cm3. Alveolar and deposited tracheobronchial surface area doses ranged from 1.3 × 102 to 8.7 × 103 mm2, and 2.6 × 101 to 1.3 × 103 mm2, respectively. Resulting inhalable and respirable fraction and metallic elements were below limit values set by Italian legislation. A variable concentration of metallic elements was detected in the different fractions of UFPs in relation to the sampling site, the emission source and the size range. This data could be useful in order to increase the knowledge about occupational exposure to fine and ultrafine particles and to design studies aimed to investigate early biological effects associated with the exposure to particulate matter in the foundry industries.
    Electronic ISSN: 2075-4701
    Topics: Mechanical Engineering, Materials Science, Production Engineering, Mining and Metallurgy, Traffic Engineering, Precision Mechanics
    Published by Molecular Diversity Preservation International
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  • 9
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    Exploring Perinatal Asphyxia by Metabolomics (2020)
    Molecular Diversity Preservation International
    Details
    Publication Date: 2020-04-04
    Description: Brain damage related to perinatal asphyxia is the second cause of neuro-disability worldwide. Its incidence was estimated in 2010 as 8.5 cases per 1000 live births worldwide, with no further recent improvement even in more industrialized countries. If so, hypoxic-ischemic encephalopathy is still an issue of global health concern. It is thought that a consistent number of cases may be avoided, and its sequelae may be preventable by a prompt and efficient physical and therapeutic treatment. The lack of early, reliable, and specific biomarkers has up to now hampered a more effective use of hypothermia, which represents the only validated therapy for this condition. The urge to unravel the biological modifications underlying perinatal asphyxia and hypoxic-ischemic encephalopathy needs new diagnostic and therapeutic tools. Metabolomics for its own features is a powerful approach that may help for the identification of specific metabolic profiles related to the pathological mechanism and foreseeable outcome. The metabolomic profiles of animal and human infants exposed to perinatal asphyxia or developing hypoxic-ischemic encephalopathy have so far been investigated by means of 1H nuclear magnetic resonance spectroscopy and mass spectrometry coupled with gas or liquid chromatography, leading to the identification of promising metabolomic signatures. In this work, an extensive review of the relevant literature was performed.
    Electronic ISSN: 2218-1989
    Topics: Biology , Medicine
    Published by Molecular Diversity Preservation International
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  • 10
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    A longitudinal 1H-NMR metabolomics analysis of urine from newborns with hypoxic-ischemic encephalopathy undergoing hypothermia therapy. Clinical and medical legal insights (2018)
    Public Library of Science
    Details
    Publication Date: 2018-04-18
    Electronic ISSN: 1932-6203
    Topics: Medicine , Natural Sciences in General
    Published by Public Library of Science
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