Dendritic cells (DCs) can essentially contribute to innate and adaptive immune system in various organs. A double-color immunofluorescence analysis was carried out with anti-CD11c and -HLA-DRα antibodies to detect DCs in 53 skin wounds (their postinfliction intervals: group I, 0–3 days; group II, 4–7 days; group III, 9–14 days; and group IV, 17–21 days). CD11c+HLA-DRα+ DCs were first observed in skin wounds with postinfliction intervals of 3 days, and the DC numbers were found to be elevated in skin wounds with the subsequent increase in postinfliction intervals. Semi-quantitative morphometric analyses showed that the DC number was the highest in the 12-day-old wound. More than 50 DCs were present in 8 of 10 samples (80%) in group II and 14 of 16 samples (87.5%) in group III, and there was no difference between the two groups. Thus, the presence of DCs in a skin wound was possibly estimated as postinfliction intervals of at least 3 days. Furthermore, when a skin wound contained > 50 DCs, its age would be judged as 4–14 days. Collectively, the appearance of DCs in human skin wounds may provide useful information in determining the age of a wound.

Immunohistochemical investigation of aquaporin (AQP)1 and AQP3 was performed in human skin wounds obtained from forensic autopsy cases. A total of 55 human skin wounds of different postinfliction intervals were collected as follows: group I, 0–3 days (n = 16); II, 4–7 days (n = 11); III, 9–14 days (n = 16); and IV, 17–21 days (n = 12). In uninjured skin samples, AQP1 and AQP3 could be slightly detected in dermal vessels and keratinocytes, respectively. The percentage of AQP1+ vessels and the number of AQP3+ keratinocytes were apparently elevated in accordance with wound ages. The number of AQP3+ keratinocytes was distinctly evident in groups II and III. Morphometrically, both AQP1+ vessel area and AQP3+ cell number were markedly increased in group II, compared with other three groups. With regard to forensic safety, AQP1+ vessel area of over 5% would imply wound ages of 4–12 days. Moreover, the positive area of > 15% would suggest wound age of 7–10 days. Especially, most samples of skin wounds aged 5–10 days except for only one sample (a 10-day-old wound) showed AQP3+ cell number of > 300, and the remaining other samples had that of < 300. Thus, the AQP3+ cell number of > 300 would indicate wound ages of 5–10 days. Collectively, immunohistochemical analyses of AQP1 and AQP3 in human skin wounds would support the objective accuracy of wound age determination.

BACKGROUND:Biobanks increasingly presume long-term storage of biomaterials and data that shall be used for future research projects which are today unspecified. Appropriate consent documents for sample donors must therefore explain the breadth of consent and other elements of the biobank governance framework. Recent reviews demonstrated high variability in what issues these documents mention or not and how the issues are explained. This might undermine the protection of sample donors, complicate networked biobank research, create research waste and impact on public trust. METHODS:A systematic analysis of international research guidelines and existing broad consent templates was performed. Based on this information an interdisciplinary expert group from the AKMEK (Permanent Working Party of German RECs) developed a draft template and organized a comprehensive stakeholder consultation. After revision the final template was consented by all 53 German RECs. RESULTS:This paper briefly explores the spectrum of potentially relevant issues for broad consent forms. It then elaborates the template and how it was designed to be applicable in different types of biobanks. DISCUSSION:To further improve the validity and applicability of broad consent forms in biobank and other big data research, practice evaluations are needed. We hope that in this regard the presented template supports the development of new consent forms as well as the evaluation and revision of existing ones.

Recent experimental developments in generation and detection of THz acoustical phonons have led to studies of phonon emission, propagation and absorption in solids. New results of this acoustical phonon spectroscopy concern phonon interactions with collective excitations (phonons, photons, magnons), localized excitations (resonant scattering of impurities, radiationless transitions) and electronic excitations in metals, superconductors and semiconductors. The different experimental methods and their applications to phonon interaction studies are discussed.

Endothelial progenitor cells (EPCs), a newly identified cell type, are bone marrow-derived progenitor cells that co-express stem cell markers and vascular endothelial growth factor (VEGF) receptor (Flk-1). In this study, a double-color immunofluorescence analysis was carried out using anti-CD34 and anti-Flk-1 antibodies to examine the time-dependent appearance of EPCs, using 52 human skin wounds with different wound ages (Group I, 0–1 days; Group II, 2–6 days; Group III, 7–14 days; and Group IV, 17–21 days). In wound specimens with an age of less than one day, CD34+/Flk-1+ EPCs were not detected. EPCs were initially observed in wounds aged two days, and their number was increased in lesions with advances in wound age. In morphometrical analysis, the average number of EPCs was the highest in the wounds of Group III. Especially, 20 out of 21 wounds aged 7–12 days had >20 EPCs, and all wound samples with postinfliction intervals of 14–21 days had <15 EPCs. These observations at least showed that >20 EPCs would indicate a wound age of 7–12 days. Taken together, our observations indicate the detection of EPCs would be useful for wound age determination.

Huntington's disease (HD), an autosomal dominantly inherited polyglutamine or CAG repeat disease along with somatomotor, oculomotor, psychiatric and cognitive symptoms, presents clinically with impairments of elementary and complex visual functions as well as altered visual-evoked potentials (VEPs). Previous volumetric and pathoanatomical post-mortem investigations pointed to an involvement of Brodmann's primary visual area 17 (BA17) in HD. Because the involvement of BA17 could be interpreted as an early onset brain neurodegeneration, we further characterized this potential primary cortical site of HD-related neurodegeneration neuropathologically and performed an unbiased estimation of the absolute nerve cell number in thick gallocyanin-stained frontoparallel tissue sections through the striate area of seven control individuals and seven HD patients using Cavalieri's principle for volume and the optical disector for nerve and glial cell density estimations. This investigation showed a reduction of the estimated absolute nerve cell number of BA17 in the HD patients (71,044,037 ± 12,740,515 nerve cells) of 32% in comparison with the control individuals (104,075,067 ± 9,424,491 nerve cells) (Mann-Whitney U-test; P < 0.001). Additional pathoanatomical studies showed that nerve cell loss was most prominent in the outer pyramidal layer III, the inner granular layers IVa and IVc as well as in the multiform layer VI of BA17 of the HD patients. Our neuropathological results in BA17 confirm and extend previous post-mortem, biochemical and in vivo neuroradiological HD findings and offer suitable explanations for the elementary and complex visual dysfunctions, as well as for the altered VEP observed in HD patients.

We performed immunohistochemical study combined with morphometrical analyses in order to examine the expression of matrix metalloproteinase-2 (MMP-2) and MMP-9 using 55 human skin wounds of different ages: group I, 0–3 days (n = 16); II, 4–7 days (n = 11); III, 9–14 days (n = 16); and IV, 17–21 days (n = 12). Immunopositive reactions for MMP-2 were observed in all human skin specimens including uninjured skin as control. The number of MMP-2+ macrophages was significantly increased in accordance with wound ages. In contrast to MMP-2, no MMP-9+ signals were detected in uninjured and wound specimens aged less than 1 day. However, the number of MMP-9+ macrophages profoundly appeared in groups II and III. Morphometrically, in all of wound samples aged 9–12 days, MMP-2+ cell number was more than 20. On the contrary, most of the remaining samples had <20 positive cells. However, only one sample (a 7-day-old wound) showed 21 positive cells. Thus, with regard to practical applicability with forensic safety, MMP-2+ macrophages of >20 would indicate a wound age of 7–12 days. Additionally, 10 out of 12 wound specimens aged 9–12 days showed the MMP-2+ cell number of >25, implying that MMP-2+ cell number of >25 would indicate the wound age of 9–12 days. On the contrary, all wound samples aged 3–14 days except for only one sample had MMP-9+ cell number of >30, indicating that MMP-9+ cell number of >30 would indicate the wound age of 3–14 days. Collectively, MMP-2 seemed to be more distinct marker, compared with MMP-9.

BACKGROUND:The angiotensin converting enzyme (ACE) has been repeatedly discussed as susceptibility factor for major depression (MD) and the bi-directional relation between MD and cardiovascular disorders (CVD). In this context, functional polymorphisms of the ACE gene have been linked to depression, to antidepressant treatment response, to ACE serum concentrations, as well as to hypertension, myocardial infarction and CVD risk markers. The mostly investigated ACE Ins/Del polymorphism accounts for ~40%-50% of the ACE serum concentration variance, the remaining half is probably determined by other genetic, environmental or epigenetic factors, but these are poorly understood. MATERIALS AND METHODS:The main aim of the present study was the analysis of the DNA methylation pattern in the regulatory region of the ACE gene in peripheral leukocytes of 81 MD patients and 81 healthy controls. RESULTS:We detected intensive DNA methylation within a recently described, functional important region of the ACE gene promoter including hypermethylation in depressed patients (p = 0.008) and a significant inverse correlation between the ACE serum concentration and ACE promoter methylation frequency in the total sample (p = 0.02). Furthermore, a significant inverse correlation between the concentrations of the inflammatory CVD risk markers ICAM-1, E-selectin and P-selectin and the degree of ACE promoter methylation in MD patients could be demonstrated (p = 0.01 - 0.04). CONCLUSION:The results of the present study suggest that aberrations in ACE promoter DNA methylation may be an underlying cause of MD and probably a common pathogenic factor for the bi-directional relationship between MD and cardiovascular disorders.

Immunohistochemical study combined with morphometry was carried out to examine the expression of cyclooxygenase-2 (COX-2) using 60 human skin wounds of different ages: group I, 0–4 h (n = 11); II, 8 h–2 days (n = 21); III, 3–9 days (n = 14); and IV, 12–21 days (n = 14). In wound specimens aged 2 h to 2 days, anti-myeloperoxidase-positive neutrophils observed at the wound site expressed immunopositive reaction to COX-2. In wound specimens of more than 3 days, CD68-positive macrophages as well as neutrophils were positively immunostained with anti-COX-2. In group II, all 21 wound samples had COX-2-positive ratios of >40 %, and 15 out of them showed >50 %. In group III, only three wound samples with the postinfliction intervals of 3 days showed positive ratios of 40–50 % and the remaining 11 cases less than 40 %. In groups I and IV, all 25 wound specimens had COX-2-positive ratio of <40 %. With regard to the practical applicability with forensic safety, these observations suggested that a COX-2-positive ratio of >40 % indicated a wound age of 8 h to 3 days. Moreover, COX-2-positive ratios, considerably exceeding a ratio of 50 %, indicate a wound age of 8 h to 2 days. Collectively, COX-2 would be a useful marker for the determination of early wound age.

100 randomised cases where a person lived and died in isolation in Munich were analysed. Factors such as social background, living situation, education, physiological and psychological state of health were evaluated. Personal isolation ( =seclusion) seems to depend on various social, financial, psychological or physical reasons. Lack of contact with other people not only leads to psychological problems, but isolation also contributes to increased illness and early death. In order to improve the present social situation in Munich preventive social measures are necessary to achieve increase in health status for the elder and a decrease in mortality rate.

Brain serotonin (5-HT) neurotransmission plays a key role in the regulation of mood and has been implicated in a variety of neuropsychiatric conditions. Tryptophan hydroxylase (TPH) is the rate-limiting enzyme in the biosynthesis of 5-HT. Recently, we discovered a second TPH isoform (TPH2) in vertebrates, including man, which is predominantly expressed in brain, while the previously known TPH isoform (TPH1) is primarly a non-neuronal enzyme. Overwhelming evidence now points to TPH2 as a candidate gene for 5-HT-related psychiatric disorders. To assess the role of TPH2 gene variability in the etiology of psychiatric diseases we performed cDNA sequence analysis of TPH2 transcripts from human post mortem amygdala samples obtained from individuals with psychiatric disorders (drug abuse, schizophrenia, suicide) and controls. Here we show that TPH2 exists in two alternatively spliced variants in the coding region, denoted TPH2a and TPH2b. Moreover, we found evidence that the pre-mRNAs of both splice variants are dynamically RNA-edited in a mutually exclusive manner. Kinetic studies with cell lines expressing recombinant TPH2 variants revealed a higher activity of the novel TPH2B protein compared with the previously known TPH2A, whereas RNA editing was shown to inhibit the enzymatic activity of both TPH2 splice variants. Therefore, our results strongly suggest a complex fine-tuning of central nervous system 5-HT biosynthesis by TPH2 alternative splicing and RNA editing. Finally, we present molecular and large-scale linkage data evidencing that deregulated alternative splicing and RNA editing is involved in the etiology of psychiatric diseases, such as suicidal behaviour.

PurposeDeciding about the limitation of life-sustaining treatment (LST) is a major challenge for intensive care medicine. The aim of the study was to investigate the practices and perspectives of German intensive care nurses and physicians on limiting LST.MethodsWe conducted an anonymous, self-administered questionnaire survey among the 268 nurses and 95 physicians on all 10 intensive care units of the Munich University Hospital, Germany.ResultsThe response rate was 53%. Of all respondents, 91% reported being confronted with the topic at least once a month. Although all reported limiting cardiopulmonary resuscitation, almost no one reported limiting artificial hydration. Half of nurses and junior physicians felt uncertain about the decision-making process. Junior physicians were most dissatisfied with their training for this task and expressed the highest fear of litigation. Nurses were less satisfied than physicians with the communication process. Both nurses and relatives were not routinely involved in decision making. There is no standardized documentation practice, and many notes are not readily accessible to nurses.ConclusionsLimiting LST is common in German intensive care units. The major shortcomings are team communication, communication with the patient's family, and documentation of the decision-making process.

The immune response against prostate cancer seems to be inefficient although tumour cells show an over-expression of tumour-associated antigens suggesting that regulatory networks inhibit immune cell function locally. To address this proposition, lymphocytes within prostate cancer-inflicted tissue were analysed for the expression of markers associated with negative regulatory function and exhaustion.Prostate cancer, benign prostatic hyperplasia and healthy prostate tissues were investigated by immunohistology for CD25, FOXP3, PD-1 and B7-H1.We had previously documented that prostate cancer islets are surrounded by clustered accumulations of CD3(+) lymphocytes, which lack perforin and interferon-gamma (IFN gamma) expression, thus are apparently quiescent. Here, we report that these clusters contain numerous CD25(+) and FOXP3(+) cells. These markers are associated with regulatory T cells, and their presence in lymphocyte clusters near prostate cancer regions indicates an environment with negative impact on immune response against cancer cells. Consistent with this hypothesis, cells expressing PD-1 and its ligand B7-H1, which are markers associated with exhaustion of lymphocyte function, were also detected in the lymphocyte clusters.Expression of molecules associated with inhibition and exhaustion of lymphocytes may reflect events contributing to ineffective immune responses against cancer cells. (C) 2009 Elsevier Ltd. All rights reserved.

Im Januar 2008 wurden die Ergebnisse der Laenderstudie 2006 von A. Slemeyer und G. Schoknecht in der Zeitschrift Blutalkohol publiziert (ITRD-Nummer D362075), die den Beweiswert der AAK-Analyse auch im strafrechtlich relevanten Konzentrationsbereich ab 1,1 Promille Blutalkoholkonzentration (BAK) beziehungsweise 0,55 mg/l Atemalkoholkonzentration (AAK) belegen sollen. Die Dokumentation weist jedoch einige Auffaelligkeiten und Maengel auf, die erklaerungsbeduerftig sind und diskutiert werden. Vor allem faellt die hohe Ausfallquote von 25 Prozent der ueber 3.500 gemeldeten Datensaetze auf, die einige Fragen offen laesst. Ferner wurde unter anderem hinsichtlich eines Trinkendes keine Auswertung vorgenommen. Die Untersuchungsergebnisse des Muenchener Teilkollektivs der Studie, zum Beispiel mit 90 Prozent verwertbaren Datensaetzen, werden vorgestellt und die Daten verglichen. Der aus den vorgestellten (Teil-)Ergebnissen der Dokumentation abgeleitete Anspruch einer Gleichstellung von AAK- und BAK-Analyse im strafrechtlich relevanten Konzentrationsbereich ist bisher nicht ausreichend zu begruenden, vor allem angesichts der beschriebenen Benachteiligung von 95 Prozent der BAK-Faelle gegenueber AAK bei dem unterstellten Konversionsfaktor von 2.000 und speziell im Grenzwertbereich. (A ) ABSTRACT IN ENGLISH: In January 2008, Slemeyer A. and Schoknecht G. published the results of their nationwide study from 2006 in the journal Blutalkohol (ITRD number D362076), which was performed to prove the strenght of evidence of BrAC-analyses in concentration ranges of statutory limits with penal relevance from 1, 1 per mil BAC respectively 0,55 mg/1 BrAC and more. However, the documentation exhibits some conspicuous things and deficits, which are at least in need of an explanation and discussion. First of all high rate of dropouts of 25 percent of more than 3,500 reported data impresses and leaves some open questions. Furthermore, there was no evalation with respect to the end of drinking. The results of the Munich's part of this study, where e.g. 90 percent of the data could be evaluated, are compared with the collective study. The demand derived from the study, that BrAC-analyses may be used equally in a concentration range relevant in penal law is not justifiable in a sufficient manner, especially in view that BAC cases are disadvantaged in 95 percent compared to simultaneous BrAC if a conversion factor of 2,000 is used as here and especially in the range of the statutory limits. (A)

Fibrocytes, a newly identified cell type, are bone marrow-derived mesenchymal progenitors that coexpress hematopoietic cell antigens and fibroblast products. In this study, a double-color immunofluorescence analysis was carried out using anti-CD45 and anti-collagen type I antibodies to examine the time-dependent appearance of fibrocytes, using 53 human skin wounds with different wound ages (group I, 0-3 days; group II, 4-7 days; group III, 9-14 days; and group IV, 17-21 days). In wound specimens with an age of less than 3 days, CD45(+)/collagen type I+ fibrocytes were not detected. The fibrocytes were initially observed in wounds aged 4 days, and their number increased in lesions with advances in wound age. In a semiquantitative morphometrical analysis, the average number of fibrocytes was highest in the wounds of group III. These findings imply that human skin wounds containing fibrocytes are at least 4 days old. Moreover, a fibrocyte number of over 10 indicates a wound age between 9 and 14 days (i.e., group III). Based on the average number of fibrocytes in each group, a fibrocyte number of over 15 more strongly suggests a wound age of 9-14 days. Together, our observations indicate the participation of fibrocytes in wound healing of human skin inducing the accumulation of extracellular matrix components, and therefore, detection of fibrocytes could be a useful marker for wound age determination.

Eine Studie in Deutschland aus dem Jahr 2001 zeigte, dass 39% der 202 untersuchten Freizeitbodybuilder missbräuchlich anabole androgene Steroide verwendet haben sollen. Anabole androgene Steroide führen neben dem erwünschten Muskelaufbau zu diversen Nebenwirkungen, die sowohl das äußere Erscheinungsbild betreffen, als auch zu krankhaften Veränderungen der inneren Organe führen. Anhand von 15 Sektionsfällen des Instituts für Rechtsmedizin der Ludwig-Maximilians-Universität zu München werden typische pathologische Obduktionsbefunde dargestellt, die eine Verdachtsdiagnose begründen können. Auffallend waren insbesondere schwerwiegende arteriosklerotische und auch thrombotische Befunde am Herzen, z. T. exzessive Verfettungen der Leber sowie eine Hodenatrophie.