Lewis typing of human bloodstains by enzyme-linked immunosorbent assay (ELISA) using monoclonal anti-Le(a) and anti-Le(b)

The Lewis blood grouping of human dried bloodstains could be determined by an enzyme-linked immunosorbent assay (ELISA) using monoclonal anti-Le(a) and anti-Le(b) antibodies with an avidin-biotin complex (ABC). The bloodstains aged 1 year were used as samples, and approximately 1 mg of the stains was enough to type each Lewis antigen reliably by this method. The Lewis substances of 106 individual stains were correctly typed regardless of their ABO blood group system.     

Determination of ABO blood grouping from human oral squamous epithelium by the highly sensitive immunohistochemical staining method EnVision+

Using the highly sensitive immunohistochemical staining method EnVision+, which employs a dextran polymer reagent for the secondary antibody, the detection of the ABH antigens was attempted in the oral squamous epithelium. This new technique uses monoclonal antibody as a primary antibody and it takes about three hours for staining. The time is much shorter than conventional absorption-elution testing or absorption-inhibition testing for the determination of ABO blood grouping. Secretor saliva samples were stained at strong intensity by the antibody, which corresponded to its blood group and anti-H. On the one hand, nonsecretor saliva samples were stained at strong intensity only by the antibody that corresponded to its blood group, and at weak intensity only by anti-H. Since human oral squamous epithelium antigens were stained specifically by this method, we can examine the ABO blood group of saliva samples and perform cytodiagnosis at the same time. Our research suggested that the EnVision+ Method is a useful technique for ABO blood grouping of saliva in forensic cases.     

Fatal subarachnoid hemorrhage complicating actinomycotic meningitis

Actinomycosis is caused by Gram-positive Actinomyces species that are part of the normal oral flora with low virulence. We describe a rare case of sudden death of a 48-year-old man with actinomycotic basilar meningitis that was complicated by fatal subarachnoid hemorrhage. Autopsy revealed meningitis at the basilar region of the brain, and histological examination revealed characteristic bacterial aggregates with extensive leukocyte infiltration and severe vasculitis of arteries of this region. Rupture of an artery by severe arteritis was thought to be the cause of the subarachnoid hemorrhage. The probable primary source of infection was found in the left lung. To the best of our knowledge, the complication of subarachnoid hemorrhage has not been reported previously in actinomycotic meningitis.     

Dependability of ABO findings in stored blood samples

Report on the occurrence of "acquired" A or B in stored blood samples. This bacterial alteration is of importance when an indirect experimental investigation with the absorption-elution technique is needed in advanced cases of hemolysis. One has to consider this disturbing factor in identification tests (alcoholic blood samples). In dried blood stains we did not notice this problem, but it has to be taken into account in genitals stains.     

Information from penile swabs in sexual assault cases

A survey has been made to assess the evidential value of tests carried out on 660 casework penile swabs. Most were from suspects in sexual assaults and were examined to see if the donor had had recent anal, oral or vaginal intercourse. The swabs were tested for one or more of the following: blood, faeces, saliva, vaginal secretions, semen. Blood was seldom found, it was usually weak and insufficient for grouping. Faeces were only identified on a pair of swabs from a dead homosexual showing that proof of buggery by this means is rare. Amylase, suggestive of saliva and oral intercourse, was occasionally detected. Glycogen-rich epithelial cells were sometimes present indicating vaginal intercourse. Semen was frequently found but its presence may not result from a recent sexual act. An ABO group different from the donor was obtained from a fifth of the swabs typed. Grouping in other blood group systems was rarely attempted or successful. Penile swabs provided a means of detecting a victim's ABO blood group on a suspect when it would not have been possible to demonstrate the suspect's group on samples from the victim. They also had value in assaults involving more than one offender. The main limitation of penile swabs was the paucity of material on them and the sampling site affected the interpretation of the results.     

Orosomucoid (ORM) typing by isoelectric focusing: description of two new alleles in a German population and thermostability in bloodstains

The genetic polymorphism of serum orosomucoid (ORM) was studied in 168 unrelated German individuals using isoelectric focusing followed by immunoprinting. Two new alleles, tentatively designated ORM1*14 and ORM2*13, were identified. The method was successfully applied to demonstrate ORM1 types in dried bloodstains. Each type of ORM1 was also correctly determined in bloodstains heated at 130 degrees C for 30 min. The results indicated that ORM1 is a new powerful genetic marker system for the grouping of bloodstains.     

A case of Ludwig angina: a case report and review of the literature

Ludwig angina is a rapidly progressing submaxillary, submandibular, and sublingual necrotizing cellulitis of the floor of the mouth that can have lethal consequences due to airway obstruction. Various aerobic and anaerobic microorganisms, and less often fungi, have been implicated to cause Ludwig angina, including oral flora such as streptococci and staphylococci. Early recognition and the use of parenteral antibiotics can prevent mortality and morbidity. We report a case of a 25-year-old white man who was admitted to the hospital by his dentist after being diagnosed with Ludwig angina secondary to periodontal abscesses involving teeth #17 and #32. Although antibiotics were administered, while in the hospital, the decedent had difficulty swallowing and was drooling. He suddenly began to have seizure-like activity thought to be anoxic myoclonus. The decedent was aggressively resuscitated and taken to the operating room for neck exploration and a tracheostomy. Neck exploration revealed severe necrotizing acute inflammation of the deep soft tissues and musculature of the neck. He remained on life support for 7 days until he was declared brain dead. Ludwig angina is a progressive cellulitis that often results in death by asphyxia. Ludwig angina can be complicated by subsequent deep neck infection. The underlying etiologies and common scenarios are examined, and significant autopsy findings and dissecting procedures are discussed. The pathophysiology of Ludwig angina is studied with a review of the current literature.     

Detection of ethyl glucuronide in blood spotted on different surfaces

This study aims to show that sensitive detection of ethyl glucuronide in dried blood spotted onto various surfaces after a period of 24h is feasible. At present, there is insufficient information how tightly ethyl glucuronide (EtG) binds to various materials and how easily it can be eluted. 4ml aliquots of blood samples obtained from seven volunteers after consumption of alcoholic beverages were applied to six different surfaces. After drying and a 24h-storage at 20±2°C the samples were re-dissolved in water, and EtG was subsequently analyzed by a LC-MS Paul-type ion trap. A comparison was made between dried and corresponding fluid samples. EtG was detectable in all subjects' samples following consumption of alcohol. EtG was also detectable after a storage time of four weeks at 4°C in whole blood that had been preserved with EDTA. EtG was detectable in all samples dried on different surfaces and its concentration remained relatively constant irrespective of the particular condition of the material. Detection of EtG in blood spots from the scene may indicate recent alcohol consumption in cases where collection of blood remained undone or could not be performed.     

GC-MC法测定血液中丙泊酚

目的 建立血液中丙泊酚的气相色谱-质谱联用(GC-MS)分析方法.方法 血液以2,4-二甲基-6-叔丁基苯酚为内标,用乙醚进行液液萃取,离心后取有机层,水浴下挥干,GC-MS检测.结果 血液中丙泊酚与内标分离良好,在0.02～10 μg/mL范围内线性良好,线性方程为y=0.3136 x-0.0068,相关系数为0.9...     

A sandwich enzyme-linked immunosorbent assay for ABO blood typing of semen by using anti-p 84 monoclonal antibody as a marker of blood group substance in semen

A blood group substance (BGS), a protein with ABH antigenic activity, was isolated from human seminal plasma and designated as p 84 (Sato, 1995). We have developed a method for determining the ABO blood type of semen by performing a sandwich enzyme-linked immunosorbent assay (ELISA) in which p 84 is captured with an anti-p 84 monoclonal antibody, and evaluated the specificity and sensitivity of this method. Although BGS activity was detected in semen sensitively by this method, it was not detected in saliva, urine, breast milk, blood or vaginal secretions. Since the concentration of p 84 in semen was independent of the secretion status, the status can be determined as non-secretor when p 84 but not BGS activity was detected. To determine the stability of BGS activity on p 84, dried stains of semen on filter paper were kept at 4, 26, and 37 degrees C for 8 months, 2 years and 1 month, respectively, and their BGS activities were examined. After 8 months at 4 degrees C, over 60% of the original BGS activity was recovered from the stain. The activity could be detected even from a square as small as 0.25 by 0.25 cm. After 1 month at 37 degrees C and 2 years at 26 degrees C, 31 and 20% of the BGS activity, respectively, still remained. It could be detected from the pieces of 1.0 by 1.0 cm and 0.5 by 0.5 cm squares, kept for 1 month at 37 degrees C and 2 years at 26 degrees C, respectively. Finally, semen was mixed with saliva or blood at varying volumetric ratios and used for the sources of dried stains. The BGS activity of p 84 could be detected in the stains until the ratio between semen and saliva or blood reached 1:4. We conclude that this sandwich ELISA offers a more sensitive and specific method for determining the ABO blood type of semen samples obtained from sexual assault victims than existing methods, such as the conventional absorption-elution and classical hemagglutination-inhibition tests.     

Specimen preparation for ABO determination of various kinds of blood stains

Trying to optimize the preparation of blood stains, we found methanol fixation not to produce very good results for the determination of ABO blood group antigens. It is advantageous to transfer blood stains before testing to cotton cloth. This transfer is also of practical use if blood stains are to be saved on a smooth surface for lateral determination. We testet on 35 different carrier materials, on which blood stains in casework often were found, whether blood grouping gave better results on either the original material or after transfer. Results are shown on a table. The test revealed, that solubility of the stain in aqua dest is a good sign for a successful transfer. Blood stains on pine-wood soil, soil and loam were not suited for ABO grouping.     

The ABO blood grouping of a minute hair sample by the immunohistochemical technique

The unlabeled antibody (PAP) immunoperoxidase technique was applied to the ABO blood grouping of human scalp hairs. Hair samples were subjected to longitudinal- or cross-sectioning, thus obtaining suitable samples for subsequent immunostaining. The immunostaining was carried out using rabbit anti-A and anti-B sera as the primary antibodies. With this technique, the group-specific staining which is revealed as a dark brown precipitate was clearly observed within the medullae of the hair shaft, and depending on the presence or absence of these precipitates, respective blood groups of unknown hair samples were determined. At the hair root, on the other hand, positive stainings were observed not only in medullary cells but also in some cortical cells of the keratogenous zone. From the present study, it can be safely said that this technique is of practical use for the ABO blood grouping from a minute (less than 3 mm) hair sample.     

Fatal salmonella aortitis with mycotic aneurysm rupture

Salmonellae most commonly cause uncomplicated cases of gastroenteritis but have a predilection for damaged blood vessels, especially those damaged by atherosclerosis. The abdominal aorta is most frequently affected. The most serious complication of aortitis is mycotic aneurysm formation with subsequent rupture. The authors present the case of a 61-year-old man who was found unresponsive at home 3 days after discharge from the hospital for treatment of gastroenteritis with bacteremia. Postmortem examination revealed a ruptured mycotic aneurysm with a large retroperitoneal hematoma. Numerous gram-negative rods were embedded in the wall of the aorta and surrounding inflammatory infiltrate, compatible with the patient's previously isolated. Whereas abdominal aortic aneurysm rupture is most commonly associated with atherosclerosis, the isolation of from blood cultures, coupled with radiographic evidence of gas surrounding the aorta, should raise the suspicion of infectious aortitis. Whereas fatal rupture of an aortic aneurysm secondary to atherosclerosis alone or in conjunction with aortitis will not have an impact on the manner of death, infections are reportable and thus have public health implications.     

生物检材采集与保存套管的应用

目的探讨生物检材采集与保存套管在法医学中的应用价值。方法在不同温、湿度环境下，观察悬空放置在生物检材采集与保存套管、有孔与外界相通的套管及密闭管内的湿润棉签的干燥时间30次；分别用生物检材采集与保存套管和医用棉签采集纸袋保存口腔细胞、血液、皮肤脱落细胞样本各20例，磁珠法提取DNA并进行DNA定量；用生物检材采集与保存套管采集口腔脱落细胞和血样进行DNA直接扩增。结果在温度4～30℃、相对湿度21％～90％的环境下，湿润棉签在套管内的平均干燥时间为7．89h，有孔管内的为23．30h，密闭管内观察15天仍不干燥，出现霉斑。生物检材用套管采集保存比医用棉签采集纸袋保存方式获得的DNA量显著提高，平均高达0．968倍；用套管采集口腔细胞和血样进行直接扩增，操作简单方便，成功率高。结论生物检材采集与保存套管具有快速干燥、对检材无损耗和浓缩等优点，可提高检材DNA的提取效率，且适合直接扩增。     

Ultraviolet 365 as an Alternative Light Source for Detection of Blood Serum

The use of alternative light sources (ALS) in bloodstain analysis has focused on dried (whole) blood, while information on detection of blood serum is lacking. Serum detection by ALS could provide valuable information at a crime scene, as serum may become separated from blood during clotting and cast off, especially in cases where the victim is moved. Additionally, a perpetrator may concentrate on the removal/scouring of dried blood with small amounts of serum going unnoticed, as it dries relatively clear on certain objects. In this report, the detection of human blood serum was evaluated using ultraviolet (UV) light at two different wavelengths. These results show that ultraviolet (UV) at 365 nm (UV365) was effective in the detection of even small amounts of blood plasma and serum, compared with UV at 395 nm, which was not. UV365 was also found to be useful in distinguishing blood imprints from clotting blood which had been transferred to material versus blood that had been added directly. Taken together, these results demonstrate that UV365 may be utilized as a simple, nondestructive method for blood serum detection.     

Comparison of fingernail ridge patterns of monozygotic twins

The ridge patterns on the fingernails of corresponding fingers of a pair of twins were compared microscopically and found to be readily distinguishable from one another. Based on blood grouping in six blood group systems (ABO, Rhesus, Ss, Duffy, Kidd, and Kell), the probability that the twins were monozygotic was calculated to be 89.1%.     

Evaluation of antisera for bloodstain grouping. I. ABH, MN, and Rh

Sixty-eight different commercially available blood grouping antisera and lectins with ABH, MN, and Rh D, C, E, c, and e specificities were serologically evaluated for their applicability to bloodstain antigen determination. The characteristics of the antisera were determined with red cells, with fresh bloodstains, and with series of aging bloodstains. The Rh antisera were tested under a variety of serological conditions and with bloodstains on various substrata. Additionally, studies on optimization of absorption-elution procedure variables were carried out, and some data on the storage characteristics of red cells and blood grouping antisera were gathered.     

Rapid phenotyping of the group specific component by immunofixation on cellulose acetate

Determination of the genetically controlled variants of the polymorphic Gc system was achieved by electrophoresis on cellulose acetate membranes followed by immunofixation with a specific anti-Gc antiserum. The method is applicable to plasma, whole hemolyzed blood, and dried blood. Multiple specimens can be analyzed simultaneously within 60 to 80 min. The cellulose acetate electrophoretogram of the Gc variants remains as a permanent record.