Genome Sequences of Microbacteriophages Zada and Ioannes.

Microbacteriophages Zada and Ioannes were isolated from soil and characterized. Genomes were then sequenced and annotated. This was done using the host bacterium Microbacterium foliorum Zada and Ioannes are both lytic phages with a Siphoviridae morphotype.

Coexpression of human perforin improves yeast-mediated delivery of DNA and mRNA to mammalian antigen-presenting cells.

Previous studies underlined the capacity of recombinant yeast as efficient vehicle for the targeted delivery of functional nucleic acids as well as proteinaceous antigens to mammalian antigen-presenting cells (APCs). To improve this yeast-mediated cargo transport into APCs, we investigated the impact of coexpression of the human membrane-perturbing protein perforin in comparison with bacterial listeriolysin O (LLO) on the yeast-based delivery of DNA, mRNA and proteins to mammalian APCs. In contrast to LLO, a cholesterol-dependent pore-forming toxin of Listeria, intracellular expression of human perforin in Saccharomyces cerevisiae had no impact on yeast cell viability, while its coexpression significantly increased translocation of ovalbumin and subsequent activation of ovalbumin-specific T lymphocytes. Likewise, perforin improved the expression of the model antigen enhanced green fluorescent protein after yeast-mediated DNA and mRNA delivery, whereas LLO was only able to enhance DNA delivery. Taken together, our data show that human perforin, besides bacterial hemolysins, represents a promising means to improve the yeast-mediated delivery of functional nucleic acids and proteins to mammalian APCs.

CNS Measures of Pain Responses Pre- and Post-Anesthetic Ketamine in a Patient with Complex Regional Pain Syndrome.

BACKGROUND: Previous reports have indicated that ketamine anesthesia may produce significant improvement if not complete recovery of patients with complex regional pain syndrome (CRPS). AIMS: Here we report on a patient who had CRPS affecting mainly the right side of her body who underwent functional magnetic resonance imaging (fMRI) scans prior to and in the months following apparent successful treatment with anesthetic doses of ketamine. MATERIALS AND METHODS: The patient underwent two imaging sessions: one during her pain state (CRPS+) and 1 month after her ketamine treatment in her pain-free state (CRPS-). Both spontaneous and evoked (brush, cold, and heat) pain scores decreased from 7­9/10 on a visual analog scale prior to the treatment to 0­1 immediately following and for months after the treatment. For each imaging session, the identical mechanical (brush) and thermal (cold and heat) stimuli were applied to the same location (the skin of the dorsum of the right hand). RESULTS: Comparison of CRPS+ vs CRPS- for the three stimuli showed significant changes throughout the cerebral cortex (frontal, parietal, temporal, cingulate, and hippocampus), in subcortical regions such as caudate nucleus, and in the cerebellum. In addition, resting state network analysis showed a reversal of brain network state, and the recovered state paralleled specific default networks in healthy volunteers. DISCUSSION: The observed changes in brain response to evoked stimuli provide a readout for the subjective response. CONCLUSION: Future studies of brain function in these patients may provide novel insight into brain plasticity in response to this treatment for chronic pain.

[Lidocaine patch for therapy of neuropathic and non-neuropathic pain. A clinical case series of 87 patients]. / Lidocain-Pflaster zur Therapie neuropathischer und nichtneuropathischer Schmerzen. Klinische Fallstudie an 87 Patienten.

BACKGROUND: Topical lidocaine patches (LP) reduce pain in postherpetic neuralgia and other forms of focal neuropathy. The aim of this study was to determine clinical predictors of therapeutic success. MATERIAL AND METHODS: The medical histories of 87 patients with neuropathic (NS) and non-neuropathic pain (NNS) who had received LP as an add-on to their established pain medication were retrospectively analyzed. The variables assessed were gender, age, analgesic co-medication, pain localization, adverse effects and presence of dynamic allodynia. The impact of these variables on the clinical pain-relieving effect (scored on a 5-point scale) was investigated. RESULTS: A total of 24 out of 28 patients with manifest allodynia scored the therapy with LP as beneficial, patients without allodynia (n=59, 67.8%) profited significantly less frequently with only 39% (p<0.001). The probability of profiting from LP therapy in the presence of allodynia was found to be about tenfold higher compared to patients without allodynia (odds ratio 9.14). Of the 87 patients investigated 48 were female (55.2%). Allodynia was considerably more frequent in women (39.6%) compared to men (23.1%) but this was insignificant. Of the female patients 62.5% responded to LP treatment, compared to only 43.6% of men. In more than 60% of cases rated as very good pain relief allodynia was manifest and in non-responders only in less than 10%. The variables age, pain localization and analgesic co-medication were not related with the success of therapy. DISCUSSION: Patients with manifest allodynia profited significantly more frequently from LP therapy and were less frequently non-responders. Female patients showed therapeutic success more often together with a higher rate of allodynia. CONCLUSIONS: In the presence of allodynia, in especially of neuropathic origin, LP seems to be an effective and save option for add-on therapy, this being independent from pain localization and age. Gender specific effects however need more systematic investigation.

The PTPN22gain-of-function+1858T(+) genotypes correlate with low IL-2 expression in thymomas and predispose to myasthenia gravis.

Protein tyrosine phosphatase, non-receptor type 22 (PTPN22) inhibits T-cell activation and interleukin-2 (IL-2) production. The PTPN22(gain-of-function)+1858T(+) genotypes predispose to multiple autoimmune diseases, including early-onset (non-thymomatous) myasthenia gravis (MG). The disease association and the requirement of IL-2/IL-2 receptor signaling for intrathymic, negative T-cell selection have suggested that these genotypes may weaken T-cell receptor (TCR) signaling and impair the deletion of autoreactive T cells. Evidence for this hypothesis is missing. Thymoma-associated MG, which depends on intratumorous generation and export of mature autoreactive CD4(+) T cells, is a model of autoimmunity because of central tolerance failure. Here, we analyzed the PTPN22 +1858C/T single nucleotide polymorphism in 426 German Caucasian individuals, including 125 thymoma patients (79 with MG), and investigated intratumorous IL-2 expression levels. Unlike two previous studies on French and Swedish patients, we found strong association of PTPN22 +1858T(+) genotypes not only with early-onset MG (P=0.00034) but also with thymoma-associated MG (P=0.0028). IL-2 expression in thymomas with PTPN22 +1858T(+) genotypes (P=0.028) was lower, implying weaker TCR signaling. We conclude that the PTPN22(gain-of-function) variant biases towards MG in a subgroup of thymoma patients possibly by impeding central tolerance induction.

Effects of monocyte chemoattractant protein 1 on blood-borne cell recruitment after transient focal cerebral ischemia in mice.

Monocyte chemoattractant protein 1 (MCP-1) plays an important role in inflammatory reactions following cerebral ischemia. It is known that MCP-1 overexpression leads to increased infarct volume and elevated hematogenous cell recruitment, while MCP-1-deficient mice develop smaller infarcts. It was supposed that MCP-1 dependent macrophage recruitment might be the underlying mechanism of ischemic brain damage but a precise distinction of local microglia and invading macrophages was not performed. In this study we investigated the differential role of MCP-1 on inflammatory cells in MCP-1-deficient mice, using green fluorescent protein (GFP) transgenic bone marrow chimeras. After 30-min of focal cerebral ischemia microglia was rapidly activated and was not different between MCP-1-deficient mice and wild type controls. Activated microglia outnumbered GFP-positive macrophages over the study period. Furthermore, macrophage infiltration was significantly reduced at day 7 in MCP-1-deficient animals (31.2+/-20.1 cells/mm(2)) compared to MCP-1 wild type mice (131.5+/-66.7 cells/mm(2), P<0.001). Neutrophils were also significantly reduced in MCP-1-deficient mice (62% on day 4% and 87% on day 7; P<0.001). This is the first investigation in cerebral ischemia showing that MCP-1 is necessary for recruiting blood-borne cells to the injury site whereas it does not affect the microglia activation and migration. However, the remarkable predominance of activated microglia and the additional attenuation of invading macrophages suggest that different mechanisms than macrophage recruitment are responsible for the MCP-1-mediated neuroprotective effects after experimental stroke.

A Tactile Seat for Direction Coding in Car Driving: Field Evaluation.

This in-traffic, field study examined the merit of using a car seat instrumented with tactile stimulation elements (tactors) to communicate directional information to a driver. A car seat fitted with an 8 times 8 matrix of tactors embedded in the seat pan was used to code eight different directions (the four cardinal and four oblique directions). With this seat mounted in a car, a field study was conducted under both smooth road and brick road vibratory conditions. The primary performance measures were directional accuracy and reaction time, measured under both alerted and simulated surprise conditions. Overall, the results show that the tactile chair seat provides a promising and robust method of providing directional information. The percentage of correct directional responses was very high (92 percent of all trials), and incorrect responses were typically just one location segment (45 degrees) off.

Obliterating intracranial vasculopathy mimicking multiple sclerosis.

BACKGROUND: The first ever diagnosis of multiple sclerosis (MS) requires consideration of both diagnostic criteria and differential diagnosis. Clinicians are particularly challenged by rare conditions which may mimic MS symptoms and relapses. CASE REPORT: We report the case of a young female patient who presented with relapsing left hemispheric symptoms that were highly suspicious of MS but were caused by an idiopathic occlusive angiopathy of the circle of Willis. CONCLUSION: Occlusive disease of the great cerebral arteries in young patients is a rare but important differential diagnosis of MS. It has to be considered in patients presenting with the first symptoms suspicious of MS as substantial treatment consequences will arise.

Monocyte phagocytosis of viable Staphylococcus aureus is impaired by barbiturates, but not by propofol.

BACKGROUND: Barbiturates and propofol are used for deep sedation of patients with elevated intracranial pressure refractory to standard therapeutic regimens. Such patients often suffer from bacterial infections, which are most commonly caused by Staphylococcus aureus. Various interactions of anesthetics with components of the host defense have been documented, but very little is known about the influence on monocytes, which are a first-line defense against bacterial invasion. Therefore, we studied the effects of thiopental, methohexital, and propofol on monocyte phagocytosis using an in vitro whole blood model of viable S. aureus. MATERIALS AND METHODS: Whole blood samples were preincubated with different concentrations of thiopental, methohexital, and propofol. Phagocytosis was stopped at different time points after addition of viable S. aureus. Monocytes then were stained with monoclonal antibodies for flow cytometric analysis of monocyte recruitment (ratio of ingesting monocytes). Furthermore, the fluorescence intensity of ingested bacteria served as semiquantitative measurement of phagocytosis activity. RESULTS: Both barbiturates inhibited monocyte recruitment and phagocytosis activity concentration-dependently, whereas propofol did not affect any of the investigated parameters. At concentrations of 7.6 x10(-3) M thiopental or 1.1 x 10(-3) M methohexital and greater, monocyte recruitment and phagocytosis activity were significantly inhibited. The calculated half-maximum inhibitory concentration (IC50) of thiopental was 8.4 x 10(-3) M for monocyte recruitment and 8.6 x 10(-3) M for phagocytosis activity. The corresponding values for methohexital were 4.1 x 10(-3) M and 1.1 x 10(-3) M, respectively. CONCLUSION: The two barbiturates induce concentration-dependent inhibition of monocyte phagocytosis, whereas propofol is without effect. In combination with previously described effects on granulocyte function, these findings suggest that defense against bacterial infection might be reduced by barbiturates.

Filling a cervical spine cage with local autograft: change of bone density and assessment of bony fusion.

To date, it remains debatable whether cervical spine fusion cages should be filled with any kind of bone or bone substitute. Using a bone substitute would produce additional costs, using an autologous bone graft from the iliac crest would make the use of the cage at least questionable. As an alternative, cortical and subcortical bone from the anterior osteophytes of the segment in which the disc has been removed could be used to fill the cage: higher costs and complications at the iliac crest could both be avoided and the cage could be filled. However, the fate of these bone chips made from the anterior osteophytes is unclear as well as whether fusion will occur using this technique. The objective of the current study was to investigate possible changes in the bone density of this local autograft in the cage within the first 12 months after surgery by means of computed tomography. A second objective was to assess segmental bony fusion using this technique. 21 patients, suffering from degenerative disc disease of the cervical spine, were included into this prospective study. They all underwent anterior decompression, cage insertion and plate stabilisation. The cage (Rabea, Signus Medizintechnik, Alzenau, Germany), was filled with bone chips made from the anterior osteophytes of the segment that underwent discectomy. On the third day after surgery as well as three, six and 12 months after surgery, an axial computed tomography scan through the cage was taken and density within the apertures of the cage was measured in a standardised manner. Flexion-extension lateral radiographs were taken to investigate segmental fusion. Statistical significance was assumed to be at a 95 % level of significance. 23 cages were implanted. The mean value of the bone density obtained by computed tomography was 505 (+/- 119) HU on day three, 635 (+/- 156) HU after three months, 769 (+/- 162) HU after six months, and 814 (+/- 198) after 12 months. There was a significant difference when the values after 12 months were compared to those obtained after three days (p < 0.001) and after three months (p = 0.004). Bony fusion was seen in 21 out of 23 segments (91.3 %) after 12 months. It may be concluded that this technique could be an alternative to the current treatment options.

[Effects of reduced shear stress on inflammatory reactions in vitro. Effects of pathological flow conditions on leukocyte-endothelial interactions and monocyte tissue factor expression in human cell cultures]. / Einfluss verminderter Scherkräfte auf Entzündungsreaktionen in vitro Effekte pathologischer Strömungsbedingungen auf Leukozyten-Endothel-Interaktionen und monozytäre "Tissue-factor-Expression" in humanen Zellkulturen.

BACKGROUND: During malperfusion and inflammation leukocyte adhesion is common. The purpose of this study was to examine the effects of reduced shear stress on leukocyte-endothelial interactions and subsequent inflammatory reactions such as up-regulation of tissue factor. METHODS: Isolated neutrophils and monocytes were co-incubated with human umbilical venous endothelium at 0-3 dynes/cm(2) in a flow chamber. Adhesion and tissue factor expression on adherent leukocytes were examined at various flow conditions. RESULTS: At 2-3 dynes/cm(2) adhesion occurred only on TNFalpha-activated endothelium. Below 1 dyne/cm(2) similarly increased adhesion was also observed on non-activated endothelium. As was observed for leukocyte adhesion, these shear stress-dependent cell interactions also resulted in an up-regulation of tissue factor on adherent monocytes from non-activated co-cultures. CONCLUSION: Apart from additional activators of inflammation, reduced shear forces may directly contribute to inflammation.

[Intelligence, attention, and memory in patients with myasthenia gravis]. / Intelligenz, Aufmerksamkeit und Gedächtnis bei Patienten mit Myasthenia gravis.

BACKGROUND: In patients with myasthenia gravis (MG), poor performance on cognitive tests has been found. Performance on memory tasks has been reported to correlate with disease activity, but memory in MG patients was also recently found to be unimpaired. PATIENTS AND METHODS: Cognitive functioning was examined in 23 patients with MG and 23 healthy controls. The patients were assessed for IQ, memory, attention, and motor performance. Immunoglobulin G antiacetylcholine receptor autoantibody titers were determined. Event-related potentials were generated for patients and controls. RESULTS: Mean IQs of patients were at average. Memory and attention were not more impaired in patients than controls. Event-related potentials were not delayed in either group. Patients with elevated autoantibody concentrations similarly showed unimpaired neurocognitive and motor functioning. CONCLUSION: In this study, no evidence of neuropsychological impairment was found in MG, arguing against the involvement of higher cortical functions. Sleep abnormalities rather than central mechanisms may explain the memory impairments reported by some studies.

Synaptic congenital myasthenic syndrome in three patients due to a novel missense mutation (T441A) of the COLQ gene.

Congenital myasthenic syndromes (CMS) with deficiency of endplate acetylcholinesterase (AChE) are caused by mutations in the synapse specific collagenic tail subunit gene (COLQ) of AChE. We identified a novel missense mutation (T441A) homozygously in three CMS patients from two unrelated German families. The mutation is located in the C-terminal region of the ColQ protein, which initiates assembly of the triple helix, and is essential for insertion of the tail subunit into the basal lamina. Density gradient analysis of AChE extracted from muscle of one of the patients revealed the absence of asymmetric AChE. All patients were characterized by an onset of disease in childhood, exercise-induced proximal weakness, absence of ptosis and ophthalmoparesis, a decremental EMG response, and deterioration in response to anticholinesterase drugs. However, age at onset, disease progression, disease severity, and functional impairment varied considerably among the three patients. As adults, two siblings from one family experience only mild impairment, while the third patient requires a wheelchair for most of the day and assisted ventilation at night.

["Problem-based learning--inflammation and transplantation": an integrated, subject-centred course in the clinical section of the medical curriculum at the medical school of the University of Muenster]. / Problemorientiertes Lernen--Entzündungsmedizin & Transplantation: ein integrierter, themenzentrierter Kurs im klinischen Studienabschnitt an der Medizinischen Fakultät der Westfälischen Wilhelms-Universität (WWU) Münster.

Conventional medical curricula present information pertinent to chronic inflammatory diseases, infectious diseases and transplantation, via systematic lectures and courses in medical specialties without any integrated approach. The authors report on a 3-week model course that attempts to provide students with an overview of clinical presentation, diagnostics, and therapy of representative disease entities with particular emphasis on the interdisciplinary approach to these problems in hospital practice. In addition to problem-based learning in small groups, the model course comprises interdisciplinary concept lectures, practical demonstrations of specific diagnostic procedures, and bedside teaching. In the meantime, the course "Problem-Based Learning--Inflammation and Transplantation" has been held twice successfully as a mandatory course in the clinical part of the curriculum at the Muenster Medical School.

[Aortocaval compression syndrome]. / Aortokavales Kompressionssyndrom.

Aortocaval compression syndrome (supine hypotensive syndrome) represents a common complication mainly of late pregnancy, although the syndrome has been described to occur as early as 16 weeks of gestation. The nature and severity of symptoms range from unspecific complaints to severe maternal hypotension, loss of consciousness, cardiovascular collapse, and consecutive fetal depression. Predominantly, the syndrome is provoked by placing the parturient supine. Since supine positioning is required for diverse diagnostic and therapeutic procedures in obstetrics, these involve increased risk of aortocaval compression. For the anesthetist, cesarean section is most relevant, because of the coincidence of several risk factors. The following article begins by reviewing the pathophysiology of the syndrome, known risk factors and anesthesiological procedures that predispose to the syndrome. The second part is concerned with prophylactic measures and therapeutic options, together with the discussion of a clinically practicable algorithm.

1.9- m and 2.0- m laser diode pumping of Cr(2+) :ZnSe and Cr(2+) :CdMnTe.

Efficient diode pumping at wavelengths of 1.9 and 2.0microm of a Cr(2+): ZnSe laser with an output power of 105 mW and a slope efficiency of 35% with respect to the absorbed pump power is presented. In addition, Cr(2+): CdMnTe has been laser diode pumped as well as operated in the continuous-wave regime, to the best of our knowledge for the first time.

Phantom limb pain: a report of two cases.

The efficacy of pre-emptive analgesia for phantom limb pain is still unclear. It is generally accepted that pre hyphen;amputation pain increases the incidence of phantom and stump pain, even if pre-emptive analgesia is performed before and during surgery and in the postoperative period. Two cases of traumatic upper limb amputations are described here with no pre-existing pain. Both received similar antinociceptive treatment by continuous block of the brachial plexus through infusion of ropivacaine 0.375% at 5 ml/h for 10 days. Treatment of case 1 was initiated immediately after surgery; however, this amputee developed intensive phantom limb pain which persisted at 6 months. Early use of the prosthesis after surgery was not possible for this patient. The intensity of phantom limb pain in case 2 decreased significantly after 6 months, even though brachial plexus blockade was not started until 5 weeks post-trauma. This patient used a functional prosthesis intensively beginning early after amputation. Serial magnetoencephalographic recordings were performed in both patients. Only case 2 showed significant changes of cortical reorganization. In case 1 markedly less cortical plasticity was found. A combination of relevant risk factors such as a painful neuroma, behavioural and cognitive coping strategies and the early functional use of prostheses are discussed as important mechanisms contributing to the development of phantom pain and cortical reorganization.

Rapid response of identified resident endoneurial macrophages to nerve injury.

Macrophages play a central role in the pathogenesis of peripheral neuropathy but the role of resident endoneurial macrophages is undefined because no discriminating markers exist to distinguish them from infiltrating hematogenous macrophages. We identified and characterized resident endoneurial macrophages during Wallerian degeneration in radiation bone marrow chimeric rats created by transplanting wild-type Lewis rat bone marrow into irradiated TK-tsa transgenic Lewis rats. In such animals, resident cells carry the transgene, whereas hematogenous cells do not. As early as 2 days after sciatic nerve crush and before the influx of hematogenous macrophages, resident transgene-positive endoneurial macrophages underwent morphological and immunophenotypic signs of activation. At the same time, resident macrophages phagocytosing myelin were found, and proliferation was detected by bromodeoxyuridine incorporation. Continuous bromodeoxyuridine feeding revealed that resident endoneurial macrophages sequentially retracted their processes, proliferated, and expressed the ED1 antigen, rendering them morphologically indistinguishable from hematogenous macrophages. Resident endoneurial macrophages thus play an early and active role in the cellular events after nerve lesion before hematogenous macrophages enter the nerve. They may thus be critically involved in the pathogenesis of peripheral neuropathy particularly at early stages of the disease and may act as sensors of pathology much like their central nervous system counterparts, the microglial cells.