Not Everything Dies: An Autopsy & A Body

The forensic pathology is a field of study which is the most accurate in determining the causes of death. The modern techniques allow to estimate almost the exact time of death, sadly in some circumstantial situation it is impossible to pronounce and estimate the exact time of demise. An autopsy reveals not only some infections and diseases but also the cause which might be non or accidental. We must remember forensic pathology concerns itself with the intersection of the legal world and the pathologic world. There is a bias of forensic pathology toward the pathology of injury. As opposed to illness as injury is, it is frequently involved in civil and criminal litigation. Not to mention that it is of great importance to the legal world. Many of illnesses are of importance of forensic pathology. There is always a treat of exposure to injurious agent. The causes of death are bias. The pathology report defines the cause of death, yet, it does not give a definition of death. The investigative techniques of forensic pathology are equally applicable in the determination of the cause of death which was proceeded by injury, disease and the case whether the person was dead or alive. Pathology to the mid-nineteen century was a single specialty, then the other branches of the science emerged such as [1]anatomic pathology, [2]clinical pathology, and [3]forensic pathology. The main and predominant function of forensic medicine is the examination of the dead body. The unusual or suspicious death must be investigated in some legal ways, having a particular protocol to follow.  It is a judge who is empowered to investigate thus inquire, for that reason there is an inquisitional system of administration of justice. In majority of deaths a physician who took care of a deceased while alive is acknowledged to certify the death. The physician should be confident the person is deceased and he or she died of illness or disease. The cause of death is apparent and obvious.  When the physician is not confident, the case is directed to the category of non-certifiable death. The appropriate authorities are acknowledged about this type of death e.g. coroner or medical examiner. The law officer decides if the case may be signed out as a natural death or it requires further investigation and autopsy examination. Why the post mortem examination is important?

It determines the causes of death. The naming might be a bit confusing, its meaning comes from Greek words ''auto'' – self and ''opsy'' – look. Well it is not exactly the self look at the body, We have different notions that describe it more accurately such as: ''nercopsy'' which means  - ''necros'' – death and ''opsy'', or ''biopsy'' which stands for ''bio'' – life and opsy.  Autopsy if someone does not perceive its accuracy in naming may be called – a post mortem examination.  It is detecting, describing, and interpreting any signs of injury or disease that may be present but are useful in understanding as much about how the person came to be deceased as possible. Forensic pathology is involved in the pathology of trauma or injury it is distinguished as homicidal, accidental, suicidal, in cases, where the bodily damage and injury may be the major finding.

Every type of disease is concerned as important, the death may seem natural, yet, some causes my be termed as ‘injurious’ as in the chest tumors associated with asbestos exposure or the bladder tumors associates with  [4]aniline dyes. Most of the cases are injuries based on mechanical trauma such as shooting, stabbing, punching,  road, rail and air accidents, suicide. The medicolegal officer is about to determine in four part system which death it was: homicidal, accidental, suicidal or natural. The natural deaths are all those without any violence or injury. Accidental deaths are those that result  from injury where was no intent to harm the person, homicidal  and suicidal deaths  are those resulting from some intent to cause an injury or death. In homicide the subject is a person other than a deceased. In suicide the subject is a deceased. Autopsy gives  a time frame within the death probably could not happen and the possibility it did.

“The pathologist has some certain pattern to follow; the overall procedures of undergoing an autopsy are worldwide the same, the main purpose is to look for injures, wounds, abrasions that directly or indirectly led to demise”.

The beginning starts of evaluation of the identity of the person next of  body surface itself. The identification of the person gives us some kind of insight how the person lived and did just before he or she died. The social background is very important. Before the external procedure starts the body is measured; it is very important to estimate or rule out the following factors:

-          if small assailant could have damage a large victim;

-          if the height of injuries above the ground of stabbing, gunshot and transportation are at issue;

-          it determines the physical characteristics of the deceased when alive.

The clothes and any personal belongings are thoroughly examined at the criminal laboratory, clothing is checked to detect presence of blood, vomitus, semen, fecal material. The general overlook of the body whether it is clean or not; it may also directly show self-neglect,  child neglect, mental disorders, drug dependency, and senile dementias. The dirt and skin infections may indicate all these factors.

The state of the body regarding stiffness should be noted. The state of rigor mortis is important in the moment the body is found because it indicates the possible time of death. The photography is a routine, the body is photographed. The X- Ray is performed in case of a child’s demise a complete skeletal X-Ray is taken.

In case of traffic accident numerous samples are taken to be analyzed such as: glass fragments from windshields, fibers, blood, semen, extraneous hairs, skin fragments, fingernails … etc. Again all is cataloged and photographed. The toxicological test of DNA and [5]PCR are taken to gather a material which indicates a possible assailant from submicroscopic amounts of tissue.

The internal autopsy is begun by ‘Y’ incision, it is done to avoid any visibility of the incision when the body is viewed and to preserve the large arteries in the neck for use to inject embalming fluids. It is also possible to make an incision on the front of the neck up to the larynx. It enables to examine the contents of the head, neck, thorax,  and abdomen.  The brain is examined by cutting the scalp over the vertex of the head from behind the ear, the cut traversing the upper part of the scalp after that the scalp is reflected back from the skull, the top of the skull is sewed and removed exposing the brain for examination.  After a complete autopsy the body is sewn . The fact that the autopsy was done is not discernable by persons viewing the clothed body.

“Blood samples are taken in all deaths.”

Taking samples of stomach content, of bile content, of uterus content, of blood, and other body fluids is a must! The significance of an [6]ocular fluid is a core autopsy procedure. Unpleasant to the eye as it is, it does a value of ‘tempus fugit’ – it gives an almost accurate and proximate indication of the time of death.

The full autopsy report is unbiased. It states and describes the cause of death. It doesn’t give any opinions or reflections. When toxicological analysis is done and the result can be viewed by the family of the deceased, it  is added to the post mortem examination report approximately three weeks later. The life function of the body may long gone, yet, in modern forensic medicine not everything dies, and the preliminary function of law – enforcement members is to find out the cause and effect. Be it as it may We do ask You; do not let Yourself be hurt this time!

Bibliography:

1.     
Criminal Investigations – Crime Scene Investigation.2000

2.      Forensic Science.2006

3.      Techniques of Crime Scene Investigation.2012

4.      Forensics Pathology.2001

5.      Pathology.2005  

6.       „A Short History of the Polymerase Chain Reaction". PCR Protocols. Methods in Molecular Biology.

7.      Molecular Cloning: A Laboratory Manual (3rd ed.). Cold Spring Harbor, N.Y.: Cold Spring Harbor Laboratory Press.2001

8.      "Antibodies as Thermolabile Switches: High Temperature Triggering for the Polymerase Chain Reaction". Bio/Technology.1994

9.       "Thermostable DNA Polymerases for a Wide Spectrum of Applications: Comparison of a Robust Hybrid TopoTaq to other enzymes". In Kieleczawa J. DNA Sequencing II: Optimizing Preparation and Cleanup. Jones and Bartlett. 2006

10.   "Microscale chaotic advection enables robust convective DNA replication.". Analytical Chemistry. 2013

11.  Human Physiology. An Integrate. 2016

Acknowledgements: 

The Police Department; 

https://www.politie.nl/mijnbuurt/politiebureaus/05/burgwallen.html and a Chief Inspector – Mr. Erik Akerboom                                               ©

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[1]  It is a medical specialty that is concerned with the diagnosis of disease based on the macroscopic, microscopic, biochemical, immunologic and molecular examination of organs and tissues. Over the last century, surgical pathology has evolved tremendously: from historical examination of whole bodies (autopsy) to a more modernized practice, centered on the diagnosis and prognosis of cancer to guide treatment decision-making in oncology. Its modern founder was the Italian scientist Giovan Battista Morgagni. Anatomical pathology is one of two branches of pathology, the other being clinical pathology, the diagnosis of disease through the laboratory analysis of bodily fluids and/or tissues. Often, pathologists practice both anatomical and clinical pathology, a combination known as general pathology. Similar specialties exist in veterinary pathology.

[2] Clinical pathologists work in close collaboration with clinical scientists (clinical biochemists, clinical microbiologists, etc.), medical technologists, hospital administrators, and referring physicians to ensure the accuracy and optimal utilization of laboratory testing.Clinical pathology is one of the two major divisions of pathology, the other being anatomical pathology. Often, pathologists practice both anatomical and clinical pathology, a combination sometimes known as general pathology. Similar specialties exist in veterinary pathology. Clinical pathology is itself divided into subspecialties, the main ones being clinical chemistry, clinical hematology/blood banking and clinical microbiology and emerging subspecialties such as molecular diagnostics and proteomics. Many areas of clinical pathology overlap with anatomic pathology. This overlap includes immunoassays, flow cytometry, microbiology and cytogenetics and any assay done on tissue. Overlap between anatomic and clinical pathology is expanding to molecular diagnostics and proteomics as we move towards making the best use of new technologies for personalized medicine. The analysers, by the association of robotics and spectrophotometry, allowed these last decades a better reproducibility of the results of proportionings, in particular in medical biochemistry and hematology. The companies of in vitro diagnosis henceforth try to sell chains of automats, i.e. a system allowing the automatic transfer of the tubes towards the various types of automats of the same mark. These systems can include the computer-assisted management of a serum library. These analysers must undergo daily controls to guarantee a result just possible, one speaks about quality control. These analysers must also undergo daily, weekly and monthly maintenances. Microscopic analysis is an important activity of the pathologist and the laboratory assistant. They have many different colourings at their disposal. Immunofluorescence, cytochemistry, the immunocytochemistry are also used in order make a correct diagnosis. This stage allows the pathologist to determine the character of the liquid: “normal”, tumoral, inflammatory even infectious. Indeed, microscopic examination can often determine the causal infectious agent, in general a bacterium, a mould, a yeast, or a parasite, more rarely a virus. A big part of the examinations of clinical pathology, primarily in medical microbiology, use culture media. Those allow, for example, the description of one or several infectious agents responsible of the clinical signs.

[3] Forensic pathology is pathology that focuses on determining the cause of death by examining a corpse. A post mortem is performed by a medical examiner, usually during the investigation of criminal law cases and civil law cases in some jurisdictions. Coroners and medical examiners are also frequently asked to confirm the identity of a corpse. It is a part of forensic medicine.

[4] Aniline is an organic compound with the formula C₆H₅NH₂. Consisting of a phenyl group attached to an amino group, aniline is the prototypical aromatic amine. Its main use is in the manufacture of precursors to polyurethane and other industrial chemicals. Like most volatile amines, it possesses the odour of rotten fish. It ignites readily, burning with a smoky flame characteristic of aromatic compounds. Aniline is a planar molecule. The amine is nearly planar owing to conjugation of the lone pair with the aryl subsistent. The C-N distance is correspondingly shorter. In aniline, the C-N and C-C distances are close to 1.39 Å, indicating the π-bonding between N and C.

Industrial aniline production involves two steps. First, benzene is nitrated with a concentrated mixture of nitric acid and sulfuric acid at 50 to 60 °C to yield nitrobenzene. The nitrobenzene is then hydrogenated (typically at 200–300 °C) in the presence of metal catalysts. The largest application of aniline is for the preparation of methylene dianiline and related compounds by condensation with formaldehyde (as discussed above). The principal use of aniline in the dye industry is as a precursor to indigo, the blue of blue jeans. Aniline is also used at a smaller scale in the production of the intrinsically conducting polymer polyaniline.

[5] Polymerase chain reaction (PCR) is a technique used in molecular biology to amplify a single copy or a few copies of a segment of DNA across several orders of magnitude, generating thousands to millions of copies of a particular DNA sequence. It is an easy, cheap, and reliable way to repeatedly replicate a focused segment of DNA, a concept which is applicable to numerous fields in modern biology and related sciences. The vast majority of PCR methods rely on thermal cycling, which involves exposing the reactants to cycles of repeated heating and cooling, permitting different temperature-dependent reactions—specifically, DNA melting and enzyme-driven DNA replication—to quickly proceed many times in sequence. Primers (short DNA fragments) containing sequences complementary to the target region, along with a DNA polymerase, after which the method is named, enable selective and repeated amplification. As PCR progresses, the DNA generated is itself used as a template for replication, setting in motion a chain reaction in which the original DNA template is exponentially amplified. The simplicity of the basic principle underlying PCR means it can be extensively modified to perform a wide array of genetic manipulations. PCR is not generally considered to be a recombinant DNA method, as it does not involve cutting and pasting DNA, only amplification of existing sequences. Typically, PCR consists of a series of 20–40 repeated temperature changes, called cycles, with each cycle commonly consisting of two or three discrete temperature steps (see figure below). The cycling is often preceded by a single temperature step at a very high temperature (>90 °C (194 °F)), and followed by one hold at the end for final product extension or brief storage. The temperatures used and the length of time they are applied in each cycle depend on a variety of parameters, including the enzyme used for DNA synthesis, the concentration of bivalent ions and dNTPs in the reaction, and the melting temperature (Tm) of the primers.

It is critical to determine a proper temperature for the annealing step because efficiency and specificity are strongly affected by the annealing temperature. This temperature must be low enough to allow for hybridization of the primer to the strand, but high enough for the hybridization to be specific, i.e., the primer should bind only to a perfectly complementary part of the strand, and nowhere else. If the temperature is too low, the primer may bind imperfectly. If it is too high, the primer may not bind at all. A typical annealing temperature is about 3–5 °C below the Tm of the primers used. Stable hydrogen bonds between complementary bases are formed only when the primer sequence very closely matches the template sequence. During this step, the polymerase binds to the primer-template hybrid and begins DNA formation.  PCR allows isolation of DNA fragments from genomic DNA by selective amplification of a specific region of DNA. This use of PCR augments many ways, such as generating hybridization probes for Southern or northern hybridization and DNA cloning, which require larger amounts of DNA, representing a specific DNA region. PCR supplies these techniques with high amounts of pure DNA, enabling analysis of DNA samples even from very small amounts of starting material. Other applications of PCR include DNA sequencing to determine unknown PCR-amplified sequences in which one of the amplification primers may be used in Sanger sequencing, isolation of a DNA sequence to expedite recombinant DNA technologies involving the insertion of a DNA sequence into a plasmid, phage, or cosmid (depending on size) or the genetic material of another organism. Bacterial colonies (such as E. coli) can be rapidly screened by PCR for correct DNA vector constructs. PCR allows for rapid and highly specific diagnosis of infectious diseases, including those caused by bacteria or viruses. PCR also permits identification of non-cultivatable or slow-growing microorganisms such as mycobacterium, anaerobic bacteria, or viruses from tissue culture assays and animal models. The basis for PCR diagnostic applications in microbiology is the detection of infectious agents and the discrimination of non-pathogenic from pathogenic strains by virtue of specific genes.

[6] Postmortem analysis for chemical constituents of the blood can be difficult unless the blood is collected very soon after death and prior to coagulation.  However, ocular fluids (aqueous and vitreous humor) and the retina can be useful for the diagnosis of several pathologic conditions, or exposure to various chemicals, for some time after death has occurred. Vitreous (and in some cases aqueous) humor can be used to estimate the time of death and can be useful as an aid in the diagnosis of renal disease, nitrate poisoning, hypomagnesemicsyndromes, calcium status and salt poisoning. In all species that have been examined (cattle, dogs, swine, and rabbits), urea nitrogen and creatinine concentrations in ocular fluid correlate very closely with serum concentrations for up to 24 hours (8h in rabbits) after death at body temperature (37°C). At room temperature (20 to 24°C) or refrigerated (4°C), they can be stable for longer periods of time.

Therefore, postmortem vitreous humor urea nitrogen concentrations can be useful to diagnose antemortem renal disease. Retina can be useful for postmortem diagnosis of organophosphate poisoning and for evaluation of antemortem exposure to certain illegal drugs such as clenbuterol.  Since it is a neural tissue, retina contains a large amount of cholinesterase which is inhibited by organophosphate insecticides.  This inhibition can be measured in the same manner as it is in brain or blood. Measurement of retina cholinesterase inhibition has been successfully used to diagnose organophosphate exposure up to 24h postmortem. For postmortem analysis of retina, the whole eyeball should be enucleated and submitted frozen.  ADDL personnel will then remove the retina form the interior of the eye.