Center for Forensic Medicine, Medical University of Vienna, Sensengasse 2, A-1090 Vienna, Austria.
Division of Livestock Sciences (NUWI), University of Natural Resources and Life Sciences, Gregor-Mendel-Straße 33, A-1180 Vienna, Austria.
Leg Med (Tokyo). 2023 May;62:102214. doi: 10.1016/j.legalmed.2023.102214. Epub 2023 Feb 15.
The unambiguous diagnosis of asphyxiation is still a major challenge for the forensic pathologist, especially in terms of highly advanced decomposed corps.
In order to demonstrate asphyxiation particularly in profoundly putrid bodies we hypothesized that hypoxic stress is basically responsible for generalized fatty degeneration of visceral organs which can be detected by histological examination using a special staining technique referred to as Oil-Red-O Stain (Sudan III-red-B-stain). To test this hypothesis we examined different tissues (myocardium, liver, lung and kidney) of 107 people divided into 5 groups. These are: (i) 71 case-victims who were found in a truck and died most likely due to asphyxiation, whereby any other violent or natural cause of death was ruled out by postmortem examination; (ii) 10 barely decomposed positive-control-victims; (iii) 6 non-decomposed positive-control-victims; iv) 10 drowning non-decomposed positive-control victims, and v) 10 negative-control-victims. Apart from general histological special staining methods, an immunohistochemically approach as a case-control-study on lung tissues of same individuals was carried out by means of using two polyclonal rabbit-antibodies against (i) HIF-1-α (Hypoxia Inducing Factor-1 alpha) and (ii) SP-A (pulmonary surfactant-associated protein A) to detect both the transcription factor and pulmonary surfactants. The positive proof of already either of them gives evidence of death caused by hypoxia.
Histological examination of myocardium, liver and kidney of the 71 case-victims and the 10 positive-control-victims using Oil-Red-O Stain showed a fatty degeneration of small droplet type; there was no evidence for fatty degeneration in tissues of the 10 negative-control-victims. These findings strongly indicate a causal association between oxygen deficiency and generalized fatty degeneration of viscera due to insufficient oxygen supply. In terms of methodology, this special staining technique seems to be very informative, even applicable on decomposed corps. Results of immunohistochemistry indicate that on the one hand the detection of HIF-1α is not possible to perform on (advanced) putrid bodies, whereas the verification of SP-A is still feasible on the other.
Positive Oil-Red-O staining and the immunohistochemical detection of SP-A can serve as a serious hint for the diagnosis of asphyxia on putrid corpses, considering other circumstances of death that have been determined.
对于法医病理学家来说，明确窒息的诊断仍然是一个主要挑战，尤其是在高度先进的腐烂尸体方面。
为了特别在高度腐烂的尸体中证明窒息，我们假设缺氧应激基本上是导致内脏器官普遍脂肪变性的原因，这可以通过使用特殊染色技术（即油红 O 染色（苏丹 III-红-B-染色））进行组织学检查来检测。为了验证这一假设，我们检查了 107 个人的不同组织（心肌、肝脏、肺和肾脏），分为 5 组。这些是：（i）71 名在卡车上发现的病例受害者，他们很可能因窒息而死亡，通过尸检排除了任何其他暴力或自然原因的死亡；（ii）10 名轻度腐烂的阳性对照受害者；（iii）6 名非腐烂的阳性对照受害者；（iv）10 名非腐烂的溺水阳性对照受害者，和（v）10 名阴性对照受害者。除了一般的组织学特殊染色方法外，还通过使用针对（i）HIF-1-α（缺氧诱导因子 1 阿尔法）和（ii）SP-A（肺表面活性剂相关蛋白 A）的免疫组织化学方法对相同个体的肺组织进行了病例对照研究，以检测转录因子和肺表面活性剂。其中任何一种的阳性证明都证明了缺氧引起的死亡。
对 71 名病例受害者和 10 名阳性对照受害者的心肌、肝脏和肾脏进行油红 O 染色的组织学检查显示出小滴类型的脂肪变性；10 名阴性对照受害者的组织中没有脂肪变性的证据。这些发现强烈表明，缺氧和内脏普遍脂肪变性之间存在因果关系，这是由于供氧不足所致。就方法学而言，这种特殊的染色技术似乎非常有信息量，甚至适用于腐烂的尸体。免疫组织化学的结果表明，一方面，在（高级）腐烂的尸体上无法进行 HIF-1α 的检测，而另一方面，仍然可以对 SP-A 进行验证。
阳性油红 O 染色和 SP-A 的免疫组织化学检测可以作为在腐烂尸体中窒息诊断的重要提示，同时考虑到已经确定的其他死亡情况。
