An advanced method for the immunohistochemical detection of nitric oxide synthase (NOS) in the female genital tract.

The expression of nitric oxide synthase (NOS) in male and female urogenital tissues has been investigated by using conventional light microscopical immunoperoxidase staining. We present an improved immunohistochemical method for the specific and simultaneous detection of endothelial and neuronal NOS (eNOS/nNOS) in vaginal tissue. Specific antibodies have been used in combination with the tyramide signal amplification method. We found a subepithelial meshwork of varicose nerve fibers. A subpopulation of fibers presented immunoreactivity specific for nNOS. Epithelial cells also showed cytoplasmatic labeling for nNOS. Arteries presenting signals for eNOS in their endothelial layer were found in close proximity to nNOS-positive nerve fibers.

[The patient has died now what? : Step by step-what happens with patients after their death?] / Patient tot was nun? : Schritt für Schritt ­ was passiert mit dem Patienten nach seinem Tod?

The determination of a person's death, the subsequent post-mortem examination and the creation of the death certificate are part of regular medical work. The post-mortem examination, which is exclusively a medical duty, must be carried out immediately after the determination of death and includes in particular the definition of the cause of death and the type of death, which, in the case of a non-natural or unexplained death, entails further investigations by the police or public prosecutor and sometimes also forensic examinations. This article aims to shed more light on the possible processes after the death of a patient.

[Post-mortem examination prior to cremation--an instrument to verify the quality of medical post-mortems and uncover non-natural deaths?]. / Die Krematoriumsleichenschau--Instrument zur Qualitätskontrolle der ärztlichen Leichenschau und Aufdeckung nichtnatürlicher Todesfälle?

The external post-mortem examination, its deficient quality and possible causes have been the subject of numerous political and professional discussions. The external post-mortem examination is the basis for the decision whether further criminal investigations are required to clarify the cause of death. It is thus an essential instrument to ensure legal certainty. Before cremation, a second external post-mortem examination is performed by a public medical officer to make sure that errors of the first post-mortem are corrected. In the present study, cases were retrospectively analyzed in which a forensic autopsy had been ordered on the basis of the results of the post-mortem examination performed before cremation. The entries on the death certificate regarding the manner and cause of death were compared with the autopsy results. Between 1998 and 2007, 387 autopsies were ordered after external examination before cremation. In 55 cases (14.2%), the autopsy revealed a non-natural death, although a natural death had been attested on the death certificate. In descending order, a wrong manner of death was attested by clinicians, general practitioners and emergency physicians. With regard to the place where the first external post-mortem had been performed the lowest error rate was seen in nursing homes. Concerning the cause of death, discrepancies between the first post-mortem and autopsy were found in 59.4% of the cases. In this respect, general practitioners and clinicians were ranking first, whereas in nursing homes the cause of death was wrongly assessed in over 70% of cases. At present, the medical post-mortem does not meet the required quality standards, especially with regard to legal certainty. Determination of the cause of death on the basis of the external post-mortem examination is a challenging task even for the experienced medical examiner. As to the categorization of the manner of death it has to be stated that non-natural deaths are often not recognized or that the possibility to certify a death as unclear is not sufficiently used. As a result, it seems important to demand intensive, qualified, additional training in external post-mortem examinations for physicians.

Expression and distribution of cyclic AMP- and cyclic GMP-binding protein kinases in the human vagina- an immunohistochemical study.

INTRODUCTION: In contrast to research findings describing the localization of nitric oxide synthases (NOS), guanylyl cyclases, and cyclic adenosine monophosphate (cAMP)- and cyclic guanosine monophosphate (cGMP)-degrading phosphodiesterase isoenzymes in the human vagina, the distribution of proteins known as major targets for cyclic nucleotides has not yet been evaluated. cAMP- and cGMP-dependent protein kinases (cAK, cGKI) have been identified as important receptors for cyclic nucleotides downstream the signaling cascades. AIM: To investigate, by means of immunohistochemistry, the expression of cAK and cGKI in relation to endothelial NOS (eNOS), vasoactive intestinal polypeptide (VIP), and protein gene product 9.5 (PGP 9.5) in the human vagina. MAIN OUTCOME MEASURES: Expression and distribution of cAK and cGKI(alpha,beta) in relation to eNOS, VIP, and PGP 9.5 in human vaginal tissue. METHODS: Immunohistochemical techniques were applied to sections of human vaginal full wall specimens in order to evaluate the presence of cAK and cGKI(alpha,beta) in relation to VIP, PGP 9.5, and eNOS, respectively. Western blot analyses were conducted using cytosolic supernatants of homogenized specimens of the vaginal wall and epithelium. RESULTS: Immunostaining specific for cGKIbeta was observed in vascular and nonvascular smooth muscle of the vagina. In the endothelial layer, cGKIbeta was found colocalized with eNOS. In contrast, no signals indicating cGKIalpha were registered. cAK-positive subepithelial vessels were found to be innervated by a dense meshwork of PGP-containing varicose nerve fibers, some of which presented expression of VIP. The expression of cAK and cGKIbeta was confirmed by Western blotting. CONCLUSIONS: Our results demonstrate the expression of cAK and cGKIbeta in the human vagina. The colocalization with VIP and eNOS underlines the significance of both the cAMP and GMP pathway in the control of human vaginal vascular and nonvascular smooth muscle.

Urinary collecting system invasion reflects adverse long-term outcome and is associated with simultaneous metastatic spread at the time of surgery and with multilocular dissemination during postsurgical follow-up in renal cell cancer.

OBJECTIVES: To assess the prognostic implication of urinary collecting system invasion (CSI) in renal cell cancer (RCC). METHODS: Surveying a mean follow-up of 85 months, we investigated a cohort of 834 patients after radical (n = 710) or partial (n = 124) nephrectomy. At the time of surgery, 63 patients (7.6%) suffered from metastatic RCC. Various histopathologic parameters were analysed, and cancer specific survival (CSS) curves were individualized for each parameter. Furthermore, multivariate analysis was accomplished. RESULTS: Collecting system invasion was independently associated with a significant decline in CSS and was associated with simultaneous metastatic spread at the time of surgery and multilocular (involvement of at least two different organ systems) dissemination. CONCLUSIONS: The prognostic implication of CSI in RCC appears to be more complex than expected. Therefore, pathologists should report on CSI to enable selection of patients to be investigated in prospective studies which are needed to clarify the prognostic role of CSI in RCC.

[Aquatic fatalities--a systematic retrospective analysis]. / Ertrinkungstod--eine systematische retrospektive Untersuchung.

In a retrospective analysis of the autopsy material (n = 5,767) of the Institute of Legal Medicine of the Hanover Medical School covering the period of 1998-2007, all aquatic fatalities were evaluated, categorized and systematically compared under epidemiological and forensic criteria. The total of 156 cases of death by drowning (2.7 % of all autopsies) included 38 bathtub drownings and 28 deaths in the water for which no pathological anatomical cause of death could be reliably demonstrated. A control group (n = 221) was investigated for the presence of aqueous liquid in the sphenoid sinuses and compared with the findings of the drowning cases without signs of putrefaction. About 16 % of the control cases had fluid in the sphenoid sinuses compared with 57.6 % in the drowning group. Most of the drowning victims were men (60.9 %), whereas in the group of bathtub drownings the majority were women. More than half of the drowning cases (n = 89) could be classified as accidents. The individual groups showed a different incidence of findings associated with drowning.

Age-, tumor-, and metastatic tissue-associated DNA hypermethylation of a T-box brain 1 locus in human kidney tissue.

BACKGROUND: While a considerable number of tumor-specific hypermethylated loci have been identified in renal cell cancer (RCC), DNA methylation of loci showing successive increases in normal, tumoral, and metastatic tissues could point to genes with high relevance both for the process of tumor development and progression. Here, we report that DNA methylation of a locus in a genomic region corresponding to the 3'UTR of the transcription factor T-box brain 1 (TBR1) mRNA accumulates in normal renal tissues with age and possibly increased body mass index. Moreover, a further tissue-specific increase of methylation was observed for tumor and metastatic tissue samples. RESULTS: Biometric analyses of the TCGA KIRC methylation data revealed candidate loci for age-dependent and tumor-specific DNA methylation within the last exon and in a genomic region corresponding to the 3'UTR TBR1 mRNA. To evaluate whether methylation of TBR1 shows association with RCC carcinogenesis, we measured 15 tumor cell lines and 907 renal tissue samples including 355 normal tissues, 175 tissue pairs of normal tumor adjacent and corresponding tumor tissue as well 202 metastatic tissues samples of lung, bone, and brain metastases by the use of pyrosequencing. Statistical evaluation demonstrated age-dependent methylation in normal tissue (R = 0.72, p < 2 × 10-16), association with adiposity (P = 0.019) and tumor-specific hypermethylation (P = 6.1 × 10-19) for RCC tissues. Comparison of tumor and metastatic tissues revealed higher methylation in renal cancer metastases (P = 2.65 × 10-6). CONCLUSIONS: Our analyses provide statistical evidence of association between methylation of TBR1 and RCC development and disease progression.

[Causes of death among pilots: "acute myocardial infarction"--are the present examination methods for airworthiness sufficient?]. / Todesfälle bei Piloten: "Der akute Myokardinfarkt"--Sind die derzeitigen Flugtauglichkeitsuntersuchungen ausreichend?

The European Union plans to harmonize the aviation requirements, in particular the flight crew licensing requirements. On 23 May 2007, the German Federal Ministry of Transport, Building and Urban Affairs published the Flight Crew Licensing Requirements, which are based on the Joint Aviation Requirements, Flight Crew Licensing 3, Amendment 5. These guidelines also list the examination methods to be used for testing the medical fitness of pilots. In this Amendment some examinations which were part of the routine tests before JAR-FCL3 became effective have been deleted, e.g. the exercise ECG. This article presents two cases from the autopsy material of the Institute of Legal Medicine in Hanover and discusses the problems associated with the new examination guidelines.

[First report of lethal gamma-hydroxybutyrate (GHB) intoxication after consumption of gamma-butyrolactone (GBL) in Lower Saxony]. / Erster Todesfall durch gamma-hydroxybuttersäure (GHB) nach Aufnahme von gamma-Butyrolacton (GBL) in Niedersachsen.

A 25-year-old drug addict died from an overdose of GBL after mistaking it for water in preparing a dilution. He realized his error and wrote a call for help in the chat room of a drug forum, so that we know the time, type and approximate amount of drug consumed. The deceased had written numerous contributions in this drug forum before, which provide further information on his drug habits and the duration of the abuse of GHB, GBL and other drugs. According to the Criminal Investigation Department of Lower Saxony, this is the first death due to GHB/GBL reported in Lower Saxony.

Ethanol-induced malfunction of neutrophils respiratory burst on patients suffering from alcohol dependence.

BACKGROUND: Polymorphonuclear, neutrophil granulocytes (PMN) play a major role in the control of infections, and people who abuse alcohol are susceptible to infections. Resistance against infections ensues intracellularly following initial phagocytosis of microorganisms with the oxygen-dependent respiratory burst, the key enzyme of which is the respiratory burst oxidase, whereby oxygen radicals are produced for microbial destruction. To date there is insufficient information available in connection with the process of impaired defence against infection in patients suffering from alcohol dependence. Therefore, our investigation was carried out to determine the influence of alcohol exposition on the formation of oxygen radicals and the respiratory burst. METHODS: 4.5 ml of whole blood was taken from 10 healthy adults and 10 patients suffering from alcohol dependence. An additional 3.5 ml of whole blood was taken from the alcoholic patients for determination of the blood alcohol concentration. The respiratory burst of PMN was tested using the Four-Colour-Continuous Flow Cytometer. Each experimental procedure consisted of 4 test samples [negative controls, Escherichia coli, FMLP-supplement (N-formyl-l-methionyl-l-leucyl-l-phenylalanin), PMA-supplement (phorbol-12-myristate-13-acetate)]. Differing concentrations of ethanol were also introduced to each of the tests performed (0.20 to 4.00 g/l). RESULTS: Ethanol revealed a marked decrease of burst activity in those patients suffering from alcoholism with increased alcohol concentration. A dependence between the burst activity and the ethanol concentration was seen to be statistically significant. This effect was only evident after stimulation with E. coli and FMLP in those patients with alcohol dependence. CONCLUSION: The results presented in this study show an impairment in the function of PMN in those patients addicted to alcohol due to the decrease in burst activity. In view of the results of the different stimuli, the second-messenger effects were not evident. A clarification of this phenomenon could well be assumed as an allosteric receptor effect on the burst oxidase, namely, a direct effect on the phagocytosis interaction between circulating granulocytes and causative organisms.

[Postmortem dismemberment/mutilation--medicolegal and criminalistic evaluation of the autopsies performed by the Institute of Legal Medicine at the Hanover Medical School]. / Leichenzerstückelungen/-verstümmelungen --differenzierte rechtsmedizinische und kriminalistische Aufarbeitung des Sektionsgutes des Institutes für Rechtsmedizin der Medizinischen Hochschule Hannover.

Among all the autopsies performed by the Institute of Legal Medicine at the Hanover Medical School between 1978 and 1998. 16 cases of postmortem dismemberment/mutilation were evaluable under criminalistic and medicolegal aspects. In the present study, particular attention was paid to the psychosocial circumstances of the perpetrator's and victim's lives and also to the classification of the different types of postmortem dismemberment/mutilation illustrated by case reports. Apart from an unfavourable personal life situation at the time of the offence, essential predisposing factors were: poor integration in society and family, occupational problems, drug abuse and mental diseases. As the interactions between these factors differ in every single case and may be combined with other factors, the analysis of such homicides should always be based on the assessment of the individual case.

[Practicability of the mobile one-finger scanner Cross Match MV5 in fingerprinting of corpses: are mobile fingerprinting scanners suitable for use in mass disasters?]. / Praktikabilität des mobilen Einzelfingerscanners Cross Match MV5 in der Leichendaktyloskopie: Sind die mobilen Einzelfingerscanner für den Einsatz bei Massenkatastrophen geeignet?

Dactyloscopy is a special field in the police records department and a suitable means for identifying unknown dead persons as well as solving crimes by taking fingerprints from living persons. Apart from the conventional methods of dactyloscopy, mobile and more compact instruments are to facilitate taking prints of fingertips and palms also at the scene of mass disasters. In the presented study, living persons and corpses were examined to find out the possible uses and limits of mobile one-finger scanners. The concrete issue of the investigation was whether the mobile one-finger scanner Cross Match MV5 is suitable for application in mass disasters. The device was used in 12 corpses aged 5 weeks to 76 years (mean postmortem interval 5.5 days) and in 28 living persons aged 6 weeks to 87 years. In summary, the scanner produced qualitatively good prints in all age groups of the living individuals. In the corpses, the prints were only partly evaluable. In particular, fingers and fingertips with soot blackening and livid discoloration were difficult to analyse. Postmortem rigidity also complicated the handling of the scanner. In fresh corpses, the scanner can be recommended without reservation. Even if the epidermis was detached, the scanner was able to produce evaluable prints of the dermis of the hypothenar.

Adiposity and age are statistically related to enhanced RASSF1A tumor suppressor gene promoter methylation in normal autopsy kidney tissue.

Age, adiposity, and smoking are risk factors for the development of renal cell carcinoma. Hypermethylation of the RAS association domain family 1A gene (RASSF1A) promoter belongs to the most frequently detected epigenetic alterations in human cancers including renal cell carcinoma. RASSF1A is functionally involved in cell cycle control in normal cells and depletion promotes a number of cellular changes increasing the risk for neoplastic growth. We investigated the hypothesis that age, modulated by the factors adiposity and anthracosis as a surrogate for smoking, is a predictor of RASSF1A promoter methylation in normal kidney tissue. Using a cross-sectional study design, we quantitatively analyzed RASSF1A methylation in 78 normal autopsy kidney tissues by quantitative combined bisulfite and restriction analysis and bisulfite sequencing, and statistically evaluated the degree of relative methylation for a relationship with the predictor age and study factors adiposity and state of anthracosis. Statistical analysis showed that age (regression analysis; P < 0.001), adiposity (univariate analysis; P = 0.016), and state of anthracosis (t test; P = 0.005) are each significantly associated with an increase of RASSF1A promoter methylation in normal kidney tissue. However, only age (P = 0.008) and adiposity (P = 0.008) were identified as independent predictors of RASSF1A promoter methylation using covariance analysis. This study provides statistical evidence that the common cancer risk factors age and adiposity enhance RASSF1A promoter methylation in nonmalignant kidney tissue.

Anatomy of the sural nerve in a computer-assisted model: implications for surgical minimal-invasive Achilles tendon repair.

BACKGROUND: Sural nerve injuries are an evident risk especially of minimal-invasive surgical Achilles tendon repair. However, detailed anatomical studies focusing on the relationship of the sural nerve with the Achilles tendon at various levels are scarce, even pending in two planes. AIM: To determine the position and course of the sural nerve in relation to the Achilles tendon in two planes after trans-section and computer-assisted determination. METHODS: The exact course of the sural nerve was determined in 10 cadavers (55.3 years, 19-89 years), using a computer-assisted method in two planes (transversal/sagittal). RESULTS: The sural nerve crossed the Achilles tendon at 11 (8.7-12.4) cm proximal to the tuber calcanei. The distance between the lateral crossing and the proximal musculotendineus junction was 35 (20-58) mm. Starting from the tuber calcanei, the distance was 2/2 mm (transversal/sagittal plane) at 11 cm proximal to the tuber calcanei, 4/4 mm at 10 cm proximal, 5/6 mm at 9 cm, 8/10 mm at 5 cm and 11/18 mm at the tuber calcanei. CONCLUSION: In the lateral crossing region of the sural nerve and the lateral proximal Achilles tendon 9-12 cm proximal to the tuber calcanei, a close relationship of both anatomical structures can be visualised using computer-assisted measurements; caution is suggested to prevent sural nerve entrapment in either open or percutaneous Achilles tendon repair.

The white pulp in the setting of the septic spleen caused by different bacteria: a comparative morphometric study.

In the past little attention has been paid to histomorphologic changes accompanying the phenomenon of the septic spleen, thus indirectly reinforcing the old axiom that the spleen is an organ of mystery. It is especially noteworthy that the relationship between different causative bacteria and histopathologic abnormalities of the white pulp has not been investigated. In this study morphometric analysis was performed on the white pulp of 30 spleens obtained at autopsy from individuals with premortal sepsis. A strictly defined age- and sex-matched control group was analyzed for statistical comparison. Our findings demonstrate a significant depletion of B- and T-areas in the septic spleen, accompanied by a significant tendency towards reactive germinal center hyperplasia regardless of the type of bacteria responsible. However, depletion of splenic B-areas was shown to be significantly pronounced in the setting of premortal enterococcemia in comparison with a panel of gram-negative flagellated bacteria. It is felt that certain bacterial virulence factors (e.g. flagellation and/or structural components of the cell wall) might be pathogenetically involved in the observed changes, reflecting a partially different activation of splenic lymphocytes in the setting of the septic spleen.

A switch from epithelial to mesenchymal properties correlates with lymphovascular invasion in squamous cell carcinoma of the penis.

The purpose of the study was to assess the incidence and prognostic role of epithelial-to-mesenchymal-transition (EMT) in squamous cell carcinoma of the penis (SCCP). Sixty tumor specimens of surgically treated SCCP patients characterized by a central histopathologic review were stained with antibodies against E-cadherin, N-cadherin, ß-catenin, and vimentin. Staining profiles were scored by two independent raters, and correlated with pertinent clinical and pathological parameters and cancer-specific mortality (CSM; median follow-up: 34 months, interquartile range: 6-60 months). Correlation statistics proved good interobserver agreement in staining evaluation (K-values between 0.62 and 1.00, all p<0.001). Based on consensus decision between both raters, 36 study cases (60%) showed a switch from E-cadherin to N-cadherin (as a hallmark of EMT), which correlated with the presence of lymphovascular invasion (ρ=0.287, p=0.026) while failing to interfere with CSM. Although cadherin switch was correlated with a loss of membranous ß-catenin expression (ρ=0.629, p<0.001), none of the study cases showed nuclear ß-catenin expression, and only three EMT cases (8.3%) had tumor buds revealing vimentin expression. Our data suggest that roughly half of surgically treated SCCP cases exhibit EMT, a parameter correlating with lymphovascular invasion. However, further studies are clearly needed to validate the so far unresolved possible role of cadherin switch in terms of predicting nodal spread in patients with SCCP. Moreover, the apparently complex mechanisms driving EMT in SCCP should be explored by future studies, as knowledge about these might provide a so far unexploited basis for the development of novel targeted therapies against SCCP with metastatic dissemination.

[An unusual case of double death]. / Ein ungewöhnlicher Doppeltodesfall.

The report presents the case of an alleged double homicide or a homicide followed by suicide of a couple, in which the court-ordered autopsy of both victims revealed that the homicide had been committed by the husband immediately before he died himself from cardiac failure. The article gives a short summary of the pathophysiological relations between emotional tension (stress) and physical strain and the effects on the cardiovascular system. The case report describes a homicide by strangulation immediately followed by the perpetuator's death from a natural cause.

[Fatalities after injuries by wild animals]. / Todesfälle nach Verletzungen durch Wildtiere.

The article summarises three fatalities after attacks by wild animals. The first case describes a 90-year-old woman who died as a result of pneumonia after a bear fell on her and caused multiple chest fractures. The second case deals with a 76-year-old woman who was hit in the middle face by the hoof of a camel and, thereafter, died of myocardial infarction. The third case describes a 27-year-old biologist who died from severe blunt trauma after an attack of a wild living elephant. The article gives a summary of typical injury patterns of selected wild animals and outlines potential reasons of death as a result of the injuries.

[Spontaneous rupture of the urinary bladder]. / Spontane Harnblasenruptur.

The report presents the case of a 33-year-old man under the primary assumption of homicide in which the court-ordered autopsy revealed death as a result of bleeding due to a spontaneous rupture of the urinary bladder. The article gives a short summary of the pathomorphological findings and their respective causes, for example the typical location of the rupture at the posterior wall and roof of the bladder and the common concomitant drunkenness. Furthermore, obligatory forensic investigations--beside the autopsy--are mentioned and discussed.