We conduct research that aims at improving the diagnostic forensic casework. We also participate in preclinical research on tissue turnover, and carry out studies on pathophysiological mechanisms behind various ̽»¨¾«Ñ¡ conditions caused by disease and injury states. Several projects involve collaboration with the Swedish National Board of Forensic Medicine, and with researchers at KI and several other universities.
We are running several projects in parallel representing both basic and translational research, particularly aiming at developing methods to improve the postmortem diagnosis of severe ̽»¨¾«Ñ¡ conditions. Although having different focus, the projects have several points of contacts.
A core facility at ̽»¨¾«Ñ¡, KI Donatum, was established in 2005, and offers postmortem material to researchers. Swedish regulations regarding the use of material from deceased subjects are very strict, and certain conditions must be fulfilled. For donation for transplantation, collection and handling of tissues have been organized at different sites in Sweden. According to Swedish law all citizens may also indicate their wish to donate tissue after their death for research purposes. KI Donatum was formed to meet these needs and helps research leaders to find suitable postmortem tissues from well characterized donors for various analyses. KI Donatum offers support in writing applications for ethical approval, and may give suggestions about study design. Further, KI Donatum provides the practical service regarding continuous surveillance of donors, control of inclusion criteria, and accurate collection and processing of tissue samples according to the needs of each research project. The prospective strategy implies that extensive and project specific ̽»¨¾«Ñ¡ information and pertinent background data regarding each donor can be provided.
Contact: Henrik Druid. henrik.druid@ki.se
Traumatic brain injury (TBI) is caused when significant force is applied to the head. This may be due to an impact by blunt force, by the penetration of an object, or a rotational acceleration or deceleration of the head. TBI is one of the leading causes of death among young individuals, and the success in management in the clinical setting is dependent on an accurate characterization of the distribution and severity of the brain injuries. Hence a number of biomarkers in serum and liquor have been evaluated, and introduced, to assist in the assessment of injury severity and prognosis, and which serve as important tools for the choice of actions taken to treat the patients. In the postmortem setting, forensic pathologists focus on the detection, distribution and amount of contusions and secondary hemorrhages to assess the injuries. However, in many cases of fatal TBI the bleedings occupy a very small proportion of the total brain volume, and may as such not explain the death, but rather serve as indirect evidence of neuronal, and particularly axonal injuries, which may disrupt vital parts of the brain neurocircuitry. In the early phase, axonal injuries are however very difficult to visualize upon conventional microscopic examination.
Our studies will focus on the detection, distribution and amount of axonal injuries to help in the assessment of injury severity. To this end we will use select morphological methods such as light sheet microscopy, fluorescent correlation spectroscopy, stereology and biochemical analyses of tissue homogenates, serum and liquor. We also plan to use proximity ligation assay to identify transient protein interactions occurring at different time windows to estimate the age of the injuries, since this information is important for the police in obscure cases. The studies include experimental studies on rats with models of both rotational force and blunt force impact as well as postmortem human brain investigations.
PI: Henrik Druid, henrik.druid@ki.se
In mass disasters and in many homicides the identification of the victims is an important, but challenging task. Countries around the world participate in collaborative networks to provide reference information of missing persons to facilitate the identification casework. However, when there is no clue as to the identity of the victim, the sex and the age constitute the most important variables to limit the search for possible matches. The sex can usually be determined by either anthropological methods or with DNA analyses, whereas the age of the person remains a challenge to all professionals. We have developed a feasible protocol for analysis of bomb-pulse generated C-14 levels in dental enamel of teeth, which can provide an estimate of the year of birth with high accuracy. These analyses are now performed on a regular basis upon the request by pathologists and the police. By adding the age at death of the person, as estimated e.g. by aspartic acid racemization analysis, the approximate year of death can also be determined. Analysis of C-13 can be performed in parallel and provide clues to the geographical origin of the decedent.
We have now started to perform studies on human bones, since many skeletal finds lack teeth. The analyses include bomb-pulse generated C-14, and aspartic acid racemization ratio in different fractions of select types of bones, and also DNA methylation of select genes. Since different components of bone are exchanged at different rates during life, we will apply mathematical modeling to provide appropriate prediction of year of birth and age at death, respectively. The plan is also to analyze a few more stable isotopes to improve prediction of provenance and hopefully we will be able to track migration of subjects.
PI: Kanar Alkass. kanar.alkass@ki.se
Some homicides may be difficult to discover, such as homicidal poisonings with agents that may escape detection upon the regular toxicological screening, and certain forms of asphyxial deaths. Furthermore, dismembering of homicide victims can leave the police and forensic pathologist with only some parts of the body, making it difficult or impossible to determine the cause of death. These studies aim at retrospectively analyze cases where the perpetrator have used uncommon methods to kill, dismembered and/or disposed of the body by various means, or even made efforts to mislead the investigators. The ambition is to identify signs upon postmortem investigation that can provide clues as to the mode of killing, the method of dismembering and means of removal of the body.
PI: Brita Zilg, brita.zilg@ki.se
The aim of these studies is to elucidate the pathophysiological and pharmacological mechanisms behind opiate toxicity. Opioids are potent painkillers, and therefore extensively used to treat severe pain. However, opioids are also widely abused due to their reinforcing effects. Due to substantial tolerance development, opioid users have to gradually increase the dose to achieve pain relief or euphoria, and this increases the risk of respiratory depression. However, the time frame for development and loss of tolerance for the different effects conveyed by opioids are not fully characterized. We are using genomic, structural and biochemical methods to study opioid tolerance and toxicity mechanisms. In particular we want to elucidate the pathophysiology behind opioid-induced respiratory depression and lung edema. A large majority of the victims of acute opioid toxicity death show massive lung edema and froth in the airways at autopsy, but these features are rarely seen in subjects who survive to the emergency departments, suggesting a very rapid course.
PI: Lydia Bennedich Kahn, Lydia.bennedich@ki.se
In collaboration with preclinical research groups at CMB, ̽»¨¾«Ñ¡, postmortem samples from autopsy cases have been collected, processed and subjected to a sensitive C-14 analysis in order to offer an age determination of different tissues. The strategy takes advantage of the changes of C-14 in the atmosphere caused by the above-ground test detonations of nuclear weapons 1955-1963. Since C-14 is incorporated into living matter in relation to the content in the atmosphere, levels in nuclear DNA that are different from those at birth of the individual can be used to detect regeneration of cell populations. Using cell sorting, and isolation of carbon from chromosomal DNA, the specific age of cardiomyocytes in the heart ventricular wall, and of neuronal and non-neuronal cell populations in select brain regions, has been estimated. These studies have provided evidence of a continuous turnover of cardiomyocytes, and of neurons in hippocampus and striatum. The project also includes cell lineage studies to identify the origin of different specialized cells. Studies are also carried out to evaluate the possible influence of various intrinsic and environmental factors on the renewal rates of discrete cell populations. These studies take advantage of the access to well-characterized human postmortem tissue samples procured and processed by KI Donatum.
PI: Henrik Druid, henrik.druid@ki.se
Clinical chemistry results are often decisive in the clinical management of both hospitalized patients and in out-patient care for decisions regarding management and treatment measures. Postmortem changes make some of these analyses either impossible to carry out or make the results difficult to interpret. Hence, there is a need for postmortem reference intervals for a large number of analyses which could assist in identifying severe ̽»¨¾«Ñ¡ conditions. At the department of forensic medicine in Stockholm, we have collected vitreous fluid upon arrival of deceased to the morgue and analyzed the samples with a blood gas instrument. Using this strategy, we have established reference levels for vitreous glucose, sodium and chloride that can be used to identify fatal hyperglycemia, and serious salt imbalances. We have also developed a model for time of death estimation using vitreous potassium levels:
We plan to evaluate the postmortem applicability of select clinical chemistry analyses which can be performed with regular instrumentation, but also as rapid tests, hence the designation deadside analysis. We will take advantage of the access to the internal organs, allowing for analysis of homogenates of tissues where possible pathologies exist. This strategy will add diagnostic information that can be available for the pathologist´s decision-making during the autopsy and provide guidance in the postmortem casework.
PI: Henrik Druid, henrik.druid@ki.se
In collaboration with the Department of Forensic Toxicology, National Board of Forensic Medicine, postmortem femoral blood concentrations of pharmaceutical drugs and illicit drugs in certified fatal intoxication cases and postmortem controls are evaluated according to a standardized protocol. Using this strategy, fatal cases are classified as A) intoxications with one drug only, B) multiple drug intoxications; and C) postmortem controls consisting of strictly selected cases where the subject with certainty died from another cause of death and was not incapacitated by drugs at the time of death. All collection, handling, and analysis of the samples follow standardized procedures established at all forensic medicine units in Sweden since 1991. This strategy has resulted in several publications presenting reference concentrations of pharmaceutical drugs in postmortem blood. This reference information is critical in order to correctly interpret such results in obscure postmortem cases.
A website, where the database will be available for other researchers and for professionals conducting forensic medicine and toxicology casework, is under construction. A collaboration has been established with Victorian Institute of Forensic Medicine in order to expand the reference database. The ambition is to include additional laboratories to increase the panorama of substances so that the database becomes more universally useful. The plan is also to include clinical data, particularly regarding toxicological results found in driving-under-the-influence cases and in non-lethal intoxications.
PI:s: Henrik Druid and Anna Jönsson. anna.jonsson@rmv.se
One of the most common and important questions in forensic death investigations is the time when injuries have been sustained. A deceased subject, either a victim of homicide, suicide or a fatal accident, may have died immediately, or several minutes, hours or even a day or more, after the injuries were sustained. We evaluating certain immunohistochemistry markers in terms of their capacity to predict the post-infliction time interval. We are also taking advantage of the proximity ligation assay technique, using combinations of antibodies to proteins that are assumed to interact in different time phases after a vessel injury. Further, RNA expression will be analyzed in samples from the same wounds. Different non-invasive sample techniques and methodological protocols that may allow for analysis of both mRNAs and protein interactions in wounds of living subjects will also be evaluated.
PI: Henrik Druid, henrik.druid@ki.se
This project aims at improving the diagnosis of drowning (or to lend support for the notion that drowning had not occurred). Diatoms in the water that enter the lungs may cross the alveolar-capillary barrier through drowning-induced ruptures and can then be distributed to, and trapped in the capillaries of remote tissues via the blood circulation. The diatoms can be extracted from tissue samples and analyzed microscopically. In collaboration with Stockholm University the extracts are studied using light microscopy with either bright field, dark field, phase contrast or polarization. We have also used Scanning Electron Microscopy for better identification of family/species. We are currently using protein K for digestion of the organic material. Although the method is today available in the routine casework, we continue to analyze more samples from drownings and non-drownings to obtain larger sets of data to provide a more firm information about sensitivity and specificity. Our plan is also to evaluate alternative extraction methods, and to investigate possible rapid screening options.
PI: Henrik Druid, henrik.druid@ki.se
Certain minor projects are also running in parallel, and are typically related to some extent to the major projects. Epidemiological studies using the Swedish forensic pathology database and forensic toxicology database are carried out, aiming at improving diagnostics in forensic medicine casework, but also to provide other agencies with certain results that may have impact on their work and responsibilities.
One important issue is the surveillance of drug-related deaths, for which we have developed a three-step protocol for rapid evaluation allowing for early reporting. We believe this is an important asset to the monitoring of the drug abuse situation, given the high numbers of opioid toxicity deaths.
Experimental studies include characterization of patterns of postmortem changes in normal tissues and in tissues with certain pathologies using both histochemical stainings and immunohistochemistry protocols. Preliminary studies have revealed that degradation and putrefaction can produce changes in normal tissue that resembles certain pathological conditions, and that various pathological changes can gradually disappear. Certain immunohistochemistry markers are promising to assist in reaching a correct diagnosis.
Methods for segmental hair analysis of drugs have been developed at the department of forensic toxicology in Linköping. These have been particularly useful in order to determine previous opioid exposures in individuals who have died of acute opioid intoxication, since the degree of tolerance, as estimated from the hair analysis, apparently is a critical factor for the fatal outcome. The possibility of development of pharmacodynamic tolerance to the effects of pharmaceutical drugs has not been explored much, and hence a justified line of research is to develop methods for hair analysis of certain groups of pharmaceuticals that are regularly encountered in fatal intoxications. Such studies may assist in the interpretation of postmortem femoral blood drug concentrations.
Studies on the diagnosis of hypothermia have been launched with the aim to evaluate biomarkers and to study the pathophysiological mechanisms. To this end both a mice model of hypothermia and postmortem human tissues are studies. In particular we are interested in the reactivity pattern of the podocytes in the kidney and the activation of platelets in the spleen.
Finally, a large project regarding diagnosis of infections in deceased subjects, still at the planning stage, will tentatively be launched during 2021. This will include development of methods to identify infections that otherwise may go undetected by conventional postmortem examinations.
PI: Henrik Druid, henrik.druid@ki.se
Shinya Kasai, Ass. Prof (2017-2019)
Ahmed Mousa, PhD (2012-2016)
Sasikala K. Nil, Professor (2004)
Akira Hayakawa, MD, PhD (2021-2022)
Kie Horioka PhD (2020)
Tara Wardi LeMaitre, PhD (2014-2018)
Lydia Kahn MD, PhD(2010-2014)
Hisako Saitoh MD(2010-2011)
Olof Bendel, PhD (2008-2009)
Fredrik C. Kugelberg, PhD (2005-2006)
Gopalakrishnan Dhanabalan
Lotta Walz
Carl Söderberg
Brita Zilg
Kanar Alkass
Joakim Strandberg
Sofi Bäcklin
Madeleine Granlind
Lars-Eric Bergman
Supervisor Henrik Druid unless otherwise stated
Anna Kawaguchi (2023)
Julia Mielcarz (2023)
Deni Rashid; supervisor Kanar Alkass (2023)
Klaske Osinga (2023)
Deni Rashid; supervisor Kanar Alkass (2022)
Malin Persson; supervisor Lydia Kahn (2022)
Emma Molander; supervisor Brita Zilg (2022)
Sebastian Gode (2022)
Nicelle Myhrberg (2022)
Sofia Andersson (2022)
Elin Haupt; supervisor Brita Zilg (2022)
Amanda Jonsson; supervisor Lydia Kahn (2022)
Paula Tolsa Ortega (2022)
Constanze Kohout; supervisor Kanar Alkass (2022)
Natalie Avigdor (2021)
Sofia Riehl (2021)
Panagiota Tselekidou (2021)
Jasmijn Maljaars; supervisor Kanar Alkass (2021)
Cecilia Magnusson; supervisor Brita Zilg (2021)
Linnea Aurich; supervisor Lydia Kahn (2021)
Hanna Tengelin; supervisor Lydia Kahn (2021)
Tobias Björkman; supervisor Lydia Kahn (2021)
Erik Alexandersson, supervisor Lydia Kahn (2021)
Isabel Auer; supervisor: Lydia Kahn (2020)
Yakub Eriksson Hydara; supervisor Brita Zilg (2020)
Simon Ström supervisor Brita Zilg (2020)
Gustav Wärn supervisor Brita Zilg (2020)
Kristoffer Hahn; supervisor: Lydia Kahn (2019)
Evelyn Maitz, supervisor Kanar Alkass (2019)
Dominika Begolli; supervisor: Kanar Alkass (2019)
Johan Pieniowski (2019)
Sibon Kabir (2019)
Hanna Hedlund (2019)
Frida Soneborg (2019)
Alice Ljungström (2018)
Michael Moridi; supervisor: Brita Zilg (2018)
Cassandra Cieslak; supervisor Kanar Alkass (2017)
Carolina Goncalves (2017)
Emma Gustafsson (2016)
Caroline Ytterman (2016)
Meta Byström (2015)
Hampus Fabricius (2015)
George Hamnström (2014)
Sabria Belarbi (2014)
Marina Montmany (2014)
Niklas Sjökvist (2014)
Sandra Koskinen (2013)
Joelle Vos (2013)
Erik Bivehed (2013)
Joan Raouf (2011)
Ann Ståhle (2011)
Jenny Rönnlund (2011)
Evan Reed (2010)
Sara Hassanzaddeh (2010)
Annika Andersson (2009)
Stephanie Harney (2009)
Hampus Janko (2009)
Anke Böhmer (2008)
Siv Andersson (2008)
Stina Norlin (2008)
Nina Granberg (2008)
Lisen Bredberg (2006)
Midia Hussein (2006)
Brita Zilg (2006)
Aziz Bitar (2005)
Anna Gustavsson (2005)
Elias Malko (2005)
Cecilia Hult (2004)
Peggy Törn (2004)
Elias Palm (2004)
Sofia Hausel (2002)
Isabella Cereceda (2003-2009)
