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  • 1
    Electronic Resource
    Electronic Resource
    Low molecular weight factor in bovine caudal epididymal fluid that stimulates calcium uptake in caput spermatozoa (1988)
    New York, NY : Wiley-Blackwell
    Gamete Research 20 (1988), S. 343-352 
    Details
    ISSN: 0148-7280
    Keywords: calcium uptake ; caudal fluid ; caput sperm ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: Secretions from the mammalian epididymis contain proteins that bind to developing sperm and are presumed to play a role in sperm maturation. The biochemical functions in sperm of most of these proteins are not known. In this report we describe the presence of a low molecular weight compound in bovine caudal epididymal luminal fluid (CF) that has a potent stimulatory effect on calcium (45Ca2+) uptake in immature caput epididymal spermatozoa. The studies were initially undertaken to characterize the effect of the protein caltrin, present in bovine seminal plasma (BSP), on calcium uptake into caput spermatozoa. Caltrin is known to block calcium influx into mature bovine sperm. Unexpectedly, the kinetics of calcium uptake into caput sperm showed a biphasic response when treated with BSP, namely, a stimulation of uptake at 1 to 5 min and inhibition of uptake after this time. Since caudal sperm do not show this biphasic response, we reasoned that BSP contained a factor derived from CF that must interact with developing sperm before the binding of caltrin to sperm can prevent further calcium uptake. We first demonstrated that preincubation of caput sperm with CF eliminated the biphasic calcium uptake effect induced in caput sperm by BSP and that caudal fluid alone had a potent stimulatory effect on calcium uptake in caput sperm. Half-maximal stimulation (fivefold over control) occurred at a caudal fluid protein concentration of 0.27 mg/ml. Partial purification of the factor indicates that it is of low molecular weight (MW ∼ 1,000), but further chemical characterization has not been carried out and its epididymal site of origin is not known. The results indicate that the regulation of intracellular calcium levels in sperm differs in immature and mature bovine sperm in that an epididymal factor promotes calcium uptake during epididymal maturation, and the seminal fluid protein caltrin prevents it at ejaculation.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120200309
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  • 2
    Electronic Resource
    Electronic Resource
    The involvement of methyl transfer reactions and S-adenosylhomocysteine in the regulation of bovine sperm motility (1985)
    New York, NY : Wiley-Blackwell
    Gamete Research 12 (1985), S. 399-409 
    Details
    ISSN: 0148-7280
    Keywords: protein carboxylmethylation ; S-adenosylhomocysteine ; sperm motility ; Life and Medical Sciences ; Cell & Developmental Biology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology
    Notes: The methylation of proteins and phospholipids has been demonstrated in mammalian sperm. The role of these methylation reactions in the regulation of sperm motility is, however, unclear. A combination of agents [adenosine, L-homocysteine and erythro-9-(2-hydroxyl-3-nonyl) adenine (EHNA)] known to increase the level of S-adenosylhomocysteine (AdoHcy), a competitive inhibitor of S-adenosylmethionine (AdoMet)-mediated methylations, has been shown to inhibit rabbit sperm motility. The level of AdoHcy in response to these agents has, however, not been measured. Therefore, enzymatic protein carboxylmethylation (PCM) and the consequences of its inhibition by elevated intrasperm AdoHcy levels on motility were studied using bovine sperm. Incubation of bovine cauda epididymal sperm with L-[methyl-3H]methionine resulted in a linear increase in PCM for up to 2 h. Treatment of sperm with adenosine (0.5 mM), L-homocysteine (0.5 mM), and EHNA (0.25 mM) produced a dramatic increase in AdoHcy levels after 15 min and a 97% inhibition of PCM after 0.5 h, followed by an inhibition of sperm motility (58% and 97%) after 1 and 2 h. Similar but less marked results were obtained with L-homocysteine (0.5 mM) alone. Individually, cycloleucine (20 mM), an inhibitor of AdoMet synthetase, and adenosine (0.5 mM) also inhibited PCM (87% and 70%, respectively) after 2 h but had no effect on sperm motility or AdoHcy levels. These studies show for the first time that agents that elevate AdoHcy levels inhibit sperm motility. This, however, leaves open the question of whether protein carboxylmethylation or other methyl transfer reactions, eg, involving phospholipid, might be critical to motility inhibition by AdoHcy. Thus, the mechanism by which AdoHcy inhibits sperm motility remains unclear.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/mrd.1120120407
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  • 3
    Electronic Resource
    Electronic Resource
    The use of biotinylated DNA probes in parentage testing: Non-isotopic labeling and non-toxic extraction (1988)
    Weinheim : Wiley-Blackwell
    Electrophoresis 9 (1988), S. 359-368 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: With a few exceptions DNA probing techniques require the use of radioisotopes and toxic DNA extraction techniques which render the method expensive, potentially hazardous and time-consuming. Most isotopic labeling techniques use the isotope 32P and require 3-10 days to visualize bands after hybridization. An alternative approach is based on the use of non-isotopic detection methods. The available nonisotopic techniques were assessed and their practicality tested. All probes analyzed were tested on samples extracted with a non-toxic extraction procedure using 6 M NaCl as the substitute for phenol and isochloroform. By manipulating probe sizes, blocking agents, selection of membrane and detection system, it is feasible to use non-isotopic labeling and detection in routine parentage testing. Reproducible results were obtained with labeling a variety of DNA probes of various sizes, plasmids and inserts. With an absence of waste disposal costs, probes that are stable for over two years and a staining procedure which takes 3-5 h versus days the technique is well suited for a normal laboratory setting. The next key to the acceptability of DNA testing will be the commercial availability of DNA probes for widespread use.
    Additional Material: 10 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150090803
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  • 4
    Electronic Resource
    Electronic Resource
    Gene frequency distribution of the B subunit of factor XIII (F XIIIB) in Minnesota Whites, Blacks and Amerindians (1985)
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 399-401 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Serum specimens from local Whites, Blacks and Amerindians were phenotyped for the B subunit of Factor XIII. Isoelectric focusing of desialylated samples on agarose gels at a pH 4-7 gradient followed by an immunoblotting technique was used for band identification. Gene frequencies for the three common alleles in Whites, F XIII*1, F XIIIB*2 and F XIIIB*3 were 0.776, 0.088 and 0.136, respectively, and were consistent with reports on this genetic marker system in European Whites. The Blacks and Amerindians were clearly differentiated from the Whites with F XIIIB*1,2 and 3 frequencies of 0.286, 0.635 and 0.079, respectively, for Blacks and 0.500, 0.034 and 0.466 for Amerindians. Based on the available population data on Factor XIIIB, this genetic system should prove to be a valuable marker for anthropological genetics and parentage testing.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150060809
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  • 5
    Electronic Resource
    Electronic Resource
    Transferrin and plasminogen phenotyping of aging blood stains using isoelectric focusing and silver stain (1985)
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 90-93 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A three-month study was conducted on blood stains to determine the reliability and viability of the transferrin (Tf) and plasminogen (PLG) genetic markers upon storage under varying conditions. Stains stored at room temperature (19-22 °C), 4 °C and -70 °C were tested at weekly intervals. Isoelectric focusing on agarose gels was followed by fixation and staining with Coomassie Brilliant Blue R-250. Gels were subsequently treated with silver stain if band intensity was too faint for phenotyping. Stains stored at room temperature and 4 °C showed a gradual decrease in band intensity. By using silver stain, it was possible to phneotype blood stains stored for at least twice the length of time as compared to those stained with Coomassie Blue. This study demonstrates that isoelectric focusing and silver staining is reliable for determining Tf and PLG markers in forensic investigations.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150060207
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  • 6
    Electronic Resource
    Electronic Resource
    Properdin factor B (Bf) distribution in North and Central American populations (1981)
    Weinheim : Wiley-Blackwell
    Electrophoresis 2 (1981), S. 320-323 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A study of properdin factor B (Bf) in 22 North and Central American populations demonstrated gene clusters delineating Old World Caucasian, New World and Black Ancestoral groups. Result of gene frequencies were comparable to data in the literature of European Caucasian and African Blacks. New World indigeneous population were clearly separated from other racial groups with hybrid population clustering in between the major genetic contributions.
    Additional Material: 2 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150020512
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  • 7
    Electronic Resource
    Electronic Resource
    Transferrin (Tf) subtypes in US Amerindians, Whites and Blacks using thin-layer agarose gels: Report on a new variant TfC8 (1982)
    Weinheim : Wiley-Blackwell
    Electrophoresis 3 (1982), S. 162-164 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Using 0.5 mm agarose gels for Tf subtyping by isoelectric focusing we were able to identify ten variants at the Tf locus in US Amerindians, Whites and Blacks. The variant TfC4 was found only in the Amerindian populations. This population specific variant had gene frequncies ranging from 0.079 in the Walapi to 0.179 in the Apache. The Black sample studied had a lower incidence of TfC3 than in whites and was the only population with the marker TfD1. Thin agarose gels provided good separation of TfC3 from TfC1. Thin agarose gels provided good separation of TfC3 from TfC1 enabling us to demonstrate a new variant, TfC8, migrating between TfC1 and TfC3.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150030309
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  • 8
    Electronic Resource
    Electronic Resource
    FXIIIA Phenotyping by isoelectric focusing and immunoblotting: Gene frequencies in a population of US Whites and Blacks (1985)
    Weinheim : Wiley-Blackwell
    Electrophoresis 6 (1985), S. 521-523 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: Plasma specimens were phenotyped for FXIIIA in local Whites and Blacks using agarose isoelectric focusing in the range pH 4-7, followed by immunoblotting. Gene frequencies for the alleles FXIIIA*1, FXIIIA*2 and FXIIIA*4 in Whites were 0.774, 0.225 and 0.001, respectively. In Blacks FXIIIA*1 was observed at a frequency of 0.771 and FXIIIA*2 at 0.229. The rare phenotype FXIIIA 3-1, which by conventional electrophoresis demonstrates two bands, was found to clearly show three bands using isoelectric focusing and immunoblotting. The technique used in this study is inexpensive, easy to perform and can differentiate the gene products of all the currently known variants of FXIIIA.
    Additional Material: 1 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150061011
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  • 9
    Electronic Resource
    Electronic Resource
    Comparison of rare PGM1 variants by isoelectric focusing and conventional electrophoresis: Identification of five new variants (1981)
    Weinheim : Wiley-Blackwell
    Electrophoresis 2 (1981), S. 323-326 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A comparison of specimens tested in our laboratory for the red cell enzymes sub-system PGM1 by isoelectic focusing and conventional electrophoriesis demonstrated 11 rare allotypes. In addition to the allotypes PGM13, PGM16, PGM16 MAL, 16 KADAR, 17 and 18, we indengified five new variants: 1a5, 1a6, 1a7, 1a8, 1a9. Of the five new variants only 1a6 and 1a9 could be separated from common PGM1 variants on conventional elehoresis. Because of the unique banding patterns for the variants observed, it was found necessary to use both isoelectic focusing and conventional methods for proper identification. Due to the presence for several different momeclatures for naming PGM variants, it is felt that a universal nomenclature should be defined to deal with PGM variants using both methods of analysis.
    Additional Material: 7 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150020513
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  • 10
    Electronic Resource
    Electronic Resource
    Routine phenotyping of phosphoglucomutase (PGM1) by thin-layer focusing: Isoelectric points of 14 different variants (1982)
    Weinheim : Wiley-Blackwell
    Electrophoresis 3 (1982), S. 165-168 
    Details
    ISSN: 0173-0835
    Keywords: Chemistry ; Biochemistry and Biotechnology
    Source: Wiley InterScience Backfile Collection 1832-2000
    Topics: Biology , Chemistry and Pharmacology
    Notes: A routine technique for phenotyping the PGM1 marker system was developed using thin-layer isoelectric focusing on agarose. Resolution of PGM1 variants by thin-layer isoelectric focusing on 0.5 mm gels was found to be superior to isoelectric focusing on ≥ 1 mm gels. Isoelectric points were determined for 14 different PGM1 variants and should serve as a useful tool for comparing variants between laboratories.
    Additional Material: 8 Ill.
    Type of Medium: Electronic Resource
    URL: http://dx.doi.org/10.1002/elps.1150030310
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