Smoking behavior and cytology of the cells of smokers

Cytological examinations of lung impression preparations from 131 smoker lungs revealed that the content of smoker cells within lung tissue increases up to a daily consumption of 40 cigarettes. Additional cigarette consumption does not raise the number of smoker cells further. Determination of the nuclei content in the smoker cells of groups with different consumption rates showed that the number of macrophages with more than two nuclei increases in proportion to the number of cigarettes smoked. If more than 50 cigarettes a day were smoked, many multinucleated giant cells were observed. Anamnestic inquiries proved that these cytological changes in the lungs were caused exclusively by the smoking habits of the deceased. Not only the number of cigarettes used per day, but also the manner of inhalation and peculiarities of cigarette smoking are reflected in the morphological changes of lung tissue. For the forensic pathologist, examination of lung impression preparations from smoker lungs makes it possible to note the quantity of daily cigarette consumption of a dead person to help to identify the deceased.     

The nature and the occurrence of birefringent material in different organs in fatal drug addiction

Insoluble birefringent tablet filler materials commonly found in tablets used in solution by drug addicts as intravenous injections were investigated microscopically. The following filler materials were investigated: talc, potato- and maize-starch, microcrystalline cellulose, magnesium stearate and siliciumoxid. The morphological characteristics of the different materials are described. Tissue sections (lung, liver, spleen, heart, bone-marrow, kidney, lymph-nodes and endocrine glands) from 33 consecutive fatality cases of intravenous drug addicts autopsied at the University Institute of Forensic Medicine in Copenhagen were studied with special reference to the occurrence and nature of birefringent material. Birefringent material was most often demonstrated in lung tissue (94%), followed by spleen (76%), liver (55%), lymph-nodes (portal: 39%) and bone-marrow (24%). The material was always localized intracellularly. Granulomatous reaction was only seen in the lungs. Except for one case, talc was the only foreign material seen in other organs than the lungs, undoubtedly due to its smaller size. The presence of insoluble foreign material in lung tissue of drug addicts indicates a habit of intravenous administration and the amount of the material indicates whether the addict usually injects tablets or only does so occasionally. The presence of birefringent material in the organs have only rarely any obvious clinical implications.     

Morphometric investigation of emphysema aquosum in the elderly

We compared the lungs of six drowned and six non-drowned persons over 70 to determine whether there is evidence of acute overinflation in drowning that can be distinguished from senile lung emphysema. All left lungs underwent intrathoracic formalin fixation to preserve their state of insufflation. To assess the effects of lung collapse, all right lungs were sectioned as usual and then immersed in formalin. After histological processing, microphotography and image processing of 12 specimens per corpse, 50 binary images of each specimen were measured by computerized morphometry. Intrathoracic fixation resulted in significantly less tissue area (and more airspace) in the left than in the right lungs of both groups. Comparing both groups' left lungs revealed that the interalveolar septa were thinner and the area occupied by connecting nodes smaller in drowning; these single nodes also tended to be smaller. There was a tendency for less alveolar tissue area per image in drowning than in control lungs and for narrow tissue structures to comprise a higher percentage of both the total tissue area and total tissue perimeter per image. We conclude that there is morphological evidence of acute overinflation even in senile lungs, but this is masked by postmortem lung collapse as suggested by the overlapping values of the right lungs. Support to the diagnosis of drowning among drowned elderly patients can only be available in lungs subjected to intrathoracic postmortem fixation.     

Longitudinal evaluation of immunohistochemical findings of milk aspiration: an experimental study using a murine model

To examine the longitudinal change of pathological findings of the lung and other organs in milk aspiration, an experimental study using a murine model was carried out. Either 0.5 or 1.0 ml cow's milk was instilled into the trachea of rats. From immediately after to 14 days after instillation, the animals were sacrificed, and the lungs, liver, kidneys, and spleen were removed. The results of immunostaining with anti-human α lactalbumin antibody indicated that not only the lung but also the kidney and spleen showed a positive reaction against the antibody over time. Experimentally aspirated milk was detectable in alveoli until 2 days after instillation. It was also detectable in renal tubules from 1 to 6h after instillation. Macrophages containing granules of aspirated milk were observed in splenic red pulp from 3h to 14 days after instillation. Detection of aspirated milk in other organs except the lung would be clear evidence of intravital milk aspiration and would suggest previous or recurrent milk aspiration.     

Ultrastructure of acute ammonia toxicity in the human lung

A tanker truck carrying anhydrous ammonia (NH3) fell off a freeway, releasing a dense cloud of NH3 gas, killing several people. The driver was dead upon impact. To our knowledge, pulmonary NH3 toxicity in humans has not been studied previously by electron microscopy (EM). Therefore, in two cases, the paraffin-embedded tissue blocks of lung were deparaffinized and reembedded in plastic for 1-mu sections and EM examination. The lung tissue of a third case, the truck driver, was similarly processed as a control. Light-microscopic pulmonary findings in the acute NH3 deaths included denudation of the tracheobronchial epithelium, edema of the lamina propria, and marked alveolar edema, congestion, and hemorrhage. In contrast, in the truck driver's lungs, the bronchial epithelium was intact, and there was no gross odor of NH3. Massive pulmonary hemorrhages in his lungs were attributed to trauma rather than NH3 inhalation. EM examination of the lungs of the truck driver showed no discernible toxic alterations in either the capillary endothelial cells or the Type I or II alveolar epithelial cells, and alveolar and capillary basement membranes were intact. In contrast, EM study of the lungs from two individuals dying acutely of NH3 inhalation showed marked swelling and imbitional edema of Type I alveolar epithelial cells; however, alveolar basement membranes and capillary endothelial cells appeared as usual. These electron-microscopic findings demonstrate the Type I epithelial cell to be the target cell of acute alveolar wall injury in NH3 inhalation.     

Increased lung weight in fatal intoxications is not unique to opioid drugs

Fatal intoxications with opioids are known to be associated with an increased lung weight, as well as with brain and pulmonary edema and urinary retention. However, there is evidence to suggest that fatal intoxications with non-opioid substances are also associated with increased lung weight; however, the latter aspect has not been comprehensively analyzed. To determine to what extent opioid and non-opioid substances are associated with increased lung and brain weight, we studied these organs in cases where the cause of death was attributed to intoxication with a single agent. Using data from cases autopsied at the National Board of Forensic Medicine (NBFM) in Sweden from 2009 through 2019 where the cause of death was attributed to a single substance, we created models of combined lung weight and brain weight. The models used age and sex as predictors as well as nested varying effects for the specific intoxicant and category of intoxicant. Suicidal hanging with negative toxicology cases served as controls. The population majority was male among both intoxications (68%) and controls (83%). The most common single substance group was opioids. All tested substances were associated with heavier lungs than controls, with the largest effect in the opioid group. Our findings show that several substances are associated with increased lung weight and that among intoxication deaths there is no difference in expected brain weight between substances. Hence, heavy lungs, without a reasonable explanation, should prompt a broad toxicological screening.     

Differential accumulation of pulmonary and cardiac mast cell-subsets and eosinophils between fatal anaphylaxis and asthma death: a postmortem comparative study

The distribution profile of infiltrated mast cell-subpopulations and eosinophils in the lung and heart sections of the patients who died of severe allergic hyperresponsiveness, was investigated. Four study groups were designed comprising 9 cases who died in systemic anaphylaxis (Group I), 10 asthmatic individuals whose death were assigned to acute and severe bronchial asthma (Group II), 10 asthmatic cases who died from non-immunological diseases (Group III). Twenty consecutive autopsies of non-allergic subjects who died of unnatural causes (Group IV) served as control group in this study. Utilizing antibodies against human tryptase and chymase and a double immunohistochemical staining method, we distinguished successfully all three subsets of mast cells (MC), MC-TC (containing both tryptase and chymase), MC-T (containing only tryptase) and MC-C (containing only chymase) types, subdivided on the basis of the protease compositions of their secretory granules. In order to immunostaining eosinophils, we used antibody to major basic protein as a marker. We also measured postmortem blood tryptase, specific and total serum IgE. The intriguing finding of this study was the marked differences of cellular composition in the lung between fatal anaphylaxis and asthma death. Significant augmentation of MCs infiltrated in lung and heart sections of anaphylaxis patients and drastic infiltration of bronchial eosinophils in asthmatic death and consequent release of their related inflammatory mediators might explain the differential expression of the associated symptoms in these two groups. The anaphylactic deaths did show neither emphysema nor significant mucous bronchial secretions whereas all asthmatic deaths did. The degree of pulmonary congestion and edema was also more severe in anaphylaxis. This corresponded with the histological findings and the location and number of mast cell-subsets and eosinophils in the different compartments of the lungs. We have demonstrated that the third type of mast cell MC-C is only found in the lungs in anaphylactic deaths. The practical consequence of our study will be that it is now possible to confirm a suspicion of anaphylaxis death not only by measurements of serum mast cell tryptase, but also by immunohistochemical methods.     

Immunocytochemical study on the ultrastructural localization of human-type ABO (H)-blood group activities in a macaque (Macaca irus)

The immunocytochemical study on the ultrastructural localization of human-type ABO(H)-activities in a crab-eating macaque (Macaca irus) was carried out by using postembedding and immuno-gold staining method. The tissue specimens examined were the esophagus, stomach (St), small intestine (Si), large intestine, liver, kidney, and pancreas. The specimens from these organs and submandibular gland (Sg) of a human (O-group) were used as staining reaction controls. Primary and secondary antibodies were commercially obtained mouse monoclonal anti-A, -B, -H (IgM), and goat anti-mouse IgM labeled with colloidal gold particles (luminal diameter 20 nm), respectively. The results were as follows: (1) In macaque specimens, only A-activity could be observed as the location of gold particles on the peripheral rim of serous secretory granules (Sg) and of epithelial cells (esophagus), the mucous droplets in epithelial cells and brush border (St, Si), the intracellular secretory canaliculi [ISC (St)] and the zymogen granules and secretory ducts (pancreas). Gold particles could be also noted at the Golgi apparatus and nascent secretory granules. (2) By periodic acid-thiocarbohydrazide-osmium tetroxide (PA-TCH-OS) reaction, hexose-rich neutral mucopolysaccharides were noted on the peripheral rim of serous secretory granules (Sg), the mucous droplets (St, Si), the ISC (St), and the brush border (Si). Such a distributional pattern corresponded well with that of gold particles, indicating that the substances were responsible for ABO(H)-activities.     

Determination of MDMA, MDA, MDEA and MBDB in oral fluid using high performance liquid chromatography with native fluorescence detection

This paper describes the analytical methodology for the determination of MDMA, MDA, MDEA and MBDB in oral fluid. After a liquid–liquid extraction, the analysis was carried out by high performance liquid chromatography (HPLC), with fluorescence detection. The detector wavelength was fixed at 285 nm for excitation and 320 nm for emission. The mobile phase, a mixture of phosphate buffer (pH = 5) and acetonitrile (75:25), and the column, Kromasil 100 C8 5 μm 250 mm × 4.6 mm, allowed good separation of the compounds in an isocratic mode in only 10 min. The method was validated and showed good limits of detection (2 ng/mL) and quantitation (10 ng/mL) for all the amphetamine derivatives. No interfering substances were detected. A stability study of these compounds in oral fluid stored at three different temperatures (−18, 4 and 20 °C) over 10 weeks was conducted, showing a time-dependent degradation of the four compounds.     

Spectral enhancement of leucocrystal violet treated footwear impression evidence in blood

The results presented demonstrate the capacity for spectral enhancement to substantially improve the forensic examination of footwear impressions in blood treated with leucocrystal violet (LCV). The UV-Vis absorption spectra were generated of (i) an aqueous solution of leucocrystal violet, (ii) leucocrystal violet in 3% H(2)O(2), (iii) LCV working solution and (iv) whole blood added to LCV working solution. The resultant fluorescence emission spectra were subsequently generated (lambda(ex)=630nm, lambda(em)=661-900nm). The results indicate that the UV-Vis absorption spectra of an unbuffered solution of whole blood with LCV working solution produces a strong absorbance curve with a maxima at 630nm. Subsequent excitation at this wavelength and generation of the emission spectrum in the fluorescence mode indicates that a solution of whole blood added to LCV working solution is an extremely weak fluorophore. Therefore, to enable an adequate and timely enhancement of blood impression evidence treated with LCV utilising either visible fluorescence or infrared luminescence requires (i) selection of the most appropriate excitation wavelength (lambda(ex)) and emission wavelength (lambda(em)) with extremely narrow band pass filters, which in the absence of substrate matrix interference is excitation at 630nm producing the emission maxima at 665nm and (ii) a visual enhancement system such as a CCD colour IR video camera with image integration.     

3种甲苯胺蓝异染肥大细胞方法的比较

目的比较3种甲苯胺蓝异染肥大细胞方法的应用性。方法采用甲苯胺蓝水溶液与高锰酸钾配比染色（方法一）、甲苯胺蓝酒精溶液和氯化钠溶液配比（方法二）、甲苯胺蓝水溶液染色、冰醋酸分色（方法三）3种方法对人和小鼠肺组织冰冻切片进行染色,比较染色结果、并评价方法稳定性及实用性。结果方法一染色肺组织切片,结果切片蓝色背景清晰,肥大细胞胞质内紫红色颗粒鲜明,与背景蓝色对比度高,易辨识,且多次重复染色效果稳定;方法二及方法三肺组织的蓝染背景、和肥大细胞胞浆颗粒的紫红色染之间的对比度均不及方法一清晰,稳定性亦次之。结论甲苯胺蓝水溶液与高锰酸钾配比染色法异染肺组织冰冻切片,结果清晰、操作简便且稳定性好,适合在相关检验中选用。     

Microchip-based cell lysis and DNA extraction from sperm cells for application to forensic analysis

The current backlog of casework is among the most significant challenges facing crime laboratories at this time. While the development of next-generation microchip-based technology for expedited forensic casework analysis offers one solution to this problem, this will require the adaptation of manual, large-volume, benchtop chemistry to small volume microfluidic devices. Analysis of evidentiary materials from rape kits where semen or sperm cells are commonly found represents a unique set of challenges for on-chip cell lysis and DNA extraction that must be addressed for successful application. The work presented here details the development of a microdevice capable of DNA extraction directly from sperm cells for application to the analysis of sexual assault evidence. A variety of chemical lysing agents are assessed for inclusion in the extraction protocol and a method for DNA purification from sperm cells is described. Suitability of the extracted DNA for short tandem repeat (STR) analysis is assessed and genetic profiles shown. Finally, on-chip cell lysis methods are evaluated, with results from fluorescence visualization of cell rupture and DNA extraction from an integrated cell lysis and purification with subsequent STR amplification presented. A method for on-chip cell lysis and DNA purification is described, with considerations toward inclusion in an integrated microdevice capable of both differential cell sorting and DNA extraction. The results of this work demonstrate the feasibility of incorporating microchip-based cell lysis and DNA extraction into forensic casework analysis.     

Visualization of latent fingerprints using fluorescence lifetime imaging on paper emitting strong fluorescence

Latent fingerprints were successfully visualized using fluorescence lifetime imaging (FLIM) on paper which emits strong fluorescence with a lifetime close to that of fingerprints and thus from which it is difficult for time-resolved spectroscopy to visualize fingerprints. Latent fingerprint samples on paper were excited using a 450 nm or 532 nm nanosecond pulsed-laser, and time-resolved fluorescence images were obtained at a delay time of 6–16 ns in intervals of 1 ns, to the excitation pulse. The excitation beam was expanded using a lens, and the fluorescence from the fingerprints was captured using an intensified CCD camera. Because of the large fluorescence intensity of the background paper of approximately two to four orders of magnitude larger than that of the fingerprint, the fingerprint was not visualized on each fluorescence image by time-resolved spectroscopy. However, the fingerprint was visualized in a FLIM image constructed using a series of the fluorescence images for the case with the fluorescence intensity of the background paper being four orders of magnitude larger than that of the fingerprint. The difference in fluorescence lifetime in the FLIM image of the visualized fingerprint and background paper was in the order of 0.1 ns, which was an order of magnitude smaller than the inherent fluorescence lifetime of a few nanoseconds for the fingerprints and paper. It was demonstrated that, at a background fluorescence intensity with a certain order of magnitude larger than that of fingerprints, FLIM has the potential to visualize latent fingerprints which cannot be visualized by time-resolved spectroscopy.     

Death associated with volatile substance inhalation--histologic, scanning electron microscopic and energy dispersive X-ray spectral analyses of lung tissue

The investigation of deaths due to the inhalation of volatile substances may be complicated by a lack of scene and autopsy findings. Mechanisms of death may not be determinable at autopsy, and there may be very few markers of inhalant abuse. A 21-year-old man is reported who died from the combined effects of methadone toxicity and toluene inhalation. Histological examination of the lungs revealed congestion and edema, as well as particles of blue, pigmented material within the interstitium and in macrophages. Scanning electron microscopy was undertaken, revealing that the particles contained granules that measured 0.15-0.2microm in diameter, within the range of mean particle sizes for inorganic paint pigments. Energy dispersive X-ray spectral analysis of the granules demonstrated a significant percentage of titanium (12%) confirming their origin from paint. Ancillary investigations such as electron microscopy and X-ray spectral analysis in cases of possible lethal volatile inhalation may prove useful adjuncts in determining the type of substance inhaled and in providing evidence of previous non-lethal episodes.     

Current strategic approaches for the detection of blood doping practices

Aerobic sport performance may be strongly influenced by the number of red blood cells available for transport and delivery of oxygen from lungs to muscles. Often, athletes search for an acute increase in red blood cells by means of blood transfusions. This paper reviews the possibilities for detecting such prohibited practice. Flow cytometry methods are able to detect a double population of red blood cell membrane surface antigens, thus revealing an allogeneic transfusion. Other ingenious approaches for total hemoglobin mass measurements or to test for the metabolites of blood bag plasticizers in urine are new trends for facing the detection of autologous transfusions. Steady increase of red blood cell number may be obtained also by erythropoietic stimulant agents such as erythropoietin, analogs and mimetics. The challenge of detecting those substances has stimulated the development of indirect markers of altered erythropoiesis, leading to the consequent development of the hematological blood passport approach, which is gaining legal acceptance.     

Immunohistochemical study of blood group activities in severely burned human tissue

The blood, livers and lungs obtained from donors or cadavers of known blood groups were experimentally burned, while the temperatures inside the tissue specimens were automatically measured. All the distinguishable portions with various thermo-changes in each burned tissue specimen were examined for their blood-group activities A, B, Lea, Leb and P1 by means of the immunofluorescence and immunoperoxidase techniques. In the layer immediately before charring of the blood masses (tissue temperature: ca. 200-250 degrees C), the activities A, B, Lea, Leb and P1 could be specifically demonstrated on the erythrocyte membranes. In case of the burned livers and lungs, the A- and B-activities could be detected in the small blood vessels in the layer immediately before charring (ca. 200-250 degrees C), though the layer was so severely thermo-changed, that their original tissue structures could be no more observed. In the severely thermo-coagulated, porous and hardened layer of both organ tissues, the A- and B-activities could be demonstrated on the endothelial cells of the hepatic sinus and small blood vessels (especially in the alveolar walls). The simply thermo-coagulated inner portions well retained their original tissue structures and the A- and B-activities remained on the epithelial cells of the alveoli and bronchioles of the lung as well as the cells described above. The Lewis blood-group activities could be demonstrated on the epithelial cells of the bronchioles in the simply thermo-coagulated inner portion of the lung. The P1-activity could not be demonstrated in the burned tissues of the livers and lungs.     

人体组织中ABH物质分布的研究

本研究采用特异性红细胞粘附试验(SRCA 试验)方法,系统地研究了11例已知 ABO 血型及分泌状态尸体的38种组织器官 ABH 物质的定位与分布,发现粘膜、粘液腺及前列腺中均含有丰富的 ABH 物质,并受分泌状态控制。血管内皮细胞、复层上皮细胞、胰腺腺泡上皮及汗腺中也含有较多的 ABH 物质,但不受分泌状态的影响。新发现肺泡上皮、肝小胆管粘膜上皮亦含有 ABH 物质。其它组织器官除自身的血管内皮及红细胞含有 ABH 物质外,均未测出 ABH 物质。采用 SRCA 试验,室温放置13天的皮肤组织亦能正确地测出其 ABO 血型。     

Prevalence of nicotine consumption in drug deaths

One hundred consecutive drug death victims autopsied at the Institute of Forensic Medicine, University of Freiburg, between 1995 and 1997 were studied retrospectively as to whether the drug users had also consumed nicotine. The study included histological examination of the lung tissue for smoker cells and radioimmunological as well as GC-MS assays of the urine for cotinine, the primary metabolite of nicotine. It was found that 98 out of 100 drug victims had consumed nicotine in addition to illicit drugs or replacements. Yellowish-brown discolorations on the middle and index fingers were discernible in 44 drug victims, whereas fresh or scarred burns due to glowing cigarettes were found in six deceased drug consumers. Diseases of the bronchial system typical of heavy smokers were seen in 35 cases. Siderophages could be demonstrated in 17 of the 100 drug deaths.     

低温荧光法显现汗潜指纹初探

目的　探索汗潜指纹显现的新方法。方法　利用低温荧光摄影方法加强指纹沉积物的固有荧光强度 ,来提高汗潜指纹显现的成功率。结果　在 4 1 5nm波段激发下用黄滤色镜拍摄指纹的显现效果最好。结论　低温荧光法是一种有效的潜指纹显现的技术手段     

The reaction of lung mast cells to acute barbiturate and bromureide poisoning (author's transl)]

Guinea pigs were poisoned by large oral doses of hypnotics in order to study the effect of bromureides on the mast cell contents of the lungs in comparison to the influence of barbiturate intoxication. The dose of the hypnotics chosen was thus, that all animals died within the first three hours after the beginning of feeding. Then the degranulation of mast cells with decrease of the mast cell number in the lung tissue was merely small in the barbiturate poisoned animals, but very extensive in the animals intoxicated by bromureides. The difference is highly significant. The possible influence of this effect upon the different clinical course of both kinds of intoxication is discussed.