G protein-coupled receptor 40 expression in human melanoma - correlation with tumour thickness, AJCC stage and survival.

BACKGROUND: In melanoma, preclinical data suggest a possible role of polyunsaturated fatty acids inhibiting cell growth. A new target molecule for free fatty acids, the G protein-coupled receptor GPR40, was identified in melanoma cells. OBJECTIVES: The aim of this study was to investigate GPR40 expression in human melanocytic tissues and to evaluate its potential as a prognostic marker. METHODS AND RESULTS: A total of 114 tissue sections of naevi, primary melanoma and melanoma metastasis were immunohistochemically stained with anti-GPR40. The staining was evaluated, using the immunoreactivity scoring system. Compared to naevi, primary melanoma and melanoma metastasis showed significantly higher levels of GPR40 (P < 0.05). In primary melanoma, GPR40 expression positively correlated with tumour thickness (P = 0.044) and AJCC level (P = 0.017) and in melanoma metastasis with AJCC level (P = 0.035). Primary melanoma patients with high levels of GPR40 had a significantly poorer overall survival (P = 0.004) and shorter disease-free survival (0.040). CONCLUSION: The present study identified GPR40 as a novel target molecule in melanoma. First evidence for a potential role of the receptor in tumour progression and metastases was found, and it could be demonstrated that GPR40 expression is negatively correlated with patient's survival.

Formation of n- and p-type regions in individual Si/SiO2 core/shell nanowires by ion beam doping.

A method for cross-sectional doping of individual Si/SiO2 core/shell nanowires (NWs) is presented. P and B atoms are laterally implanted at different depths in the Si core. The healing of the implantation-related damage together with the electrical activation of the dopants takes place via solid phase epitaxy driven by millisecond-range flash lamp annealing. Electrical measurements through a bevel formed along the NW enabled us to demonstrate the concurrent formation of n- and p-type regions in individual Si/SiO2 core/shell NWs. These results might pave the way for ion beam doping of nanostructured semiconductors produced by using either top-down or bottom-up approaches.

Commensurate germanium light emitters in silicon-on-insulator photonic crystal slabs.

We report on the fabrication and characterization of silicon-on-insulator (SOI) photonic crystal slabs (PCS) with commensurately embedded germanium quantum dot (QD) emitters for near-infrared light emission. Substrate pre-patterning defines preferential nucleation sites for the self-assembly of Ge QDs during epitaxial growth. Aligned two-dimensional photonic crystal slabs are then etched into the SOI layer. QD ordering enhances the photoluminescence output as compared to PCSs with randomly embedded QDs. Rigorously coupled wave analysis shows that coupling of the QD emitters to leaky modes of the PCS can be tuned via their location within the unit cell of the PCS.

Role of surface-segregation-driven intermixing on the thermal transport through planar Si/Ge superlattices.

It has been highly debated whether the thermal conductivity κ of a disordered SiGe alloy can be lowered by redistributing its constituent species so as to form an ordered superlattice. By ab initio calculations backed by systematic experiments, we show that Ge segregation occurring during epitaxial growth can lead to κ values not only lower than the alloy's, but also lower than the perfect superlattice values. Thus we theoretically demonstrate that κ does not monotonically decrease as the Si- and Ge-rich regions become more sharply defined. Instead, an intermediate concentration profile is able to lower κ below both the alloy limit (total intermixing) and also the abrupt interface limit (zero intermixing). This unexpected result is attributed to the peculiar behavior of the phonon mean free path in realistic Si/Ge superlattices, which shows a crossover from abrupt-interface- to alloylike values at intermediate phonon frequencies of ∼3 THz. Our calculated κ's quantitatively agree with the measurements when the realistic, partially intermixed profiles produced by segregation are considered.

ALD grown bilayer junction of ZnO:Al and tunnel oxide barrier for SIS solar cell.

Various metal oxides are probed as extrinsic thin tunnel barriers in Semiconductor Insulator Semiconductor solar cells. Namely Al2O3, ZrO2, Y2O3, and La2O3 thin films are in between n-type ZnO:Al (AZO) and p-type Si substrates by means of Atomic Layer Deposition. Low reverse dark current-density as low as 3×10-7 A/cm2, a fill factor up to 71.3%, and open-circuit voltage as high as 527 mV are obtained, achieving conversion efficiency of 8% for the rare earth oxide La2O3. ZrO2 and notably Al2O3 show drawbacks in performance suggesting an adverse reactivity with AZO as also indicated by X-ray Photoelectron Spectroscopy.

Two-dimensional microrheology of freely suspended liquid crystal films.

Smectic liquid crystals form freely-suspended, fluid films of highly uniform structure and thickness, making them ideal systems for studies of hydrodynamics in two dimensions. We have measured particle mobility and shear viscosity by direct observation of the gravitational drift of silica spheres and smectic islands included in these fluid membranes. In thick films, we observe a hydrodynamic regime dominated by lateral confinement effects, with the mobility of the inclusion determined predominantly by coupling of the fluid flow to the fixed boundaries of the film. In thin films, the mobility of inclusions is governed primarily by coupling of the fluid to the surrounding air, as predicted by Saffman-Delbrück theory.

Detecting artery occlusion and critical flow diminution in the case of an acute ischemic stroke--methodological pitfalls of common vascular diagnostic methods.

PURPOSE: None of the vascular emergency diagnostic methods commonly used in the case of acute ischemic stroke, i. e. CTA, color-coded duplex sonography (CCDS), MRA, and DSA, is free of restrictions due to physical and physiological characteristics. As a result, misleading results initiating an inappropriate acute therapeutic intervention or hampering a promising one cannot be excluded. We aimed to assess the type and frequency of methodological pitfalls occurring in this situation. MATERIALS AND METHODS: We retrospectively analyzed data of 269 consecutive patients admitted to our stroke unit with a clinical syndrome of an acute stroke. All patients underwent one or more vascular emergency diagnostic methods on a routine basis. RESULTS: 37 patients were excluded because of a final diagnosis other than ischemic stroke. 76 of 232 ischemic stroke patients underwent emergency diagnostic methods with two or more vascular examination techniques. Controversial results occurred in 20 patients and related to the detection and localization of large artery occlusion and its differentiation from a low/slow flow situation and the identification of critical cerebral flow diminution distal to large artery occlusion/severe stenosis. Methodological pitfalls were able to be most reliably resolved by CCDS. Within the whole cohort of ischemic stroke patients, vascular constellations susceptible to misinterpretation were diagnosed in 40 (17.2 %) patients. CONCLUSION: We recommend providing several techniques including CCDS in an emergency stroke setting and applying techniques with respect to diagnostic findings.

Regulation of specific cell-mediated cytotoxic response against SV40-induced tumor associated antigens by depletion of suppressor T cells with cyclophosphamide in mice.

When cyclophosphamide was administered to mice before immunization with syngeneic SV40 transformed cells, the specific immune response elicited, as was measured by in vitro 51Cr release assay was stronger and lasted longer when compared to the response generated in noncyclophosphamide-treated mice. The augmentation effect of the drug was dependent on cyclophosphamide concentration being optimal at 100 mg/kg and on the time of drug administration in relation to antigen immunization being optimal at 2 d before antigen administration. Transfer of T cells from normal syngeneic mice to drug-treated animals abolished the cyclophosphamide-induced augmentation of immune response. These results implied that cyclophosphamide sensitive T cells suppressed the in vivo generation of specific effector T cells against SV40-induced tumor-associated antigens.

Visualization of domain formation in the inner and outer leaflets of a phospholipid bilayer.

Large vesicles (5-10-micron in diameter) were formed in the presence of phospholipids fluorescently labeled on the acyl chain and visualized using a fluorescence microscope, charge-coupled-device camera and digital image processor. When such vesicles contained a fluorescent phosphatidic acid (PA) and were exposed to 2 mM CaCl2 or 0.5 mM PrCl3, it was possible to visualize PA-enriched domains within the vesicles. Calcium-induced domain formation was reversible in the presence of 4 mM EGTA. Vesicles were formed containing fluorescent PA on either the inner or outer leaflet of the bilayer and the patching and dissolution of patching were studied under conditions where calcium was present on the outside of the vesicle and where calcium was distributed across the bilayer. In addition, vesicles were formed with two different fluorescent PA's, one on the inner leaflet and a different one on the outer leaflet of the bilayer. The results of the experiments show that in vesicles formed primarily with naturally occurring phospholipids such as egg phosphatidylcholine or brain phosphatidylethanolamine, there was no coordinate action of the two leaflets of the bilayer. An exception to this was found, however, if the vesicles were formed in the presence of primarily dioleoyl phospholipids (greater than 95 mol %). In these vesicles there was a coordinate or coupled response to calcium by the two leaflets of the bilayer. In most cases, however, the two leaflets of the bilayer showed independent or uncoupled domain formation.

[Marginal protection of retinal cells by bisperoxovanadium : Appropriate therapy in the model of retinal ischemia?] / Marginale Protektion retinaler Zellen durch Bisperoxovanadium : Geeignete Therapie im Modell der retinalen Ischämie?

BACKGROUND: Ischemic processes usually lead to the destruction of retinal cells and therefore play a key role in a multitude of eye diseases. OBJECTIVE: The aim of this study was to investigate whether bisperoxovanadium has a potential neuroprotective effect in an ischemia/reperfusion animal model. MATERIAL AND METHODS: Initially, ischemia was induced in one eye of an ischemia/reperfusion model and 3 days later, a 14-day medication-based treatment was initiated. Bisperoxovanadium was administered intraperitoneally every 3 days. Subsequently, the number of ganglion cells, the rate of apoptosis, amacrine cells, macroglia, microglia, and their activation state, as well as photoreceptors were determined by histological and immunohistochemical analyses. RESULTS: In comparison to the control group, a significant retinal ganglion cell loss, a significant reduction of the inner layers as well as a decrease in photoreceptor and amacrine cell numbers could be determined in the ischemic eyes. In addition, there was an increase in the number of microglia in these animals. The rats treated with bisperoxovanadium did not exhibit a significant neuroprotective effect regarding the number of ganglion cells, the rate of apoptosis, macroglia, amacrine cells, or photoreceptors; however, a low structural degeneration of photoreceptors could be observed as an effect of the treatment. Additionally, fewer microglia and activated microglia were observed after bisperoxovanadium treatment. CONCLUSION: Bisperoxovanadium seems to have only a marginal neuroprotective effect on ischemic retinae. It needs to be examined whether earlier therapy onset, higher dose or different route of administration would significantly improve the results or whether this therapeutic approach is unsuitable.

Downbeat nystagmus: aetiology and comorbidity in 117 patients.

OBJECTIVES: Downbeat nystagmus (DBN) is the most common form of acquired involuntary ocular oscillation overriding fixation. According to previous studies, the cause of DBN is unsolved in up to 44% of cases. We reviewed 117 patients to establish whether analysis of a large collective and improved diagnostic means would reduce the number of cases with "idiopathic DBN" and thus change the aetiological spectrum. METHODS: The medical records of all patients diagnosed with DBN in our Neurological Dizziness Unit between 1992 and 2006 were reviewed. In the final analysis, only those with documented cranial MRI were included. Their workup comprised a detailed history, standardised neurological, neuro-otological and neuro-ophthalmological examination, and further laboratory tests. RESULTS: In 62% (n = 72) of patients the aetiology was identified ("secondary DBN"), the most frequent causes being cerebellar degeneration (n = 23) and cerebellar ischaemia (n = 10). In 38% (n = 45), no cause was found ("idiopathic DBN"). A major finding was the high comorbidity of both idiopathic and secondary DBN with bilateral vestibulopathy (36%) and the association with polyneuropathy and cerebellar ataxia even without cerebellar pathology on MRI. CONCLUSIONS: Idiopathic DBN remains common despite improved diagnostic techniques. Our findings allow the classification of "idiopathic DBN" into three subgroups: "pure" DBN (n = 17); "cerebellar" DBN (ie, DBN plus further cerebellar signs in the absence of cerebellar pathology on MRI; n = 6); and a "syndromatic" form of DBN associated with at least two of the following: bilateral vestibulopathy, cerebellar signs and peripheral neuropathy (n = 16). The latter may be caused by multisystem neurodegeneration.

Progesterone synthesized by Schwann cells during myelin formation regulates neuronal gene expression.

Previously, progesterone was found to regulate the initiation and biosynthetic rate of myelin synthesis in Schwann cell/neuronal cocultures. The mRNA for cytochrome P450scc (converts cholesterol to pregnenolone), 3beta-hydroxysteroid dehydrogenase (3beta-HSD, converts pregnenolone to progesterone), and the progesterone receptor were found to be markedly induced during active myelin synthesis. However, the cells in the cocultures responsible for these changes were not identified. In this study, in situ hybridization was used to determine the localization of the enzymes responsible for steroid biosynthesis. The mRNA for cytochrome P450scc and 3beta-HSD were detected only in actively myelinating cocultures and were localized exclusively in the Schwann cells. Using immunocytochemistry, with minimal staining of the Schwann cells, we found the progesterone receptor in the dorsal root ganglia (DRG) neurons. The progesterone receptor in the neurons translocated into the nuclei of these cells when progesterone was added to neuronal cultures or during myelin synthesis in the cocultures. Additionally, a marked induction of the progesterone receptor was found in neuronal cultures after the addition of progesterone. The induction of various genes in the neurons was also investigated using mRNA differential display PCR in an attempt to elucidate the mechanism of steroid action on myelin synthesis. Two novel genes were induced in neuronal cultures by progesterone. These genes, along with the progesterone receptor, were also induced in cocultures during myelin synthesis, and their induction was blocked by RU-486 (a progesterone receptor antagonist). These genes were not induced in Schwann cells cultured alone after the addition of progesterone. These results suggest that progesterone is synthesized in Schwann cells and that it can indirectly regulate myelin formation by activating transcription via the classical steroid receptor in the DRG neurons.

Multicolor-FISH applied to resolve complex chromosomal changes in a case of T-ALL (FAB L2).

We report on a patient with a clinically diagnosed acute lymphoblastic leukemia (ALL) with partial unrecorded complex translocation events especially involving chromosomes 5, 9 and 18. At the GTG-band level the karyotype was abnormal in 20% of the analyzed cells. The complex karyotype was studied in more detail by spectral karyotyping (SKY) and multicolor banding (MCB) to characterize it in more detail. Thus, the karyotype could be described very accurately and in summary three different clones were detected, reflecting a high rate of karyotypic evolution in this patient.

Secondary cell-mediated cytotoxic response to challenge of rats with syngeneic Gross virus-induced lymphoma.

Secondary cell-mediated cytotoxicity generated in vivo against a syngeneic Gross virus-induced lymphoma [(C58NT)D] in WF rats was detected by the 4-hour 51Cr release assay. At 30 days or more following primary tumor cell inoculation, after the tumors had regressed, lymphoid cells had little or no detectable direct cytotoxic reactivity. At rechallenge with tumor cells, high levels of cytotoxicity were detected in the peritoneal exudate, peripheral blood, mesenteric lymph node, and spleen cells. The secondary cellular immune response after challenge developed earlier, reached higher levels, and lasted longer than the primary immune response. The secondary cytotoxic reactivity was shown to be immunologically specific by the use of various tumor cells both as target and inhibitor cells. Treatment of immune spleen cells with specific antiserum to rat T-cells and complement abolished their cytotoxic reactivity, whereas removal of complement receptor-bearing cells or phagocytic cells did not reduct the cytotoxicity. These data demonstrated that specific-memory T-cells persisted for long periods in the lymphoid organs of immune rats and could rapidly become cytotoxic from rechallenge with the tumor.

In vitro inhibition of cell-mediated cytotoxicity against syngeneic Friend virus-induced leukemia by immunoregulatory alpha globulin.

Immunoregulatory alpha-globulin (IRA) derived from normal human plasma decreased cytotoxic reactivity as measured by an in vitro 5-iodo-2'-deoxyridine release assay of immune mouse lymphocytes against the syngeneic Friend virus-induced leukemia, FBL-3. This inhibitory effect depended on the dose of IRA used and was not due to the cytotoxic effects of IRA on the effector cells or target tumor cells. We also found elevated levels of serum alpha-gloubins in FBL-3 tumor-bearing mice as compared to normal mice. These data and the demonstration of decreased specific cytotoxic reactivity in FBL-3 tumor-bearing mice suggest that IRA functions in the suppression of the host's immune response against tumors.

Inhibition of cell-mediated cytotoxicity against tumor-associated antigens by suppressor cells from tumor-bearing mice.

Spleen cells from mice bearing primary tumours induced by Moloney strain of murine sarcoma virus (M-MuSV) strongly inhibited the in vitro generation of specific secondary cell-mediated cytotoxic response of spleen cells from M-MusV regressor mice. These suppressor cells were resistant to treatment with anti-theta serum and complement or to X-irradiation. It appeared that suppressor cells may have had a role in limiting the host's immune response against tumor growth.

Natural cytotoxic reactivity of rat lymphocytes against syngeneic Gross virus-induced lymphoma.

Lymphoid cells from many normal W/Fu rats reacted in a 51Cr release cytotoxicity assay against (C58NT)D, a syngeneic Gross leukemia virus-induced tumor. The reactivity was maximal in young rats at 5-8 weeks of age and rapidly declined thereafter. Within reactive rats, the cytotoxicity was widely distributed among the various lymphoid organs. Since immunization of W/Fu rats with (C58NT)D was shown to elicit specific cell-mediated cytotoxic reactivity, studies were done to compare the characteristics of the natural reactivity with those of the immune reactivity. The specificity of both types of reactivity was analyzed in detail by an inhibition assay. The natural and the immune reactivities appeared directed against antigens associated with rat endogenous type-C viruses. The major differences between the natural reactivity and immune reactivity were the nature of the effector cells. Whereas immune reactivity was T-cell dependent, normal reactivity was not affected by treatment with antisera against T cells plus complement. Natural effector cells were not adherent and did not have macrophage properties. The active cells also did not have receptors for Ig or complement. The absence of detectable cell-surface markers on the natural effector cells was seen in studies of natural cytotoxic reactivity of mice, and it is proposed that the natural cytotoxicity in both systems is mediated by a unique subpopulation of lymphoid cells, tentatively designated "N" cells.

In vitro studies on the cell-mediated immune response to human brain tumors. I. Requirement for third-party stimulator lymphocytes in the induction of cell-mediated cytotoxic responses to allogeneic cultured gliomas.

For study of those properties of human gliomas that might contribute to their ability to escape cell-mediated immune attack, cultured human glioma cells were examined for their ability to elicit allogeneic cytolytic lymphocyte responses in vitro. Of 9 glioma lines, 5 were unable to elicit allogeneic cytolytic lymphocyte responses in mixed lymphocyte--tumor cultures although the concentration of stimulating glioma cells was varied over a fortyfold range. However, lymphocytes specifically cytolytic for 4 of the nonstimulatory lines could be generated if irradiated, third-party stimulator lymphocytes were added to cultures containing responder lymphocytes and glioma cells. The specific cytolytic lymphocytes produced in these cultures were inactivated by treatment with the monoclonal anti-T-cell antibody OKT3 plus complement and were thus identified as T-cells. However, nonspecific, non-T-lytic effectors were also generated. The results of these experiments demonstrated that certain cultured gliomas possessed a defect in immunogenicity that can be overcome by "help" from an allogeneic mixed lymphocyte reaction. The possible nature of this help and the potential implications of these results for the immunotherapy of human gliomas are discussed.

Phase I trial of temozolomide using an extended continuous oral schedule.

Temozolomide, a methylating imidazotetrazinone, has antitumor activity against gliomas, malignant melanoma, and mycosis fungoides and is presently administered as a 5-day oral schedule every 4 weeks. This Phase I study aimed to determine the maximum tolerated dose of temozolomide administered as a single oral daily dose for a continuous 6- or 7-week period, evaluate the plasma pharmacokinetics on this schedule, and compare total plasma exposure over 7 weeks with the conventional 5-day regimen. Twenty-four patients with varying tumor types (17 of 24 gliomas) received temozolomide. All had clinically evaluable, refractory disease; normal renal, hepatic, and bone marrow function; and WHO performance status < or = 2. Temozolomide was administered at 50 mg/m2/day, increasing by 25 mg/m2/day/cohort until at 100 mg/m2/day grade 4 myelotoxicity forced dose reductions to 85 mg/m2/day, then 75 mg/m2/day. At 75 mg/m2/day the regimen was extended to 7 weeks, allowing the future potential combination with irradiation for primary gliomas. Patient responses (standard Union International Contre Cancer criteria; for gliomas objective response) and toxicity were assessed. Temozolomide plasma pharmacokinetics were determined on day 1 and at the beginning of the final week of administration (n = 5). The most frequent toxicities were myelosuppression and grades 1 and 2 nausea and vomiting. Grade 4 leucopenia and thrombocytopenia occurred in one of four patients receiving 100 mg/m2/day temozolomide and in one of seven patients receiving 85 mg/m2/day. These hematological toxicities did not exceed grade 2 in 10 patients receiving 75 mg/m2/day temozolomide. One of 4 malignant melanoma patients and 7 of 17 glioma patients (41%) demonstrated tumor responses. The overall response rate for this prolonged schedule was 33% (objective response, 7 of 24 patients; partial response, 1 of 24 patients); also, 6 of 17 glioma patients maintained SD. Peak plasma temozolomide concentrations were obtained 30-90 min after oral administration. Elimination in plasma was best described by a monoexponential equation with an elimination half-life of 96 +/- 16 min. No plasma accumulation of temozolomide occurred. Toxicity was greatest in higher dose cohorts, with a resultant maximum tolerated dose of 85 mg/m2/day, whereas lower dose cohorts tolerated the schedule well. The area under the temozolomide plasma versus time curve was noncumulative between the first and last week of the schedule. Temozolomide administration of 75 mg/m2/day over a 7-week period permits a 2.1-fold greater drug exposure/4 weeks in comparison with the 5-day schedule of 200 mg/m2/day repeated every 28 days. The overall response rate was 33% (glioma patients, 41% and a further 25% SD). Temozolomide (75 mg/m2/day) for 7 weeks is the recommended starting dose for further assessment of this schedule.