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PubMedDate not supplied

Demonstration of placental aminopeptidase isozymes by disc electrophoresis and its application to the forensic pregnancy test from blood stains.

Serum and placental aminopeptidase isozymes were characterized by means of disc electrophoresis. Human pregnancy sera exhibited three bands with leucine aminopeptidase (LAP) activity. The fastest moving LAP band was also present in nonpregnancy and foetal sera. The other two cystine aminopeptidase bands (CAP1 and CAP2), characteristic of pregnancy, were demonstrated in lysosomal extracts and hence suggested to be derived from the lysosomes of the placenta. These two CAP bands of lysosomal origin appeared in sera of pregnant women at different times during the course of gestation; CAP1 band in the 3rd month and CAP2 band in the 6th month. The information obtained from serum samples was applied to the medicolegal diagnosis of pregnancy from blood stains. Excess haemoglobin in blood stain extracts could be eliminated by subjecting them to a P-cellulose column. The results and their implications were discussed. This technique proved to be of practical use in the examination of blood stains up to periods of about 2 weeks after stain formation.

Aminopeptidases↗
PubMed1977

The degradation of Romanowsky-type blood stains in methanol.

The oxidative demethylation of Romanowsky-type stains in methanol has been examined quantitatively with respect to its effect upon the staining of blood smears. Spectral changes in bound dye, observed through two color filters, have been measured for the nuclei and cytoplasm of segmented neutrophils and monocytes utilizing the LARC automated differential analyzer. Stain decomposition in methanol results in a large loss in staining intensity with little change in color. The loss in intensity has been correlated with the observed spectral changes in the degraded stain. High-performance liquid chromatographic analysis of degraded stain samples has shown the products of methanolic degradation to be different from those obtained in aqueous polychroming reactions. To maintain a stain of defined thiazine dye composition and thus defined staining properties, refrigeration is recommended.

Blood Cells↗
PubMed1996

PCR-based human leukocyte antigen (HLA) DQ alpha typing of blood stained light and dark blue denim fabric.

Obtaining typable PCR products from DNA purified from blood stained blue denim has been difficult since inhibitors of PCR in blue denim apparently co-purify with the DNA. Organic and chelex extraction methods were tested for their ability to purify typable DNA from either light or dark blue denim fabric both stained with blood. DNA purified from the light blue denim using either method was successfully used in obtaining correct HLA-DQ alpha typing results. The chelex, but not the organic, procedure was able to yield typable DNA when the dark blue denim was the substrate. Therefore, the chelex method may be more effective than the organic method in preventing compounds that inhibit PCR from co-purifying with the DNA.

Blood Stains↗
PubMed1995

DNA extraction and amplification from Giemsa-stained blood smears.

DNA extraction was attempted from Giemsa-stained blood smears on glass slides that had been stored for several years. High molecular weight DNA bands were clearly visible after electrophoresis when DNA was extracted from specimens stored up to 2 years. Specimens more than 4 years old demonstrated a smeared pattern, suggesting degeneration of the DNA, but they could be rescued by PCR amplification using primers of HLA-DQA1 genes. The recovery could be pursued even in 11-year-old specimens.

Azure Stains↗
PubMed1979

[Typing ACP isoenzymes in blood stains: test limits and temperature dependancy (author's transl)].

Two series of test blood stains of known red-cell acid phosphatase types A, AB, AC, B, BC, and C are applied to two different types of stain carriers (glass and cotton) and stored at different temperature ranges: (a) 37 degrees C at high air humidity, (b) 22 degrees C, (c) 4 degrees C, and (d) - 20 degrees C. Electrophoresis is carried out either on cellulose-acetate-foils or on thin agarose layers. Visualisation of the isoenzyme sites is achieved using umbelliferyl phosphate as a substrate. The most important results are: Electrophoresis on "Cellogel"-foils turns out advantageous compared to agarose since (a) less stain material is required, (b) reproducibility is rather reliable, and (c) electrophoretic separation is clearer. Blood stains on glass are identified after longer periods of storage than stains on cotton. If stains are stored at room temperature the time limits of demonstration are in the range of 3 to 6 weeks. The size of the sample necessary for demonstrating varies widely. Approximately twenty to twenty-five per cent of a blood drop are needed. The method described is recommended as practicable and reliable.

Acid Phosphatase↗
PubMed1991

Genetic studies of human apolipoproteins. XIX. Apolipoproteins E and A-IV phenotyping from whole blood and blood stains.

Apolipoproteins E and A-IV reveal common, genetically determined, structural polymorphisms, which can be detected in serum or plasma, using isoelectric focusing and immunoblotting techniques. In the present study, we have investigated the possibility of detecting the gene products of these two polymorphic loci in whole blood and blood stains. A total of 68 plasma samples and their corresponding whole blood samples were screened. Phenotypic patterns obtained in whole blood were identical to those obtained in plasma for both the APO E and APO A-IV polymorphisms. The signal-to-noise ratio in the APO E patterns was higher in whole blood than in fresh plasma. The gene products of both the APO E and APO A-IV were found to be very stable in several months old blood stains. The detection of apolipoprotein polymorphisms in whole blood and blood stains signifies their potential applications, on a wider scale, to several areas including forensic serology, paternity testing, clinical and population genetic studies.

Apolipoproteins A↗
PubMed1990

Routine phenotyping of native and desialyzed alpha 2HS-glycoprotein from blood stains.

The application of a polyacrylamide gel isoelectric focusing (PAGIEF) and immunoblotting procedure for the identification of native alpha 2HS-glycoprotein (AHSG) in routine casework blood stains has produced reportable results on 57.2% of samples. This reporting rate is lower than that for group specific component (GC) (83.8%) and phosphoglucomutase (PGM 1) (72.8%) phenotyping of the same samples. Blood stain samples were desialyzed with 1 U/ml neuraminidase, overnight at room temperature prior to PAGIEF in gels containing pharmalyte pH 5-6 and 2.5 M urea. Simple AHSG patterns were developed by immunoblotting. This procedure was five times as sensitive as the native AHSG method and desialyzation was reproducible over a range of incubation times and neuraminidase concentrations. The application of the desialyzed AHSG analysis to routine casework samples has resulted in a significant increase in the number of reportable results (762 reported out of 1027 samples). This reporting rate (74.2%) compares favourably with that for GC (79.1%) and PGH 1 (69.6%) phenotyping of the same samples. The three AHSG alleles (AHSG*1, 2 and 3) are clearly resolved after sample desialyzation and separation in gels containing pharmalyte pH 5-6 and 2.5 M urea. The sensitivity of desialyzed AHSG phenotyping approaches that of GC and this technique is worthy of inclusion in blood stain screening protocols of forensic laboratories in regions where the population has a limited range of rare AHSG alleles.

Blood Proteins↗
PubMed2006

Penetrating keratoplasty combined with vitreoretinal surgery for severe ocular injury with blood-stained cornea and no light perception.

OBJECTIVE: To evaluate the effects of treatment of severe ocular injury with blood-stained cornea and no light perception by combined penetrating keratoplasty and vitreoretinal surgery, and to analyze the relevant factors. METHODS: Records of 7 severely injured eyes of 7 patients with blood-stained cornea and no light perception who underwent penetrating keratoplasty combined with vitrectomy using a temporary keratoprosthesis were evaluated retrospectively. The preoperative visual acuity was no light perception in all injured eyes with a mean intraocular pressure of 3 mm Hg and a range from 2 to 5 mm Hg. The average interval from emergency wound closure to vitrectomy was 18 days with a range from 12 to 21 days. The mean follow-up was 28 months with a range from 26 to 30 months. RESULTS: The postoperative visual acuity was better than light perception in 5 eyes with the best corrected visual acuity from light perception to 0.06. The retina was attached in 5 eyes. The postoperative intraocular pressure ranged from 5 to 15 mm Hg with a mean of 12 mm Hg; it was significantly higher than the preoperative one (p < 0.05). The postoperative complications mainly included temporary intraocular elevation (1 eye), corneal neovascularization (4 eyes), corneal rejection (4 eyes), retinal detachment (2 eyes) and ocular atrophy (2 eyes). CONCLUSION: Penetrating keratoplasty combined with vitrectomy using a temporary keratoprosthesis is a safe and effective method in treating severe ocular injury with blood-stained cornea and no light perception.

Adult↗
PubMed1987

Spontaneous abortion following mid-trimester amniocentesis. Clinical significance of placental perforation and blood-stained amniotic fluid.

The clinical significance of placental perforation and blood-stained amniotic fluid was studied in a group of 7238 Danish women undergoing mid-trimester amniocentesis for prenatal diagnosis under ultrasound guidance. The risk of spontaneous abortion was significantly increased both in pregnancies where the placenta was perforated and in those with blood-stained amniotic fluid. The risk estimate nearly doubled after placental perforation and more than doubled with a bloody tap. It is concluded that for women at relatively low risk of a fetal genetic abnormality, the indication of the amniocentesis should be reconsidered if a placental perforation is unavoidable.

Abortion, Spontaneous↗
PubMed1989

Human identification and sex determination of dental pulp, bone marrow and blood stains with a recombinant DNA probe.

Recombinant DNA hybridizing specifically to a 300 nucleotide repeat DNA sequence (BLUR8) of human specificity and to human repeat DNA sequence (pHY10) on the Y chromosome was used for human identification and sex determination of degraded DNA samples of blood stains, dental pulp, and bone marrow. This radioactive technique enabled reliable and sensitive human and sex determination from blood stains that were more than 80 years old. Less than 1 piece of 0.5 cm length thread of blood stain was enough for both tests. DNA from relatively fresh dental pulp and bone marrow was clearly identified. The human identification test, which could recognize up to 0.3 ng DNA correctly, was 3 to 5 times more sensitive than the sex determination test.

Adolescent↗
PubMedDate not supplied

[Sex determination in blood stains through identification of Y-chromatin: medico-legal application].

The Y-chromatin is visualized in human interphase nuclei, corresponding to the distal portion of the Y-chromosome, which shows marked fluorescence after staining with quinacrine. This report describes the results of sex determination on blood smears fixed in methanol and blood stains left at room temperature for 13 weeks (1st report), and for 10 months (2nd report). Blind trials showed that a reliable sex determination of blood stains on glass left for at least six months is possible. The application of this method in forensic practice is discussed.

Blood Stains↗
PubMed1987

Identification in blood stains through DNA typing with C4 and HLA-DR probes.

A restriction fragment length polymorphism analysis using double digestion of DNA preparations with XbaI and BglII restriction enzymes and hybridization with C4 and HLA-DR probes is described. The typing conditions selected reveal extensive individual variation in both C4 and DR gene regions. In our panel of 46 unrelated individuals, 37 different phenotypic patterns were recognized when both probes were used, and preliminary discriminative power values of 0.865 and 0.914 were calculated for C4 and DR beta, respectively. The probability of a chance match using both systems is probably about 1.5.10(-2). The potential of this method for individual identification of blood stains was demonstrated on DNA prepared from 6-month-old dried blood stains from seven panel individuals. The seven individuals were all identified when comparing stain DNA patterns with panel control patterns. No RFLP pattern changes were observed following storage of blood stains. Based on these experiments with C4 and DR beta DNA typing under laboratory conditions, it is concluded that DNA typing with such probes may become a powerful tool in future stain identification analyses.

Blood Grouping and Crossmatching↗
PubMed1980

[Determination of adenylate kinase (AK) phenotypes in blood stains].

The author examined blood spots, prepared experimentally with known types of the enzyme adenyl kinase (AK) 1-1 and 2-1 on the most frequently met in the practice materials-carriers: cloth, paper, wood, knife, glass and stone. The studies were performed by means of electrophoresis on cellulose acetate folio. She found that the types of AK could well be determined with spot duration of 5-6 months, saturating the material (cloth, paper), and over 1 year in the presence of crusts of dried blood, regardless of the material-carrier (including cloth and paper). She recommends that the system AK should be introduced in the practice during examination of blood spots at experts' reports according to material evidences. In view of phenotype frequencies of AK in our country the theoretical probability for two blood spots, randomly taken, to differ only by AK is 12.85%.

Adenylate Kinase↗