Subretinal electrical stimulation of the rabbit retina with acutely implanted electrode arrays

Background Subretinal implants intend to replace photoreceptor function in patients suffering from degenerative retinal disease by topically applying electrical stimuli from the subretinal space. This study intended to prove the feasibility of a newly developed transchoroidal surgery and, furthermore, of a subretinal electrode array, which closely resembles envisioned human implants to electrically stimulate the visual system in rabbits.Methods Five rabbits (ten eyes) were implanted with a 4×2-electrode array via a transchoroidal access to the subretinal space. The electrodes were connected to an arbitrary stimulus generator to apply voltage pulses. Retinae were accessed by light microscopy after stimulation with various intensities.Results The stimulating foil could be introduced into the subretinal space in all eyes. In seven of ten eyes electrically evoked cortical potentials following subretinal electrical stimulation could be elicited. Threshold voltages ranged from less than 0.1 to 2.38 V with a corresponding threshold charge of approximately 1.0 nC per electrode or 10 µC/cm². Histology revealed localized retinal damage over some of the electrodes succeeding stimulation strengths of 2 V and consistent damage over all electrodes succeeding voltages of 3 V.Conclusions The study demonstrates the feasibility of the transchoroidal surgical access to place subretinal implants in rabbit eyes and provides proof of successful cortical activation following subretinal electrical stimulation by an electrode array envisioned for human implantations.     

Anti-cancer and antibacterial trioxacarcins with high anti-malaria activity from a marine Streptomycete and their absolute stereochemistry

The ethyl acetate extract from the Streptomyces sp. isolate B8652 delivered the trioxacarcins A to approximately C (2a to approximately 2c) and additionally three new derivatives designated as trioxacarcins D to approximately F (2d to approximately 2f). All trioxacarcins showed high anti-bacterial and some of them high anti-tumor and anti-malaria activity. The structures of the new antibiotics were derived from mass, 1D and 2D NMR spectra and confirmed by comparison of the NMR data with those of known derivatives. The absolute configuration of the trioxacarcins is deduced from the X-ray analysis of gutingimycin (2g) and from the known stereochemistry of the L-trioxacarcinoses A and B.     

Selective processing of food words during insulin-induced hypoglycemia in healthy humans

Rationale Hypoglycemia leads to undernutrition of the brain. Favoring selective processing of food stimuli would be an adaptive cognitive strategy. However, hypoglycemia is known to impair several aspects of cognitive function, and it is unknown whether selective cognitive processing of food stimuli occurs during insulin-induced hypoglycemia.Methods In a single-blind repeated measures design, healthy young adults (n=12, six female, mean age 28 years; mean body mass index 22.5 kg/m²) performed a standard Stroop word-color test, as well as a variant with food words designed to detect selective processing of food cues. Two sessions were scheduled with a 4-week interval. In each session, a hyperinsulinemic clamp method produced a normoglycemic (plasma glucose: 4.7 mmol/l) period, followed on 1 day by a hypoglycemic (2.7 mmol/l) testing period, and on the other day a second normoglycemic testing period (counterbalanced order).Results Color naming verbal reaction time (RT) increased during hypoglycemia (P<0.0001). The extent of the Stroop cognitive interference was independent of plasma glucose level. The key finding is that RT for food words increased more than for non-food control words (P<0.004), and this effect was not predicted by hunger ratings.Conclusions Our data provide new evidence that during hypoglycemia, attention is directed selectively to food-relevant stimuli. The results are discussed in terms of adaptation.     

Pharmacokinetic herb-drug interactions: are preventive screenings necessary and appropriate?

Pharmacokinetic interactions often occur as a result of activity changes of drug-metabolizing and transporting proteins, especially cytochrome P450 (CYP) isoenzymes and P-glycoprotein (P-gp). The activity of these enzymes and drug transporters can be enhanced or inhibited by synthetic drugs as well as by natural products. Since the number of herb-drug interactions has increased in recent years, systematic in vitro screenings and more clinical studies to identify such interactions were proposed for herbal medicinal products. However, previous results regarding this issue are not only contradictory but also of less predictability. One reason for the discrepancies could be the lack of validation of the recommended in vitro tests. Furthermore, it has to be considered that pharmacokinetic drug interactions are not only mediated by herbal medicines but also by several foods, beverages and life-style products. Since herbal medicines are considered to have a broad therapeutic range, a preventive risk assessment for pharmacokinetic drug interactions should first be realized for synthetic drugs with a narrow therapeutic index. Efforts to identify all possible interactions will lead to limitless investigations and to inconsistent decisions.     

Genomic DNA-chip hybridization reveals a higher incidence of genomic amplifications in pancreatic cancer than conventional comparative genomic hybridization and leads to the identification of novel candidate genes

Genomic analyses aimed at the detection of high-level DNA amplifications were performed on 13 widely used pancreatic cancer cell lines and 6 pancreatic tumor specimens. For these analyses, array-based comparative genomic hybridization (Matrix-CGH) onto dedicated microarrays was used. In comparison with chromosomal CGH (eight amplifications), a >3-fold number of DNA amplifications was detected (n = 29). The most frequent amplifications mapped to 7p12.3 (three pancreatic cancer cell lines and three pancreatic tumor specimens), 8q24 (four pancreatic cancer cell lines and one pancreatic tumor specimen), 11q13 (three pancreatic cancer cell lines and three pancreatic tumor specimens), and 20q13 (four pancreatic cancer cell lines and three pancreatic tumor specimens). Genes contained in the consensus regions were MYC (8q24), EGFR (7p12.3), and FGF3 (11q13). In six of seven pancreatic cancer cell lines and pancreatic tumor specimens with 20q13 amplifications, the novel candidate gene NFAT C2, which plays a role in the activation of cytokines, was amplified. Other amplifications also affected genes for which a pathogenetic role in pancreatic carcinoma has not been described, such as BCL10 and BCL6, two members of the BCL family. A subset of amplified genes was checked for overexpression by means of real-time PCR, revealing the highest expression levels for BCL6 and BCL10. Thus, Matrix-CGH allows the detection of a high number of amplifications, resulting in the identification of novel candidate genes in pancreatic cancer.     

mRNA expression of leukemia-associated antigens in patients with acute myeloid leukemia for the development of specific immunotherapies

Specific immunotherapies for patients with acute myeloid leukemia (AML) using leukemia-associated antigens (LAA) as target structures might be a therapeutic option to enhance the graft-vs.-leukemia effect observed after allogeneic stem cell transplantation or to prolong a complete remission (CR) achieved by chemotherapy. Significant mRNA expression of LAA is a prerequisite for such immunotherapies. Here, previously characterized antigens associated with solid tumors (TAA) and newly characterized LAA were investigated for their expression in up to 60 AML patients and in leukemia cell lines. To investigate their specificity for leukemic blasts, the mRNA expression was also characterized in PBMN and CD34 positive cells of healthy volunteers and in a panel of normal tissues. The following antigens showed high mRNA expression in AML patients: MPP11 was detected in 43/50 (86%), RHAMM in 35/50 (70%), WT1 in 40/60 (67%), PRAME in 32/50 (64%), G250 in 18/35 (51%), hTERT in 7/25 (28%) and BAGE in 8/30 (27%) of AML patients. Real-time RT-PCR showed a tumor-specific expression of the antigens BAGE, G250 and hTERT, as well as highly tumor-restricted expression for RHAMM, PRAME and WT1. The antigen MPP11 was overexpressed. These antigens might be candidates for immunotherapies of leukemia patients and, because of their simultaneous expression, also for polyvalent vaccines.     

Conception of heavy ion beam therapy at Heidelberg University (HICAT)

The ion beam therapy facility HICAT presently under construction at the Heidelberg University Clinic will be the first clinical irradiation facility for heavy ions in Europe. Its capacity should enable the treatment of 1000 patients per year. The use of different ion species ranging from protons to oxygen under identical conditions should clarify the question of which particle species is best suited in terms of indication. A synchrotron will accelerate the particles to energies corresponding to water-equivalent ranges from 2 cm to 30 cm. An intensity-controlled raster scanning technique will be used to optimize the use of the favorable depth dose distribution of ions. The planned heavy-ion gantry will be the first world-wide. The facility should be complete in 2006.     

Candidate genes upregulated in density dependent growth inhibition of lung cancer cells

Prognosis of lung cancer remains poor despite the recent development of new chemotherapeutic agents. Novel therapeutic strategies therefore need to be developed. The search for factors inhibiting tumor growth in a paracrine/autocrine fashion might result in a well-tolerated adjuvant tumor therapy. In this study we aimed to identify candidate genes for such inhibitors of tumor cell growth. Native and heat-inactivated supernatants of confluent, slow growing H460 tumor cell cultures and of sparse (non-confluent), fast growing H460 tumor cell cultures were tested in proliferation assays. We observed that native supernatant of confluent H460 and A549 cells contain proteins inhibiting tumor cell growth of NSCLC cell lines. Microarray gene expression analysis of sparse and confluent H460 cells exhibited overexpression of 7 candidate genes in confluent, slow growing cells. The products of these genes possess cell growth inhibitory function and also exist in the extracellular compartment. The increased expression level of these genes was verified using real-time RT-PCR analysis. Our results show that especially components of IGF pathway appear to be involved in exogenous growth inhibition of confluent cells. Further investigations of these factors may result in the identification of autocrine/paracrine tumor cell growth inhibitory proteins for future use in clinical applications.     

Signal transduction in anaplastic large cell lymphoma cells (ALCL) mediated by the tumor necrosis factor receptor CD30

Expression of the cytokine receptor CD30 is a characteristic feature of anaplastic large cell lymphoma (ALCL). Reports regarding CD30-mediated signaling in ALCL cells are highly controversial, especially with respect to the regulation of cell survival. In this study, we stimulated 6 ALCL-derived cell lines with immobilized anti-CD30 antibody. CD30-induced cell death was investigated by Western blot and FACS analysis. CD30-dependent cell proliferation and activation was analyzed by applying the trypan blue exclusion method and a luciferase-based ATP assay. The expression of cell cycle relevant proteins and the activation of mitogen-activated protein (MAP) kinases were also examined. We demonstrated that activation of CD30 did not lead to the cleavage of pro-caspase-3. FACS analysis confirmed that in all examined cells cell death was not mediated by CD30. Cell growth was strongly inhibited in 2 of the 6 cell lines and restrained cell growth was accompanied by expression of the cell cycle inhibitor p21(WAF1/CIP1). Furthermore, stimulation of CD30 led to the activation of the p38 MAP kinase but not of the extracellular signal-regulated kinase (ERK) or the jun N-terminal kinase (JNK). Interestingly, activation of CD30 induced a strong synergistic reduction of cell activity, if the p38 MAP kinase activity was blocked by SB203580. The aim of the study was to elucidate CD30-induced signaling in different ALCL-cells. Our results suggest that CD30-mediated apoptosis is not a common feature in this cell type and that p38 MAP kinase is involved in CD30-mediated singal transduction.     

Three-dimensional visualization of the nasal cavity and paranasal sinuses. Clinical results of a standardized approach using multislice helical computed tomography

OBJECTIVE: The aim of this study was to introduce 3-dimensional (3D) visualization of the nasal cavity and paranasal sinuses as a diagnostic tool and to examination the clinical results of its standardized application. METHODS: One hundred sixty patients with chronic or acute nasal symptoms underwent helical computed tomography scanning and fiberoptic endoscopy of the sinuses, and 120 of them underwent endoscopic sinus surgery. The 3D images were compared with the axial and multiplanar reconstructed images using a checklist comprising important anatomic landmarks. After qualitative assessment of the representation of anatomic structures in healthy subjects, the method was applied to pathologic cases. The 3D images of these patients were correlated with the preoperative and intraoperative findings. RESULTS: Six hundred virtual endoscopies (VEs) were performed. The VE views allowed a realistic illustration of the various pathologic findings, except from cases with highly obstructive sinonasal disease. The correlation between the preoperative fiberoptic endoscopy and the intraoperative findings was significant (r = 0.83, P = 0.001). CONCLUSION: The standardized clinical application of this method serves as an important supplement to 2-dimensional slices and conventional endoscopy.