Single Nuclei Sequencing Reveals Novel Insights Into the Regulation of Cellular Signatures in Children With Dilated Cardiomyopathy.

BACKGROUND: Dilated cardiomyopathy (DCM) is a leading cause of death in children with heart failure. The outcome of pediatric heart failure treatment is inconsistent, and large cohort studies are lacking. Progress may be achieved through personalized therapy that takes age- and disease-related pathophysiology, pathology, and molecular fingerprints into account. We present single nuclei RNA sequencing from pediatric patients with DCM as the next step in identifying cellular signatures. METHODS: We performed single nuclei RNA sequencing with heart tissues from 6 children with DCM with an age of 0.5, 0.75, 5, 6, 12, and 13 years. Unsupervised clustering of 18 211 nuclei led to the identification of 14 distinct clusters with 6 major cell types. RESULTS: The number of nuclei in fibroblast clusters increased with age in patients with DCM, a finding that was confirmed by histological analysis and was consistent with an age-related increase in cardiac fibrosis quantified by cardiac magnetic resonance imaging. Fibroblasts of patients with DCM >6 years of age showed a profoundly altered gene expression pattern with enrichment of genes encoding fibrillary collagens, modulation of proteoglycans, switch in thrombospondin isoforms, and signatures of fibroblast activation. In addition, a population of cardiomyocytes with a high proregenerative profile was identified in infant patients with DCM but was absent in children >6 years of age. This cluster showed high expression of cell cycle activators such as cyclin D family members, increased glycolytic metabolism and antioxidative genes, and alterations in ß-adrenergic signaling genes. CONCLUSIONS: Novel insights into the cellular transcriptomes of hearts from pediatric patients with DCM provide remarkable age-dependent changes in the expression patterns of fibroblast and cardiomyocyte genes with less fibrotic but enriched proregenerative signatures in infants.

Estimates of Non-Alcoholic Food-Derived Ethanol and Methanol Exposure in Human.

Food-derived alcohol is almost not in question due to its low concentration. Nevertheless, could it pose a problem for some risk groups and forensic cases? To answer this, we aimed to simultaneously evaluate ethanol and methanol ingredients of a variety of non-alcoholic foods in two different countries and estimate their possible health and forensic consequences. Alcohols in foods were analysed by headspace gas chromatography (HS-GC). Human average acute daily food consumptions and food-derived blood alcohol concentrations (BAC) were determined by using the data of the EFSA Nutrition Survey. Methanol and ethanol ingredients of similar foods varied between the two cities. Most foods produce higher methanol concentrations than the Maximum Allowable Dose Level (23 mg). Especially fruit juices lead to the critical level of ethanol for children (6.0 mg/kg bw). Based on the results, adult daily intake of selected food groups does not bear ethanol that exceeds the legal limit of BAC or the limit not allowed by the religious and does not lead to acute alcohol toxicity. But these low levels of ethanol and methanol consumed via non-alcoholic foods for life can raise the vulnerability to chronic health problems (cancer, liver cirrhosis, Alzheimer's disease, autism, ocular toxicity, alterations in fetal development), and may lead to positive ethanol metabolite results (e. g. Ethyl glucuronide) when a low cut-off level is used. Therefore, studies on the alcohol contents of various natural and processed non-alcoholic foods along with their effects on humans, and new regulations on labeling the food products and conscious food consumption are in particular importance. It would also be important to consider unintentional alcohol consumption via non-alcoholic foods in the evaluation of clinical and forensic cases.

[National and international aspects of the medico-legal examination of injured witnesses and defendants in criminal proceedings].

Depending on the type of offence, the medico-legal examination of victims of violence for the purpose of collecting evidence in criminal proceedings requires prompt physical examination of the victim, the exact description and, wherever possible, photographic documentation of the injuries and their forensically correct interpretation. Examination, documentation and assessment have to meet minimum standards. In addition it must be ensured that the examiner or expert has an independent organisational, administrative and financial status. An independent expert must not be an organisational part of the police force, the public prosecution and the court. Legal regulations have to make sure that the expert cannot be influenced "through official channels". His or her financial independence must make attempted bribery at least improbable. The independence of the expert is particularly important, if persons become victims of violence while they are under the custody of the state, especially the police, in prisons and after compulsory commitment to a psychiatric hospital.

Articular Joint Lubricants during Osteoarthritis and Rheumatoid Arthritis Display Altered Levels and Molecular Species.

BACKGROUND: Hyaluronic acid (HA), lubricin, and phospholipid species (PLs) contribute independently or together to the boundary lubrication of articular joints that is provided by synovial fluid (SF). Our study is the first reporting quantitative data about the molecular weight (MW) forms of HA, lubricin, and PLs in SF from cohorts of healthy donors, patients with early (eOA)- or late (lOA)-stage osteoarthritis (OA), and patients with active rheumatoid arthritis (RA). METHODS: We used human SF from unaffected controls, eOA, lOA, and RA. HA and lubricin levels were measured by enzyme-linked immunosorbent assay. PLs was quantified by electrospray ionization tandem mass spectrometry. Fatty acids (FAs) were analyzed by gas chromatography, coupled with mass spectrometry. The MW distribution of HA was determined by agarose gel electrophoresis. RESULTS: Compared with control SF, the concentrations of HA and lubricin were lower in OA and RA SF, whereas those of PLs were higher in OA and RA SF. Moreover, the MW distribution of HA shifted toward the lower ranges in OA and RA SF. We noted distinct alterations between cohorts in the relative distribution of PLs and the degree of FA saturation and chain lengths of FAs. CONCLUSIONS: The levels, composition, and MW distribution of all currently known lubricants in SF--HA, lubricin, PLs--vary with joint disease and stage of OA. Our study is the first delivering a comprehensive view about all joint lubricants during health and widespread joint diseases. Thus, we provide the framework to develop new optimal compounded lubricants to reduce joint destruction.

Molecular identification of fungi found on decomposed human bodies in forensic autopsy cases.

To investigate which fungi can be found during forensic autopsies, a PubMed literature review was done in regard to fungal growth on decomposed human bodies. Unfortunately, the existing data is limited and not all fungi were identified to the species level. We, therefore, collected skin samples with macroscopically visible fungal growth from 23 autopsy cases in Germany and identified the fungi to the species level by molecular methods. The identified species included Aspergillus fumigatus and Candida albicans, which pose an allergenic risk, especially to persons with underlying lung diseases. Because safety standards are lacking, we recommend the use of respiratory protection during exhumations and forensic autopsies, when fungal growth is noted. With regard to the future, a database was set up which could possibly be used as a forensic tool to determine the time of death.

[Cause and manner of death in the autopsy material of the Institute of Forensic Medicine, Justus Liebig University Giessen, from 2002-2006]. / Todesursachen und Todesarten im Obduktionsgut des Instituts für Rechtsmedizin der JLU Giessen in den Jahren 2002 bis 2006.

A retrospective analysis of the autopsy material of the Institute of Forensic Medicine, Justus Liebig University Giessen, in the years 2002 to 2006 showed that the Giessen material is similar to that of comparable studies but contains a relatively large share of lethal intoxications and cases with a concrete suspicion of poisoning in which the cause of death was not definitely clarified by further investigations. On the basis of our own results it is demonstrated that additional chemical, toxicological and histological examinations are helpful to determine initially unclear causes of death. In view of the already very low autopsy rates in Germany it is strange that more detailed diagnostic measures are often omitted.

[Intentional poisoning of two wives by their husband?]. / Vorsätzliche Vergiftung zweier Frauen durch ihren Ehemann?

The authors report on the death of two women who were married to the same man one after the other. Exhumation and toxicological investigation of the first wife, who had died 7 years before, did not produce any conclusive evidence of homicide. With regard to the circumstances of death of the second wife the husband made different statements. According to the result of the chemical and toxicological investigations death was caused by acute intoxication with the beta-blocker metoprolol. The man was found guilty of killing on request (which is a criminal offence in Germany) by administering the beta-blocker metoprolol through a transnasal gastric tube.

[Forensic assessment of suspected medical malpractice and outcome of preliminary proceedings against medical practitioners]. / Rechtsmedizinische Begutachtung bei Behandlungsfehlerverdacht und Ausgang arztstrafrechtlicher Ermittlungsverfahren.

In a retrospective study, all autopsies performed at the Institute of Forensic Medicine of the Justus Liebig University of Giessen between 1968 and 2007 for suspected medical malpractice with fatal outcome were evaluated and analysed with regard to the reason for initiating preliminary investigations and the medical specialties involved. In 109 cases, the prosecution files could be inspected as to the outcome of the proceedings. As there are hardly any comparable studies in the German-speaking countries, the data were compared with the results of two older studies. In many cases, medical malpractice could be excluded directly at the end of the post-mortem examination. Most of the charges were dropped according to Section 170 subs. 2 Code of Criminal Procedure.

An unusual occupational accident: fall into a sewage plant tank with lethal outcome.

Occupational accidents, often presenting with lethal outcomes, are a rarely reported issue in forensic literature. However, these incidents are part of medicolegal casework with special regard to reconstruction, liabilities and insurance law-related issues, respectively. We report on a lethal occupational accident in a metropolitan sewage plant. When performing routine controls, a technician fell into an overflow sewer and was immediately pulled into a 30 cm diameter drain. Rescue efforts were initiated immediately, but had to be terminated due to gas warning. Rescue teams continued the search, however, the body remained undiscoverable. Forty-eight hours later, the cadaver was found in an adjacent digester tank, from where it was finally rescued. It was concluded, that the body had been transported between the overflow sewer and the digester tank through a 120 m pipeline with several 90 degrees bendings and branch connections with a minimum diameter of 25 cm at the discharge valve. On medicolegal examination, the cadaver showed marked signs of advanced decomposition caused by anaerobic microorganisms in the 37 degrees C biomass environment. Moreover, as a consequence of the passage of the pipeline system, signs of massive trauma (several comminuted and compound fractures) were disclosed at autopsy. To us, this is the first report on a lethal occupational accident in a sewage plant; our observations demonstrate the rapid progress of putrefaction in a warm anaerobic bacterial environment and the massive trauma sustained.

Heroin-associated nephropathy.

Since the first reports in the late 1960s and early 1970s there have been numerous studies describing the clinical and pathological features of renal diseases associated with chronic parenteral abuse of heroin, cocaine, morphine, amphetamine, and other narcotic and hallucinogenic drugs, including several adulterants. The past 35 years have witnessed an explosive growth in illicit drug use in many parts of the world. Meanwhile, drug addict nephropathy constitutes an important cause of end-stage renal disease. The term heroin-associated nephropathy' includes different morphological findings following chronic drug abuse. Up to now it still remains ambiguous as to whether or not heroin/morphine itself, adulterants, other diseases like hepatitis B and C infection, or HIV, lead to a spectrum of morphologically described heroin-associated' findings in the kidneys. As a measure of prevention it appears that the purity of heroin plays an important role.

[Medicolegal aspects of witnessed suicide due to gunshot to the head. Part 1: Circumstances and psychopathology]. / Rechtsmedizinische Aspekte des Suizides durch Kopfschuss vor Zeugen. Teil I: Umstände und psychopathologische Befunde.

In the USA the frequency of witnessed suicide, i.e. suicide committed in the presence of at least one further individual, is reported to range between 5 and 15%. As up to now no detailed analysis of this special issue has been given in the medicolegal literature, this article presents and discusses a number of cases including 8 males and 1 female (age 19-58 years, mean 41.2 years) who had all committed suicide by inflicting a gunshot to the head. In 3 cases post-mortem blood alcohol concentrations of 1.73, 1.88 and 2.23 g/kg respectively were observed. Toxicological tests produced negative results. 5 cases had a medical history of psychiatric disorder with endogenous depression in 3, chronic alcohol abuse and drug abuse with concomitant psychosis in one case each. As far as the motives were known, domestic arguments were of prevailing importance. With one exception the suicides were committed at home. In 6 cases one witness was present, in 2 cases 2 individuals and in one case 22 persons witnessed the suicide. The reported cases are discussed in comparison with the psychiatric-psychological classification according to McDowell et al. with the aim to provide a solid and comprehensive medicolegal method to distinguish between homicide and suicide. Furthermore aspects of psychiatric sequelae and psychological support of the witnesses, which are also of importance for the forensic pathologist called to the scene, are discussed.

[Medicolegal aspects of witnessed suicide due to gunshot to the head. II. Legal medicine aspects and examination of the firing hand]. / Rechtsmedizinische Aspekte des Suizides durch Kopfschuss vor Zeugen. Teil II: Rechtsmedizinische Befunde und Schusshanduntersuchung.

The second part of the paper on suicides by gunshots to the head in the presence of witnesses focuses on relevant morphological autopsy findings such as entrance site, signs of close range or contact shots, bullet path etc. and also discusses selected aspects of ascertaining gunshot residues. For the identification of the shooter an integral medicolegal assessment of all the facts including the investigation results and the autopsy findings is essential. However, the morphological findings alone do not allow safe diagnosis, as for example in a homicide the temporal region, which was affected in all our cases, may have been deliberately chosen by the perpetrator as a localization typical of suicide. Thus methods to ascertain gunshot residues on the firing hand (by means of adhesive films and the polyvinyl-alcohol collection method--PVAL) are of great practical importance. In seven cases adhesive films and/or the polyvinyl-alcohol collection method were used. In one case the gunshot residues (GSR) were analysed by means of tape lifts and subsequent scanning electron microscopy (SEM). It was found that especially the combined application of topographical (adhesive tape/PVAL) and cumulative (SEM) methods allowed for the doubtless identification of the shooter, thus usually confirming the suicide. By the example of one case it is demonstrated that without the immediate collection of evidence at the scene objective reconstruction of the event becomes impossible. On the basis of the reported cases recommendations are finally given for a differentiated approach in the medicolegal evaluation of alleged witnessed suicide by gunshot (to the head).