Search

Currently, in routine diagnostics, most molecular testing is performed on formalin-fixed, paraffin-embedded tissue after a histomorphological assessment. In order to find the best possible and targeted individual therapy, knowing the mutational status of the tumour is crucial. The “AVENIO Millisect” system Roche introduced an automation solution for the dissection of tissue on slides. This technology allows the precise and fully automated dissection of the tumour area without wasting limited and valuable patient material. In this study, the digitally guided microdissection was directly compared to the manual macrodissection regarding the precision and duration of the procedure, their DNA concentrations as well as DNA qualities, and the overall costs in 24 FFPE samples. In 21 of 24 cases (87.5%), the DNA yields of the manually dissected samples were higher in comparison to the automatically dissected samples. Shorter execution times and lower costs were also benefits of the manual scraping process. Nevertheless, the DNA quality achieved with both methods was comparable, which is essential for further molecular testing. Therefore, it could be used as an additional tool for precise tumour enrichment.

BACKGROUND Immune checkpoint inhibitors (ICIs) are among the most promising treatment options for melanoma and non-small cell lung cancer (NSCLC). While ICIs can induce effective anti-tumor responses, they may also drive serious immune-related adverse events (irAEs). Identifying biomarkers to predict which patients will suffer from irAEs would enable more accurate clinical risk-benefit analysis for ICI treatment and may also shed light on common or distinct mechanisms underpinning treatment success and irAEs. METHODS In this prospective multi-center study, we combined a multi-omics approach including unbiased single-cell profiling of over 300 peripheral blood mononuclear cell (PBMC) samples and high-throughput proteomics analysis of over 500 serum samples to characterize the systemic immune compartment of patients with melanoma or NSCLC before and during treatment with ICIs. FINDINGS When we combined the parameters obtained from the multi-omics profiling of patient blood and serum, we identified potential predictive biomarkers for ICI-induced irAEs. Specifically, an early increase in CXCL9/CXCL10/CXCL11 and interferon-γ (IFN-γ) 1 to 2 weeks after the start of therapy are likely indicators of heightened risk of developing irAEs. In addition, an early expansion of Ki-67+ regulatory T cells (Tregs) and Ki-67+ CD8+ T cells is also likely to be associated with increased risk of irAEs. CONCLUSIONS We suggest that the combination of these cellular and proteomic biomarkers may help to predict which patients are likely to benefit most from ICI therapy and those requiring intensive monitoring for irAEs. FUNDING This work was primarily funded by the European Research Council, the Swiss National Science Foundation, the Swiss Cancer League, and the Forschungsförderung of the Kantonsspital St. Gallen.

Christina Schmitt,1 Anna Lechanteur,2 François Cossais,1 Coralie Bellefroid,2 Philipp Arnold,1 Ralph Lucius,1 Janka Held-Feindt,3 Geraldine Piel,2 Kirsten Hattermann1 1Institute of Anatomy, University Kiel, Kiel D-24098, Germany; 2Laboratory of Pharmaceutical Technology and Biopharmacy (LTPB), CIRM, University of Liège, Liège, Belgium; 3Department of Neurosurgery, University Medical Center Schleswig-Holstein UKSH, Kiel D-24105, GermanyCorrespondence: Kirsten HattermannInstitute of Anatomy, University Kiel, Olshausenstr. 40, Kiel D-24098, GermanyTel +49 431 880 2460Fax +49 431 880 1557Email k.hattermann@anat.uni-kiel.deIntroduction: The polyphenolic spice and food coloring ingredient curcumin has beneficial effects in a broad variety of inflammatory diseases. Amongst them, curcumin has been shown to attenuate microglia reaction and prevent from glial scar formation in spinal cord and brain injuries.Methods: We developed a protocol for the efficient encapsulation of curcumin as a model for anti-inflammatory drugs yielding long-term stable, non-toxic liposomes with favorable physicochemical properties. Subsequently, we evaluate the effects of liposomal curcumin in experimental models for neuroinflammation and reactive astrogliosis.Results: We could show that liposomal curcumin can efficiently reduce the reactivity of human microglia and astrocytes and preserve tissue integrity of murine organotypic cortex slices.Discussion and Perspective: In perspective, we want to administer this curcumin formulation in brain implant coatings to prevent neuroinflammation and glial scar formation as foreign body responses of the brain towards implanted materials.Keywords: glial scar, neuroinflammation, nanoparticles, brain implants

Glioblastoma multiforme (GBM) is a poorly curable disease due to its profound chemoresistance. Despite recent advances in surgery, radiotherapy and chemotherapy, the efficient treatment of GBMs is still a clinical challenge. Beside others, AT101, the R-(−) enantiomer of gossypol, and demethoxycurcumin (DMC), a curcumin-related demethoxy compound derived from Curcuma longa, were considered as possible alternative drugs for GBM therapy. Using different human primary GBM cell cultures in a long-term stimulation in vitro model, the cytotoxic and anti-proliferative effects of single and combined treatment with 5 µM AT101 and 5 µM or 10 µM DMC were investigated. Furthermore, western blots on pAkt and pp44/42 as well as JC-1 staining and real-time RT-PCR were performed to understand the influence of the treatment at the molecular and gene level. Due to enhanced anti-proliferative effects, we showed that combined therapy with both drugs was superior to a single treatment with AT101 or DMC. Here, by determination of the combination index, a synergism of the combined drugs was detectable. Phosphorylation and thereby activation of the kinases p44/42 and Akt, which are involved in proliferation and survival processes, were inhibited, the mitochondrial membrane potential of the GBM cells was altered, and genes involved in dormancy-associated processes were regulated by the combined treatment strategy. Combined treatment with different drugs might be an option to efficiently overcome chemoresistance of GBM cells in a long-term treatment strategy.

Brain implants are promising instruments for a broad variety of nervous tissue diseases with a wide range of applications, e.g. for stimulation, signal recording or local drug delivery. Recently, graphene-based scaffold materials have emerged as attractive candidates as neural interfaces, 3D scaffolds, or drug delivery systems due to their excellent properties like flexibility, high surface area, conductivity, and lightweight. To date, however, there is a lack of appropriate studies of the foreign body response, especially by glial cells, towards graphene-based materials. In this work, we investigated the effects of macroscopic, highly porous (>99.9%) graphene oxide (GO) and reduced graphene oxide (rGO) (conductivity ∼1 S m−1) scaffolds with tailorable macro- and microstructure on human astrocyte and microglial cell viability and proliferation as well as expression of neuroinflammation and astrogliosis associated genes in an indirect contact approach. In this in vitro model, as well as ex vivo in organotypic murine brain slices, we could demonstrate that both GO and rGO based 3D scaffolds exert slight effects on the glial cell populations which are the key players of glial scar formation. These effects were in most cases completely abolished by curcumin, a known anti-inflammatory and anti-fibrotic drug that could in perspective be applied to brain implants as a protectant.

Ingredients of brown seaweed like fucoidans are often described for their beneficial biological effects, that might be interesting for a medical application. In this study, we tested an extract from Dictyosiphon foeniculaceus (DF) to evaluate the effects in glioblastoma and uveal melanoma, looking for a possible anti-cancer treatment. We investigated toxicity, VEGF (vascular endothelial growth factor) secretion and gene expression of tumor and non-tumor cells. SVGA (human fetal astrocytes), the human RPE (retinal pigment epithelium) cell line ARPE-19, the tumor cell line OMM-1 (human uveal melanoma), and two different human primary glioblastoma cultures (116-14 and 118-14) were used. Tests for cell viability were conducted with MTS-Assay (3-(4,5-Dimethylthiazol-2-yl)-5-(3-carboxymethoxyphenyl)-2-(4-sulfophenyl)-2H-tetrazolium), and the proliferation rate was determined with cell counting. VEGF secretion was assessed with ELISA (enzyme-linked immunosorbent assay). The gene expression of VEGF receptor 1 (VEGFR1), VEGF receptor 2 (VEGFR2) and VEGF-A was determined with real-time qPCR (quantitative polymerase chain reaction). DF lowered the cell viability of OMM-1. Proliferation rates of ARPE-19 and OMM-1 were decreased. The VEGF secretion was inhibited in ARPE-19 and OMM-1, whereas it was increased in SVGA and 116-14. The expression of VEGFR1 was absent and not influenced in OMM-1 and ARPE-19. VEGFR2 expression was lowered in 116-14 after 24 h, whereas VEGF-A was increased in 118-14 after 72 h. The extract lowered cell viability slightly and was anti-proliferative depending on the cell type investigated. VEGF was heterogeneously affected. The results in glioblastoma were not promising, but the anti-tumor properties in OMM-1 could make them interesting for further research concerning cancer diseases in the human eye.

Localized therapy of the highly malignant brain tumor glioblastoma multiforme (GBM) could help to drastically improve the treatment efficiency and increase the patient’s median survival. Here, a macroscopic PDMS matrix composed of interconnected microchannels for tailored drug release and localized GBM therapy is introduced. Based on a simple bottom-up fabrication method using a highly versatile sacrificial template, the presented strategy solves the scaling problem associated with the previously developed microchannel-based drug delivery systems, which were limited to two dimensions due to the commonly employed top-down microfabrication methods. Additionally, tailoring of the microchannel density, the fraction of drug-releasing microchannels and the macroscopic size of the drug delivery systems enabled precise adjustment of the drug release kinetics for more than 10 days. As demonstrated in a long-term GBM in vitro model, the release kinetics of the exemplarily chosen GBM drug AT101 could be tailored by variation of the microchannel density and the initial drug concentration, leading to diffusion-controlled AT101 release. Adapting a previously developed GBM treatment plan based on a sequential stimulation with AT101, measured anti-tumorigenic effects of free versus PDMS-released AT101 were comparable in human GBM cells and demonstrated efficient biological activity of PDMS-released AT101.

Background The treatment of glioblastomas (GBM) is still a clinical challenge. Current GBM therapeutic plans focus on the development of new strategies for local drug administration in the tumor cavity to realize an efficient long-term treatment with small side-effects. Here, different amounts of residual GBM cells and healthy brain cells define the microenvironment of the tumor cavity after individual surgical GBM resection (complete or incomplete). Methods We evaluated available in vivo data and determined the required amounts and numerical ratios of GBM and healthy brain cells for our in vitro (in)complete resection dual co-culture model. We applied a generic two-drug treatment [Temozolomide (TMZ) in combination with AT101, followed by single AT101 treatment] strategy and analyzed the results in comparison with appropriate mono-culture systems to prove the applicability of our model. Results We established a suitable GBM dual co-culture model, mimicking the complete and incomplete resection in vitro, giving stable and reliable results on drug testing. Both dual co-culture conditions protectively influenced on cell death and growth rates of primary GBMs when treated with TMZ + AT101/AT101, although the treatment strategy per se was still efficient. Cell death of astrocytes correlated with amounts of increasing GBM cell numbers in the incomplete resection model upon drug treatment, and probably GBM-released chemokine and cytokines were involved in this interplay. Conclusions Our results suggest that this dual co-culture model provides a biologically relevant platform for the discovery and compound screening of local GBM treatment strategies.

Highly malignant gliomas are characterized by pronounced intra‑ and intertumoral heterogeneity. On the genetic level, this heterogeneity may be caused by spontaneous mutation events, but recent studies have reported distinct mutational signatures that may be caused by an enzyme family with cytidine desaminase activity, the apolipoprotein B mRNA editing enzyme catalytic polypeptide‑like (APOBEC) proteins. Among these, APOBEC3B contributes to tumor progression in a variety of types of tumor, including breast cancer. In the present study, the expression of APOBEC3B was detected at the mRNA and protein levels in solid human glioma tissue and human glioma cell lines. In vitro, treatment with temozolomide, the most commonly used chemotherapeutic in glioma therapy, induced APOBEC3B expression. Furthermore, the knockdown of APOBEC3B by clustered regularly interspaced short palindromic repeats/CRISPR associated protein 9 resulted in reduced proliferation and enhanced chemosensitivity of glioma cells. Thus, APOBEC3B contributes to glioma progression and may be a future target for therapeutic intervention.

Glioblastoma multiforme (GBM) is a poorly curable disease due to its heterogeneity that enables single cells to survive treatment regimen and initiate tumor regrowth. Although some progress in therapy has been achieved in the last years, the efficient treatment of GBMs is still a clinical challenge. Besides the standard therapeutic drug temozolomide (TMZ), quinoline-based antimalarial drugs such as hydroxychloroquine (HCQ) and BH3 mimetics such as AT101 were considered as possible drugs for GBM therapy. We investigated the effects of sequentially applied single and combined TMZ, HCQ and AT101 treatments in a long-term stimulation GBM in vitro model. We performed all investigations in parallel in human astrocytes and two differentially TMZ-responsive human GBM cell lines and adjusted used drug concentrations to known liquor/plasma concentrations in patients. We determined amounts of dead cells and still remaining growth rates and depicted our results in a heatmap-like summary to visualize which sequential long-term treatment schedule seemed to be most promising. We showed that sequential stimulations yielded higher cytotoxicity and better tumor growth control in comparison to single TMZ treatment. This was especially the case for the sequences TMZ/HCQ and TMZ + AT101/AT101 which was as effective as the non-sequential combination TMZ + AT101. Importantly, those affected both less and more TMZ-responsive glioma cell lines, whilst being less harmful for astrocytes in comparison to single TMZ treatment. Sequential treatment with mechanistically different acting drugs might be an option to reduce side effects in long-term treatment, for example in local administration approaches.

This paper advocates a perspective on metaphors in audio-visual media that conceives of these as processes of meaning making, i.e., as dynamic embodied conceptualizations, constitutively bound to the flow of experience. This involves experience in a double sense: as immediate affection through an audio-visually orchestrated form of movement experience and as sensory-motor experiences of metaphoric source domains. Drawing on an interdisciplinary (linguistic and film analysis) method, this study presents an analysis of a German political TV report on winners and losers of the financial crisis. The goal of this case study is twofold: to reconstruct the complexity of metaphoric meaning making in audio-visual media and to illustrate a theoretical claim: that metaphors in audio-visual compositions emerge dynamically from sensory and affective experiences.

Honing is mostly the last step in the production process for precise elements and thus has to comply with high standards for the resulting quality regarding form, measurement and surface quality. Tolerances are currently even further reduced. Crucial for the ability of the honing process to reach the high quality is the process control. A new approach to a further improvement of the honing process could be the use of vibration analysis during the machining process. Vibration analysis for process control is already successfully used, e.g. for grinding. The paper presents an experimental setup for the measurement of vibrations during the honing process and shows first results of conducted experiments.

The finishing of precise bores is an important machining step, especially in the automotive industries. Bores can vary in diameter and form, from bigger piston bores in motor cylinders to rather small bores in fuel injection systems. Quality parameters for these bores are form- and geometric accuracy as well as surface roughness and structure. All requirements can economically be met by using the honing process as the final finishing operation for readymade bores. Honing is an abrasive machining process using abrasive grains usually made of diamond or Cubic Boron Nitride (CBN) in metallic bonding. During the process three movement components, namely an axial movement along the bore axis, a rotational movement around the bore axis and the feeding movement of the honing stone, are combined. The feeding movement of the honing stone is induced by an axial movement of the feeding cone, which is translated into a radial movement of the honing stone at the interface between honing stone and work piece. The feeding movement is decisive for the results of the honing process. For a regulation of the process that enables high process stability and repeatability as well as good quality results from the first part it is important to know about the forces in the feeding system of the tool. The following paper describes the static and dynamic correlation of the process forces in order to give a basis for a new regulation approach especially for bores with smaller diameters (below 20 mm) where the direct measurement of the process forces with a tool- integrated sensor is difficult. Measured signals are investigated for their dynamic influences. The theoretical approach is then compared to signals measured during the process.

Honing is a machining process that can economically produce very exact bores regarding form, geometry and surface quality. It is mainly used as the final finishing operation for ready made parts and typically conducted on the inner surface of a cylinder. Due to their specific surface structure which has a good ability to keep lubricants and a high wear resistance, honed parts usually serve as functional surfaces to guide moved parts. Today, the main fields for applications of honing technology can be found in most sectors of the metal working industry, e.g. the automotive industries, hydraulic and pneumatic manufacture and aircraft industries. During the long-stroke honing process the tool, equipped with one or several abrasive honing stones, is performing three overlaying movement components. These are a rotational movement, an oscillating stroke movement in axial direction and the radial feed movement of the honing stone. This feed movement is one of the decisive factors for the results of the honing process. It can be realized through feed control or, in a more recent approach, through force control strategies. By feed controlled honing, the position of the honing stone is changed in defined invariable steps in certain time intervals. In contrast, by force controlled honing the force of the honing stone against the wall of the workpiece is measured indirectly by a force sensor, which is located within the feed unit of the honing machine. A required force value is given and, if necessary, the honing stone is fed outwards to reach and maintain a constant cutting force throughout the honing process. This constant force is supposed to produce better results of the honing process in terms of geometric and form accuracy as well as surface structure. The presented research studies aim at improving process know-how to further increase the production accuracy and process repeatability. In a comparison between parts honed with the two different feeding strategies for the honing stone, force components and moments were analyzed and correlated to the phases of the process and several influencing parameters. In addition to the forces and moments measured during the honing process, the quality parameters of the honed parts have also been analyzed and compared.

Ethylene influences many processes in Arabidopsis (Arabidopsis thaliana) through the action of five receptor isoforms. We used high-resolution, time-lapse imaging of dark-grown Arabidopsis seedlings to better understand the roles of each isoform in the regulation of growth in air, ethylene-stimulated nutations, and growth recovery after ethylene removal. We found that ETHYLENE RECEPTOR1 (ETR1) is both necessary and sufficient for nutations. Transgene constructs in which the ETR1 promoter was used to drive expression of cDNAs for each of the five receptor isoforms were transferred into etr1-6;etr2-3;ein4-4 triple loss-of-function mutants that have constitutive growth inhibition in air, fail to nutate in ethylene, and take longer to recover a normal growth rate when ethylene is removed. The patterns of rescue show that ETR1, ETR2, and ETHYLENE INSENSITIVE4 (EIN4) have the prominent roles in rapid growth recovery after removal of ethylene whereas ETR1 was the sole isoform that rescued nutations. ETR1 histidine kinase activity and phosphotransfer through the receiver domain are not required to rescue nutations. However, REVERSION TO SENSITIVITY1 modulates ethylene-stimulated nutations but does not modulate the rate of growth recovery after ethylene removal. Several chimeric receptor transgene constructs where domains of EIN4 were swapped into ETR1 were also introduced into the triple mutant. The pattern of phenotype rescue by the chimeric receptors used in this study supports a model where a receptor with a receiver domain is required for normal growth recovery and that nutations specifically require the full-length ETR1 receptor.

Accurate monitoring of the t(14;18) translocation is regarded as highly desirable in patients with follicular lymphoma (FL) as absence of bcl-2/IgH positive cells has been correlated with a reduced risk of recurrence and, more recently, pre-treatment t(14;18) load with response to rituximab (R; Blood 2004;103:4416-23). With the arrival of R clinical and molecular remission rates for various lymphoma entities improved significantly, creating the need to carefully review and reassess the role of PCR negativity for clinical outcome, specifically when considering the prolonged presence of the drug as compared to chemotherapy. To determine the rate of molecular clearance achieved by the addition of different doses of R 16 newly diagnosed, t(14;18) positive patients with FL (Ann Arbor stages III/IV) were investigated before, during and after primary chemotherapy with six cycles of CHOP combined with varying (1, 3 or 6) cycle numbers of R (varR1-, varR3- or varR6-CHOP, respectively) regarding molecular remission status. For this purpose classic nested PCR as well as a newly established RQ-PCR employing juxtaposed hybridisation probes were employed to assess molecular remission prior, during and post therapy. Interestingly, administering just a single cycle of R (varR1-CHOP) in combination with a standard regimen of CHOP resulted in rapid and effective clearance of t(14;18) carrying cells from the peripheral blood of 4/5 patients in this treatment group. 6/8 (6/8) varR1-/varR3-CHOP patients were fully cleared and 8/8 (7/8) var6-CHOP patients as assessed by RQ- (nested) PCR. This indicates the high clearance capacity of this combination therapy approach even when adding a very low cycle number of R (1 and 3, respectively) to CHOP in primary therapy of FL. In summary, the relationship established between molecular clearance and minimal residual disease/risk of recurrence may have to be redefined in the times of R. Upcoming large prospective trials will have to elucidate to what degree the clearance of peripheral blood from t(14;18) positive cells can still be regarded as informative regarding absence of minimal residual disease, remission status and/or risk of recurrence.

This study assessed prevalence, frequency, age and gender distribution and breakpoint locations, and detection method validity for the bcl-2/IgH rearrangement in 204 healthy individuals. For this purpose, both classic two-step, nested, semi-quantitative PCR as well as a newly established sequence-specific, hybridization probe-based real-time quantitative PCR (RQ-PCR) were employed and tested for their sensitivity and specificity for detecting t(14;18) positive cells in healthy blood donors. Interestingly, almost a quarter (24%; 39/204) of all healthy individuals carried the translocation, confirming data of one large prior report [Summers KE, Goff LK, Wilson AG, Gupta RK, Lister TA, Fitzgibbon J. Frequency of the Bcl-2/IgH rearrangement in normal individuals: implications for the monitoring of disease in patients with follicular lymphoma. J Clin Oncol 2001;19(2):420-4]. Regarding presence as well as frequency of the translocation, no correlation to age (mean frequency 2.0:10(4), with a median ofl:10(4), for40 years, and mean frequency 1.9:10(4), with a median ofl:10(4) for individualsor=40 years) nor gender was detected. Comparing the two PCR approaches, a 95.1% concordance (194/204) regarding t(14;18) detection was determined for nested and RQ-PCR, with nested PCR being slightly more sensitive (reproducible detection limit l:10(5) cells versus 1:10(4); maximum detection limit l:10(6) versus 1:10(5)). Sequence analysis confirmed individual breakpoints for all samples analyzed (29/29), indicating detection validity for both PCR approaches and ruling out contamination. The breakpoint location distribution pattern appeared to be comparable to the pattern seen with follicular lymphoma (FL) patient collectives. In conclusion, clonal bcl-2/IgH rearrangements are indeed a very frequent observation in healthy individuals, and appear to be independent of age and gender in regard to presence and frequency. This represents a conflicting finding in context of potential biological significance, and presents a potential disruptive factor for minimal residual disease (MRD) monitoring in FL patients. Prospective future trials will have to clarify the biological significance of this important observation.

Fludarabine in combination with cyclophosphamide is an effective treatment for newly diagnosed as well as relapsed follicular lymphoma. The anti-CD20 antibody rituximab has been employed successfully for the same indications. No such data were available on a combined use of these agents. Therefore, we conducted a phase II study to evaluate the safety and efficacy of a combination of rituximab (375 mg/m2), fludarabine (4 x 25 mg/m2) and cyclophosphamide (1 x 750 mg/m2), for the treatment of relapsed follicular lymphoma. An unexpected, severe hematologic toxicity with significant, prolonged thrombocytopenias WHO grade III/IV in 6 (35%) of 17 patients treated in total occurred, leading to early termination of the trial. Cytologic and serologic analyses point toward a direct toxic effect. Older patients (mean age 64.7 vs. 56.5 yr) were significantly (P = 0.02) more likely to suffer from this toxicity, whereas no other clinical or hematologic parameter differed statistically between the patients suffering from thrombocytopenia and those who did not. The addition of rituximab to fludarabine/cyclophosphamide employed at doses given above in relapsed follicular lymphoma may have led to this increase in thrombocytopenias. Therefore, caution should be exercised when combining these drugs for the treatment of patients with relapsed follicular lymphoma, especially when treating older patients.

BACKGROUND Ocular prevalence is defined as an unequal weighting of the eyes in the directional perception of stereo objects. Opinions differ as to the cause and relevance of ocular prevalence. Hans-Joachim Haase suggested that ocular prevalence is due to fixation disparity, brought about by incomplete compensation of heterophoria. He further suggested that prismatic spectacles determined by his "measuring and correcting methodology" (MKH) could restore bicentral fixation and thus establish a perceptual balance between both eyes. METHODS We examined 10 non-strabismic subjects with a visual acuity of > or = 1.0 in both eyes. It turned out that all 10 had a "fixation disparity type II", characterised according to Haase by a "disparate retinal correspondence". All subjects underwent the automatic Freiburg Ocular Prevalence Test, without and with MKH prisms. In addition we examined ocular prevalence under forced vergence and compared ocular prevalence with stereoacuity. RESULTS Spontaneous ocular prevalence ranged between 1 and 69 %. Averaged over all 10 subjects, ocular prevalence without and with the MKH prisms were not significantly different. Statistical evaluation of single subjects revealed only in one of the 10 a significant difference (Bonferroni-corrected p = 0.001). In the subgroup of 5 subjects who underwent forced vergence, ocular prevalence remained unaltered between 0 and 18 Delta base out. The stereoscopic threshold of all 10 subjects ranged between 1.5 and 14.5 arcsec. There was no correlation between ocular prevalence and stereoscopic threshold (r = - 0.2, p = 0.5). CONCLUSION Our results indicate that ocular prevalence is largely independent of phoria correction and vergence stress. The excellent stereoacuity of all subjects suggests that ocular prevalence is abandoned for the sake of optimal resolution when very small differences in depth have to be judged.

Hintergrund. Seit Ogle (1949) unterscheidet man die dissoziierte von der assoziierten Heterophorie. Dissoziierte Heterophorie zeigt sich bei vollstandiger Trennung des Binokularsehens, assoziierte Heterophorie bei prismatischer Annullierung einer “Fixationsdisparitat” (hier definiert als Vergenzfehlstellung bei Trennung des Binokularsehens nur im Gesichtsfeldzentrum). In der Literatur wird kontrovers diskutiert, ob die Prufung auf dissoziierte und assoziierte Heterophorie zu gleichen Ergebnissen fuhrt und ob einer der beiden Werte als Anhalt fur eine Prismenbrille zu bevorzugen ist. Methode. Es wurden 43 Probanden mit dem weisen Maddox-Zylinder auf dissoziierte und mit dem Kreuztest nach Hans-Joachim Haase auf assoziierte Heterophorie untersucht. Berucksichtigt wurde jeweils nur die horizontale Komponente. Bei beiden Tests hatten die Probanden die Aufgabe, jegliche Abweichung kontinuierlich mithilfe eines Herschel-Prismas zu korrigieren. Das Resultat wurde nach 1 min abgelesen. Ergebnis. Die Korrelation zwischen dissoziierter und assoziierter Heterophorie betrug r=0,89 (p

BACKGROUND The "Measuring and Correcting Methodology" after H.-J. Haase (MKH) aims at converting "fixation disparity" into bicentral fixation, using prismatic spectacles. In the context of the MKH, fixation disparity is diagnosed by a series of subjective tests. According to H.-J. Haase, a long-standing fixation disparity can lead to "disparate correspondence" between the central areas of both retinae, which consolidates the fixation disparity and gradually converts a "young" into an "old fixation disparity". In "old fixation disparity" it is thought that bicentral fixation does not occur anymore, so that stereoacuity is impaired. However, prismatic spectacles can, according to H.-J. Haase, restitute bicentral fixation and consequently improve stereoacuity, even in some cases of "old fixation disparity". METHODS Ten non-strabismic subjects with a visual acuity of >/= 1.0 in both eyes were examined. It turned out that all ten had, according to MKH, a "disparate correspondence", 5 subjects with a "young" and 5 with an "old fixation disparity". According to the MKH, a correcting prism was determined. All 10 subjects underwent the automatic Freiburg Stereoacuity Test, without and with the MKH-prism. RESULTS Without the MKH-prism, the stereoscopic threshold ranged between 1.5 and 14.5 arcsec. With the MKH-prism, the values were not significantly different. CONCLUSION Stereoacuity ranged between good and excellent in the 5 subjects with "young" as well as in the 5 subjects with "old fixation disparity". The MKH-prism did not improve the stereoacuity in any of the subjects. These results cast doubt on Haase's assertion that an "old fixation disparity" implies a reduced stereoacuity. Hence, the premise for a benefit of the MKH-prism with respect of stereoacuity is not substantiated. In the 5 subjects with a "young fixation disparity", the good stereoacuity is consistent with Haase's theory, so that a benefit of the MKH-prism for stereoacuity was not expected. In previous studies, stereoacuity was found to be better with the MKH-prism than without it. These studies are questionable since learning with repeated testing was not taken into account. We conclude that there is no sound evidence for the assumption that the MKH-prism can improve stereoacuity.

Objects are usually segregated from ground by several visual dimensions. We studied texture segregation in checkerboards defined by gradients in spatial frequency, orientation or both frequency and orientation, using Gabor-filtered noise patterns. Saliency was measured electrophysiologically using the visual evoked potential (VEP) associated with texture segregation (‘tsVEP’) (an associated component in the visual evoked potential), and psychophysically by a 2AFC task. Spatial frequency and orientation stimuli evoked percepts of texture segregation and tsVEPs in all 11 subjects. The tsVEPs to combined stimuli were larger than those to each dimension alone, but smaller (74%) than the algebraic sum of tsVEPs to both individual dimensions. Psychophysical detection rates differed significantly between all conditions (P

The optical response of doubly charged sodium clusters Nan+2++ was measured for n = 20, 40, and 58 valence electrons, for which the jellium model predicts spherical clusters. A new experimental scheme was developed which allows to separate doubly charged clusters of even mass from the singly charged with half the mass. The optical spectra are dominated by a plasmon-like resonance which is blue shifted and narrower than that of the singly charged clusters. The smallest doubly charged cluster observed was Na9++. The photo ionization cross section for singly charged clusters was found to be typically 2.6·10-19cm2 per Na atom for photon energies of around 6 eV, which is a factor of 400 smaller than the maximum in the plasmon absorption in the region of hω=2.6 eV.

The optical absorption of small, open sodium cluster ions (Na{sup +}{sub {ital n}},{ital n}=4, 7, and 11) exhibit an unexpectedly large temperature dependence. Clearly separated absorption lines observed for cold clusters ({ital T}{similar_to}35 K) are interpreted as transitions between electronic states of the Na{sup +}{sub {ital n}} molecule. At the highest temperature ({ital T}{gt}380 K), however, the clusters are liquid, and broad absorption peaks are observed whose energy positions can be explained for {ital n}{gt}7 by the model of nearly free electrons oscillating in a spheroidal container.

The aim of this study was to investigate the role of NBS1 in the pathogenesis of malignant melanoma of the skin. To exclude the common 657del5 founder mutation, a total of 376 melanoma patients from Southern Germany were analyzed for sequence alterations in exon 6 of NBS1 by direct sequencing. Analyses revealed one 657del5 mutation and three nonsynonymous sequence variations in exon 6 of NBS1 (V210F, R215W, and F222L). Analysis of an additional sample of 629 melanoma patients and 604 controls revealed no F222L mutation, indicating that this newly identified sequence alteration is not a common polymorphism. In a case-control association study including 632 melanoma patients and 615 cancer-free control participants from Southern Germany, three publicly known single nucleotide polymorphisms located in the NBS1 gene region were analyzed. No significant associations between single nucleotide polymorphisms (rs9995, rs867185 and rs1063045) or referring calculated haplotypes and melanoma risk were identified. These results suggest that NBS1 does not play a major role in predisposition to melanoma in the Southern German population but that alterations of this gene might contribute to the risk of this cancer.

There is strong evidence to suggest that the peroxisome proliferator-activated receptor (PPAR)-gamma, a member of the nuclear receptor family of transcriptional regulators, mediates tumor suppressive activities in a variety of human cancers. Recently, PPARgamma agonists were found to inhibit growth of melanoma cell lines. Here, we tested the possibility that variations in the gene encoding PPARgamma (PPARG) influence melanoma risk. Two variations of PPARG (P12A[rs1801282] and C161T [rs3856806]) were investigated in two independent case-control studies with a total of 832 melanoma patients and 790 control individuals. In the first study, homozygous carriers of the rare *T allele of the C161T polymorphism in exon 6 of PPARG were significantly more common among patients with melanoma than among healthy subjects (6.0 vs. 2.0%; P=0.0096) and this association was independent of clinical risk factors such as skin type and nevus count (odds ratio 5.18; 95% confidence interval 1.68-15.96; P=0.0041). This finding, however, could not be replicated in the second case-control study. We therefore conclude that the investigated PPARG polymorphisms are not likely to constitute a significant risk factor for the development of melanoma among German Caucasians.

The absolute photoabsorption cross sections of [Formula: see text] and [Formula: see text] have been measured at two temperatures (evaporative ensemble, 115 K). The maximum of the observed resonance is nearly independent of the temperature and agrees within 5% with the Mie plasmon resonance as calculated from the bulk electron density of cesium. This is in contrast to the sodium case, where the plasmon is redshifted by about 20%. For n=9 and 21 the integrated oscillator strength amounts to 89% [Formula: see text] and 73% [Formula: see text] per valence electron in the energy range 1.5 eV

Fludarabine in combination with cyclophosphamide is an effective treatment for newly diagnosed as well as relapsed follicular lymphoma. The anti-CD20 antibody rituximab has been employed successfully for the same indications. No such data were available on a combined use of these agents. Therefore, we conducted a phase II study to evaluate the safety and efficacy of a combination of rituximab (375 mg/m2), fludarabine (4 x 25 mg/m2) and cyclophosphamide (1 x 750 mg/m2), for the treatment of relapsed follicular lymphoma. An unexpected, severe hematologic toxicity with significant, prolonged thrombocytopenias WHO grade III/IV in 6 (35%) of 17 patients treated in total occurred, leading to early termination of the trial. Cytologic and serologic analyses point toward a direct toxic effect. Older patients (mean age 64.7 vs. 56.5 yr) were significantly (P = 0.02) more likely to suffer from this toxicity, whereas no other clinical or hematologic parameter differed statistically between the patients suffering from thrombocytopenia and those who did not. The addition of rituximab to fludarabine/cyclophosphamide employed at doses given above in relapsed follicular lymphoma may have led to this increase in thrombocytopenias. Therefore, caution should be exercised when combining these drugs for the treatment of patients with relapsed follicular lymphoma, especially when treating older patients.

Very recently, BRAF mutations were found in about 2/3 of malignant melanomas and at lower frequencies in other human cancers. The BRAF gene codes for a protein in the mitogen-activated protein kinase (MAPK) pathway. All mutations identified to date are within the kinase domain, with a single missense mutation (V599E) accounting for 80%. We investigated the hypothesis that this common somatic BRAF mutation (V599E) would contribute to melanoma predisposition in familial and polygenic malignant melanoma if occuring as a germ-line mutation. We performed comprehensive mutational screening of exon 15 of BRAF using DHPLC (denaturing high-performance liquid chromatography) and DNA sequencing techniques. No V599E mutation could be detected in 172 melanoma patients comprising 46 familial cases, 21 multiple melanoma patients and 106 cases with at least one first-degree relative suffering from other cancers. We therefore conclude that the common somatic BRAF mutation V599E does not contribute to polygenic and familial melanoma predisposition. © 2003 Wiley-Liss, Inc.

Ocular prevalence is defined as an unequal weighting of the eyes in the directional perception of stereo objects. Opinions differ as to the cause and relevance of ocular prevalence. Hans-Joachim Haase suggested that ocular prevalence is due to fixation disparity, brought about by incomplete compensation of heterophoria. He further suggested that prismatic spectacles determined by his "measuring and correcting methodology" (MKH) could restore bicentral fixation and thus establish a perceptual balance between both eyes.We examined 10 non-strabismic subjects with a visual acuity ofor = 1.0 in both eyes. It turned out that all 10 had a "fixation disparity type II", characterised according to Haase by a "disparate retinal correspondence". All subjects underwent the automatic Freiburg Ocular Prevalence Test, without and with MKH prisms. In addition we examined ocular prevalence under forced vergence and compared ocular prevalence with stereoacuity.Spontaneous ocular prevalence ranged between 1 and 69 %. Averaged over all 10 subjects, ocular prevalence without and with the MKH prisms were not significantly different. Statistical evaluation of single subjects revealed only in one of the 10 a significant difference (Bonferroni-corrected p = 0.001). In the subgroup of 5 subjects who underwent forced vergence, ocular prevalence remained unaltered between 0 and 18 Delta base out. The stereoscopic threshold of all 10 subjects ranged between 1.5 and 14.5 arcsec. There was no correlation between ocular prevalence and stereoscopic threshold (r = - 0.2, p = 0.5).Our results indicate that ocular prevalence is largely independent of phoria correction and vergence stress. The excellent stereoacuity of all subjects suggests that ocular prevalence is abandoned for the sake of optimal resolution when very small differences in depth have to be judged.

Background: Fixation disparity has been widely used as an indicator for vergence accuracy under prismatic stress. However, the targets used for measuring fixation disparity contain artificial features in that the fusional contours are thinned out. We considered that stereoacuity might be a preferable indicator of vergence accuracy, as stereo targets represent natural viewing conditions. Methods: We measured fixation disparity with a computer adaptation of Ogle's test and stereoacuity with the automatic Freiburg Stereoacuity Test. Eight subjects were examined under increasing base-in and base-out prisms. Results: The response of fixation disparity to prismatic stress revealed the curve types described by Ogle and Crone. All eight subjects reached a stereoscopic threshold below 10 arcsec. In seven subjects the stereoscopic threshold increased before double vision occurred. Conclusion: Our data suggest that stereoacuity is suitable to assess the range of binocular vision under prismatic stress. As stereoacuity bears the advantage over fixation disparity in that it can be measured without introducing artificial viewing conditions, we suggest exploring whether stereoacuity under prismatic stress would be more meaningful in the work-up of asthenopic patients than is fixation disparity.