Distribution of internal carotid artery plaque locations among patients with central retinal artery occlusion in the Eagle study population.

PURPOSE: Arterial emboli in the internal carotid artery (ICA) mainly cause cerebral ischemia; only 10 % of emboli reach the retinal arteries. Computational blood flow studies suggest that plaques situated in the ICA siphon may be a source of embolism to the ophthalmic artery (OA). To validate these calculated probabilities in patients with central retinal artery occlusion (CRAO), we reanalyzed digital subtraction angiography (DSA) images from the Multicenter Study of the European Assessment Group for Lysis in the Eye (EAGLE) study, a multicenter randomized study in patients with nonarteritic CRAO. METHODS: A reevaluation of 34 DSA studies was done from the interventional arm of the EAGLE study with regards to distribution of arterial plaques at specific ICA siphon locations and ICA stenosis. A comparison was made of plaque distribution to calculated probabilities for emboli reaching the OA from a computational fluid dynamics (CFD) model of a patient-specific ICA siphon. RESULTS: Most of the ICA plaques near the OA's origin were located in the cavernous ICA portion (31.3%). Of these, 12.5 % had plaques in the curvature opposite the OA origin, a location carrying the highest risk for embolization into the OA (according to the CFD model 12.6-13.2 % probability of embolisation into the OA). Also, 15.6 % had plaques in the paraclinoid ICA portion distal to the OA origin. CONCLUSIONS: There were 40.6% of the patients that had plaques in the cavernous and clinoid ICA portions presenting possible sources for embolic material generating RAO.

[Are there differences between internal carotid artery and aortic arch plaques among patients with retinal artery occlusion and anterior ischaemic optic neuropathy?]. / Bestehen Unterschiede bei Plaques der Arteria carotis interna und des Aortenbogens bei Patienten mit retinalen arteriellen Verschlüssen und anteriorer ischämischer Optikusneuropathie?

INTRODUCTION: Retinal artery occlusion (RAO) and non-arteritic anterior ischaemic optic neuropathy (NAION) cause painless vision deterioration and visual field defects. Associations with arteriosclerotic risk factors, hyperhomocysteinaemia, hyperfibrinogenaemia and plaques of the internal carotid artery are well kown. Results of transoesophageal echocardiography with respect to plaques in the aortic arch and duplex sonography of the internal carotid artery were evaluated and a statistical analysis was performed. MATERIAL AND METHODS: 110 patients with RAO and 49 patients with NAION were retrospectively analysed. Statistical significant differences were analysed with the Mann-Whitney test (U test) using SPSS 13.0 (IBM). RESULTS: RAO patients have statistically significantly more often plaques in the aortic arch, medium-sized plaques of the internal carotid artery, a history of stroke and hyperhomocysteinaemia. NAION patients have statistically significantly more often small plaques of the internal carotid artery. All other parameters do not show statistically significant differences between both diseases. CONCLUSIONS: RAO patients have a higher median age and seem to have more often arteriosclerotic plaques in the aortic arch and larger plaques in the internal carotid artery than NAION patients, although both diseases show comparable associations with arteriosclerotic risk factors.

Complex expression pattern of Wnt ligands and frizzled receptors in human placenta and its trophoblast subtypes.

Canonical Wingless (Wnt) signalling provoked by exogenous and endogenous Wnt ligands was recently shown to play a crucial role in the invasive differentiation of human trophoblasts. To gain insights into the expression pattern of the developmental regulators, we analysed all human Wnt ligands and their frizzled (FZD) receptors in the human placenta and different trophoblast model systems using semi-quantitative PCR. Fourteen out of 19 Wnt ligands and 8 out of 10 FZD receptors were detectable in placental tissues, however, expression patterns varied with gestational age and between different trophoblast subtypes suggesting cell-specific functions. Besides Wnt ligands acting through the canonical pathway, non-canonical ligands such as Wnt-5a, which may also activate alternative Wnt signalling pathways or inhibit canonical Wnt signalling, could be identified. Western blot analyses revealed secretion of Wnt-5a from primary trophoblast cultures and trophoblastic cell lines. To evaluate the potential role of Wnt-5a, SGHPL-5 trophoblast cells were transfected with luciferase reporter plasmids harbouring eight T-cell factor (TCF) DNA-recognition sequences which are exclusively activated through the canonical Wnt signalling pathway. Luciferase assays revealed that Wnt-3a-induced reporter activity was repressed by recombinant Wnt-5a indicating an antagonistic role in trophoblasts. The data suggest that a complex network of Wnt ligands and FZD receptors may regulate developmental processes of the human placenta.

Cdc25A phosphatase suppresses apoptosis induced by serum deprivation.

The phosphatase Cdc25A was shown to be a target of the transcription factor c-Myc. Myc-induced apoptosis appeared dependent on Cdc25A expression and Cdc25A over-expression could substitute for Myc-triggered apoptosis. These findings suggested that an important downstream component of Myc-mediated apoptosis was identified. However and in contrast, we recently reported that during TNFalpha-induced apoptosis, which required c-Myc function, Cdc25A was down-regulated in a human carcinoma cell line. We now provide evidence that Cdc25A rendered the non-transformed rat embryonic cell line 423 refractory to apoptosis, which was induced by serum deprivation and in absence of detectable c-myc levels. The survival promoting activity of cdc25A was abolished upon infection of cells with a full-length cdc25A antisense construct. To identify the signaling proteins mediating the survival function of the phosphatase, cdc25A- and akt- over-expressing pooled clones were exposed to selected chemicals, which inhibit or activate key proteins in signaling pathways. Inhibition of apoptosis by SU4984, NF023 and Rapamycin placed Cdc25A and Akt function downstream of FGF.R, PDGF.R, and compensated G-protein- and PP2A- activity. Interestingly, upon treatment with LY-294002, cdc25A- and akt- over-expressing clones exhibited similar apoptotic patterns as control cells, which indicates that neither Akt- nor Cdc25A-mediated survival functions are dependent on PI.3 kinase activity in rat 423 cells. In cdc25A-overexpressing cells increased levels of serine 473 phosphorylated Akt were found, which co-precipitated with Cdc25A and Raf1. Since activation of proteins requires dephosphorylation of particular residues in addition to site-specific phosphorylation, the anti-apoptotic effect of Cdc25A might derive from its participation in a multimeric protein complex with phosphoAkt and Raf1, two prominent components of survival pathways.

Subcellular localisation of Cdc25A determines cell fate.

Cell division cycle 25A (Cdc25A) was shown to colocalise both with nuclear and cytoplasmic proteins. Recently, we have demonstrated that overexpressed Cdc25A promoted the survival of rat 423 cells through indirect activation of PKB-protein kinase B. Using a Cdc25A:ER fusion protein, which can be shuttled from the cytoplasm into the nucleus, the present investigation evidences that the antiapoptotic effect of Cdc25A was restricted to its cytoplasmic localisation in rat 423 cells. In contrast, nuclear Cdc25A overexpression caused dephosphorylation and nuclear retention of the proapoptotic transcription factor Forkhead in rhabdomyosarcoma-like 1 (FKHRL1) in human N.1 ovarian carcinoma cells. This resulted in the increased constitutive expression of the FKHRL1 targets Fas ligand and Bim, and promoted apoptosis. Thus, the Cdc25A oncogene, which was found to be frequently overexpressed in certain human cancers, can increase or decrease the susceptibility to apoptosis depending on the cell-type-specific subcellular distribution.

Maintenance of ATP favours apoptosis over necrosis triggered by benzamide riboside.

A new synthetic drug, benzamide riboside (BR) exhibited strong oncolytic activity against leukemic cells in the 5-10 microM range. Higher BR-concentrations (20 microM) predominantly induced necrosis which correlated with DNA strand breaks and subsequent depletion of ATP- and dATP levels. Replenishment of the ATP pool by addition of adenosine prevented necrosis and favoured apoptosis. This effect was not a pecularity of BR-treatment, but was reproduced with high concentrations of all trans-retinoic acid (120 microM) and cyanide (20 mM). Glucose was also capable to suppress necrosis and to favour apoptosis of HL-60 cells, which had been treated with necrotic doses of BR and cyanide. Apoptosis eliminates unwanted cells without affecting the microenvironment, whereas necrosis causes severe inflammation of surrounding tissues due to spillage of cell fluids into the peri-cellular space. Thus, the monitoring and maintenance of cellular energy pools during therapeutic drug treatment may help to minimize nonspecific side effects and to improve attempted drug effects.

[Presentation of software for collation of inpatient diurnal pressure profiles in glaucoma patients]. / Vorstellung einer Software zur Erfassung von stationären Tagesdruckprofilen bei Glaukompatienten.

The aim of diurnal pressure profiles is clarification of therapy safety and the diagnosis of glaucoma. The evaluation of intraocular pressure curves is part of the running assessment. In order to support this evaluation software has been developed which, in addition to the digital presentation of intraocular pressure curves corrected for corneal thickness also allows immediate statistical assessment. After specifying a target pressure it also allows deduction of possible therapy recommendations based on the resulting intraocular pressure values, the cup/disc ratio and the number of antiglaucoma agents given. In this study two collectives were investigated: in the first collective, in addition to the previously determined diagnoses, the intraocular pressure, number and time of measurements, predetermined target pressure, cup/disc ratio and corneal thickness-dependent correction values were evaluated. After completing a scoring system with respect to agreement of the clinical therapy suggestions with those suggestions calculated by the software, the second collective was compared after termination of the diurnal pressure profile. Concordance was found in 73 out of 88 eyes. The digital collation system for profile measurement was found to be very well suited for graphical and clinical documentation of diurnal pressure profile curves. Therapy decisions can be deduced from the available data and supported by the software.

Tumour necrosis factor-alpha impairs chorionic gonadotrophin beta-subunit expression and cell fusion of human villous cytotrophoblast.

Growth factors expressed at the fetal-maternal interface modulate hormone expression of placental trophoblasts. The aim of this study was to investigate the effects of different cytokines on hCG subunit mRNA expression in differentiating villous cytotrophoblasts. Quantitative real-time PCR revealed a 1.8- and 6.9-fold increase of hCG-alpha and hCG-beta mRNA levels, respectively, between 36 and 60 h of term trophoblast syncytialization. Compared with controls, neither interleukin (IL)-1beta, IL-2, IL-4, IL-6, IL-10, IL-13 and IL-15 nor tumour necrosis factor (TNF)-alpha significantly altered hCG-alpha mRNA expression. Similarly, the ILs did not affect hCG-beta transcript levels. In contrast, TNF-alpha suppressed hCG-beta mRNA 3.8- and 1.8-fold at 36 and 60 h of term trophoblast differentiation. Accordingly, hCG secretion was impaired by TNF-alpha but not by the different ILs. Moreover, TNF-alpha reduced luciferase expression of reporter plasmids harbouring the proximal hCG-beta5 promoter to 35 and 77%, respectively, in primary term trophoblasts and trophoblastic SHGPL-5 cells. In addition, counting of nuclei in syncytialized, desmoplakin-negative areas revealed a 1.9-fold reduction of term trophoblast fusion in the presence of TNF-alpha. Similarly, floating explant cultures prepared from first trimester-denuded villi recovered the syncytium 2.8-fold less efficiently during 72 h of cytokine treatment. Concomitantly, TNF-alpha impaired induction of endogenous and secreted hCG-beta protein levels in these cultures. The data suggest that TNF-alpha decreases hCG-beta mRNA and protein expression by reducing gene transcription and trophoblast cell fusion. Suppression of these processes by TNF-alpha could partly explain the adverse effects of the cytokine on placental function and pregnancy outcome.

Analysis of mechanisms contributing to AraC-mediated chemoresistance and re-establishment of drug sensitivity by the novel heterodinucleoside phosphate 5-FdUrd-araC.

Chemoresistance is a biological response of cells to survive toxic stress. During cancer treatment the development of chemoresistance is a major problem. The mechanisms how cells become insensitive, and which downstream pathways are affected are not completely understood. Since it has not been well analysed which and how many regulative disorders are subsummised under the term "chemoresistance", we examined and measured arabinosylcytosine (AraC)-mediated desensitation of two mechanisms relevant for tissue homeostasis, cell cycle inhibition and apoptosis induction. MCF-7 cells harbouring ectopic mutated p53 were suitable for this investigation because they activated these mechanisms subsequently and became insensitive to AraC with regard to cell cycle inhibition and apoptosis induction. The major causal mechanism of acquired resistance against AraC was most likely through the inhibition of the first step of AraC phosphorylation within the cell, which is rate limiting for its activation. With regard to cell cycle inhibition AraC-resistant cells were also resistant against 5-fluorodeoxyuridine (5-FdUrd), but fully responsive to 5-FdUrd-induced apoptosis, evidencing that cell cycle and apoptosis are independent of each other. Apoptosis correlated with AIF-activation and was independent of Caspase 7, whereas cell cycle inhibition correlated with cyclinD1 expression but not with induction of p21 or p27. The phosphate conjugated 5-FdUrd-araC heterodimer (5-Fluoro-2'-desoxyuridylyl-(3'-->5')-Arabinocytidine), which is a prodrug of AraC-monophosphate, reactivated AIF and down-regulated cyclin D1 in AraC-resistant cells and circumvented resistance to apoptosis and to cell cycle inhibition. Also, cells which were resistant to 5-FdUrd or doxorubicin were sensitive to 5-FdUrd-araC. This investigation demonstrates that chemoresistance affects apoptosis induction and cell cycle inhibition independently and that detailed knowledge about the affected downstream pathways would enable the design of targeted intervention with small molecules to restore chemosensitivity.

[Comparison of preoperative retinometer values with postoperative visual acuity after surgery of epiretinal membranes]. / Vergleich des präoperativen Retinometerwertes mit dem postoperativen Visus nach Chirurgie epiretinaler Membranen.

BACKGROUND: We report about our clinical experience when comparing the preoperative retinometer values with the postoperative visual acuity after microsurgical excision of epiretinal membranes (ERM). PATIENTS AND METHODS: Pars plana vitrectomy with membrane peeling was performed in 56 eyes of 53 patients. Preoperatively, we obtained the distant and near visual acuity and the retinometer value. Postoperatively, we compared the preoperative retinometer value to the best postoperative visual acuity. RESULTS: The preoperative retinometer value was in 39 eyes (70 %) equal to the best postoperative distant visual acuity (+/- 1 line) and in 34 eyes (61 %) equal to the best near visual acuity (+/- 1 line). A difference of + 2 or - 2 lines or more from the retinometer value was found in 17 eyes (30 %) for the best postoperative distant visual acuity and in 22 eyes (39 %) for the best near visual acuity. 49 eyes (87.5 %) had a better visual acuity after pars plana vitrectomy, 6 eyes (10.7 %) had equal visual acuities and one eye (1,8 %) was worse than the preoperative visual acuity. The average improvement of vision after the operation was + 2.5 lines for the distant and + 2.4 lines for the near visual acuity. CONCLUSIONS: The preoperative retinometer examination is a precise method for obtaining the visual acuity of patients selected for pars plana vitrectomy with membrane peeling. Uncertain results can be achieved in eyes with very opaque ERM and ERM with macular edema.