The evaluation of an ELISA for semen-specific 19-OH prostaglandin F1 alpha/F2 alpha using ex-casework swabs and stains

The application of an enzyme-linked immunosorbent assay (ELISA) for 19-OH Prostaglandin F1 alpha/F2 alpha (19-OH PG F) to casework analysis of seminal contamination of swabs and stains is reported. The results are compared to those where the identification of semen was based on the presence of acid phosphatase and spermatozoa. Five hundred and one samples were analysed and there was good agreement between the results of ELISA and conventional methods. The determination of 19-OH PG F confirmed both the presence of semen where spermatozoa were absent and indicated semen was present when acid phosphatase and spermatozoa were both absent. The results indicate that 19-OH PG F represents a useful marker for the casework identification of semen and is particularly valuable where spermatozoa are absent.     

The application of a simple RIA technique for the detection of 19-OH F1 alpha/F2 alpha prostaglandin, a specific semen marker, in semen contaminated vaginal swabs: time since intercourse studies

The specificity of the 19-OH F1 alpha/F2 alpha prostaglandin antisera for the detection of semen in seminal/vaginal mixtures, has been evaluated. Using a parallel curve test we found that the antibody showed a high specificity for these seminal prostaglandins in seminal/vaginal mixtures at concentrations of between 2 pg and 40 pg/100 microliter. The precision of the assay has been improved by the use of a donkey-anti-rabbit ferritin-bound second antibody. The application of this detection system makes it possible to complete an assay within 4.5 h. A survey of 50 semen-free vaginal swabs obtained from 3 donors, taken throughout the menstrual cycle, showed no trace of these prostaglandins. They were also not detected in the vaginal secretions of two further donors who were undergoing medication. Using only 10-microliter aliquots of a seminal/vaginal swab extract, prepared in 500 microliter, it was possible to detect semen up to approx. 80 h after one sexual act.     

Preliminary study on using changes in astrocytes and neurons in the brain-stem of the rat after stab wounds for determining the time of injury

The changes in astrocytes and neurons in the brain-stem of the rat after stab wounds were studied immunohistochemically. Anti-GFAP and anti-albumin antibodies were used to detect the changes of astrocytes and neurons, respectively. The results showed that in normal control group, the numbers of GFAP-positive cells differed in different areas of brain-stem, but remained similar in the same area of different rats. No albumin-positive cells could be found. The number of GFAP-positive cells in antemortem injury groups increased significantly followed by a decrease, and then a further increase. Albumin-positive cells including neurons and glial cells were found after injury in the midbrain, pons and medulla oblongata. In the postmortem injury groups, there were no significant changes in either anti-GFAP or anti-albumin immunoreactivities in any area of brain-stem compared to the control group. These results indicated that the changes in the number of GFAP-positive cells could be used to estimate the time of injury and to diagnose early brain-stem injury. The antemortem and postmortem injuries can also be differentiated by the presence of albumin-positive cells. The authors suggest that the timing and early diagnosis of injury in the brain-stem should be carried out on the basis of comprehensive analyses of changes in both neurons and glial cells.     

大鼠死后不同时间坐骨神经雪旺氏细胞核DNA的变化

目的探讨大鼠死后不同时间坐骨神经雪旺氏细胞核DNA的变化及其与死亡时间的关系。方法应用Feulgen染色法及计算机图像分析技术,观测大鼠死后即刻至9d坐骨神经雪旺氏细胞核DNA染色变化及其图像分析参数,所得数据进行统计学分析。结果雪旺氏细胞核在死后6-18h细胞核呈紫红色、长椭圆形,轮廓清晰,其长轴方向与坐骨神经走向一致,死后24h着色变淡、着色面积变小,最终完全消失;在6h-9d内目标面积和积分光密度随死亡时间的延长而逐渐减小,且其均值均与死亡时间明显相关,并得出对应的回归方程。结论大鼠死后9d内,其坐骨神经雪旺氏细胞核DNA变化与死亡时间有一定的相关性。     

人体皮肤创伤组织细胞因子表达的时间相关性

目的探讨TGF-β1、b—FGF和VEGF在人体皮肤创伤组织中的表达变化及其与损伤时间的关系。方法收集15例机械性致伤的人体皮肤组织(受伤5—120min内死亡),常规制作组织切片后进行免疫组化染色,并统计分析TGF-β1、b—FGF和VEGF因子的表达情况。结果 TGF-β1在16—50min组表达升高,60～120min组呈阳性至强阳性表达;b—FGF在16—50min组表达升高明显,强阳性表达亦出现在60～120min组;VEGF仅在60—120min组出现阳性表达。统计学分析表明,TGF—β1和b—FGF的表达升高分别在受伤60~120min(P<0.01)和在16—50min(P<0.01)有显著性意义,VEGF表达升高只在60~120min(P<0.05)有显著性意义。结论 TGF-β1、b—FGF和VEGF的表达时间和表达强度与损伤时间有关;TGF-β1、b—FGF和VEGF的表达可为人体短存活期损伤时间的推断提供参考。     

人体皮肤切创纤维连接蛋白EDA、EDB的表达与损伤时间关系的研究

目的研究纤维连接蛋白(fibronectin,FN)可变剪接片段EDA、EDB在人体皮肤切创的表达情况,并摸索EDA、EDBmRNA在离体皮肤上最长检出时限,以期为早期损伤时间的推断提供实用的参考指标。方法采用DIG标记的反意RNA探针,原位杂交检测人体皮肤在损伤早期(从30min开始)FNEDA、EDBmRNA表达的变化。结果正常人体皮肤无EDA、EDBmRNA表达;在损伤早期(≤3h)FNEDA、EDB的表达随时间的延长而呈现一种不断上升的趋势;EDA、EDBmRNA主要出现在表皮基底层细胞;EDA、EDBmRNA在标本离体后4h就不能检测到。结论FNEDA、EDB可作为早期损伤时间推断的较理想的参考指标,但原位杂交的检测方法不适用于实际应用。     

兔静脉空气栓塞死后不同间隔时间气体变化的初步研究

目的探讨兔静脉空气栓塞死后不同间隔时间气体变化的规律,为空气栓塞致死的法医学鉴定提供参考资料。方法建立兔静脉空气栓塞动物模型,按死后不同间隔时间分为0h、12h、1d、2d、4d、8d和16d共7组,分别从右心检测气体体积及其中CH4、CO2和N2体积百分比浓度。结果实验组均收集到气体,阴性对照组除了16d组收集到0.95ml外,均未收集到气体（V〈0.1ml）。实验组中,0h、12h和8d3组间所收集气体的差异无统计学意义（P〉0.05）,1d和2d两组之间右心气体体积的差异无统计学意义（P〉0.05）,其它各组两两之间均存在统计学差异（P〈0.05）。实验组检测气体体积呈先下降后上升趋势,CO2浓度上下波动,气体成分均符合典型空气栓塞气体成分标准;16d对照组气体接近于腐败气体。结论兔静脉空气栓塞气体体积在死后16d内呈先轻微下降后上升的规律;静脉空气栓塞死后8d内检测结果准确、可靠,而8~16d的检测结果需结合其他证据综合分析,才能确定是否为静脉空气栓塞致死。     

Enzyme activities of leukocytes in skin cut wounds with various wound ages

A study was carried out on leukocyte enzyme activity from prints of skin cut wounds of guinea pigs at various time intervals after the injury. The following enzymes were examined: alkaline and acid phosphatase, succinate, lactate, and glucose-6-phosphate dehydrogenases. Cytochemical analysis revealed a rapid increase in enzyme activity in the 4th h after the wound occurred, which can be explained by the alteration in leukocyte metabolism induced by the damaging agent, i.e., the skin trauma inflicted. The rise in lactate dehydrogenase and glucose-6-phosphate dehydrogenase activity is the most characteristic feature. In accordance with these changes in enzyme activity, some conclusions can be made about certain intracellular alterations. There is a close relationship between the course and the degree of these cellular changes and the time interval between the injury and examination of the enzyme activity. These changes make it possible to draw conclusions concerning the vital character and the interval after inflicting as well.     

Radioimmunoassay detection limits for 19-OH F1 alpha/F2 alpha prostaglandin in normal, infertile and vasectomized semen stains. Analysis of saliva, sweat and urine for possible non-specific or matrix effects

The sensitivity of a simple radioimmunoassay (RIA) for the detection of 19-OH prostaglandin F1 alpha/F2 alpha (PG F) has been evaluated on a number of semen samples from vasectomized, infertile and fertile donors. The specificity of the test has been examined by testing saliva, sweat and urine from a number of male and female donors. The assay technique could readily detect the PG in semen stains prepared from 0.2 microliter of semen from normal, infertile and vasectomized donors. The detection limit of the assay system, based on the observed displacement, was calculated to be approximately 0.05 microliter semen. The assay could be conducted over a pH range of 7.5-10.5 even after the PG has been heated to 100 degrees C. The 19-OH series of PG were absent from sweat, saliva and female urine using the normal assay protocol; volumes in excess of 100 microliters of some urines particularly from women in labour and those with acute urinary tract infection showed some displacement. Low levels of PG were detected in 50% of the male urine analysed. However, urine samples from men who had engaged in recent sexual activity contained relatively high concentrations of PG which could be readily detected in 10 microliters of urine. These results emphasise the potential of these compounds as specific and sensitive markers for the presence of human semen.     

Changes in the lipid composition of latent fingerprint residue with time after deposition on a surface

The work described in this report was focused on generating increased knowledge of fingerprint chemistry, particularly the composition of a latent fingerprint at the time it is deposited, and the chemical changes in lipid components that occur over time. Fingerprints from five male donors (aged 25-34 years) were collected and aged under controlled conditions. The prints were then sampled at set intervals, solvent extracted with dichloromethane, co-derivatized with MSTFA and analyzed by gas chromatography-mass spectrometry (GC-MS). It was shown that there was loss of squalene from prints stored in the light or in the dark. Loss was more rapid in the light, with squalene in prints from some donors not detected after 9 days storage. For these same donors, squalene was still detected after 33 days storage in the dark, but at much lower levels than in fresh prints. For saturated fatty acids (tetradecanoic, palmitic and stearic acid) there was a trend towards an increase in levels of these substances during storage (up to about 20 days) followed by a decrease back to original levels or below. This was the case for samples stored in the light or in the dark. For palmitoleic acid, a similar trend was seen. For oleic acid, this trend was seen for samples stored in the dark. For samples stored in the light the general trend was a decrease in level over the storage period (up to 33 days).     

Structural changes to the skin in the area of gunshot wounds]

Microscopic changes in skin structures around gunshot and stabbed wounds, which may be used as features characterizing injury volume were studied. Factors influencing the significance of these changes were presented and mechanisms of their occurrence is discussed.     

大鼠脑外伤后脑组织MMP-9的表达与损伤时间相关性

目的观察大鼠脑外伤后脑组织MMP-9的表达规律,探讨其在损伤时间推断中的应用价值。方法健康SD大鼠75只,随机分为损伤组、假损伤组和正常对照组。损伤组采用自由落体方法建立脑损伤模型;致脑损伤后分10个时间点,采用HE染色观察损伤区的变化,采用免疫组化方法检测各时间点阳性细胞的种类和比值,并与假损伤组和对照组进行统计分析比较。结果免疫组化染色显示损伤组伤后1h可见中性粒细胞阳性着色,6h后可见神经细胞阳性染色并逐渐增强,伤后5d阳性细胞数达到峰值,而后开始减弱,14d基本消失。假损伤组和正常对照组未见阳性表达。将实验组中2h～7d阳性细胞比值与其它各时间点相比,差异有统计学意义(P<0.05);实验组各时间点与假损伤组和对照组相比,差异均有统计学意义(P<0.05)。统计分析结果见表1。结论损伤后MMP-9阳性表达的规律性,有望成为脑损伤时间判定的参考指标。     

Evaluation of the 190H analogues of prostaglandins E1, E2, F1 alpha and F2 alpha as specific markers for the identification of human semen in body fluid mixtures

The specificity of antisera raised against each of the prostaglandin series 190H E1/E2 and 190H F1 alpha/F2 alpha, produced in males, was evaluated by radioimmunoassay. Further, the ability of these antisera to detect semen specific prostaglandins in mixtures of body fluids was examined. Antisera directed against the 190H E1/E2 series cross-reacted with prostaglandin E1 and marginally with E2. Antisera raised to the 190H F1 alpha/F2 alpha series were, however, highly specific to the semen specific prostaglandins 190H F1 alpha/F2 alpha and 190H E1/E2. It was possible to detect picogramme quantities of contaminating 190H F1 alpha/F2 alpha on vaginal swabs taken up to 72 h after intercourse and on vaginal swabs stored at room temperature for up to 2 years. These prostaglandins were not detected on semen free vaginal swabs, in faecal material, saliva, urine or in a sample of human milk (stain). A limited study of casework material is also described. Detection of the 190H F series, as a group, has considerable potential in the identification of human semen at picogramme levels, eliminating the need for alternative chemical tests and extensive microscopic examination.     

Effect of different post-feeding intervals on the total time of development of the blowfly Lucilia sericata (Diptera: Calliphoridae)

By estimating the age of the immature stages of flies developing on a corpse, forensic entomologists are able to establish the minimum post-mortem interval. Blowflies, which are the first and most important colonizers, usually leave the cadaver at the end of the last larval stage searching for a pupation site. This period of development is referred as the post-feeding or wandering stage. The characteristics of the ground where the corpse was placed might be of notable importance for the post-feeding dispersal time: For pupariation the larvae prefer an environment protected from light and predators and may have a longer dispersal time in order to reach an appropriate pupation site. Hence, the dispersal time can vary and may influence the total time of development which may lead to an erroneous calculation of the post-mortem interval. This study investigates the effect of various post-feeding time intervals on the development of the blowfly Lucilia sericata at a temperature of 25°C. As larvae reached the post-feeding stage a pupariation substrate was offered at 0 and after 12, 24 and 48h. Only the larvae with a dispersal time of 24h (total time of development 325.2h; median) and 48h (total time of development 347.7h; median) showed a significantly longer total development time compared to the control group (total time of development 318.4h; median). The mortality rate did not differ between groups; however the flies that emerged from the group with a dispersal of 48h were significantly smaller indicating increased energy consumption during dispersal. The results of this study indicate that a prolonged post-feeding stage could increase the total developmental time of L. sericata which should be taken into consideration when interpreting entomological findings. The need for a serious examination of current rearing practices in forensic entomology laboratories is indicated because reference data sets for the time of development are usually produced by offering the post-feeding stage a substrate for pupariation immediately.     

Survival time, capacity of action and radiological diagnosis after gunshot wounds of the cranium (author's transl)]

Three cases of suicidal temporal gunshots. Different circumstances in the clinical course and in the ability of action.--In one case there was a bullet tract in the lower temporal region with considerable destruction of the base of the skull, followed by immediate inability of action and early death. Another case with a transtemporal gunshot without radiological signs of a laceration of the base of skull showed severe disturbance of cerebral functions for some days and remaining blindness. In the third case there was a pistol shot in the temporal region with retained missile, full ability of acting and undisturbed consciousness. Causative for the differences in the effect of the gunshots are differences of weapons and amunition and the anatomical position of the wound track in the temporal region. Discussion about the role of concussion of the brain by different rate of transgression of energy from the missile to the skull. Reference to the importance of X-ray analysis and to a radiographic documentation of the findings in such cases.     

大鼠皮肤挫裂创细胞间粘附分子-1的表达

目的 观察大鼠皮肤钝器伤后细胞间粘附分子-1(ICAM-1)表达的变化规律。方法 应用免疫组化方法,观察大鼠皮肤钝器伤后1h、3h、6h、12h、1d、3d、5d、7d、9d ICAM-1的表达情况,并用MIAS图像分析系统进行图像分析。结果 正常对照组大鼠皮肤有低水平ICAM-1表达,钝器伤后ICAM-1表达增加,伤后7d达峰值,伤后9d开始下降。结论 皮肤钝器伤可诱导ICAM-1表达增加,其时序性变化可望作为一种客观指标用于法医学损伤时间的推断。     

The macro- and micromorphological signs of the edge of stabbing-cutting weapons in skin wounds

110 stab-cut skin wounds in 34 dead bodies as well as 38 stab-cut instruments were studied. Three morphologic types of traces left by blade edges of stab-cut instruments, i.e. puncture, microrupture and microcut, were established. Gross and micromorphology of each of these traces depends on the properties of blade edges of stab-cut instruments.