The PGM1 polymorphism as revealed by ultrathin-layer isoelectric focusing in the population of Padua

The occurrence of PGM₁ phenotypes in 589 samples from the population of Padua was investigated by ultrathin-layer isoelectric focusing. All ten phenotypes were observed. Frequencies of the PGM₁ alleles (1+ = 0.6180; 1? = 0.1163; 2+ = 0.2122; 2? = 0.0535) have been compared to those found in other populations.     

Transferrin subtyping of bloodstains and semen using isoelectric focusing and immunoblotting

Transferrin (TF) subtyping was carried out on bloodstains that had been made on cotton sheeting and stored under a variety of conditions ranging from −20°C to +37°C. The time limit of detection was longer than 54 weeks after dry storage under each condition. Moreover the correlation between isoprotein types of the TF in blood and semen samples from the same individual was determined in 103 men. All three TF common types and two rare types in all semen samples correlated with the type found in the corresponding blood sample. A combination of isoelectric focusing separation and immuno-enzyme-linked detection may prove to be very useful for forensic TF subtyping.     

The esterase D polymorphism as revealed by isoelectric focusing in ultra-thin polyacrylamide gels

The polymorphism of human red cell esterase D (EsD) was studied using isoelectric focusing (pH 4-6) in ultra-thin polyacrylamide gels. Typing was possible without the EsD isozymes attaining true equilibrium focusing conditions. Using this single method, six phenotypes (EsD 1, 2-1, 2, 5-1, 5-2 and 5) could be recognized in the White population of south-east England. Family studies showed these to be controlled by three co-dominant alleles and the gene frequencies were calculated to be EsD1 0.8856; EsD2 0.0946 and EsD5 0.0198. For successful and reliable EsD typing by this method, the electrophoretic system must be carefully optimized with respect to the duration of electrophoresis and the temperature attained in the gel during the electrophoretic run.     

Gc subtypes determined by ultrathin-layer isoelectric focusing. Distribution in the Veneto population (Italy)

The distribution of Gc phenotypes in the population of Veneto was investigated by ultrathin-layer isoelectric focusing. In our sample (n = 732) the six common phenotypes, Gc 1S, 1F, 1S1F, 2, 2-1S, 2-1F and a further phenotype, GC 1S1C3, were observed and the following frequencies calculated: Gc 1S = 0.560792; GC 1F = 0.159153; Gc2 = 0.277323; Gc 1C3 = 0.002732. Our gene frequencies have been compared with those found in other populations.     

The polymorphism of plasminogen (PLG) by ultrathin-layer isoelectric focusing. Distribution in the Veneto population (Italy)

The distribution of plasminogen phenotypes in the population of Veneto was investigated by ultrathin-layer isoelectric focusing. In our sample (n = 1325), the three common phenotypes PLG1, PLG2, PLG2-1 and two further phenotypes PLG1-V and PLG2-V were, observed and the following frequencies calculated: PLG1 = 0.84038; PLG2 = 0.15811; PLGV = 0.00151. These gene frequencies are compared to those found in other populations. Analysis of 41 mother-child pairs was in agreement with an autosomal codominant inheritance.     

A method to increase the volume of sample applied to isoelectric focusing gels

Isoelectric focusing of extracts from diluted or aged bloodstains may be more successfully accomplished with larger sample volumes applied to the gel. A technique is described using teflon tubing to apply larger sample volumes (up to 100 μl) to isoelectric focusing (IEF) gels. This method is reproducible and easy to perform.     

Esterase D phenotyping of bloodstains and hair roots by low voltage isoelectric focusing

A new isoelectric focusing method is described for phenotyping of esterase D in blood stains and hair roots. It permitted easy and rapid discrimination of six phenotypes determined by ESD*1, ESD*2 and ESD*7. Experiments showed it to be practicable in forensic stain work. In addition, this technique was also usable in phenotyping of ESD 5.     

The use of isoelectric focusing (IEF) as a method for the combined phenotyping of erythrocyte acid phosphatase (EAP) and esterase D (EsD)

The simultaneous isoelectric focusing (IEF) in polyacrylamide gels (PAG) of erythrocyte acid phosphatase (EAP) and esterase D (EsD) allows the poor discriminating power (DP) of EsD to be usefully combined with a highly discriminating system EAP, such that a joint DP of 0.766 was achieved compared with PGM IEF DP 0.756. Focusing was carried out in a centrally flattened gradient containing ampholines (pH 4-6 and 6-8) and the chemical spacer 3-(N-morpholino) propanesulphinic acid (MOPS). It enabled the identification of six EsD phenotypes including the recently discovered EsD5 isozymes. The application of this method to casework bloodstains is discussed.     

Gc subtypes: demonstration using isoelectric focusing. Study of population and formal genetics

Gc-subtypes were determined by isoelectric focusing and immunfixation on samples from 492 unrelated blood donors from Berlin. The frequency of the three genes was found to be GcIF = 0.1270, GcIS = 0.6006, Gc2 = 0.2724. Analysis of 78 parents with 190 children did not show deviations from the expected mode of inheritance. Investigation of the adults from paternity cases and of their children on the other hand obtained similar results. No rare alleles were observed.     

Haptoglobin subtyping by isoelectric focusing in ultrathin-layer polyacrylamide gels. Population genetic data for Hanover and Lower Saxony

This report describes a method for subtyping haptoglobin by means of isoelectric focusing in 0.2-mm ultrathin-layer polyacrylamide gels. Haptoglobin (Hp) is purified by ion-exchange chromatography and reduced. The well-known advantages of ultrathin-layer gels combine high isoelectrophoretic resolution of the Hp subtypes with less demands for time and material and make sequential visualization by fixation and protein staining possible. The distribution of the Hp subtypes in 1500 unrelated adults from Hanover and Lower Saxony is presented. Allelic frequencies are calculated to be: Hp*2FF = 0.0030; *2FS = 0.5620; *2SS = 0.0290; *1F = 0.1537; *1S = 0.2523. Segregation analysis for 68 matings shows an autosomal codominant mode of transmission in all cases. For the population investigated the chance of isolated paternity exclusion with the subtyped Hp system amounts to 33.91%.     

Genetic study of red cell esterase D polymorphism by ultrathin layer isoelectric focusing. Distribution in the Veneto population (Italy)

Human red cell Esterase D (EsD) was analyzed by isoelectric focusing (IEF) on ultrathin-layer polyacrylamide gel with a pH range of 5.0-6.0. Hemolysates were treated with Dithiothreitol to avoid loss of activity and change of the isozyme patterns by in vitro storage effects. In our sample of 951 unrelated persons from Veneto, seven different phenotypes were observed. The following allele frequencies were calculated: EsD1 = 0.8476, EsD2 = 0.1336, EsD5 = 0.0178, and EsDV = 0.0010.     

Simultaneous diagnosis of coagulation factor XIIA (F13A) and plasminogen (PLG) phenotypes by polyacrylamide gel isoelectric focusing.

In this paper, we report a simple rapid method for simultaneous determination of Coagulation Factor XIIIA (F13A) and plasminogen (PLG) phenotypes by PAGIF with a nominal pH range of 3.5 to 10, followed by immunofixation and silver stain. Critical considerations concerning the conditions of molecular separation and detection strategies are also presented.     

The establishment of the presence and group classification of sperm in mixed stains by isoelectric focusing

The article deals with experimental data on design of the method used for detection of sperm, blood and vaginal secretions according to isoelectric points of acid phosphatases, differential diagnosis of ABO group antigens in "mixed" stains as well as detection of phenotype of red cell acid phosphatase by isoelectric focusing method.     

中国成都汉族及泰国群体D7S2846基因座的遗传多态性

研究STR基因座D7S2 846的遗传多态性 ,为法科学应用提供基础数据。应用PCR及PAG电泳技术 ,对376名中国成都汉族无关个体及 131名泰国无关个体进行了调查。两群体分别检出 8个和 7个等位基因 ,首次获得该基因座基因在两群体中的频率分布。两群体基因型频率分布均符合Hardy Weinberg平衡。家系调查证实了等位基因的传递遵循孟德尔遗传规律。该基因座在两群体中的个人识别能力 (Dp)分别为 0 85 70、 0 86 0 2 ,杂合度 (H )分别为0 6 915、 0 6 870 ,多态性信息含量 (PIC)分别为 0 6 445、 0 6 5 5 3 ,非父排除率 (PE )分别为 0 415 2、 0 40 85。D7S2 846基因座在法医学个人识别及亲子鉴定中具有较高的实用价值。     

中国汉族群体人类补体C8A多态性

采用免疫沉淀、SDS-聚丙烯酰胺凝胶电泳 (SDS- PAGE)、被动转印及酶免分析 ,研究了人类补体 C8A等位基因频率在成都地区汉族群体中的分布。 12 1份样本被分为 3种常见型 ,即 C8A- A、C8A- B及 C8A- AB,由两个等位共显性基因 C8A * A及 C8A* B控制 ;同时发现了 2个稀有亚型 ,即 A3亚型及新发现的 Ax亚型。等位基因频率为 C8A* A=0 .5 0 83,C8A* B=0 .4835 ,C8A*稀有型 =0 ,0 0 83。说明 C8A多态性在中国群体中具有良好的分布 ,个人识别率(DP)达到 6 1.14% ,可用于法医学个人识别及亲子鉴定     

云南汉族CRHBP基因多态性与暴力攻击行为的相关性

目的探讨云南汉族人群促肾上腺皮质激素释放激素结合蛋白(corticotropin releasing hormone-binding protein, CRHBP)基因多态性与暴力攻击行为的相关性。方法对云南汉族111例有攻击行为的服刑人员(包含53例抢劫,58例故意伤害)和189例健康对照样本采用改良的多重高温连接酶检测反应技术(improve Multiplex ligase detection reaction, iMLDR)检测CRHBP基因的4个Tag SNPs(rs10062367, rs32897, rs7718461, rs7721799)的基因型,应用SPSS 20.0和SHEsis软件对结果进行统计分析。结果 rs32897、rs7718461、rs7721799的等位基因和基因型分布在暴力组、抢劫亚组、故意伤害亚组和对照组中均无显著差异(P>0.05),rs10062367等位基因和基因型分布在暴力组、抢劫亚组和对照组中也无显著差异(P>0.05),但在故意伤害亚组与对照组中具有显著差异(P<0.05)。单倍型ATGA可使暴力的相对风险显著增高(P<0.05),单倍型GCAA可使指向他人暴力的相对风险显著增高(P<0.05)。结论 CRHBP基因rs10062367位点多态性可能与云南汉族人群针对他人的攻击行为有关,单倍型ATGA是暴力行为的风险因子,个体携带单倍型GCAA会使指向他人的躯体攻击行为风险增加。     

Orosomucoid (ORM) typing by isoelectric focusing: description of two new alleles in a German population and thermostability in bloodstains

The genetic polymorphism of serum orosomucoid (ORM) was studied in 168 unrelated German individuals using isoelectric focusing followed by immunoprinting. Two new alleles, tentatively designated ORM1*14 and ORM2*13, were identified. The method was successfully applied to demonstrate ORM1 types in dried bloodstains. Each type of ORM1 was also correctly determined in bloodstains heated at 130 degrees C for 30 min. The results indicated that ORM1 is a new powerful genetic marker system for the grouping of bloodstains.     

Polymorphism of EsD by isoelectric focusing: description of the new allele EsD*Kofu and phenotyping in bloodstains

The polymorphism of EsD was investigated in 1115 unrelated Japanese individuals by isoelectric focusing. Besides the three common phenotypes two heterozygotes EsD 7-1 and EsD 7-2 were observed. The gene frequencies were: EsD*1 = 0.6234, EsD*2 = 0.3663, and EsD*7 = 0.0103. In addition, a rare variant was detected in a probandus living in the city of Kofu. The family analysis suggested the hereditary occurrence of a new allele EsD*Kofu. The isoelectric focusing method was successfully applied to phenotyping EsD in bloodstains; each phenotype was demonstrated at 37 degrees C for up to 2 weeks, at room temperature for up to 9 weeks, and at 4 degrees C for over 20 weeks after stain formation.