Identification of fetal hemoglobin in blood stains by high performance liquid chromatography

A new method for the identification of fetal hemoglobin (Hb F) in blood stains by reverse-phase high performance liquid chromatography is described. Differentiation between fetal and adult blood stains is based on the existence of gamma-chain peaks which are characteristics of Hb F. Very few gamma chains appeared on chromatograms of all the adult blood stains examined. The level of Hb F could be determined by measuring the total of chromatogram gamma-globin chain areas, and expressing it as a percentage of total Hb. Levels in six cord blood stains on filter paper ranged from 81.1% to 91.3% and remained constant for at least 12 weeks. This method is of great value for its simplicity, sensitivity and speed, and most importantly for its reliability in the field of forensic medicine.     

Differentiation of fetal and adult bloodstains by pyrolysis-gas-liquid chromatography

A pyrolysis-gas-liquid chromatographic method has been developed for the differentiation of adult and fetal bloodstains. In a blind-coded study, five adult and three fetal bloodstains were correctly identified on the basis of the pyrograms of stain extracts. The differentiation between adult and fetal bloodstains is based on the peak height ratio of two long-retention-time peaks appearing in their pyrograms. The first of these peaks has been tentatively identified as indole derived from the pyrolysis of tryptophan, while the second peak is an as-yet unidentified molecular fragment produced by the pyrolysis of some component of the hemoglobin molecule other than the amino acids tryptophan, tyrosine, and phenylalanine.     

An efficient method to eliminate streaking in the electrophoretic analysis of haptoglobin in bloodstains

A bloodstain extraction procedure that improves the analysis of haptoglobin in dried bloodstains has been developed. The streaking of electrophoresis gels caused by deteriorated hemoglobin can be eliminated by incorporating chloroform in the bloodstain extraction procedure. The method is easier to execute than previously published techniques for eliminating the adverse effects of deteriorated hemoglobin on the analysis of haptoglobin. Bloodstains up to two years old were correctly phenotyped in haptoglobin by this method.     

A new method for typing haptoglobin in bloodstains using immobilized allo A lectin

Allo A lectin from the beetle, which is beta-D-galactose specific, reacts to haptoglobin but not to hemoglobin. The use of allo A-Sepharose for typing haptoglobin in bloodstains helped eliminate hemoglobin from the bloodstain extract and presented highly resolved haptoglobin patterns by disc gel electrophoresis. This method is simple and rapid for typing haptoglobin in bloodstains and can be easily used in forensic science laboratories.     

分光光度法检测硫化氢中毒血液中的硫化血红蛋白

目的研究生物检材中硫化氢的检验方法。方法用分光光度法测定生物检材(血液)中的硫化血红蛋白。结果硫化血红蛋白在波长612nm处有特征吸收,且其在此处的吸光度与硫化血红蛋白的浓度成正比。结论硫化氢中毒案件可通过分光光度法测定中毒者血液中硫化血红蛋白的浓度来确定中毒的程度。     

Glycated hemoglobin: a useful post-mortem reference marker in determining diabetes

Glycated hemoglobin (HbA(1c)) has been demonstrated to be a useful marker for long-term glucose control in diabetes. This parameter characterizes each non-enzymatic fixation of glucose on hemoglobin. It is a useful test in addition to periodic glycemia controls since it reflects the mean glycemia of the past 60 days. We studied the conservation of HbA(1c) at 4 degrees C as a function of time with different anti-coagulants and preservatives (3, 6 months, 1 year). A total of 106 tests were performed using the high performance liquid chromatography (HPLC) method dedicated to the semi-automatic analysis of HbA(1c) (Bio-Rad) and we applied the method in forensic cases. Conservation at 4 degrees C was good for as long as 3 months in blood samples collected with fluoride and 6 months in samples collected in a dry or in a heparinized tube. In non-diabetic subjects, HbA(1c) reference values obtained from forensic samples were identical to those of living controls (3.5-6.25% of total hemoglobin). All positive HbA(1c) results were confirmed by a medical evaluation. This method was successfully applied to five forensic cases. In cases of increased acetonemia, acetone or isopropanol are easily measured. However, in some unexplained post-mortem circumstances, increased HbA(1c) permits to differentiate alcoholic or starvation ketoacidosis from the diabetic cases. Glycated hemoglobin should, therefore, be considered the forensic marker of choice in the post-mortem diagnosis of a diabetic disorder and demonstrates its usefulness in post-mortem validation.     

Determination of the age of subdural hematomas from the concentration of hemoglobin

The photocolorimetric method was employed to measure hemoglobin levels in 46 traumatic subdural hematomas of different age. The data thus obtained were used to construct a logarithmic regression model for the determination of the age of subdural hematomas from the concentration of hemoglobin. The model allows to determine injury time points and intervals at any desired level of confidence probability. The fraction of dispersion of hematoma age values attributable to regression was estimated at 41% which accounts for 74.7% of the maximally possible magnitude. Results of the study can be used in practical work of a forensic-medical experts.     

Discrimination Between Infant and Adult Bloodstains Using Micro‐Raman Spectroscopy: A Preliminary Study

In the present study, we used micro‐Raman spectroscopy with high‐resolution analysis to discriminate between bloodstains from infants and bloodstains from adults. Raman peaks were detected at 674, 754, 976, 1002, 1105, 1127, 1176, 1248, 1340, 1368, 1390, 1560, and 1611 cm^?1; these peaks were derived from hemoglobin, albumin, and glucose. However, a peak was obtained at 1105 cm^?1, which was assigned to histidine; this peak was observed only for bloodstains from adults. Human adult hemoglobin (HbA) is composed of an α₂β₂ tetramer structure, whereas human fetal hemoglobin (HbF) is composed of an α₂γ₂. Therefore, the lack of a Raman peak at 1105 cm^?1 in bloodstains from infants indicates the possibility of two histidine substitutions (His116Ile and His143Ser) in the γ chain of HbF. This study discriminates between bloodstains from infants and bloodstains from adults using micro‐Raman spectroscopy, with beneficial implications in forensic science.     

Identification of hemoglobin derivatives by means of electrofocusing

A method for the identification and quantitation of hemoglobin derivatives is described. Samples diluted 1 in 100 were applied to precoated polyacrylamide-gel plates containing an Ampholine gradient of pH 3.5 – 9.5 and focused for 1.5 h at a constant 40 W. All hemoglobins under investigation were characterized by specific patterns which allow them to be differentiated. Electrofocusing is recommended for screening tests, but quantitative evaluation using densitometry is possible as well.     

On the commodification of the black female body: the critical implications of the alienability of fetal tissue

Recent scientific experimentation has revealed that fetal tissue yielded from abortions has remarkable therapeutic value. This Note posits that the demand for fetal tissue likely will expand to the point where the current supply no longer satisfies it. Therefore, in order to obtain tissue from women who would not otherwise donate their abortuses, should research organizations, pharmaceutical companies, and doctors be allowed to offer women a "financial incentive" for their fetal tissue? That is, should women be allowed to sell their fetal tissue? This Note explores the question from a Critical Race Theory perspective. It analyzes the impact that a market in fetal tissue will have on Black women, who are more likely to participate in such a market due to their precarious economic situation, their higher abortion rate, and the effects of internalized oppression. The Note concludes that because Black women will be disproportionately exploited, as well as disenfranchised from the benefits produced by a market in fetal tissue, fetal tissue should not be made market alienable.     

抗人血红蛋白胶体金检测试剂条的研制

目的制备法医学检验所用的确定人血的免疫胶体金层析试剂条。方法选取抗人血红蛋白单克隆细胞株,制备其小鼠腹水,从腹水中纯化出单克隆抗体。制备胶体金并用一纯化的单克隆抗体包被,制成免疫胶体金。取玻璃纤维以免疫胶体金浸泡,烘干。在一硝酸纤维素膜上两个不同位置分别点加另一抗人血红蛋白抗体和羊抗鼠IgG。搭建试剂条并检测其灵敏度和特异性。结果制成的免疫胶体金试剂条可对稀释至20万倍的人血红蛋白溶液显示阳性,对法医学检验常见8种动物的血溶液显示阴性。结论所制备抗人血红蛋白胶体金试剂条可以应用于法医学检验。     

多重置换扩增在含抑制物检材中的应用

目的研究多重置换扩增(multiple displacement amplification,MDA)在含抑制物检材中的抗抑制能力,与磁珠法纯化检材相比较,证明其在法医学中的应用及意义。方法将不同浓度血红素和腐殖酸与样本DNA进行混合,分为MDA处理组、磁珠法处理组和空白对照组,PCR-STR单基因座D3S1358扩增联合聚丙烯酰胺凝胶电泳检测,并应用Amp F詛STR誖IdentifilerTMPlus试剂盒联合毛细管电泳检测。结果血红素质量浓度大于1 ng/μL或腐殖酸质量浓度大于0.1 ng/μL时,空白对照组经单基因座STR检测不能得到扩增产物;磁珠法处理组血红素质量浓度大于100 ng/μL或腐殖酸浓度大于1 ng/μL时,不能够得到扩增产物;MDA处理组各浓度抑制物均能成功扩增,完全不受抑制物影响。结论 MDA技术可消除血红素及腐殖酸的抑制作用,其抗抑制能力优于磁珠法纯化DNA,具有一定的法医学应用意义。     

The comparative assessment of the practical value of the currently employed methods for the recognition and species specificity of the blood

The objective of the present study was to evaluate sensitivity and specificity of the HemDirect method on test-plates (Seratec) for detecting human hemoglobin (HHb). These characteristics were compared with those of other widely used methods designed for the detection of blood traces, viz. thin layer chromatography, hemotest, spectrofluorimetry, and identification of blood species specificity (by countercurrent immunoelectrophoresis in agar and on the acetate-cellulose film). It was shown that the HemDirect test is highly specific and far more sensitive than other techniques used for the same purpose in the practical work. It can be recommended as the method of choice for the detection of blood microtraces.     

Application of mRNA Expression Analysis to Human Blood Identification in Degenerated Samples that were False‐negative by Immunochromatography,,

Forensic laboratories are often faced with cases in which methamphetamine hydrochloride‐mixed blood is unable to be identified as human blood by immunochromatography against human hemoglobin A0. The application of mRNA expression analysis to samples that showed a false‐negative with immunochromatography was investigated as an alternative approach that did not depend on the antigen–antibody reaction. Real‐time PCR was used to examine the expression levels of blood markers such as glycophorin A, spectrin beta, and hemoglobin beta. Hemoglobin beta was the only marker that was specifically detected in blood, while glycophorin A was useful for determining human specificity. Hemoglobin beta showed good detection sensitivity and was detectable in 37‐year‐old blood stains. Hemoglobin beta was exclusively detectable in methamphetamine hydrochloride‐mixed blood stains. Detergents and disinfectants did not significantly influence mRNA markers. The proposed mRNA expression analysis was suitable for human blood identification as an alternative method to immunochromatography.     

A New Approach for the Carbon Monoxide (CO) Exposure Diagnosis: Measurement of Total CO in Human Blood Versus Carboxyhemoglobin (HbCO)

The aim of the study is to present the application of a headspace–gas chromatography–mass spectrometry (HS‐GC‐MS) method for the determination of the carbon monoxide (CO) blood concentration and to compare it with carboxyhemoglobin (HbCO) saturation. In postmortem cases, the HbCO measured by spectrophotometry frequently leads to inaccurate results due to inadequate samples or analyses. The true role of CO intoxication in the death of a person could be misclassified. The estimation of HbCO from HS‐GC‐MS CO measurements provides helpful information by determining the total CO levels (CO linked to hemoglobin (HbCO) and CO dissociated from hemoglobin). The CO concentrations were converted in HbCO saturation levels to define cutoff blood CO values. CO limits were defined as less than 1 μmol/mL for living persons, less than 1.5 μmol/mL for dead persons without CO exposure, and greater than 3 μmol/mL for dead persons with clear CO poisoning.     

Sex determination in the subadult ilia: an indirect test of Weaver's nonmetric sexing method

The sexing of subadult remains has been an ongoing problem in physical anthropology for many years. This is due in part to the scarcity of subadult collections of known age and sex which are large enough to be used to develop and test analytical methods. Several methods have been devised but few have produced reliable results. In 1980, Weaver presented a method for sexing subadult ilia using a nonmetric trait (the raised versus nonraised auricular surface), which has an accuracy of 75% in fetal females and 92% in fetal males. His method has not been tested for reliability on a different subadult sample. An indirect test of Weaver's method was made on a sample of subadult South Dakota Arikara Indian ilia by comparing the ratio of raised to nonraised auricular surfaces with an expected 1:1 sex distribution. Bimodal sex distributions in the Arikara formed unrealistic sex ratios, following an age-related shift from a 6:1 raised/nonraised ratio in newborns to a 1:4 ratio in young adolescence. Significant age correlations were found both in the present study and in Weaver's published results. The age-to-sex correlations indicated no confounding in the present study. The results of this test suggest that auricular surface morphology is not sex specific in subadult ilia, but may be related to aspects of shape and morphology in pelvic growth.     

Evaluation of the placenta in a stillborn fetus to estimate the time of death

The authors present a case of a presumably unknown pregnancy, in which the newborn was found dead in the bathroom. The child was considered stillborn and the death due to natural causes. The literature on estimation of the time of fetal death is reviewed. It has been reported that certain placental histological changes are useful in predicting the time of death in stillborn fetuses, but little has been published about their potential role. Furthermore, parameters evaluated by literature in examining the placenta for determining time of fetal demise in cases of stillbirth are not uniform in the few studies that have been published. We emphasize that any attempt to estimate the time of fetal death without an adequate knowledge of placental morphological changes is futile and forensically unjustifiable, and that a comprehensive placental, external, and histological examination of the fetus is still the method of choice to gain a reliable forensic answer.     

Green pigments in epidermal blisters of decomposed cadavers

A spectrophotometrical method for discrimination between sulfhemoglobin (SHb) and choleglobin was applied to analyses of the green pigments in epidermal blisters of decomposed cadavers. SHb appeared to be a predominant constituent of the pigments, although significant amounts of choleglobin, one third to one sixth of SHb, were also demonstrated. The present results indicate that the greenish discoloration of decomposed cadavers is due to formation of a mixture of SHb, choleglobin and possibly other denatured hemoglobin derivatives.     

The fetus as a patient: emerging rights as a person?

Dramatic scientific breakthroughs in medical technology have revolutionized the physician's diagnostic prowess in the art of obstetrics. Scientific procedures now reveal previously undetectable secrets about the womb's tiny inhabitants. In the last few years, perinatologists have not only demonstrated the ability to discern fetal abnormalities of an extraordinary variety, but also have become increasingly successful in correcting many of these defects in utero. This article identifies the potential medicolegal conflicts that may arise as fetal surgery becomes an accepted medical practice. It begins by surveying the legal rights of unborn persons with a particular emphasis on the role of viability in determining those rights. The article will then examine the concept of viability as developed by the Supreme Court in Roe v. Wade and later abortion decisions and concludes that the current judicial deference to the medical community in determining viability is adequate for balancing rights in the abortion context. However, conflicts among physicians and between the other and her unborn child that may arise in the fetal surgery context suggest that viability may be an adequate benchmark for resolving such conflicts. The article concludes with a recommendation to reform the current method of resolving the critical question of when a fetus becomes viable.     

Identification of fetal blood stains by radioimmunoassay of alpha 1-fetoprotein.

Radioimmunoassay of alpha 1-fetoprotein(AFP) for medico-legal identification of fetal blood stains using a commercial kit is described. The AFP content in fetal blood stains on filter paper ranged from 21--320 ng/9 mm2. The protein was detected in stains of adult blood and retroplacental blood in only negligible amounts. Aging of the blood stains did not influence the values up to 1 month. The method is simple and sensitive enough for application to medico-legal-practice.