Double replica electroblotting for the simultaneous phenotyping of C6 and C7, and its application to the examination of bloodstains

A new reliable and reproducible technique for the simultaneous determination of C6 and C7 types is presented, which employs double replica electroblotting after isoelectric focusing. It permitted clear discrimination of both C6 and C7 components, and the patterns were nearly comparable to those demonstrated separately. The population data obtained by this new technique fitted the genetic hypothesis. The present double replica electroblotting method was successfully applied to the combined phenotyping of C6 and C7 from bloodstains which were stored at room temperature for up to 4 weeks. The method is quite suitable for medicolegal examination of bloodstains particularly for the saving in the amount of sample.     

Direct fluorometric determination of cyanide in human materials

A detailed procedure of a new simple fluorometric assay for cyanide in human blood, stomach contents and urine is presented. This method requires neither diffusion nor distillation of cyanide from samples; small amounts of the samples were directly added to incubation mixtures, which were then heated at 80 °C for 45 min and acidified for fluorescence measurements. Our method is much more rapid than the previous ones. Some data are presented on specificity and sensitivity of the present method.     

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.     

A stability study on Gc subtyping in bloodstains: comparison by two different techniques

The limits of determination of Gc subtypes in bloodstains were compared between the immunofixation method and the sulfosalicylic acid precipitation method using isoelectric focusing on polyacrylamide gel. By the immunofixation method Gc subtyping in bloodstains was successfully made at 37 degrees C after 7 weeks, at room temperature after 17 weeks and at 4 degrees C even after 25 weeks storage. By the sulfosalicylic method Gc subtypes were no longer able to be determined a few weeks after stain formation. The superiority of the results obtained by the immunofixation method makes it the recommended method for the Gc subtyping from bloodstains in medicolegal practice.     

A practical method for the accurate determination of methemoglobin in blood containing carboxyhemoglobin

Kinetics of the oxidation of carboxyhemoglobin (HbCO) by potassium ferricyanide was studied photometrically in a weakly acid solution. An increase in the absorbance at 630 nm reached a maximum within 10 min when over a 100-fold excess of ferricyanide to hemoglobin iron was used. A slight decrease in the absorbance was observed after completion of the reaction when over a 500-fold excess of the reagent was used. In the presence of 0.4% Sterox SE, the absorbance began to decrease without complete oxidation. From these findings, a simple, rapid and accurate method for the determination of methemoglobin (Met-Hb) in blood was devised. The method was compared with two other methods, using 11 blood samples containing various amounts of HbCO, and proved to be suitable for blood containing elevated HbCO as well as for ordinary blood.     

Factor B subtyping in sera and bloodstains by isoelectric focusing and immunoblotting

The polymorphism of BF was investigated in 765 unrelated Japanese individuals by isoelectric focusing and immunoblotting. Besides five common subtypes three rare variants were observed. The allele frequencies were: BF*S = 0.8078, BF*FA = 0.1797, BF*FB = 0.0105, BF*Var. = 0.0020. The above method was successfully applied to subtyping BF in stored bloodstains. The determination limits were: at 4 degrees C 8 weeks, at room temperature 2 weeks and at 37 degrees C only 2 days after storage. The BF subtyping is of practical use in medicolegal individualization of unknown bloodstains.     

Identification of fetal bloodstains by enzyme-linked immunosorbent assay for human alpha-fetoprotein

A rapid and highly sensitive enzyme-linked immunosorbent assay (ELISA) for determination of human alpha-fetoprotein (AFP) using commercially available reagents was devised and applied to identification of fetal bloodstains. When experimentally prepared bloodstains, 1 by 2 mm in area, were submitted to analysis, only fetal bloodstains showed positive reactions in the present ELISA. The reactions did not change significantly when these bloodstains were stored at room temperature for one week. The present ELISA seems to be suitable for forensic science practice.     

The World Development Report 2008: inconsistencies,silences, and the myth of ‘win-win’ scenarios

The World Development Report 2008 (WDR-2008) on agriculture and development has been received with much expectation and controversy. This paper welcomes some aspects of the WDR-2008 that help us reinvigorate some debates on agricultural development, so far marginalised in international development policy agendas. The paper, however, focuses on some critical problems in the report and the World Bank's stance on agriculture. First, there are tensions between advocacy and research and between the World Bank's rhetoric and operational realities. Secondly, the report suffers from the usual adherence to superficial win-win scenarios that mask conflict of interest and power relations. Thirdly, the WDR-2008 is caught in a tension between neo-populist pro-small farmer views and ‘modernist’ pro-agribusiness stances. Fourthly, the analysis of agricultural development in isolation from broader development processes and especially without a systematic analysis of industrialisation and agriculture–industry relations seriously limits the analytical and empirical value of the report.     

C6 phenotyping in sera and bloodstains by isoelectric focusing followed by electroimmunoblotting

The genetic polymorphism of C6 was investigated in 329 unrelated Japanese individuals using isoelectric focusing in polyacrylamide gels followed by an electroimmunoblotting technique. Besides six common phenotypes C6 A, AB, B, AB2, BB2 and B2, six rare variants were observed. The allele frequencies were: C6*A = 0.4422, C6*B = 0.4757, C6*B2 = 0.0714, C6*A3 = 0.0015, C6*M1 = 0.0046 and C6*B3 = 0.0046. The population data confirmed that the C6*B2 allele is the third common allele characterizing Japanese. The present electroimmunoblotting technique was applied to demonstrate C6 types in dried bloodstains. The C6 types were determined from bloodstains stored at 4 degrees C for up to 10 weeks, at room temperature for up to 2 weeks and at 37 degrees C for up to 4 days. The results show that this component system offers a new powerful means for the medico-legal grouping of bloodstains.