Differentiation of mite species by using cellulose acetate and equilibrium polyacrylamide gel electrophoresis of isoenzymes

The isoenzymes of various species of medically and economically important mites were studied using cellulose acetate and equilibrium polyacrylamide gel electrophoresis. Interspecific differences in isoenzymes were found, which were species-specific. Intraspecific differences in isoenzymes were also found when individual mites were examined. The efficiency of each technique, and their use in various fields of acarology, are discussed. In addition, possible phylogenetic relationships as revealed by these techniques are suggested.     

Additional lactate dehydrogenase (LDH) isoenzymes in normal testis and spermatozoa of adult man

Summary Adult human testicular tissue contains up to six previously undescribed lactate dehydrogenase (LDH) isoenzymes in addition to the five LDH isoenzymes normally found and the sixth found in spermatogenic cells and spermatozoa, LDH-X. Additional LDH isoenzymes were also found in spermatozoa but not in seminal fluid or in serum. After electrophoresis one additional LDH isoenzyme of testicular tissue was localized between LDH-1 and LDH-2, two between LDH-2 and LDH-3, two between LDH-3 and LDH-4, and two between LDH-4 and LDH-5. These localizations indicate that the additional LDH isoenzymes are tetramers combining the A and B subunits of the five normal LDH isoenzymes and the C subunit of LDH-X. The additional LDH isoenzymes may be important in the metabolism of spermatogenic germ cells and spermatozoa.     

Esterase-D polymorphism in Assam by cellulose acetate electrophoresis

Summary With the help of a simplifed and quick method, cellulose acetate electrophoresis, the phenotypes of esterase D were determined in an Assamese population. The gene frequencies of Es D¹ were 0.7263 and 0.2737 for Es D².

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Zusammenfassung In einer Stichprobe aus Assam wurde mit Hilfe einer einfachen und schnellen Methode, der Cellulose-Acetat-Elektrophorese, die Bestimmung der Esterase D-Phänotypen durchgeführt. Die Genfrequenzen wurden für Es D¹ zu 0,.7263 und für Es D² zu 0.2737 bestimmt.

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Supported by the Deutsche Forschungsgemeinschaft, the Stiftung VW and the Fonds der Chemischen Industries.     

Two-dimensional electrophoresis on the layers of cellulose acetate membrane

A new type of two-dimensional electrophoresis for analysis of protein using cellulose acetate membrane has been developed. Prior to the separation, proteins in a sample are concentrated to a narrow zone on a strip of cellulose acetate according to “steady-state stacking” of isotachophoresis. Electroendosmotic counterflow on cellulose acetate membranes is advantageous for the isotachophoretic concentration of large sample volumes. The concentrated protein zone is then subjected to electrophoretic separation on the same strip. This first-dimensional separation including the concentrating process is named “concentrating electrophoresis.” Iso-electric focusing on several layers of cellulose acetate membrane is performed in the second-dimensional step. Many kinds of detection methods can be applied to the layers among which proteins are distributed. The novel two-dimensional electrophoresis takes only 5 h to perform.     

Beta-agonists enhance the lactic dehydrogenase (LDH) expression in serum and ventricular myocytes of mice

Beta-agonists have skeletal muscle specific protein anabolic effects and are also known to cause cardiac hypertrophy. Changed total LDH and its isozymic patterns are conveniently employed for the detection of different pathophysiological states of the tissues. The purpose of this study is to confirm total LDH and its isozymic expression in ventricular tissue and serum in mice following oral administration of single but higher dose of isoproterenol (Iso) and clenbuterol (Cl) (100 mg/kg body wt. and 20 mg/kg body wt., respectively), after 4, 8 and 20 hours of drug administration. Mice heart witnessed increased total LDH levels with time. Serum on the other hand showed decline in total LDH concentrations at the initial points of the drug treatment. No doubt, total LDH expression increased towards 20th h post-drug treatment but this increase is mainly due to anaerobic isozymes, i.e. LDH4 and LDH5. The findings of the present study suggest that tissue damage is definitely caused by two beta-agonists after giving single dose for shorter time span (20 hours) and the impact of the damage varies from drug to drug. Increase in total LDH in serum is not due to release from heart but from some other tissues having anaerobic metabolism.     

Esterase-D polymorphism in Assam by cellulose acetate electrophoresis.

With the help of a simplified and quick method, cellulose acetate electrophoresis, the phenotypes of esterase D were determined in an Assamese population. The gene frequencies of Es D(1) were 0.7263 and 0.2737 for Es D(2).     

Production of monoclonal antibodies to lactate dehydrogenase (LDH) isoenzymes for immunohistochemical study on fixed tissue section

Summary Purified lactate dehydrogenase (LDH) isoenzyme 1 (H or B subunits) and isoenzyme 5 (M or A subunits) were used to prepare monoclonal antibodies (MAb) suitable for immunohistochemical detection on formalin fixed paraffin-embedded tissue sections. In the initial fusions, screening of the antibodies was based on enzyme linked immunosorbent assay (ELISA) against the immunogens. None of the antibodies obtained was satisfactory. There were various problems related to specificity, crossreactivity, affinity and also the properties of the monoclonal antibody itself. Using a combined system involving more than one method for screening, two suitable monoclonal antibodies, MAb65 (to H-type LDH) and MAb25 (to M-type LDH) were selected. Both antibodies reacted specifically with corresponding LDH isoenzymes as shown in a series of tests. Their reactivity in sections of formalin fixed paraffin-embedded tissue indicated that both antibodies are suitable reagents for immunohistochemical studies.     

血清蛋白醋酸纤维薄膜电泳实验的改进

<血清蛋白的醋酸纤维薄膜电泳>实验采用宽膜条很好的解释了由窄膜条造成的区带弯曲问题.实验用到的一些试剂较昂贵.在保证和提高生物化学实验教学质量的前提下.以降低实验成本、减少对环境污染角度出发,采用牛血清代替人血清、改变实验条件、回收缓冲液和漂洗液等方法对实验进行了改进,改进的结果是实验稳定,重复性好,用于实验课教学可以达到预期的实验目的和要求.