[The identification of esterase D isoenzyme (EsD) phenotypes in the dental pulp by isoelectric focusing].
An identification method based on the isoelectrofocusing (IEF) assay of the EsD phenotypes in dental cavity pulp is reported. The IEF has been carried out on polyacrylamide gel (T = 5% and C = 3%), using ampholines ranging pH 4.5-5.4. A pre-focusing of 20 min at 2,000 V, 100 mA and 8 W was used; focusing conditions were: 2,000 V, unlimited current and W, for 75 min at 4 degrees C. Electropherograms were read under UV at 366 nm after reaction with 4-methylumbelliferyl acetate giving a fluorescent product. The content of a single dental cavity has been found to be the smallest amount of sample needed for the assay in alive subjects, excluding dental pathology which could interfered with the results. In bodies, excluding interferences from abnormal preservation conditions, a clear decrease in the reliability of the assay has been observed after the 6th day since the death. In any case, on the basis of the authors' experience, the dental pulp, as compared to blood, seems a preferable sample for identification purposes.
Istituto di Medicina Legale, Università di Verona.
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De Leo
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Tagliaro
D043182:Carboxylesterase; D002265:Carboxylic Ester Hydrolases / Q000032:analysis*; D003643:Death; D003782:Dental Pulp / Q000201:enzymology*; D004588:Electrophoresis, Cellulose Acetate; D005069:Evaluation Studies as Topic; D005553:Forensic Dentistry; D006801:Humans; D007525:Isoelectric Focusing / Q000379:methods; D007527:Isoenzymes / Q000032:analysis*; D010641:Phenotype; D013997:Time Factors
D007527:Isoenzymes; D002265:Carboxylic Ester Hydrolases; D043182:Carboxylesterase; C020118:ESD protein, human
Identificazione dei fenotipi dell'isoenzima esterasi D (EsD) nella polpa dentaria mediante isoelettrofocalizzazione.