Time course of the response to navigated repetitive transcranial magnetic stimulation at 10 Hz in chronic neuropathic pain.

Objective This prospective study evaluated the time to response and outcomes of navigated repetitive transcranial magnetic stimulation (TMS) at a frequency of 10 Hz in patients with chronic neuropathic pain. Methods This prospective study included patients with unilateral chronic neuropathic pain. All patients received motor cortex stimulation at 10 Hz over nine consecutive days using repetitive TMS. Outcome was evaluated over a six-week follow-up period using the visual analogue scale, the German Pain Questionnaire and time to pain reduction. Results Fifty patients (23 female, 27 male) were recruited. Two patients were excluded from analysis owing to premature discontinuation of treatment and follow-up. 31/48 patients in the cohort suffered from atypical facial pain. The pain duration ranged approximately from six months to 27 years. After six weeks, 28/46 patients reported a significant level of pain relief (P < 0.001). Conclusion Navigated repetitive TMS for chronic pain is a non-invasive modality with demonstrable clinical benefit. In particular, patients with atypical facial pain with a clear clinicoanatomical correlate responded well to high-frequency stimulation. Patients with a mean pain history of less than five years benefited significantly from this treatment, so early treatment with repetitive TMS should be encouraged.

A framework to assess landscape structural capacity to provide regulating ecosystem services in West Africa.

The Sudanian savanna landscapes of West Africa are amongst the world's most vulnerable areas to climate change impacts. Inappropriate land use and agriculture management practices continuously impede the capacity of agricultural landscapes to provide ecosystem services (ES). Given the absence of practical assessment techniques to evaluate the landscape's capacity to provide regulating ES in this region, the goal of this paper is to propose an integrative assessment framework which combines remote sensing, geographic information systems, expert weighting and landscape metrics-based assessment. We utilized Analytical Hierarchical Process and Likert scale for the expert weighting of landscape capacity. In total, 56 experts from several land use and landscape management related departments participated in the assessment. Further, we adapted the hemeroby concept to define areas of naturalness while landscape metrics including Patch Density, Shannon's Diversity, and Shape Index were utilized for structural assessment. Lastly, we tested the reliability of expert weighting using certainty measurement rated by experts themselves. Our study focused on four regulating ES including flood control, pest and disease control, climate control, and wind erosion control. Our assessment framework was tested on four selected sites in the Vea catchment area of Ghana. The outcome of our study revealed that highly heterogeneous landscapes have a higher capacity to provide pest and disease control, while less heterogeneous landscapes have a higher potential to provide climate control. Further, we could show that the potential capacities to provide ecosystem services are underestimated by 15% if landscape structural aspects assessed through landscape metrics are not considered. We conclude that the combination of adapted land use and an optimized land use pattern could contribute considerably to lower climate change impacts in West African agricultural landscapes.

Biplanar Fluoroscopy-Guided Percutaneous Lead Implantation for Spinal Cord Stimulation: Technical Note.

INTRODUCTION: The correct positioning of spinal cord stimulator leads is assessed radiographically during their percutaneous implantation for trial stimulation. Usually the C-arm is repositioned several times to allow imaging in different planes, which may extend the total duration of surgery. The study aimed to evaluate whether the concurrent intraoperative use of 2 C-arms could safely reduce the duration of surgery. MATERIALS: This retrospective study included cases of percutaneous implantation of a spinal cord stimulation (SCS) lead for trial neurostimulation between 2006 and 2011. The duration of the surgical intervention was recorded, along with the duration of the preparation stage in the operating room. In addition, total radiation exposure time per case was recorded. RESULTS: Ninety-four patients underwent percutaneous implantation of an SCS lead (72 thoracolumbar, 22 cervical). In 73 cases 2 C-arms were used, with 21 cases performed with a single C-arm. In both the cervical and thoracolumbar groups, a biplanar configuration was associated with significant reduction in the mean length of the surgical phase, by 29 minutes (P = 0.017) and 14 minutes, respectively (P = 0.016), albeit while increasing the duration of the preoperative preparation stage. There was no significant difference in the total duration in the operating room or in the total radiation exposure time between groups. CONCLUSIONS: Here we present a technical note on the use of a biplanar fluoroscopy configuration for percutaneous implantation of SCS leads. This arrangement correlated with a reduction in surgery duration without increasing total radiation exposure, representing a practical and safe adjustment to current practice.

Regionalisation of climate change sensitive forest development types for potential afforestation areas.

This paper describes how to use sectoral planning information from forestry to predict and up-scale information on Climate Change sensitive forest development types for potential afforestation areas. The method was developed and applied in the frame of the project RegioPower with focus on the case study region 'Oberes Elbtal-Osterzgebirge'. The data for our study was taken from forest management planning at level of the Federal State of Saxony, Germany. Here, a silvicultural system is implemented, which describes best practices to develop our actual forests into Climate Change adapted forest development types. That includes the selection of drought resistant tree species, a broad range of tree species mixtures per eligible forest development type and the tending, harvesting and regeneration strategies to be applied. This information however, exists only for forest areas and not for areas which could be potentially afforested. The eligibility of the forest development types within the actual forest areas depends on site information, such as nutrient potential, exposition and hydrological soil parameters. The regionalisation of the forest development types to landscape scale had to be based on topographical parameters from the digital elevation model and hydrological soil parameters from soil mapping. In result, we could provide maps for regional planning and decision making with spatially explicit information on the eligible forest development types based on forest management planning information. These maps form a valuable input for testing and optimising afforestation areas with regard to improving the ability of our case study region to mitigate Climate Change effects such as water erosion or drought.

Assessment of the effects of forest land use strategies on the provision of ecosystem services at regional scale.

This paper presents results of a case study in Middle Saxony, Germany, where the impact of conversion, afforestation and alternatively introduction of short rotation coppice areas on the provision of ecosystem services was tested in a spatially inexplicit and a spatially explicit way to formulate recommendations for regional planning. While the spatially inexplicit testing did not lead to clear results regarding to what degree forests or short rotation coppice areas are desirable and applicable, the spatially explicit testing revealed that an increase in the forest area or area with short rotation coppice by 29.7% in unstructured agriculturally dominated Loess regions, 14.4% in more topographically structured parts in the North-East of the model region and 23.6% in its mountainous parts would be beneficial. Potentially resulting losses in the provision of bioresources and regional economy can be considerably reduced by replacing afforestation areas with short rotation coppice. In summary, we found that the spatially explicit analysis of land use scenarios in combination with a more detailed land use classification and including an assessment of changes in land use pattern gave us an improved basis for assessing different possible planning strategies and to enhance the communication between forest management planners and regional planners.

Reduced immune effector cell NKG2D expression and increased levels of soluble NKG2D ligands in multiple myeloma may not be causally linked.

BACKGROUND: There is limited understanding of the dysregulation of the innate immune system in multiple myeloma (MM). We analysed the expression of the activating receptor NKG2D on NK cells and T cells of MM patients and investigated the impact of soluble versus membrane-bound NKG2D ligands on the expression of NKG2D. DESIGN: NKG2D expression on NK cells and CD8+ alphabeta T cells from patients with MM or monoclonal gammopathy of uncertain significance and healthy controls was examined flow-cytometrically. Sera from patients and controls were analysed for soluble NKG2D ligands (sNKG2D ligands). RESULTS: Significantly fewer NK cells and CD8+ alphabeta T cells from patients expressed NKG2D compared to healthy controls (NK cells: median 54% interquartile range (IQR) 32-68 versus 71% IQR 44-82%, P = 0.017, CD8+ alphabeta T cells: median 63% IQR 52-81 versus 77% IQR 71-90%, P = 0.018). The sNKG2D ligand sMICA was increased in patients [median 175 (IQR 87-295) pg/ml] versus controls [median 80 (IQR 32-129) pg/ml, P < 0.001], but levels of sMICA did not correlate with NKG2D expression on effector cells. To elucidate the mechanism of NKG2D down-regulation, we incubated lymphocytes from healthy donors in the presence of sNKG2D ligands or in co-culture with MM cell lines. sNKG2D ligands in clinically relevant concentrations did not down-regulate NKG2D expression, but co-culture of effector cells with myeloma cells with high surface expression of NKG2D ligands reduced NKG2D expression significantly. CONCLUSIONS: These results indicate that MM is associated with a significant reduction in NKG2D expression which may be contact-mediated rather than caused by soluble NKG2D ligands.

Pancreas carcinoma antigen fused to invariant chain elicits T-cell response and tumor growth inhibition.

OBJECTIVES: The major histocompatibility complex class II chaperone invariant chain (Ii) is widely used as a carrier for inserted antigenic sequences and their introduction into the class II processing pathway. The tumor-associated antigen core 2beta 1,6 N-acetylglucosaminyltransferase (C2GnT), a glycosyltransferase present in human pancreatic tumor cells, is not expressed by normal pancreatic tissues. METHODS: A set of expression vectors was engineered where the class II binding region of Ii was replaced by C2GnT-derived sequences. We investigated in vitro whether dendritic cells transfected with Ii-C2GnT constructs were capable to stimulate proliferation of CD4 T cells. We also tested whether vaccination with Ii-C2GnT would protect mice from tumor development. RESULTS: Invariant chain-C2GnT fusion proteins bind to human DR1, DR3, DR4 and to mouse I-A molecules. Our results demonstrate that the plasmid DNA encoding the C2GnT epitope embedded in Ii induces tumor-specific T-cell responses. Mice immunized with the Ii constructs showed reduced growth of Panc02 pancreatic tumor cells. CONCLUSIONS: Therefore, Ii clipped with the tumor-associated antigen C2GnT shows promise for the treatment of pancreatic cancer.

Cytokine-induced killer cells are type II natural killer T cells.

BACKGROUND: Until now, cytokine-induced killer (CIK) cells were assumed to be part of the type I natural killer T (NKT) cell population, but it was not yet investigated if this is correct. METHODS: For analysis, CIK cells were generated by various culture conditions. Human type I NKT cells express a T cell receptor (TCR) composed of an invariant V alpha 24-J alpha Q chain combined with one of several V beta chains. The V alpha 24 is a reliable marker for the presence of these TCRs. RESULTS: While comparing cultures stimulated with different substances, we observed the lack of any V alpha 24 on the surface of CIK culture cells. CONCLUSION: We conclude that CIK cells do not belong to the type I NKT cells.

[Health behaviour of men. Health and illness in private letters, 1800-1900]. / Gesundheitsverhalten von Männern. Gesundheit und Krankheit in Briefen, 1800-1950.

Current research shows that men generally have a lower awareness of health and health prevention than women. However, differences in illness behaviour can be observed in terms of how to deal with diseases, in the awareness of complaints and in the use of physicians. In this chapter, private correspondences of men from different social classes (aristocrats, scientists, artisans and journeymen) are presented for the period between 1800 and 1950. The questions addressed are the following: What expectations do men have from medicine and how do they experience treatment given by doctors? What actions do they perceive as dangerous for their health and how do they handle them? The health strategies of men are closely linked to the respective concepts of masculinity in different historical epochs. The point in this investigation is to reveal that the experiences of illness and of health are composed of individual as well as of social components: both reflect the history, situation and way of living of one specific person.

Anti-tumoral capabilities of effector cells after IFN-alpha or CpG-motif treatment of cocultured dendritic cells.

INTRODUCTION: Ex vivo expansion of monocyte-derived dendritic cells (mDCs) and subsequent coculture with autologous cytokine-induced killer (CIK) cells is an established system to create specific and non-specific anti-tumoral immunity. mDCs constitute the most frequently applied DC subset in clinical studies. One recently published approach to optimize the immunological functions of the DC/CIK cell system is the replacement of interleukin (IL)-4 by interferon (IFN)-alpha in the maturation process of the DCs. MATERIALS AND METHODS: The expressions of relevant surface antigens of IL-4-DCs and IFNalpha-DCs by flow cytometry and the anti-tumoral activation of effector cells cocultured with both types of DCs using cytotoxicity assays were compared. In addition, short-term coculture experiments with both types of DCs and IFNgamma-LAK effector cells were performed and compared with standard CIK cell coculture experiments. RESULTS: Regarding the expressions of functionally relevant surface markers, no differences could be detected for CD80, CD83, and HLA-DR between IFNalpha-DCs and IL-4-DCs, whereas the mean fluorescence intensities of CD40, CD86, CD54, and HLA-ABC were decreased and the expression of CD14 was increased for IFNgamma-DCs. Moreover, no enhancement of cytotoxicity of cocultured CIK cells against tumor cell lines (A498 and SW480) was detected by the use of IFNalpha-DCs. Additionally, coculture experiments with IFNgamma-LAK cells were performed and unexpectedly higher lysis rates in comparison with the established IL-4-DC/CIK coculture model was observed. Early incubation of the mDCs with several CpG-ODNs failed to increase the anti-tumoral cytotoxicity of the cocultured IFNgamma-LAK cells. CONCLUSIONS: These results demonstrate that in the mDC/CIK cell system, IFNalpha-DCs are not superior in inducing anti-tumoral cytotoxicity and even moderately inferior regarding the expression of functionally relevant surface markers compared with IL-4-DCs.

Patients' expectations, motivation and multi-dimensional subjective and objective socio-medical success in medical rehabilitation measures.

This report describes the subjective and objective socio-medical rehabilitation success of a cohort of patients with a primary musculoskeletal or cardiovascular diagnosis 1 year after medical rehabilitation. Furthermore, the predictive value of patients' motivation, expectations and subjective need for rehabilitation, at time of application, on rehabilitation success are examined. Data were derived from a German study, comparing different methods of social medical assessments for the selection of patients for inpatient medical rehabilitation. The results discussed refer to a sample of 352 patients with primary orthopaedic or cardiovascular disease who replied to the 1-year follow-up survey. Rehabilitation subjectively had positive effects for the majority and a positive socio-medical outcome (for at least 80% of the participants); although there was lower subjective success concerning job-related problems and working ability. Multiple regression analysis showed that somatic impairments, functional disabilities in general life as well as health and coping-related expectations/motivation are significant predictors of rehabilitation success. Generally, the explained variance by the regression model was low and further potential criteria of rehabilitation success should be investigated.

SAP and SLAM expression in anti-CD3 activated lymphocytes correlates with cytotoxic activity.

Signalling lymphocyte activation molecule (SLAM)-associated protein (SAP) is a small protein that is mutant in humans with X-linked lymphoproliferative (XLP) disease. Patients with XLP disease are affected by fatal EBV infection and malignant B-cell lymphomas. The increased risk for B-cell lymphomas is suggested to result from impaired immunosurveillance of B-cell proliferation by T cells. In this study, we investigated the role of SLAM and SAP in activation of effector cells with cytotoxic activity, cytokine-induced killer (CIK) cells, which are generated by non-specific stimulation of the TCR and addition of exogenous IL-2. Agonistic TCR activation 1 day after preparation (day +1) resulted in cell activation, with a peak of SLAM on day +6 visible at both the protein and mRNA level as well as membrane detectable SLAM. This increase in SLAM expression correlated significantly with SAP expression at the mRNA level as well as at the protein level. Cytotoxic activity peaked 1 day after the observed SAP and SLAM peaks. At that point in time, IL-10 secretion, which was high during the early days of culture, decreased. In conclusion, activation of peripheral blood cells with agonistic anti-CD3 antibody and exogenous IL-2, as used for generation of CIK cells, results in significant SLAM and SAP activation 5 days after TCR stimulation. This peak correlates with cytotoxic activity against tumour cells. Expression of SLAM and SAP seems to be important in the activation of cytotoxic effector cells.

Mechanisms of thymidine kinase/ganciclovir and cytosine deaminase/ 5-fluorocytosine suicide gene therapy-induced cell death in glioma cells.

Suicide gene transfer using thymidine kinase (TK) and ganciclovir (GCV) treatment or the cytosine deaminase (CD)/5-fluorocytosine (5-FC) system represents the most widely used approach for gene therapy of cancer. However, molecular pathways and resistance mechanisms remain controversial for GCV-mediated cytotoxicity, and are virtually unknown for the CD/5-FC system. Here, we elucidated some of the cellular pathways in glioma cell lines that were transduced to express the TK or CD gene. In wild-type p53-expressing U87 cells, exposure to GCV and 5-FC resulted in a weak p53 response, although apoptosis was efficiently induced. Cell death triggered by GCV and 5-FC was independent of death receptors, but accompanied by mitochondrial alterations. Whereas expression of Bax remained unaffected, in particular, GCV and also 5-FC caused a decline in the level of Bcl-2. Similar findings were obtained in 9L and T98G glioma cells that express mutant p53, and also underwent mitochondrial apoptosis in both the TK/GCV and CD/5-FC system. Upon treatment of 9L cells with 5-FC, Bcl-xL expression slowly declined, whereas exposure to GCV resulted in the rapid proapoptotic phosphorylation of Bcl-xL. These data suggest that TK/GCV- and CD/5-FC-induced apoptosis does neither require p53 nor death receptors, but converges at a mitochondrial pathway triggered by different mechanisms of modulation of Bcl-2 proteins.

Biology of weed pollen allergens.

Weeds represent a heterogeneous group of plants, usually defined by no commercial or aesthetic value. Important allergenic weeds belong to the plant families Asteraceae, Amaranthaceae, Urticaceae, Euphorbiaceae, and Plantaginaceae. Major allergens from ragweed, mugwort, feverfew, pellitory, goosefoot, Russian thistle, plantain, and Mercurialis pollen have been characterized to varying degrees. Four major families of proteins seem to be the major cause of allergic reactions to weed pollen: the ragweed Amb a 1 family of pectate lyases; the defensin-like Art v 1 family from mugwort, feverfew, and probably also from sunflower; the Ole e 1-like allergens Pla l 1 from plantain and Che a 1 from goosefoot; and the nonspecific lipid transfer proteins Par j 1 and Par j 2 from pellitory. As described for other pollens, weed pollen also contains the panallergens profilin and calcium-binding proteins, which are responsible for extensive cross-reactivity among pollen-sensitized patients.

A neuroblastoma-selective suicide gene therapy approach using the tyrosine hydroxylase promoter.

In this study, selective expression of therapeutic transgenes was evaluated in neuroblastoma cells. Promoter fragments of the genes for neuron-specific enolase (NSEp), tyrosine hydroxylase (THp), and dopamine-beta-hydroxylase (DBHp) were studied in neuroblastoma and nonneuronal cell lines by transient transfection experiments using fluorescence-activated cell sorting (FACS) analysis of enhanced green fluorescent protein (egfp) and luciferase (luc+) assay. Both reporter gene assays revealed a neuroblastoma-selective expression mediated by NSEp and THp, whereas DBHp was active only in a murine neuroblastoma cell line. Reporter gene expression by NSEp in neuroblastoma cells was markedly higher than expression by THp, but NSEp also showed considerable background activity in nonneuronal cells. THp-driven expression of egfp was 35-fold higher in human neuroblastoma MHH-NB11 compared with nonneuronal HeLa cells. Thus, THp was chosen for a neuroblastoma-selective suicide gene therapy approach using the herpes simplex virus type 1 thymidine kinase (HSV-tk)/ganciclovir (GCV) system. A retrovirus vector that contained an expression cassette of a HSV-tk/egfp fusion gene and THp in antisense orientation was generated. Stably transduced human neuroblastoma cells and nonneuronal cell lines were generated, and HSV-tk/egfp expression was measured by FACS and GCV cytotoxicity assay. There was a 2.2-fold difference in green fluorescence and a 1.4-fold difference in cell killing between the human neuroblastoma MHH-NB11 and HeLa cells after HSV-tk/egfp gene transfer. The overall difference in THp-HSV-tk/egfp-mediated cell killing between neuroblastoma and nonneuronal tumor cell lines was statistically significant (P = 0.001). In conclusion, the present study demonstrated the feasibility of a neuroblastoma-selective gene therapy approach using the THp/HSV-tk/egfp expression cassette.

Differential cytotoxicity and bystander effect of the rabbit cytochrome P450 4B1 enzyme gene by two different prodrugs: implications for pharmacogene therapy.

The time course of cytotoxicity induction and the bystander effect of the rabbit cytochrome P450 4B1 (cyp4B1)/4-ipomeanol (4-IM) or 2-aminoanthracene (2-AA) pharmacogene therapy systems were investigated and compared with the herpes simplex virus type 1 thymidine kinase/ganciclovir (HSV-tk/GCV) system. Experiments were performed in rat 9L gliosarcoma cells stably expressing cyp4B1 (9L-4B1), HSV-tk (9L-tk), or their egfp (enhanced green fluorescent protein) fusion genes. Cyp4B1-mediated activation of 2-AA showed a high cell killing efficiency within only 48 hours with an onset after already 15 minutes of prodrug exposure. Residual 9L-4B1 cells were mostly damaged sublethally upon 2-AA treatment showing an S phase arrest by cell cycle analysis. 4-IM treatment of 9L-4B1 cells generated an overall weaker cell killing, especially after prodrug exposure times of less than 48 hours. Residual cells surviving 4-IM treatment showed a G2/M arrest and restarted proliferation after prodrug treatment was stopped. HSV-tk/GCV pharmacogene therapy resulted in a slower cytotoxicity induction than cyp4B1/2-AA treatment with a significantly lower cell killing efficiency after 24 and 48 hours. HSV-tk/GCV-mediated cytotoxicity was widely similar to the cytotoxicity induced by cyp4B1/4-IM with the exception of a continuous 48-hour prodrug exposure where 4-IM treatment showed a significantly higher cell killing rate. Cells surviving HSV-tk/GCV suicide gene therapy were not viable and showed an S-phase arrest. Whereas HSV-tk/GCV induced a strong bystander effect, only moderate bystander cell death depending on cell-to-cell contact was demonstrated in 9L/9L-4B1 cocultures upon 2-AA treatment and was even absent with 4-IM, thereby contrasting with earlier reports. The absence of a strong bystander effect may limit, on one hand, the overall utility of the cyp4B1 systems for cancer gene therapy. On the other hand, the weak bystander effect together with the fast induction of cytotoxicity may provide marked advantages for the use of the cyp4B1 systems as biosafety enhancers for gene marking or replacement studies and donor lymphocyte infusions after allogeneic bone marrow transplantation.