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PubMed1989

[Quantitative determinations of Hp, Pi, Tf and GC in stored blood stains].

Quantitative analysis of serum proteins in blood stains by rocket immunoelectrophoresis showed the following results: Long elution times in processing blood stains do not produce a significantly higher yield of serum proteins in stains, and they increase the risk of structural alteration of protein molecules. The amount of proteins detected decreases with increasing age of blood stains. Some proteins are already altered in stains after a short storage time and are no longer useful for phenotyping. Our results confirm that blood stain material should be processed as soon as possible.

Blood Grouping and Crossmatching↗
PubMed2004

[Determination of AB0 blood group system from degraded blood stains on serological and molecular genetic level].

The AB0 blood group system typing remains one of the basic laboratory tasks in a forensic practice. However, problems arise when the analysed samples are seriously degraded. We took blood samples from six volunteers (three men, three women) and made blood stains on pieces of sterile cotton cloth. Blood stains were incubated at three different temperatures (22 degrees C, 37 degrees C, 56 degrees C) for various periods of time (1 day, 1 week, 14 days, 1 month, 3 months, 6 months, 1 year). For blood stains degraded at 22 degrees C we also analysed the samples after 3.5 hours of incubation. Moreover, we tried to determine the AB0 blood group system after thermal degradation at high temperature, accurately at 200 degrees C for 10 min. For the AB0 blood group system typing a Polymerase Chain Reaction method was used to amplify glycosyltransferase gene, when DNA had been isolated from artificially created blood stains, followed by their subsequent artificial thermal degradation. For serological AB0 typing the mixed agglutination and the Therkelsen method were used. The DNA analysis seemed to solve problems with seriously degraded blood stains but we found out that classical serological methods were even better in some cases.

ABO Blood-Group System↗
PubMed1979

[Changes of various proteins in older blood stains].

26 unequally old groups of blood stains up to 180 days old were examined for the presence of albumin, Ig A, Ig G immunoglobulins, beta 1 C globulin and transferrin. To ensure different external conditions, each group was composed so as to contain blood stains of the same age but preserved under different conditions and on different types of underlying material. Albumin and Ig G immunoglobulin proved to be among the most stable ones. However, due to variations in the results the practical uses of this type of blood stain age estimation appear to be problematic.

Beta-Globulins↗
PubMed1985

Blood staining of the cornea. Light microscopic and ultrastructural features.

Eleven blood-stained corneas were examined by light and transmission electron microscopy at intervals ranging from one month to seven years after initial staining occurred. Blood staining in each case was associated with focal loss of endothelial cells or endothelial degenerative changes and elevated intraocular pressure. Globules of erythrocytic breakdown products penetrated the discontinuous endothelium and intact Descemet's membrane. Large deposits, primarily extracellular, displaced but did not interrupt the stromal lamellae. Keratocytes in blood-stained corneas contained erythrocytic breakdown products and hemosiderin, and were remarkable for extensive degenerative changes in contrast to keratocytes in areas of cleared cornea, which contained smaller amounts of hemosiderin and were relatively normal. After one year, clearing could be seen to involve peripheral and posterior stroma, and to a lesser degree, the anterior stroma. We found no evidence to support the contention that blood-derived macrophages play a role in the clearing of erythrocyte debris. The stereotyped pattern of peripheral, posterior, and anterior stromal clearing observed seems to be consistent with diffusion of hemoglobin breakdown products out of the cornea as the primary mechanism of clearing.

Blood↗
PubMed1986

Corneal blood staining. An animal model.

Corneal blood staining was established in the rabbit cornea by injecting autologous, citrate-buffered blood in the anterior chamber. Increased intraocular pressure was maintained above 30 mm of Hg by self-sealing trans-limbal injections repeated every 12 hours. Typically, corneal edema developed in 3 days, followed several days later by a red discoloration that turned brown about 2 days later. Histopathologically, the edematous cornea disclosed endothelial swelling and attenuation with marked stromal edema. Histochemically, the red-stained cornea disclosed only extracellular hemoglobin particles; the brown-stained corneas showed extracellular and intracellular hemoglobin particles as well as intracellular hemosiderin in keratocytes. Spectrophotometric analysis of the keratectomy specimens suggested the presence of porphyrins in all stages of blood staining, including the edematous cornea. Oxyhemoglobin was found in red-stained corneas, while methemoglobin was present in the brown-stained corneas. It was concluded that endothelial degeneration uniformly accompanied corneal blood staining in this model and that keratocytes are actively involved in hemoglobin degradation.

Animals↗
PubMed1979

Preparation of anti-alpha 2-macroglobulin using Canavalia lineata DC lectin for differentiating species-specificity of blood stains.

To differentiate species-specificity of blood stains, anti-alpha 2-macroglobulin was raised in rabbits against Canavalia lineata DC lectin-serum complex (LSC). Adsorption of anti-LSC with human lipoprotein resulted in antiserum specific for alpha 2-macroglobulin. It was confirmed by Ouchterlony test that the antiserum adsorbed successively with monkey serum or anti-LSC adsorbed directly with monkey serum reacted with only human serum but not with mammalian ones. Immunoelectrosyneresis and anti-LSC consumption test could identify species-specificity of blood stains kept for up to two years and for up to several years, respectively. It is indicated that anti-LSC is quite effective for differentiating species-specificity of blood stains.

Adsorption↗
PubMed1985

[Stability of beta-HCG in old blood stains].

Pregnancy was diagnosed from old blood stains by means of radioimmunoassay of beta-HCG. Up to 45 days, the age of the blood stains did not influence values, and extraction by ultrasound did not improve the results.

Adolescent↗
PubMed1989

Species identification of blood stains using a biotin-labeled DNA probe.

In an attempt to identify blood stains of human origin, DNA was extracted from human and animal dried blood stains and examined using recombinant DNA techniques. As a DNA probe, biotin-labeled or radioisotopically labeled human repetitive sequence 2.3 Kb Hind III fragment (pH 12) in pBR 322 was used. The probe clearly hybridized with human DNA, but not with DNA extracted from frog, fish, mouse, rat, rabbit, chicken, cat, dog, and sheep. DNA from a Japanese monkeies and from a chimpanzee also hybridized with comparable efficiency. The minimum amount of the blood for the species identification was estimated to 0.4 microliter of fresh blood, 2.1 microliters of 2 weeks old blood stain and 3.8 microliters of 3 months old blood stains, indicating that only one drop of human blood is sufficient for species identification. There was no difference of "signal" between isotopic and non-isotopic procedures for DNA analysis. The advantages of the non-isotopic method for species identification of blood stains in forensic science is briefly discussed.

Animals↗
PubMed1983

The typing of group-specific component (Gc protein) in human blood stains.

It is known that the typing of group-specific component (Gc protein) in human blood stains is difficult since Gc protein of the extracts of blood stains migrates more anodally to the alpha 1-globulin region in agar-gel immunoelectrophoresis, while Gc protein in liquid blood normally migrates to the alpha 2-globulin region. We have reported that the Gc protein found in the alpha 1-region is the result of binding of actin to Gc protein (Shinomiya, K., Kimura, H., Yoshida, K., and Shinomiya, T., J. Biochem., 92 (1982) 1163-1171, which renders it difficult to determine the Gc-phenotypes in the blood stains. On the basis of the above findings, we developed the method of phenotyping the Gc protein of human blood stains by agar-gel immunoelectrophoresis. Since the binding activity of actin to Gc protein is lost after treatment with a high concentration of guanidine HCl, the extracts of blood stains were treated with 4 M guanidine HCl to dissociate Gc protein and actin and then dialyzed to remove guanidine HCl. By this method we are able to determine the phenotypes of Gc protein in blood stains. The method we have developed is a useful tool in the forensic laboratory.

Alpha-Globulins↗
PubMedDate not supplied

Sex determination of blood stains with a recombinant DNA probe: comparison with radioactive and non-radioactive labeling methods.

A recombinant DNA probe hybridizing specifically to human repeat DNA sequence (pHY10) of which about 3000 copies are present on the Y chromosome was used for sex determination of degraded DNA samples of blood stains. Human blood stains of male and female origin were readily differentiated with the pHY10 DNA probe. This radioactive technique enabled reliable and sensitive sex determination from blood or dried blood stains greater than 20 years old. Less than 1 microliter of blood or 1 piece of 0.5 cm length thread of blood stain from cotton fabric was sufficient for the test using dot blot hybridization. Compared with the radioactive labeling method, the photobiotin labeling method showed one thirtieth to one fiftieth lower sensitivity and presented some problems which are expected to be resolvable.

Blood Stains↗
PubMed1980

Microspectrophotometric studies of Romanowsky stained blood cells. I. Subtraction analysis of a standardized procedure.

This paper describes a microspectrophotometric study of blood smears stained by a simple, standardized Romanowsky technique, using only the dyes azure B and cosin. Absorbance spectra are presented for twenty-two classes of cellular object, and for the two dyes in solution, together with tabulations of spectral maxima, and suitable wavelengths for use in automated image processing. The colours of objects stained with azure B/eosin are discussed in terms of absorbance spectra. By a spectral subtraction technique, it is shown that the differential colouration of various cell structures may be explained satisfactorily in terms of the varying proportions of only four dye components. These are the monomers and dimers of azure B and eosin. Polymerization was found to occur both in solution and on binding to biopolymers. A similar analysis of a conventional Romanowsky stain would present much greater difficulties, due to the greater number of dye components, which, however, contribute little to the colours observed.

Azure Stains↗
PubMed1996

Trabeculectomy and manual clot evacuation in traumatic hyphaema with corneal blood staining.

BACKGROUND: The management of traumatic hyphaema with raised intraocular pressure and corneal blood staining is difficult. Residual blood clots after anterior chamber washout are responsible for sustained postoperative elevation of intraocular pressure, even after trabeculectomy and clot evacuation. METHODS: Thirty-five patients with traumatic hyphaema, elevated intraocular pressure and varying degrees of corneal blood staining underwent a combined trabeculectomy with manual clot evacuation from the anterior chamber in a general hospital. RESULTS: The postoperative control of intraocular pressure was found to be adequate in all patients at the end of two months. Examination of the posterior segment was made possible earlier. Although the procedure is more complex, no significant complications were encountered. CONCLUSION: In patients presenting with traumatic hyphaema, secondary glaucoma and corneal blood staining, trabeculectomy with manual extraction of the clot through a large incision appears to be a safe and reliable procedure where medical therapy fails to control the intraocular pressure.

Adolescent↗
PubMed1979

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.

Blood Stains↗
PubMed1980

[Determination of subgroups A1-A2 in blood stains].

The results obtained in the determination of subgroups A1, A2 in blood stains using lectines anti-A1 and anti-H demonstrate the advantages of the absorption elution or mixed agglutination techniques over satiation method. In blood stains of up to 6 months of age, the lectine-aided determination of subgroups was reliable for periods of up to 3 months from the appearance of the blood stains.

ABO Blood-Group System↗
PubMed1989

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.

Abortion, Illegal↗
PubMed1993

Prenatal diagnosis by in situ hybridization on uncultured amniocytes: reduced sensitivity and potential risk of misdiagnosis in blood-stained samples.

Maternal cell contamination was assessed in 18 macroscopically blood-stained amniotic fluid samples from male fetuses. The samples were analysed by double-target fluorescent in situ hybridization (ISH) with Y and X chromosome-specific probes. The only sample with an aberrant karyotype (47,XY, + 18) was also analysed by hybridization with a chromosome 18-specific probe. An interpretation of extensive maternal cell contamination was made in two samples, one of which was the sample with trisomy 18. ISH with the chromosome 18-specific probe on this latter sample showed that the sensitivity of the ISH method for chromosome enumeration of uncultured amniotic fluid samples may be reduced in blood-stained samples. It was calculated that by using ISH for chromosome enumeration of the two extensively contaminated samples, a case of trisomy 21 might have been overlooked in both samples, while a case of trisomy 18 might only have been overlooked in one of the samples. It is concluded that ISH should not be used for chromosome enumeration of uncultured amniotic fluid samples that are macroscopically blood-stained without further technical developments.

Amniotic Fluid↗