Genetic polymorphism of human C1R subcomponent of the first complement component in the Japanese population

Genetic polymorphism of the C1R subcomponent of human complement component C1 has been investigated in neuraminidase treated EDTA plasma samples of 440 healthy Japanese individuals living in Tokyo by means of thin-layer polyacrylamide gel isoelectric focusing (PAGIEF) at pH 3.5-9.5 in the presence of 8.0 M urea followed by an electroblotting with enzyme immunoassay. Three common and three rare alleles were detected in the Japanese population. Of these, two common alleles were identical to C1R*1 and C1R*2 and other new alleles were tentatively designated C1R*3, C1R*4, C1R*5 and C1R*6, respectively. The results of the family studies suggested that the genetic model for C1R polymorphism assumed autosomal codominant Mendelian inheritance. The allele frequencies were estimated as C1R*1 = 0.4216, C1R*2 = 0.3602, C1R*3 = 0.2068, C1R*4 = 0.0091 and C1R*R(C1R*5 and C1R*6) = 0.0023, respectively. The distribution of allotypes fitted the Hardy-Weinberg equilibrium. The C1R system provides a useful genetic marker for human genetics, anthropologic studies and forensic science.     

An agarose gel electrophoretic method for typing phosphoglucomutase-1, esterase D, or glyoxalase I

A conventional agarose gel electrophoretic method was described for typing phosphoglucomutase-1, esterase D, or glyoxalase I as single systems. Bloodstain extracts were absorbed into 1-mm-thick agarose gels via an application mask. The electrode wick distance was 12 cm and electrophoresis was carried out at 400 V at 6 degrees C. The electrophoretic run times were 30 min for glyoxalase and 1 h for esterase D or phosphoglucomutase. This method is reliable and produces highly resolved band patterns. Additionally, the shorter separation times as a result of the increased voltage gradient permitted typing of more samples in a given time period compared with presently used methods. This technique requires little technical expertise and can be incorporated into the laboratory at a minimal cost.     

An improved method for post-PCR purification for mtDNA sequence analysis

Mitochondrial DNA (mtDNA) analysis of forensic samples typically is performed when the quantity and quality of DNA are insufficient for nuclear DNA analysis or when maternal relatives may be the only reference source. Many of the steps required in the analytical process are both lengthy and labor intensive. Therefore, improvements in the process that reduce labor without compromising the quality of the data are desirable. The current procedure requires purification of the amplicons by centrifugal filtration after PCR and prior to cycle sequencing. Because this method requires several manipulations to perform, alternate cleanup procedures were investigated. These include the use of 1) Qiagen QlAquick PCR Purification columns, 2) Concert Rapid PCR Purification columns, and 3) ExoSAP-IT reagent. When the yield of purified amplicons, quality of the sequence profile, and ease of assay were evaluated, the use of ExoSAP-IT reagent for post-amplification purification was chosen to replace the filtration method.     

An improved means of enzyme typing of hair roots using isoelectric focusing.

An improved method of grouping hair, based on the alleles of PGM observed by isoelectric focusing, has been described. The increased discriminating power of this system (0.77) compared to that obtained by the starch gel technique (0.55) provides a new and more sensitive means of typing hair.     

An improved method for subtyping Pi in old bloodstains.

Immunofixation procedures were used for detecting alpha-1 antitrypsin protease inhibitor (Pi) phenotypes in bloodstains. Neuraminidase elution of bloodstains, together with isoelectric focusing, immunofixation, and silver staining techniques, makes possible Pi subtyping in old bloodstains. No extra bands appear when the storage time is no longer than three months.     

An improved method for detection of diatoms (author's transl)]

An improved method for detection of diatoms in case of drowning is reported. The sample is digested with nitric acid. In order to avoid destruction of diatoms as well as losses by centrifugation etc. the time for wet digestion was reduced and the digest solution is filtered through a membrane filter. Fatty material is removed by alternate washing the filter with 2-Propanol and Petroleum ether. Following the wet digestion of the filter, aliqutos of this digest solution are filtered and the filter is examined microscopically. As the recommended procedure is less time consuming and yields almost complete recovery of diatoms the method has great probative value in case of death by drowning.     

An improved method for restoring the initial appearance of injuries on the skin

Presents a modified method for restoring the shape and size of wounds on skin preparations. The advantages of the modification over the routine A. N. Ratnevski?'s method are enumerated.     

An improved method for semen alpha-L-fucosidase typing--distribution in the Wuhan population of China

An improved method for the separation of the genetic variants of alpha-L-fucosidase in human semen is described. The method involves, isoelectric focusing in ultrathin-layer PAG containing a mixture of ampholines, pH 5-7 and pH 3.5-9.5, and separator HEPES. The Fu pattern obtained is simple, easy to interpret, and reproducible. The occurrence of fucosidase phenotypes in 189 semen samples from the population of Wuhan was investigated, and the frequencies of the Fu alleles were calculated.     

Sperm elution: An improved two phase recovery method for sexual assault samples

This report describes the validation of a two phase cell recovery technique for the elution of two common cell types, epithelia and spermatozoa, from frequently examined items submitted as part of sexual assault casework. Furthermore, separation of cell types prior to microscopic examination of cell pellets improves the scientist's confidence in observing and scoring spermatozoa that may be present. During the validation, Orchid Cellmark's Sperm Elution© method consistently recovered a greater number of spermatozoa from simulated sexual assault items and swabs taken following consensual sexual intercourse compared to a water extraction technique. On average the Sperm Elution method recovered over twice the number of spermatozoa compared to the water method. The ability to separate the cell types present allows a rapid microscope slide search for spermatozoa and faster DNA extraction protocol in comparison to Cellmark's previous preferential method.     

A microplate method for reverse ABO typing of bloodstains

A sensitive and reliable hemagglutination assay, using V-bottom microplates, is described for the detection of the ABO blood group alloantibodies in bloodstained material. When used in conjunction with an absorption-elution procedure, the microplate assay resulted in a 300% increase in the number of conclusive grouping results when compared to the Lattes crust test. The use of the microplate reverse grouping assay permits 24 specimens to be assayed conveniently on a single plate and eliminates the tedious and time-consuming microscopic examination required for the Lattes crust test.     

An improved non-destructive method for detection of latent fingerprints on documents with iodine-7,8-benzoflavone

A solution of iodine and 7,8 benzoflavone¹ is an extremely sensitive reagent for developing old latent fingerprints on porous surfaces. The new reagent solution was compared with conventional ones, e.g. iodine, iodine fixed with 7,8-benzoflavone and iodine fixed with tetrabase, and was found to be superior with regard to sensitivity, clarity and reduced background intensity. The developed blue fingerprint stains are cleared in a fairly short time by air oxidation at room temperature without altering the chemical composition of the fingerprints. This leaves open the possibility of further examination of the latent fingerprints by alternative techniques should a double check be desired or in the event of failure.     

基于等位基因特异性PCR原理建立的SNP分型新方法

目的建立一种新方法,对多个单核苷酸多态性(singlenucleotidepolymorphism,SNP)位点进行分型。方法基于等位基因特异性PCR原理,采用荧光标记复合扩增和毛细管电泳技术,根据PCR片段长度差异进行分型。选择SNP位点共11个,每个SNP位点设计两条长度不同、3’末端分别与SNP两个等位基因碱基配对的上游引物,同时为了增加特异性,在两条等位基因上游引物的3’末端第3或第4位碱基人为引入错配。在距离上游引物100～300bp范围内的合适位置,设计下游共用引物,并进行荧光标记。所有位点经过复合扩增后,PCR产物经ABIPrismTM310型遗传分析仪电泳分离,确定每个SNP的基因型。结果每个SNP位点纯合子为单一产物峰,杂合子则为长度不同的两个产物峰。不同的SNP位点扩增产物长度不同,根据产物长度和产物峰的数量进行SNP分型,一次完成11个SNP位点分型,其结果与直接测序完全一致。结论荧光标记复合扩增片段长度差异等位基因特异性PCR法是一种简单快速而有效的SNP分型新方法。     

An improved method for the histological preparation of single hairs and dust samples--morphological and immunological examination

An improved method of the serological and morphological investigation of human hair is reported. The hair was firmly fixed onto a microscopic slide with cellophane tape and observed microscopically to confirm the cuticula images and the presence of the medulla. A piece of the hair containing the medulla was dissected, embedded in paraffin, and a cross section of this hair was prepared. By treating the sample with immunohistochemistry (biotin-antibiotin ABC technique), the blood type of the hair was confirmed definitively. Dust containing shaved beard can be examined in the same way.     

An improved method for the determination of Y chromosomes in blood stains (author's transl)]

A new filter combination for fluorescence microscopy using SWP 440 interfernece filter for excitation and GG 455 as the barrier filter is described. In blind trials examining blood smears of 5 male persons and by examination of 3 weeks old blood stains a better Y chromosome demonstration has been obtained using this new technique in comparison with the "usual" filter combination: BG 12-530. In blind trials of up to 6 weeks old blood stains of one male and one female a reliable sex determination was made using the new technique.