Lugol's test reexamined again: buccal cells

Lugol's iodine staining technique was used to examine oral samples from 10 men and 10 women. Examination of saliva samples before and after extraction with water shows that the low levels (49 positive cells and 3,951 negative cells) of glycogen in buccal epithelial cells become even lower after water extraction (0 positive cells and 4,000 negative cells). In addition to the 20 samples used in this paper, 40 oral swabs extracted with water were examined under classroom conditions with much less than 1% of the epithelial cells being positive for glycogen. Furthermore, 119 saliva samples from chewed gauze in sexual assault kits were extracted with water and all of them yielded less than 1% glycogen positive cells. This paper proposes that when more than 1% of the nucleated squamous epithelial cells are glycogen positive with Lugol's test after extraction in water, it is reasonable to eliminate the mouth as a source of these glycogen positive cells.     

The examination of vaginally inserted plastic tampon applicators for genetic markers and evidence of prior sexual intercourse

Vaginally inserted plastic tampon applicators were obtained from 42 female volunteers. The applicators were examined for the presence of ABH blood group substances, phosphoglucomutase (PGM), amylase, acid phosphatase, P30, and intact spermatozoa. Each applicator was accompanied by a control blood sample, a saliva specimen, a brief sexual and menstrual history, and method of birth control of the donor. Eight of the male sexual partners of the donors submitted blood and saliva samples. One male sexual partner submitted only a saliva sample. ABH blood group substances corresponding to the donor were recovered from 36 of the 42 applicators. The remaining 6 applicators revealed a combination of the donor's and sexual partner's ABH substances. The female's PGM type was recovered from 34 of the applicators. The remaining 8 applicators failed to show PGM activity. Of the applicators, 15 indicated evidence of prior sexual intercourse by the detection of ABH substances not consistent with the applicator donor (6 samples), high levels of acid phosphatase (11 samples), or recovery of spermatozoa (8 samples) or some combination of these. All applicator samples failed to show the presence of either P30 activity or PGM factors foreign to the female.     

Background Levels of Salivary-α-amylase Plus Foreign DNA in Cases of Oral Intercourse: a Female Perspective

Saliva plus DNA from a suspect is commonly encountered in sexual assault cases on bodily swabs. However, without background knowledge, the weight of this evidence is unknown. It may indicate the presence of saliva resulting from cunnilingus, or it may represent indirect transfer. In this study, females who refrained from cunnilingus donated 43 items of underwear and 19 vaginal swabs. The samples were subjected to Phadebas^®, RSID^™-Saliva and mRNA profiling and were subsequently DNA-profiled to determine the prevalence of background saliva in the female population. The results report that 15.8% of females who refrained from cunnilingus were positive for saliva and a further 10.5% also had DNA from unknown source(s). These findings of the rate of indirect transfer were evaluated with the Bayesian approach, and it was found that the evidence of saliva plus a high foreign DNA source adds moderately strong support to the allegation of cunnilingus.     

Fatal iatrogenic BaSO4 embolism: morphological and ultrastructural findings confirmed by X-ray microanalysis and ICP-AES

A 42-year-old woman died from massive barium sulfate (BaSO(4)) lung embolism after a balloon catheter intended for elective colonography was inserted into her vagina. The vaginal insertion of the balloon catheter caused a bilateral laceration of the vaginal wall which was followed by penetration of BaSO(4) into the afferent veins and massive pulmonary embolism. Fluoroscopy performed during the fatal events and post-mortem X-rays revealed a radio-opaque substance in the vagina and uterus, the pelvic vessels and the vena cava, the right heart chambers, the lungs, and the kidneys. In addition to lungs, finely granular intravascular particles were demonstrated histologically in several organs including the brain and the glomerular capillaries. Scanning (SEM) and transmission (TEM) electron microscopy together with X-ray microanalysis, and inductively coupled plasma atomic emission spectrometry (ICP-AES) allowed the definite identification of BaSO(4) in lungs and confirmed its capacity to penetrate the pulmonary filter and to embolise via the systemic circulation in various organs.     

The diagnosis of an ABO(H) ABSe group in the traces of saliva, sperm and vaginal secretion using mixed agglutination reaction

Experimental spots of the saliva, sperm, vaginal secretion from persons with groups Ase, Bse, ABSe and Abse were studied with mixed agglutination reaction (MAR) using hemagglutinating sera anti-A and anti-B (heteroimmune, isoimmune), monoclonal antibodies. MAR with monoclonal antibodies was able to diagnose ABSe group not only in the spots of saliva, but also in the spots of sperm and vaginal secretion where heteroimmune sera failed.     

Is a Refrigerator Good or Evil? The Moral Evaluation of Everyday Objects

Certain objects such as family heirlooms are often treated as if they have intrinsic moral qualities, including sacredness and infinite value. Other objects such as instruments of torture are often seen as inherently repellent. Do people also evaluate more mundane objects such as refrigerators as morally good or bad? Here we explore the nature and scope of moral object evaluation through two experiments that asked participants to rate how morally good or bad a large set of familiar objects were. We find that (a) everyday objects tend to be seen as morally positive and (b) unlike mere liking, the moral evaluation of objects is positively linked to the age and political conservatism of the participants. These findings are discussed in relation to research on automatic evaluation, mere exposure effects, and the relationship between affective states and moral judgments.     

Hyperosmolarity Induces Armanni‐Ebstein‐like Renal Tubular Epithelial Swelling and Cytoplasmic Vacuolization

Armanni–Ebstein lesions have been considered pathognomonic for diabetes mellitus and appear as markedly swollen renal tubular epithelial cells with cytoplasmic clearing and glycogen accumulation. However, the extent to which hyperosmolarity contributes to the Armanni–Ebstein phenotype is unclear. Ten sheep were injected intravenously with 20% mannitol at 11 mOsm/kg, and subsequent histological evaluation of the kidneys showed variable degrees of osmotic nephrosis and cytoplasmic clearing of renal tubular epithelial cells similar to that seen with Armanni–Ebstein lesions. However, although morphological changes similar to Armanni–Ebstein lesions could be produced, no intracytoplasmic glycogen was demonstrated with periodic Acid–Schiff (PAS) stain. This suggests that while hyperosmolarity may contribute to the development of an Armanni–Ebstein phenotype, glycogen accumulation may result from the more complex metabolic effects of glucose on renal tubular epithelial cells. Thus, when Armanni/Ebstein‐like vacuolizations are seen at autopsy, a confirmatory PAS stain is recommended because of the potential effect of hyperosmolar states.     

The use of bacteria for the identification of vaginal secretions

We have used the 16S–23S rRNA intergenic spacer region for identifying vaginal specific bacteria. Lactobacillus crispatus and Lactobacillus gasseri were detected in vaginal secretions but not in semen, blood or saliva. Our data indicated that both L. crispatus and L. gasseri were detected in vaginal secretions from women with different levels of expression of hormonal genes including pregnant, pre- and post-menopausal women, and a woman who has had a hysterectomy. Therefore, we have demonstrated that these Lactobacilli are promising new markers for the forensic identification of vaginal secretions. We have incorporated the Lactobacilli markers into a mRNA multiplex system to produce an 11-plex assay that can identify circulatory blood, menstrual blood, saliva, semen (in the presence and absence of spermatozoa) and vaginal secretions.     

Intentional mixed buccal cell reference sample in a paternity case

We report a case where an alleged father (AF) attempted to substitute someone else's saliva sample for his reference sample in a paternity analysis. Buccal cells were collected from the AF and the child, and DNA analysis was performed using an autosomal STR loci (Identifiler). The profile from the AF showed extra peaks in some loci, as well as a much higher "X" allele peak relative to the "Y" allele peak at the amelogenin locus. After conducting reanalysis by another technician with another set of positive and negative controls, it was concluded that the only source of the mixed profile was by intentional introduction by the AF, at the time of sampling, of some foreign human biological material, most likely saliva from a woman. Owing to the inconclusive results, when the AF was called back to the lab and the peculiar results were explained to him, he admitted that he had introduced into his mouth saliva from another person in an attempt to be excluded as the father of the child. Although tampering with DNA reference samples is not common, some individuals may attempt to contaminate or otherwise adulterate specimens before DNA tests. Personnel responsible for sampling should be aware of this possibility and should try to establish procedures to avoid the problem.     

Effect of blood group active micro-organisms on the ABO grouping of human whole saliva

Three saliva samples with false positive ABO grouping results were assayed for blood group active organisms, using a variety of selective media to isolate representative strains from the salivary microflora. Eight out of 40, 8 out of 40 and 4 out of 30 strains from the three samples, respectively, showed blood group activity, which correlated well with the false positive specificities of the saliva samples. In all cases the false reaction only lasted a few days. Investigation of one of these samples before and after the appearance of the false positive activity yielded only one out of 40 blood group active organisms, using the same methods. Similar investigation of two "normal" saliva samples found none out of 40 and one out of 40 blood group active organisms, respectively. It is concluded that occasional false positive ABO grouping reactions of saliva samples are probably caused by the presence of unusually high numbers of blood group active micro-organisms, due to disturbances in the ecological balance of the salivary microflora.     

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.     

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.     

mRNA在体液斑迹鉴定与组织来源推断中的应用

体液斑迹鉴定及其组织来源推断一直是法医学研究的重要方向之一,其传统的检验方法存在诸如假阳性率高、检材易破坏等缺陷,亟需更高效的确证实验。高度分化的体细胞表达特异的m RNA分子,可推断其来源:外周血、月经血、精液、唾液、阴道分泌物、接触斑等,且灵敏度、特异度及保存时间相对较为理想,是未来用于体液斑迹鉴别的理想遗传标记。本文对m RNA在体液斑迹鉴定和组织来源推断方面的应用及其前景进行综述。     

Fatal choking in infants and children

Food asphyxiation in infants/children follows a different pattern from the adult "cafe coronary." In the absence of ethanol intoxication, infants/children are prone to mishandling nonfriable, firm, slippery foods/objects with a rounded contour. The Dade County Medical Examiner's files were searched from 1956 to mid-1983 for accidental pediatric choking deaths. Seventeen food and six foreign body asphyxiations were found. The male/female ratio was 1.4/1 and 1/1, respectively. The white/black ratio was 0.9/1 for food asphyxiation whereas no black victims were encountered choking upon foreign objects. Seventy-eight percent of all victims were between 2 months and 4 years of age. Infants/children asphyxiated on items such as a hot dog, hard candy, peanut, toy rattle, tissue paper, balloon, marble, etc. The choking event was recognized by nearby adults in most instances. Risk factors include the availability of riskful foods/objects, natural diseases with difficulty feeding, poor eating habits, and uneducated or ignorant parents/others at the scene. Although public education, package labeling, and changes in food/object design may be appropriate, the ubiquituous risk foods and small foreign objects will, on occasion, escape the eye of even the most watchful parent.     

ELISA检测精浆特异蛋白P_(30)鉴定人精斑

<正> 1978年美国Sensabaugh分离出精浆特异蛋白P30,并成功的制备出相应的抗P30血清。1987年我国也研制出抗人精浆特异蛋白P30血清,并应用该种抗血清建立了几种琼脂扩散和电泳方法鉴定人精斑。由于人精液中P30含量不高,平均为1.52±0.676mg/ml,鉴定微量精斑存在困难。为了更有效的提高方法的灵敏度。我们建立了鉴定微量人精斑的ELISA固相P30抗体法,现报告如下。     

Application of fragment analysis based on methylation status mobility difference to identify vaginal secretions

Identifying vaginal secretions attaching or adhering to a suspect’s belongings would be beneficial for reconstructing the events that have taken place during a sexual assault. The present study describes a novel approach to identify vaginal secretions by fragment analysis using capillary electrophoresis, based on the mobility differences of PCR amplicons from bisulfite-treated DNA depending on methylation status. We targeted three genome regions including each of three vaginal secretion-specific methylated CpG sites reported previously: cg25416153, cg09765089, and cg14991487. In all three genome regions, the amplicon peaks for methylated genomic DNA (gDNA) sequences were only detected in vaginal samples, whereas samples of other body fluids (blood, saliva, semen, and deposit on skin surface) only showed amplicon peaks for unmethylated gDNA sequences. In vaginal secretions, the methylation ratio of each of the three targeted regions between samples was variable, while the ratios at the three regions in each sample were similar. Furthermore, commercial vaginal epithelial cells were completely methylated at the three regions. Therefore, vaginal secretion-specific methylation may derive from vaginal epithelial cells present in the sample.In forensic cases with a limited amount of DNA, the reproducibility of a detected peak using the present method is not high due to degradation of DNA by bisulfite treatment and subsequent stochastic PCR bias. However, it was possible to detect peaks from methylated DNA sequences by performing PCR and capillary electrophoresis in triplicate after bisulfite treatment, even when bisulfite treatment was performed using 0.5 ng of gDNA from vaginal secretions. In addition, the level of methylation at each targeted region was found to be stable in vaginal secretions stored for 1 year at room temperature. Therefore, we conclude that detection of the visual peak from vaginal secretion-specific methylated DNA sequence is useful to prove the presence of vaginal secretions. This approach has the potential to analyze multiple marker regions simultaneously, and may provide a new multiplex assay to identify various body fluids.