[Talus Fractures - an Update]. / Talusfrakturen ­ ein Update.

Background: Talus fractures are rare and often result from axial trauma. As most of the talus surface is covered by cartilage, the blood supply is limited. Thus talus fractures are seen as one of the most severe fractures and often lead to significant long-term complications. Several studies suggest that the initial fracture classification can lead to correct treatment and that this can influence the long-term outcome. The aim of the current study was to investigate the importance of the initial fracture classification in respect to the radiological outcome in a large patient cohort. Patients and Methods: Over a span of 12 years, 61 patients with talus fractures were treated at our institution. Overall 45 patients were available for a retrospective analysis. Correlation analysis was performed between the initial fracture severity and the radiological outcome. Results: The average follow-up was 17.3 months (range 6-68). Significant correlations were found between the Marti-Weber Classification and Bargon Score (rs = 0.78; p < 0.0001), as well as between the Hawkins Classification and the Bargon Score (rs = 0.80; p < 0.0001). Conclusions: Precise prediction of the expected radiological outcome of talar neck and body fractures is possible through the initial fracture classification alone. Computed tomography is the accepted standard to determine the exact diagnosis and extent of injury.

Ceremonies of gratitude following the dissection course: A report on procedures in departments of anatomy in German speaking countries.

The departments of anatomy in Germany, Austria and the German-speaking part of Switzerland were sent comprehensive (18 items) questionnaires requesting details on memorial ceremonies held at the close of the dissection course in the medical curriculum, including objectives, organization, number of participants and the role of the medical students. The response rate was very high (95%). In more than 95% of instances a ceremony is held, initiated mainly after 1970. The titles of the ceremony range from commemoration ceremony (42%), service of mourning (19%) memorial service (19%) to ceremony of gratitude (7%). The number of participants exceeds 300 in 15% of these ceremonies. The invitation comes mostly from the student group organizing the ceremony (62%). The ceremony is offered mainly for the students of the course (23%), for student tutors (16%), relatives of the body donors (23%) and scientific staff (15%). The students actively participate with musical contributions (19%), gestures such as candles (17%) and flowers (12%), speeches (17%) and readings (12%). The relevance of the practical dissection course and body donation programs is also discussed. The results are compared to ceremonies in various countries with different religious backgrounds. This dissection course is unique among all courses in the medical curriculum as it obviously also has spiritual aspects.

[Saw Injuries to the Hand - Epidemiological Aspects]. / Sägeverletzungen der Hand ­ epidemiologische Aspekte.

Background: No current studies regarding saw injuries have been published in German literature for quite some time. Despite awareness measures and safety instructions, saw accidents along with crush injuries are the most common causes of severe hand injuries. Approximately 12,000 circular saw injuries occur in Germany each year. Since professional providers have increased prices due to the rising energy costs and a wide range of different home appliance saws are available, increasing use has been made of portable circular saws in the do-it-yourself market sector. Patients and methods: At our hospital, we evaluated the data of 51 male patients with saw injuries. The present study investigated factors that may contribute to accidents involving saws. Results: 80 % of the accidents occurred at home, usually on weekends or after work. 51 % of the accidents happened while patients cut firewood. In 84 % of the cases, an electric table saw was used. The majority of severe hand injuries were sustained with lower priced saws. Injuries occurred most frequently between 11 a. m. and 2 p. m., primarily with injuries to bones, tendons, blood vessels and nerves. Replantable amputations or partial amputations occured rarely. In 37 % of the patients, anatomical reconstruction using osteosynthesis and/or microsurgical techniques was performed successfully. Conclusions: Saw injuries to the hand are sustained almost exclusively by men. Serious injuries from low-priced table saws and due to the lack of protective covering are predominant. Preventive measures and mandatory training could reduce the number of saw injuries in the years to come. The results obtained by us largely confirm the data from previous publications.

Brain damage resulting from postnatal hypoxic-ischemic brain injury is reduced in C57BL/6J mice as compared to C57BL/6N mice.

Perinatal hypoxia is a critical complication during delivery and is mostly studied in animal models of postnatal hypoxic-ischemic brain injury. We here studied the effects of postnatal hypoxic-ischemic brain injury in two different sub-strains of C57BL/6 mice, i.e. C57BL/6J and C57BL/6N mice. These two sub-strains show different metabolic properties, for instance an impaired glucose tolerance in C57BL/6J mice. Genetically, this was linked to differences in their nicotinamide nucleotide transhydrogenase (Nnt) genes: In C57BL/6J mice, exons 7-11 of the Nnt gene are deleted, resulting in the absence of functional Nnt protein. The mitochondrial Nnt-protein is one of several enzymes that catalyses the generation of NADPH, which in turn is important for the elimination of reactive oxygen species (ROS). As ROS is thought to contribute to the pathophysiology of hypoxia-ischemia, the lack of Nnt might indirectly increase ROS levels and therefore result in increased brain damage. We therefore hypothesize that lesion score and lesion size will increase in C57BL/6J mice as compared to C57BL/6N mice. Surprisingly, the results showed exactly the opposite: C57BL/6J mice showed a decrease in lesion score and size, associated with a reduced number of apoptotic cells and activated microglia. In contrast, the number of cells with ROS-induced DNA modifications (detected by 8OHdG) was higher in C57BL/6J than C57BL/6N mice. In conclusion, C57BL/6J mice showed reduced ischemic consequences after postnatal hypoxic-ischemic brain injury compared to C57BL/6N mice, with the exception of the amount of ROS-induced DNA-damage. These differences might relate to the lack of Nnt, but also to a modified metabolic setting (cardiovascular parameters, oxygen and glucose metabolism, immune function) in C57BL/6J mice.

Effects of macrophage-activating lipopeptide-2 (MALP-2) on the vascularisation of implanted polyurethane scaffolds seeded with microvascular fragments.

The seeding of scaffolds with adipose tissue-derived microvascular fragments represents a promising strategy to establish a sufficient blood supply in tissue constructs. Herein, we analysed whether a single application of macrophage-activating lipopeptide-2 (MALP-2) at the implantation site further improves the early vascularisation of such microvessel-seeded constructs. Microvascular fragments were isolated from epididymal fat pads of C57BL/6 mice. The fragments were seeded on polyurethane scaffolds, which were implanted into mouse dorsal skinfold chambers exposed to MALP-2 or vehicle (control). The inflammatory host tissue response and the vascularisation of the scaffolds were analysed using intravital fluorescence microscopy, histology and immunohistochemistry. We found that the numbers of microvascular adherent leukocytes were significantly increased in MALP-2-treated chambers during the first 3 days after scaffold implantation when compared to controls. This temporary inflammation resulted in an improved vascularisation of the host tissue surrounding the implants, as indicated by a higher density of CD31-positive microvessels at day 14. However, the MALP-2-exposed scaffolds themselves presented with a lower functional microvessel density in their centre. In addition, in vitro analyses revealed that MALP-2 promotes apoptotic cell death of endothelial and perivascular cells in isolated microvascular fragments. Hence, despite the beneficial pro-angiogenic properties of MALP-2 at the implantation site, the herein evaluated approach may not be recommended to improve the vascularisation capacity of microvascular fragments in tissue engineering applications.

RpiRc Is a Pleiotropic Effector of Virulence Determinant Synthesis and Attenuates Pathogenicity in Staphylococcus aureus.

In Staphylococcus aureus, metabolism is intimately linked with virulence determinant biosynthesis, and several metabolite-responsive regulators have been reported to mediate this linkage. S. aureus possesses at least three members of the RpiR family of transcriptional regulators. Of the three RpiR homologs, RpiRc is a potential regulator of the pentose phosphate pathway, which also regulates RNAIII levels. RNAIII is the regulatory RNA of the agr quorum-sensing system that controls virulence determinant synthesis. The effect of RpiRc on RNAIII likely involves other regulators, as the regulators that bind the RNAIII promoter have been intensely studied. To determine which regulators might bridge the gap between RpiRc and RNAIII, sarA, sigB, mgrA, and acnA mutations were introduced into an rpiRc mutant background, and the effects on RNAIII were determined. Additionally, phenotypic and genotypic differences were examined in the single and double mutant strains, and the virulence of select strains was examined using two different murine infection models. The data suggest that RpiRc affects RNAIII transcription and the synthesis of virulence determinants in concert with σ(B), SarA, and the bacterial metabolic status to negatively affect virulence.

The role of IL-1ß in Pseudomonas aeruginosa in lung infection.

This mini-review examines the role of the pro-inflammatory cytokine interleukin (IL)-1ß in the interaction of Pseudomonas aeruginosa and the host immune system during lung infection. Different studies show that the reduction of the inflammatory response, especially a decrease in IL-1ß, leads to a better outcome in acute lung infection with this bacterium. This includes a higher survival rate, reduced damage to the lung tissue and, in particular, a better clearance of the airways and the tissue of the lungs from P. aeruginosa.

Mitochondrial regulation of reactive oxygen species (ROS) production-Unexpected observations in early postnatal cerebral vasculature.

Nicotinamide-nucleotide-transhydrogenase (Nnt) is a mitochondrial protein. It is altered and functionally lacking in the C57BL/6J sub-strain. This leads to the generation of more radical oxygen species than in the C57BL/6N sub-strain. During studies on the effect of Nnt in perinatal hypoxia the cerebral vasculature was investigated in postnatal day 9 mice using post mortem arterial filling with silicone rubber compounds. Surprisingly, the tiny vessels were no longer uniformly filled and a bleb-like pattern occurred in both sub-strains. Furthermore, considerably more bleb-like spots were observed in the C57Bl/6J sub-strain than in the C57Bl/6N sub-strain. These blebs might be the result of feathery vessels bursting. It remains unclear how the mechanisms in the used strains differ. Nnt might influence the vascular structure or its development and mechanisms and should be investigated further.

[Purpura fulminans of the adult--a hematological emergency]. / Purpura fulminans des Erwachsenen--ein hämatologischer Notfall.

HISTORY AND CLINICAL FINDINGS: A 51-year-old female patient with history of longterm drug abuse, was admitted to our hospital with large, stocking-shaped areas of painful, non-displaceable confluent bruising reaching up to the groin. INVESTIGATIONS: The emergency laboratory tests showed leucopenia, thrombocytopenia and anemia as well as a distinct protein C deficiency. DIAGNOSIS, TREATMENT AND COURSE: Purpura fulminans was diagnosed and treated with an initial dose of protein C. The patient survived and the skin necrosis can be treated. CONCLUSION: Purpura fulminans is an internistic and dermatological emergency situation which can lead to shock through consumptive coagulopathy. The serious course of disease can be prevented by rapid treatment with protein C.

Locally applied macrophage-activating lipopeptide-2 (MALP-2) promotes early vascularization of implanted porous polyethylene (Medpor®).

Porous polyethylene (Medpor®) is frequently used in craniofacial reconstructive surgery. Rapid vascularization of the biomaterial crucially contributes to its adequate incorporation without complications. Macrophage-activating lipopeptide-2 (MALP-2) is a toll-like receptor (TLR)-2/6 agonist with pro-angiogenic properties. Herein we analyzed whether local single-shot application of MALP-2 improves the angiogenic host tissue response to Medpor®. Medpor® (3 mm×3 mm×0.25 mm) was implanted into dorsal skinfold chambers of BALB/c mice topically exposed to different MALP-2 doses (0.1 and 0.5 µg) or vehicle (control). The vascularization of the implants and the inflammatory foreign body reaction was analyzed using intravital fluorescence microscopy, histology and immunohistochemistry over 14 days. MALP-2 treatment dose-dependently improved the vascularization of Medpor®, as indicated by a significantly higher functional microvessel density at the border and center of the implants when compared to controls. This was associated with a temporary increase of adherent leukocytes in host tissue venules during the first 3 days after implantation. At day 14, implants in MALP-2-treated chambers were surrounded by granulation tissue, which exhibited a significantly higher density of CD31-positive microvessels and number of F4/80-positive macrophages when compared to controls. Additional biomaterial-free chambers did not show any signs of angiogenesis when treated with MALP-2. This indicates that locally applied MALP-2 effectively stimulates the early vascularization of Medpor® without inducing any local or systemic side effects. Accordingly, this easy approach may further improve the rapid incorporation of this biomaterial at the implantation site.

Expression of caveolin-1 and podocalyxin in rat lungs challenged with 2-kDa macrophage-activating lipopeptide and Flt3L.

Caveolin-1 is one of the important regulators of vascular permeability in inflamed lungs. Podocalyxin is a CD34 protein expressed on vascular endothelium and has a role in podocyte development in the kidney. Few data are available on the expression of caveolin-1 and podocalyxin in lungs challenged with Toll-like receptor 2 (TLR2) agonists such as mycoplasma-derived macrophage activating lipopeptide or with immune modulators such as Fms-like tyrosine kinase receptor-3 ligand (Flt3L), which expands dendritic cell populations in the lung. Because of the significance of pathogen-derived molecules that act through TLR2 and of the role of immune modulators in lung physiology, we examine the immunohistochemical expression of caveolin-1 and podocalyxin in lungs from rats challenged with a 2-kDa macrophage-activating lipopeptide (MALP-2) and Flt3L. Normal rat lungs expressed caveolin-1 in alveolar septa, vascular endothelium and airway epithelium, especially along the lateral borders of epithelial cells but not in alveolar macrophages. MALP-2 and Flt3L decreased and increased, respectively, the expression of caveolin-1. Caveolin-1 expression seemed to increase in microvessels in bronchiole-associated lymphoid tissue (BALT) in Flt3L-challenged lungs but not in normal or MALP-2-treated lungs. Podocalyxin was absent in the epithelium and alveolar macrophages but was present in the vasculature of control, Flt3L- and MALP-2-treated rats. Compared with control and MALP-2-treated rats, Flt3L-treated lungs showed greater expression of podocalyxin in BALT vasculature and at the interface of monocytes and the endothelium. These immunohistochemical data describing the altered expression of caveolin-1 and podocalyxin in lungs treated with MALP-2 or Flt3L encourage further mechanistic studies on the role of podocalyxin and caveolin-1 in lung inflammation.

Toll-like receptor 2/6-dependent stimulation of mesenchymal stem cells promotes angiogenesis by paracrine factors.

Reconstruction of critical size bone defects represents a major challenge in orthopaedic surgery. Insufficient angiogenesis is a limiting factor for engraftment of large-scale tissue transplants. Transplantation or stimulation of local mesenchymal stem cells (MSCs) represents a potential solution to enhance angiogenesis. We recently identified angiogenic properties for the Toll-like receptor (TLR) 2/6 agonist MALP-2 and now investigated if MALP-2 could be used to stimulate MSCs in order to promote angiogenesis in vitro and in vivo. Human MSCs from the bone marrow of healthy subjects were isolated, cultured and expanded in vitro and were shown to be positive for mesenchymal stem cells markers as well as for the MALP-2 receptors TLR2 and TLR6. MALP-2 directly enhanced migration but not proliferation of human MSCs. Conditioned medium from MALP-2 stimulated MSCs significantly increased proliferation, migration and tube formation of endothelial cells. Analysis of the conditioned medium from MSCs revealed that MALP-2 stimulation enhanced the secretion of several chemokines and growth factors including vascular endothelial growth factors (VEGF) and granulocyte-macrophage colony-stimulating factor (GM-CSF). Finally, we studied MALP-2 effects on MSCs in a sheep model of tissue engineering in vivo. Therefore, MSCs were isolated from the iliac crest of black head sheep and co-cultivated with MALP-2 ex vivo. Implantation of autologous MSCs within a scaffold cylinder into the M. latissimus dorsi significantly enhanced vessel density of these constructs after 6 months. We here present the first evidence that TLR2/6-dependent stimulation of MSCs promotes angiogenesis in vitro and in vivo offering a novel strategy for therapeutic angiogenesis, e.g., for tissue engineering of bone.

[The effects of tobacco smoke on alveolar macrophages - an in vivo mouse lung model for short-term smoking]. / Die Auswirkungen von Tabakrauch auf Alveolarmakrophagen - ein In-vivo-Kurzzeitrauchmodell zur Mauslunge.

BACKGROUND: The effect of cigarette smoke (CS) on the phagocytosis of alveolar macrophages is discussed controversially on the basis of in vitro experiments. In this short report we describe the in vivo observations that we have performed. METHODS: For this purpose mice were exposed to CS for three consecutive days. One day later the fluorescent microspheres were administered intratracheally and the lung surface was investigated using long-distance fluorescence microscopy. RESULTS: We found that the numbers of neutrophils which engulfed particles was increased in the CS group as compared to controls. The overall phagocytic activity was not significantly different after CS exposure. CONCLUSIONS: In conclusion the phagocytosis of alveolar macrophages and neutrophils after short time CS exposure was not affected. Further investigations will need to look for the effects of long-term CS exposure and the phagocytosis of living bacteria.

Serum procalcitonin levels in the postmortem diagnosis of sepsis.

Procalcitonin is regarded as a valuable marker for sepsis in living persons and even in post-mortem investigations. At the Institute of Legal Medicine, 25 autopsy cases with suspected bacterial infectious diseases or sepsis were examined using the semi-quantitative PCT-Q(®)-test (B.R.A.H.M.S., Germany) in 2010 and 2011. As controls, 75 cadavers were used for which there was no suspicion of a bacterial infectious disease or sepsis. Femoral blood was cultured from the cases and from controls, and samples from the brain, heart, lungs, liver, spleen and kidneys were examined histologically for findings seen in sepsis. Twelve cases in the sepsis/infectious disease group (48%) were classifiable as sepsis following synopsis of PCT levels, autopsy results, and histopathological and microbiological findings. This study shows that the semi-quantitative PCT-Q(®)-test is a useful supplementary marker in routine autopsy investigations, capable of classifying death as due to sepsis.

An elegant technique for ex vivo imaging in experimental research-Optical coherence tomography (OCT).

Optical coherence tomography (OCT) is an elegant technology for imaging of tissues and organs and has been established for clinical use for around a decade. Thus, it is used in vivo but can also serve as a valuable ex vivo imaging tool in experimental research. Here, a brief overview is given with a focus on an ex vivo application of OCT. Image and video examples of freshly obtained murine lungs are included. The main advantage of OCT for ex vivo analysis is the non-contact, non-invasive, and non-destructive fast acquisition of a three-dimensional data set with micrometer-resolution.

All-trans retinoic acid results in irregular repair of septa and fails to inhibit proinflammatory macrophages.

All-trans retinoic acid (ATRA) is controversially discussed in emphysema therapy. We re-evaluated ATRA in the elastase model and hypothesised that beneficial effects should be reflected by increased alveolar surface area, elastin expression and downregulation of inflammatory mediators and matrix metalloproteinases (MMPs). Emphysema was induced by porcine pancreatic elastase versus saline in Sprague-Dawley rats. On days 26-37, rats received daily intraperitoneal injections with ATRA (500 µg · kg(-1) body weight) versus olive oil. Lungs were removed at day 38. Rat alveolar epithelial L2 cells were incubated with/without elastase followed by ATRA- or vehicle-treatment, respectively. ATRA only partially ameliorated structural defects. Alveolar walls exhibited irregular architecture: increased arithmetic mean thickness, reduction in surface coverage by alveolar epithelial cells type II. ATRA only partially restored reduced soluble elastin. It tended to increase the ratio of ED1(+):ED2(+) macrophages. Bronchoalveolar lavage (BAL) cells exhibited a proinflammatory state and high expression of interleukin-1ß, cytokine-induced neutrophil chemoattractant-1, tumour necrosis factor-α, nuclear factor-κB, MMP-2, MMP-9, MMP-12, tissue inhibitor of metalloproteinase (TIMP)-1 and TIMP-2 in emphysema, with ATRA exerting only few effects. MMP-7 was highly induced by ATRA in healthy but not in emphysematous lungs. ATRA reduced both MMP-2 and TIMP-1 activity in BAL fluid of emphysematous lungs. ATRA-therapy may bear the risk of unwanted side-effects on alveolar septal architecture in emphysematous lungs.

No effect in combining chondrogenic predifferentiation and mechanical cyclic compression on osteochondral constructs stimulated in a bioreactor.

Traumatic and degenerative osteochondral lesions are a common problem in orthopaedic surgery. The concept of tissue engineering represents the possibility of a promising therapeutical approach. The purpose of this study has been to improve the characteristics of osteochondral grafts consisting of a human certified collagen I-bone hybrid matrix seeded with human bone marrow stromal cells and stimulated in a custom-made biomechanoreactor. This study was undertaken as a follow-up to our prior studies. Based on our established system, we added chondrogenic growth factors (IGF-1 and TGF-beta(2)) and evaluated their effect on chondrogenic differentiation. Constructs were stimulated for 14, 21 and 28 days respectively by different protocols, including cyclic mechanical stimulation, hormonal stimulation or a combination of both. More than 70% of the cells were viable throughout the entire experimental period. Histological analysis revealed a homogeneous distribution of cells in a cartilage-like matrix organization. Immunohistological collagen II staining was positive irrespective of stimulation manner and time. Levels of DNA and glycosaminoglycans, having been normalized to DNA, did not change. Analysis of the biomechanical stiffness after 14 days showed increased stiffness in the hormonally and mechanically stimulated group compared to the static group. Stimulation time did not have a significant influence. The media supplements to foster the quality of the tissue tested here did not show any progress in our system. We conclude that cyclic compression enhances matrix stiffness, but stimulation time should be kept short and growth factors should be left out in this system with regard to clinical applicability and financial concerns.

Coincidence of different structures of mucosa-associated lymphoid tissue (MALT) in the respiratory tract of children: no indications for enhanced mucosal immunostimulation in sudden infant death syndrome (SIDS).

Mucosa-associated lymphoid tissue (MALT) is the principal inductive site for mucosal immune responses that are capable of T and B cell responses and antigen-specific responses. In previous independent studies different structures of MALT, e.g. bronchus-, larynx- and nose-associated lymphoid tissue (BALT, LALT, NALT) have been described separately in various frequencies in the human respiratory tract over life spans. Because upper respiratory tract infections are common in infants, dysregulations of mucosal immune responses might be seriously involved in the aetiology of sudden infant death syndrome (SIDS). In the present study the coincidental occurrence of the three different MALT structures in the respiratory tract within the same patients were studied, and cases of SIDS and children who had died from different traumatic and natural causes of death (non-SIDS) were compared. First, the frequency of BALT and LALT in 46 children (35 SIDS, 11 non-SIDS) with or without NALT were examined. A tendency was found of a coincidence of respiratory MALT structures. In 50 additional cases of infant death (30 SIDS, 20 non-SIDS) from the multi-centric German Study on Sudden Infant Death Syndrome (GeSID) where death had occurred in the first year of life, the coincidence was evaluated. A coincidental occurrence of BALT, LALT and NALT or BALT and LALT (each about 30%) was found in both groups, whereby the coincidence in SIDS and the control patients did not differ. Interestingly, the children with coincidental MALT were strikingly older, supporting the hypothesis of respiratory MALT formation via environmental stimulation over time.