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The discovery of the CO2-fixing enzyme Rubisco in the Archaebacteria has presented a conundrum in that they apparently lack the gene for phosphoribulokinase, which is required to generate Rubisco's substrate ribulose 1,5-bisphosphate (RuBP). However, two groups1, 2 have now demonstrated novel RuBP synthesis pathways, demystifying Rubisco's non-autotrophic and perhaps ancient role. A new CO2 fixing role for Rubisco, which is distinct from the globally dominant Calvin cycle, is providing important clues furthering our understanding of the evolution of autotrophy. This perspective is strengthened by the additional recognition in this commentary that some Rubisco-containing Archaea do also contain PRK and may represent an interesting autotrophic evolutionary transition. Supplementary material for this article can be found on the BioEssays website (http://www.interscience.wiley.com/jpages/0265-9247/suppmat/index.html). BioEssays 29:722–724, 2007. © 2007 Wiley Periodicals, Inc.

Purpose: In about 15 % of patients with SAH no causative vascular lesions can be found in acute imaging with CTA and DSA. Usually, repeat DSA is mandatory and bears the usual risk of invasive angiography. The present study attempts to assess the diagnostic impact of 3 D rotational angiography in order to avoid repeat DSA. Materials and Methods: From January 2004 to December 2012, 649 patients with an acute non-traumatic SAH were examined. 91 patients with negative initial imaging diagnostics concerning the bleeding source were included in this study. These patients underwent a second angiography scan: 61 in 4-plane technique, and 30 with 2-plane technique and additional 3 D DSA. Two cohorts were compared: patients with repeat angiography in conventional 4-plane technique from 2004 to July 2008 and 2-plane technique with additional 3 D rotational DSA from 2008 to 2012. Statistical significance was verified by means of Fisher’s exact test. Results: In the second DSA scan, 4 aneurysms in 4 patients (4/91; 4.4 %) were found and treated subsequently. Within the first 4.5 years of this study, 401 patients with SAH were treated and 61 of them underwent repeat angiography (15.2 %) compared to 30 of 248 patients (12.1 %) in the last 4.5 years of this study. In the first group we found 3 aneurysms during repeat angiography, and in the second group we found 1. No significance was reached (p = 0.29) but there was a tendency towards higher diagnostic security using 3D-DSA. Conclusion: Using 3 D rotational DSA in initial imaging workup might help to reduce false-negative results concerning the bleeding source of acute SAH. At least because of this fact, 3 D rotational DSA should be part of the diagnostic workup after acute SAH. Key Points: • In 15 % of SAH patients no causative vascular lesion can be found using conventional DSA compared to 12 % using 3 D DSA. • Patients with perimesencephalic SAH and CT-negative SAH do not require repeat DSA. • 3 D DSA might help to avoid repeat DSA in SAH patients. Citation Format: • Ringelstein A, Mueller O, Monninghoff C et al. 3D Rotational Angiography After Non-Traumatic SAH. Fortschr Rontgenstr 2014; 186: 675 – 679

Objective: Severe traumatic brain injury remains the leading cause of death in children. The present study analyses the outcome of children after severe traumatic brain injury (TBI) treated by decompressive craniectomy (DC) due to elevated intracranial pressure (ICP) in a single centre. Method: 56[for full text, please go to the a.m. URL], 64. Jahrestagung der Deutschen Gesellschaft für Neurochirurgie (DGNC)

We examined the influence of focal cerebellar lesions on working memory ( n-back task), gait, and the interaction between working memory and different gait tasks in a dual-task paradigm. The analysis included 17 young patients with chronic focal lesions after cerebellar tumor resection and 17 age-matched controls. Patients have shown mild to moderate ataxia. Lesion sites were examined on the basis of structural magnetic resonance imaging. N-back tasks were executed with different levels of difficulty ( n = 1–4) during sitting (baseline), treadmill walking, and treadmill tandem walking (dual-task conditions). Patients exhibited decreased n-back performance particularly at difficult n-back levels and in dual-task conditions. Voxel-based lesion-symptom mapping revealed that decreased baseline n-back performance was associated with lesions of the posterolateral cerebellar hemisphere and the dentate nucleus. By contrast, decreased n-back performance in dual-task conditions was more associated with motor-related areas including dorsal portions of the dentate and the interposed nucleus, suggesting a prioritization of the motor task. During baseline walking, increased gait variability was associated with lesions in medial and intermediate regions, whereas for baseline tandem gait, lesions in the posterolateral hemispheres and the dentate nucleus became important. Posterolateral regions overlapped with regions related to baseline n-back performance. Consistently, we observed increased tandem gait variability with growing n-back difficulty in the dual-task condition. These findings suggest that dual-task effects in cerebellar patients are at least partially caused by a common involvement of posterolateral cerebellar regions in working memory and complex motor tasks.

Background Occipital nerve stimulation (ONS) has been shown to be effective for selected patients with intractable headache disorders. We performed a prospective critical evaluation of complications and direct treatment costs. Methods Twenty-seven patients with chronic cluster headache (CCH, n = 24) or chronic migraine (CM, n = 3) underwent a trial phase with bilateral ONS and subsequent implantation of a permanent generator (IPG), if responsive to treatment according to predefined criteria. Procedural and long-term complications as well as direct treatment costs of neuromodulation therapy of ONS were recorded over a mean follow-up period of 20 months (range 5–47 months). Results Twenty-five of 27 patients (93%) responded to treatment. Twenty-one complications in 14 patients were identified, necessitating reoperation in 13 cases. Overall treatment costs were €761,043, including hardware-related costs of €506,019, costs for primary hospital care of €210,496, and complications related to hospitalization costs of €44,528. This results in a per case-based cost of €9445 for hospitalization and €18,741 for hardware costs, totaling €28,186. Conclusion ONS for treatment of refractory CCH and CM is a cost-intensive treatment option with a significant complication rate. Nevertheless, patients with refractory primary headache disorders may experience substantial relief of pain attacks, and headache days, respectively.

Stimulation of the greater occipital nerve has been employed for various intractable headache conditions for more than a decade. Still, prospective studies that correlate stimulation of the greater occipital nerve with outcome of patients with respect to alleviation of headache are sparsely found in literature.To identify anatomical landmarks for a reproducible stimulation of the greater occipital nerve. For the clinical implication, the individual response to therapy of patients with refractory chronic cluster headache undergoing occipital nerve stimulation was correlated with the postoperative localization of the electrodes and with the distribution of the stimulation field.Prospective observational study, approved by the local research ethics board (09-4143).University hospital, departments of neurosurgery and neurology, institute of anatomy and radiology.Ten formaldehyde fixed human cadavers were dissected to identify the passage of the greater occipital nerve through the trapezius muscle. The distance to the external occipital protuberance was triangulated measuring the distance of the nerve from the nuchal midline and the protuberance. Between December 2008 and December 2011, 21 consecutive patients suffering from chronic cluster headache underwent surgery in terms of bilateral occipital nerve stimulation, with electrodes placed horizontally at the level of C1. The postoperative x-rays were compared with the acquired landmarks from the anatomical study. The distribution of the stimulation field was correlated to the individual response of each patient to the therapy and prospectively analyzed with regard to reduction of daily cluster attacks and relief of pain intensity at 3 months and at last follow-up.The greater occipital nerve crosses the trapezius muscle at a mean distance of 31 mm below the occipital external protuberance and 14 mm lateral to the midline as found in the anatomical subjects. The electrodes were targeted at this level in all of our patients and stimulated the greater occipital nerve in all patients. Eighteen of the patients (85.7%) reported a significant reduction of the frequency of their cluster attacks and/or declined intensity of pain during the attacks. Yet, 3 of 21 patients (14.3%) did not benefit from the stimulation despite an adequate spread of the stimulation over the occiput. The spread of the stimulation-induced paraesthesias over the occiput was not correlated to a reduction of cluster attacks, to the intensity of attacks, or to the response to treatment at all.Single center non-randomized non-blinded study.From our study we conclude that a reproducible stimulation of the greater occipital nerve can be achieved by placing the electrodes parallel to the atlas, at about 30 mm distance to the external occipital protuberance. The response to the stimulation is not correlated to the field width of the paraesthesia. We, therefore, consider stimulation of the main trunk of the greater occipital nerve to be more important than a large field of stimulation on the occiput. Still, an individual response to the occipital nerve stimulation cannot be predicted even by optimal electrode placement.

Background and purpose To test the hypothesis that sporadic cerebral cavernous malformations (CCMs) are systematically associated with venous malformations (VMs) using susceptibility weighted imaging (SWI) at 7 Tesla (T) field MRI. Methods A prospective unselected series of 20 patients with symptomatic or asymptomatic sporadic CCM diagnosed using 1.5 T MRI was additionally scanned using high resolution (250 µm 2 in-plane) SWI at 7 T MRI. Imaging data were analysed to examine the presence and formation of CCM associated venous vessel structures. Interobserver agreement was assessed using kappa statistics. Results In the 20 patients harbouring 23 CCMs, a solitary or multiple venous drainage was found in all lesions. A ‘typical’ VM was found in seven lesions. In the other cases, associated abnormal venous structures were also depicted although they appeared structurally different. Excellent interobserver agreement was achieved (95% confidence interval 0.92 to 0.99). Conclusions Our data support previous assumptions that sporadic CCMs are systematically associated with local venous abnormalities involving larger outflow vessels. However, the typical appearance of a VM was not confirmed in all cases. The role of the venous environment in the pathomechanism of CCMs remains unclear.

Objective: Intracranial aneurysms (IAs) require deliberately selected treatment strategies as they are incrementally found prior to rupture and deleterious subarachnoid haemorrhage (SAH). Multiple and recurrent aneurysms necessitate both neurointerventionalists and neurosurgeons to optimize aneurysmal occlusion in an interdisciplinary effort. The present study was conducted to condense essential strategies from a single neurovascular centre with regard to the lessons learned. Method: Medical charts of 321 consecutive patients treated for IAs at our centre from September 2008 until December 2010 were retrospectively analysed for clinical presentation of the aneurysms, multiplicity and treatment pathways. In addition, a selective Medline search was performed. Results: A total of 321 patients with 492 aneurysms underwent occlusion of their symptomatic aneurysm: 132 (41.1%) individuals were treated surgically, 189 (58.2%) interventionally; 138 patients presented with a SAH, of these 44.2% were clipped and 55.8% were coiled. Aneurysms of the middle cerebral artery were primarily occluded surgically (88), whereas most of the aneurysms of the internal carotid artery and anterior communicating artery (114) were treated endovascularly. Multiple aneurysms (range 2–5 aneurysms/individual) were diagnosed in 98 patients (30.2%). During the study period 12 patients with recurrent aneurysms were allocated to another treatment modality (previously clip to coil and vice versa ). Conclusions: Our data show that successful interdisciplinary occlusion of IAs is based on both neurosurgical and neurointerventional therapy. In particular, multiple and recurrent aneurysms require tailored individual approaches to aneurysmal occlusion. This is achieved by a consequent interdisciplinary pondering of the optimal strategy to occlude IAs in order to prevent SAH.

Alterations of the intracellular ubiquitin-proteasome pathway are found in neurodegenerative and inflammatory disorders of the central nervous system, as well as in its malignancies. Inhibitory substrates of the proteasomes represent promising approaches to control autoimmune inflammations and induction of apoptosis in cancer cells. Extracellular circulating proteasomes are positively correlated to outcome prognosis in hematogenic neoplasias and the outcome in critically ill patients. Previously, we reported raised levels of proteolytic active 20S proteasomes in the extracellular alveolar space in patients with acute respiratory distress syndrome (ARDS). For the cerebrospinal fluid, we assumed that extracellular circulating proteasomes with enzymatic activity can be found, too. Cerebrospinal fluid (CSF) samples of twenty-six patients (14 females, 12 males), who underwent diagnostic spinal myelography, were analyzed for leukocyte cell count, total protein content, lactate and interleukine-6 (Il-6) concentrations. CSF samples were analyzed for concentration and enzymatic activity of extracellular 20S proteasomes (fluorescenic substrate cleavage; femtokatal). Blood samples were analyzed with respect to concentration of extracellular circulating proteasomes. Choroidal plexus was harvested at autopsies and examined with immunoelectron microscopy (EM) for identification of possible transportation mechanisms. Statistical analysis was performed using SPSS (18.0.3). In all patients, extracellular proteasome was found in the CSF. The mean concentration was 24.6 ng/ml. Enzymatic activity of the 20S subunits of proteasomes was positively identified by the fluorescenic subtrate cleavage at a mean of 8.5 fkat/ml. Concentrations of extracellular proteasomes in the CSF, total protein content and Il-6 were uncorrelated. Immunoelectron microscopy revealed merging vesicles of proteasomes with the outer cell membrane suggestive of an exozytic transport mechanism. For the first time, extracellular circulating 20S proteasome in the CSF of healthy individuals is identified and its enzymatic activity detected. A possible exozytic vesicle-bond transportation mechanism is suggested by immunoelectron microscopy. The present study raises more questions on the function of extracellular proteasome in the CSF and encourages further studies on the role of extracellular protesomes in pathological conditions of the central nervous system (tumor lesions and inflammatory processes).

Rationale and Objectives Geometrical distortion is a well-known problem in structural magnetic resonance imaging (MRI), leading to pixel shifts with variations up to several millimeters. Because the main factors of geometrical distortion are proportional to B 0 , MRI spatial encoding distortions tend to increase with higher magnetic field strength. With the increasing prospects of utilizing ultra-high-field MRI (B 0 ≥ 7 Tesla) for neuroimaging and subsequently for image-guided neurosurgical therapy, the evaluation and correction of geometrical distortions occurring in ultra-high-field MRI are essential preconditions for the integration of these data. Hence, we conducted a phantom study to determine hardware-related geometrical distortion in clinically relevant sequences for structural imaging at 7 T MRI and compared the findings to 1.5 T MRI. Material and Methods Hardware-related geometrical distortion was evaluated using a MRI phantom (Elekta, Sweden). Both applied scanner systems (Magnetom Avanto 1.5 T and Magnetom 7 T, Siemens Healthcare, Erlangen, Germany) were equipped with similar gradient coils capable of delivering 45 mT/m of maximum amplitude and a slew rate of 220 mT/m/ms. Distortion analysis was performed for various clinically relevant gradient echo and spin echo sequences. Results Overall, we found very low mean geometrical distortions at both 7 T and 1.5 T, although single values of up to 1.6 mm were detected. No major differences in mean distortion between the sequences could be found, except significantly higher distortions in turbo spin-echo sequences at 7 T, mainly caused by B 1 inhomogeneities. Conclusion Hardware-related geometrical distortions at 7 T MRI are relatively small, which may be acceptable for image coregistration or for direct tissue-targeting procedures. Using a subject-specific correction of object-related distortions, an integration of 7 T MRI data into image-guided applications may be feasible.

Myxomas are benign tumors of the mesenchymal origin and account for about half of the benign cardiac tumors. Occasionally, they occur at other locations, but the intracranial manifestation of a myxoma is exceptionally rare. As a secondary neoplasia following radiotherapy, myxoma has only been reported once in the literature.A 12-year-old girl, who was previously treated for a medulloblastoma, was diagnosed with a new lesion at the left transverse sinus in the follow-up magnetic resonance imaging (MRI). Indication for surgery was made and complete removal could be achieved.Histological examination revealed a myxoma. Further staging showed no other manifestation of the myxoma. The close relation to the radiation field of the posterior fossa makes it highly suggestive that the myxoma developed as a secondary neoplasia induced by radiotherapy. Treatment philosophy for this benign tumor entity is a completed resection of the lesion with regular follow-up MRI.

Instable upper cervical spine injuries (CSI) in very young children are rare and consecutively only few data on the treatment and operative approaches exist in the literature. Hence, we suggest an operative strategy in the case of a 15-month-old infant treated for an instable CSI at the level of C2/3 at our department. Detailed steps of the operation with special consideration to the challenging anatomy of the immature spine are presented. A toddler suffered a CSI without neurological deficits after a stair fall. Computer tomography (CT) and magnetic resonance imaging (MRI) of the cervical spine revealed an instable luxation fracture of C2/3. As repositioning in the halo vest immobilization failed, surgical fusion was indicated. Via a posterior midline approach, the lamina of C2 and C3 was conflated in a modified sublaminar wiring technique using non-resorbable sutures, sparing the ossification zones of the vertebral arches. Postoperative immobilization in a halo vest facilitated bony fusion of the laminae at C2/3 without lordotic displacement of the cervical spine. We consider the instable CSI of the immature cervical in the very young a challenging situation for every treating physician. The particular features of the growing spine require special attention to avoid damage to the growth centers of the vertebrae. To our knowledge, this is the first technical report giving detailed information of an operative approach to the severely injured immature cervical spine and postoperative management.

Cavernous malformations (cavernomas) occur in 0.4%-0.9% of the population. The most common symptoms of cavernous malformations are seizures. An improved detection of small cavernomas might possibly help to clarify cryptogenic seizures, which represent 30% of all seizures. The aim of this study was to evaluate the benefit of magnetic resonance imaging at a field strength of 7 T in the detection of cavernomas.Ten consecutive patients with known cavernomas were enrolled in this study. First a T2*-weighted gradient echo sequence was performed at 1.5 T with a slice thickness of 6 mm in axial orientation. Subsequently, the examination was repeated at 7 T. Both examinations were independently read by two blinded senior neuroradiologists. Localization and dimension of the hypointensities were recorded.One additional hypointensity was detected in one patient, which was not visible in the 1.5 T examination even retrospectively. In another patient with known cavernomatosis, innumerable new small hypointense lesions were detected at 7 T.Our small series suggests that ultra-high-field magnetic resonance imaging at 7 T improves the detection of cavernomas.

Background: Cardiac expression of the Ca-dependent inotropic protein S100A1 is decreased in human end-stage heart failure (HF) and cardiomyocyte-targeted viral-based S100A1 gene transfer rescued failing myocardium in small animal models in vivo and in vitro via improved systolic and diastolic sarcoplasmic reticulum Ca-handling. We therefore hypothesized that cardioselective AAV9-S100A1 gene therapy will improve cardiac performance in a large animal experimental HF model under clinical conditions. Methods and Results: Left ventricular (LV) posterolateral myocardial infarction (MI) was induced in pigs by occlusion of the left coronary circumflex artery and resulted in LV failure (HF) 2 weeks post-MI reflected by a 40% and 27% reduction in LV +dp/dt max. and EF, respectively, as assessed by LV catheterization and echocardiography. Post-MI HF pigs were then randomized for retroinfusion of AAV9-luciferase (luc; n=6, 1.5×10 13 total viral particles, tvp) and AAV9-S100A1 (S100A1; n=6, 1.5×10 13 tvp) driven by a cardioselective promoter via the anterior cardiac vein while the left anterior descending artery was temporarily occluded. 14 weeks after cardiac gene transfer, the S100A1-treated HF group showed significantly enhanced S100A1 protein expression (+46.7±17.9%, P Conclusions: Our translational study provides proof of concept that AAV9-S100A1 based HF gene therapy is feasible and restores cardiac function in a large animal HF model under clinical conditions. Next, certified toxicological analysis and different AAV9-S100A1 dosage protocols will be assessed to eventually advance to first phase I/II clinical studies determining therapeutic efficiency of cardiac S100A1 gene therapy in HF patients.

One unexpected feature of the human genome is the high structural variability across individuals. Frequently, large regions of the genome show structural polymorphisms and many vary in their abundance. However, accurate methods for the characterization and typing of such copy number variations (CNV) are needed. The defensin cluster at the human region 8p23.1 is one of the best studied CNV regions due to its potential clinical relevance for innate immunity, inflammation, and cancer. The region can be divided into two subclusters, which harbor predominantly either alpha- or beta-defensin genes. Previous studies assessing individual copy numbers gave different results regarding whether the complete beta-defensin cluster varies or only particular genes therein. We applied multiplex ligation-dependent probe amplification (MLPA) to measure defensin locus copy numbers in 42 samples. The data show strict copy number concordance of all 10 loci typed within the beta-defensin cluster in each individual, while seven loci within the alpha-defensin cluster are consistently found as single copies per chromosome. The exception is DEFA3, which is located within the alpha-defensin cluster and was found to also differ in copy number interindividually. Absolute copy numbers ranged from two to nine for the beta-defensin cluster and zero to four for DEFA3. The CNV-typed individuals, including HapMap samples, are publicly available and may serve as a universal reference for absolute copy number determination. On this basis, MLPA represents a reliable technique for medium- to high-throughput typing of 8p23.1 defensin CNV in association studies for diverse clinical phenotypes.

During the last decade the practice of laboratory-directed protein evolution has become firmly established as a versatile tool in biochemical research by enabling molecular evolution toward desirable phenotypes or detection of novel structure-function interactions. Applications of this technique in the field of photosynthesis research are still in their infancy, but recently first steps have been reported in the directed evolution of the CO(2)-fixing enzyme Rubisco and its helper protein Rubisco activase. Here we summarize directed protein evolution strategies and review the progressive advances that have been made to develop and apply suitable selection systems for screening mutant forms of these enzymes that improve the fitness of the host organism. The goal of increasing photosynthetic efficiency of plants by improving the kinetics of Rubisco has been a long-term goal scoring modest successes. We discuss how directed evolution methodologies may one day be able to circumvent the problems encountered during this venture.

Background Because of the high resistance rate of fibrosarcomas against cytotoxic agents clinical chemotherapy of these tumors is not established. A better understanding of the diverse modes of tumor cell death following cytotoxic therapies will provide a molecular basis for new chemotherapeutic strategies. In this study we elucidated the response of a fibrosarcoma cell line to clinically used cytostatic agents on the level of gene expression. Methods HT1080 fibrosarcoma cells were exposed to the chemotherapeutic agents doxorubicin, actinomycin D or vincristine. Total RNA was isolated and the gene expression patterns were analyzed by microarray analysis. Expression levels for 46 selected candidate genes were validated by quantitative real-time PCR. Results The analysis of the microarray data resulted in 3.309 (actinomycin D), 1.019 (doxorubicin) and 134 (vincristine) probesets that showed significant expression changes. For the RNA synthesis blocker actinomycin D, 99.4% of all differentially expressed probesets were under-represented. In comparison, probesets down-regulated by doxorubicin comprised only 37.4% of all genes effected by this agent. Closer analysis of the differentially regulated genes revealed that doxorubicin induced cell death of HT1080 fibrosarcoma cells mainly by regulating the abundance of factors mediating the mitochondrial (intrinsic) apoptosis pathway. Furthermore doxorubicin influences other pathways and crosstalk to other pathways (including to the death receptor pathway) at multiple levels. We found increased levels of cytochrome c, APAF-1 and members of the STAT-family (STAT1, STAT3), while Bcl-2 expression was decreased. Caspase-1, -3, -6, -8, and -9 were increased indicating that these proteases are key factors in the execution of doxorubicin mediated apoptosis. Conclusion This study demonstrates that chemotherapy regulates the expression of apoptosis-related factors in fibrosarcoma cells. The number and the specific pattern of the genes depend on the used cytotoxic drug. The response rates on the gene expression level, i.e. the number of genes regulated by the drugs actinomycin D, doxorubicin and vincristine, correlate to the clinical effectiveness of the drugs. Doxorubicin seems to exert its cytotoxic mechanism by regulating genes, which are involved in several different apoptosis regulating pathways. The exact knowledge of the genes affected by the drugs will help to understand the diverse modes of soft tissue sarcoma cell death in response to cytotoxic therapies.

Ultrasound computer tomography is an imaging method capable of producing volume images with high spatial resolution. The imaged object is enclosed by a cylindrical array of transducers. While one transducer emits a spherical wavefront (pulse), all other transducers are recording the radiofrequency (RF) a-scans simultaneously. Then another transducer acts as the emitter and so on. In this paper we describe the image reconstruction method and an enhanced algorithm for the a-scan preprocessing. The image reconstruction is based on a 'full aperture sum-and-delay' algorithm evaluating the reflected and scattered signals in the a-scans. The a-scans are modelled as the tissue response of the imaged object convoluted with the shape of the ultrasound pulse, which is determined by the transfer function of the transducers and the excitation. Spiking deconvolution and blind deconvolution with different parameters are used to build inverse filters of the ultrasound pulse. Applying the inverse filters to the a-scans results in sharper signals which are used for image reconstruction. Smallest scatterers of 0.1 mm size corresponding to one fifth of the used ultrasound wavelength are visible in the reconstructed images. Compared to conventional b-scans the resulting images show an approximately tenfold better resolution.

Ultrasound computer tomography is an imaging method capable of producing volume images with both high spatial and temporal resolution. The promising results of a 2D experimental setup of an ultrasound computer tomography system with at least 0.25 mm resolution encouraged us to build a new 3D demonstration system. It consists of three parts: a tank containing the sensor system, a data acquisition hardware and a computer workstation for image reconstruction and visualization. For the sensor system we developed and manufactured our own low-cost transducer array emitting or receiving ultrasound signals in three dimensions. To optimize the transducer geometry in respect to aperture angle and pressure amplitude the pressure field was simulated using the ultrasound simulation program Field II. Each transducer arrays system carries 8 emitting and 32 receiving elements with integrated amplifier and address electronics. 192 A-scans can be recorded in parallel by the data acquisition hardware. 48 multiplexing steps are needed to store all A-scans of the 1536 receiving transducers. After recording the data is transmitted to the computer workstation.

Antimicrobial peptides such as defensins provide nonspecific mucosal defense against a multitude of microorganisms. Recently, it has been shown that luminal bacteria may invade the mucosa in inflammatory bowel diseases, suggesting a defect in innate mucosal immunity. The aim of this study was to investigate the expression of human beta-defensins (HBD) in controls, Crohn's disease (CD), ulcerative colitis (UC), and unspecific inflammation. Up to 4 biopsies were taken from 103 patients (33 controls, 24 with Crohn's disease, 36 with ulcerative colitis, 10 with unspecific colitis). Mucosal mRNA was measured using real-time fluorescence temperature cycler reverse-transcription polymerase chain reaction with primers for HBD-1, HBD-2, HBD-3, tumor necrosis factor alpha, and interleukin 8. Mucosal HBD-1 expression was marginally decreased in both CD and UC. HBD-2 was increased exclusively in UC but not in CD. The expression of the novel defensin HBD-3 was strongly correlated with HBD-2 and also raised predominantly in UC. The expression of both inducible beta-defensins was enhanced in the state of inflammation. Expression of HBD-2 showed a weak correlation with interleukin 8 only in inflamed CD biopsies but not with tumor necrosis factor alpha. The missing induction of both inducible beta-defensins in CD as compared with UC may cause a defect in barrier function that predisposes to bacterial invasion.

We review the crucial properties of a phase-encoded volume holographic storage system in terms of data quality and security, which are the key issues of any bulk memory system. Two major problems which need to be tackled in holographic storage systems in terms of data quality are the hologram erasure during readout and the data encoding schemes for error-free reconstruction. We present a novel storage material, (bismuth tellurite crystals - Bi 2 TeO 5 ), which has the potential to overcome the volatility problem and avoiding the need of any further fixing. Regarding data encoding schemes, we present a general approach of gray scale modulation coding in order to improve the data capacity in comparison to normal modulation coding, while the bit error rate maintains low. Data security in a phase encoded system can be realized by exploiting its special multiplexing characteristics. We present different encryption techniques and investigate their decryption probability.

In breast cancer diagnosis, ultrasound examination provides useful additional diagnostic information. Moreover ultrasound does not harm biological tissue and can be applied frequently. But conventional ultrasound imaging methods lack both high spatial and temporal resolution. Usually, the scanner is operated manually and the tissue is deformed while getting as close as possible to the regions of interest. Therefore, image contents and image quality depend strongly on the operator. Exact measurement of tissue structures, like tumor size, is not possible. Instead of a manually controlled linear transducer array, we use ultrasound computer tomography (USCT) to image a volume directly. A few thousand ultrasound transducers are arranged in a cylindrical array around a tank containing the object to be examined coupled by water. Every single transducer is small enough to emit an almost spherical sound wave. While one transducer is transmitting, all others receive simultaneously. Afterwards a different transducer emits the next pulse. For volume reconstruction every transmitted, scattered and reflected signal is used. This new method allows reproducible image sequences with enhanced spatial and temporal resolution. For the benefit of more reconstructed 3D images per second, spatial resolution may be reduced offline. First tests with our prototype in a ring-shaped geometry have even showed nylon threads (0.4 mm) and an image quality superior to clinical ultrasound scanners.

We present several encryption techniques for holographic data storage, using orthogonal phase-code multiplexing and random phase encoding. Our system is capable of storing page-oriented data based on the selectivity of orthogonal phase-codes in a photorefractive LiNbO3 storage crystal. In order to encrypt data, random phase multiplexing is added to the system. We proof that the combination of deterministic and random phase code multiplexing is possible with low cross-talk and take advantage of this technique for an extremely secure data encryption. Moreover, the potential of phase-code multiplexing to realize arithmetic operations is exploited for data encryption purposes.© (2000) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

Phase-coded multiplexing for volume holographic memories is a promising alternative to conventional addressing methods. We present a first realization of digital data storage by means of phase-coded multiplexing. The system concept, its implementation, and its performance is described. The potential of image processing is demonstrated experimentally. Data encryption of the stored data using an additional random phase key is discussed and experimental results are shown.© (1999) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

This work describes the three-dimensional reconstruction of clustered microcalcifications based on only two digitized mammograms. First, the mammograms are examined separately to detect suspicious areas automatically. A further investigation separates microcalcifications from other structures. Based on an optimized region matching and on a specially adapted inverse discrete radon-transformation the corresponding volume is estimated from two projections and visualized by a continuously rotating object. But do two projections of a cluster carry enough information to reconstruct its three- dimensional arrangement sufficiently? We use Shannon's definition of information to estimate a lower bound of preserved information, described as the ratio of average information contained in the projections and average information contained in the volume, for simplified scenarios. Assuming two orthogonal projections of a cubic volume containing k binary representations of microcalcification positions the average information in the projections is determined by the combinatorial quantity of admissible arrangements and the size n3 of the volume. The combinatorial quantity of legal three-dimensional arrangements of microcalcification positions describes the average information carried by the volume. We showed that the amount of preserved information in the projections is more than 95% if k equals n/2 positions are found in both projections; it will exceed 98% if k equals n/4 positions are set.© (1999) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

X-ray mammography is one of the most significant diagnosis methods in early detection of breast cancer. Usually two X- ray images from different angles are taken from each mamma to make even overlapping structures visible. X-ray mammography has a very high spatial resolution and can show microcalcifications of 50 - 200 micron in size. Clusters of microcalcifications are one of the most important and often the only indicator for malignant tumors. These calcifications are in some cases extremely difficult to detect. Computer assisted diagnosis of digitized mammograms may improve detection and interpretation of microcalcifications and cause more reliable diagnostic findings. We build a low-cost mammography workstation to detect and classify clusters of microcalcifications and tissue densities automatically. New in this approach is the estimation of the 3D formation of segmented microcalcifications and its visualization which will put additional diagnostic information at the radiologists disposal. The real problem using only two or three projections for reconstruction is the big loss of volume information. Therefore the arrangement of a cluster is estimated using only the positions of segmented microcalcifications. The arrangement of microcalcifications is visualized to the physician by rotating.© (1998) COPYRIGHT SPIE--The International Society for Optical Engineering. Downloading of the abstract is permitted for personal use only.

The design and realization of a compact volume holographic memory using phase-coded multiplexing is described. We present a system capable of storing up to 480 data pages with a resolution of 640 x 480 bits in a single interaction region of a photorefractive LiNbO3 crystal. A characterization of the key components of the system is given. Firstexperimental results of high-resolution analog image storage as well as the application of the system to real-timeoptical image processing are presented.Keywords: volume holographic memory, phase-coded multiplexing, photorefractive crystal, LiNbO3 1. INTRODUCTION Optical volume holographic memories provide the potential for high storage capacity, parallel readout of wholedata pages with short access times and high data transfer rates. Multiple holographic storage in a single regionof a photorefractive crystal can be achieved using different multiplexing techniques as angular,"2 wavelength,3'4 orphase-coded5'6 multiplexing, or by combinations of these techniques (e.g.79). By combining these techniques with

CO 2 is converted into biomass almost solely by the enzyme rubisco. The poor carboxylation properties of plant rubiscos have led to efforts that made it the most kinetically characterized enzyme, yet these studies focused on 100) were active in vitro, with the fastest having a turnover number of 22 ± 1 s−1—sixfold faster than the median plant rubisco and nearly twofold faster than the fastest measured rubisco to date. Unlike rubiscos from plants and cyanobacteria, the fastest variants discovered here are homodimers and exhibit a much simpler folding and activation kinetics. Our pipeline can be utilized to explore the kinetic space of other enzymes of interest, allowing us to get a better view of the biosynthetic potential of the biosphere., Metagenomic and biochemical analysis of previously uncharacterized naturally‐occurring form‐II and II/III rubiscos leads to identification of an enzyme with the fastest CO2 fixation rate described to date.

Ribulose 1, 5-bisphosphate carboxylase/oxygenase (Rubisco) is the most abundant enzyme representing the main gateway of inorganic carbon into the biosphere by catalysing CO2-fixation during photosynthesis. In spite of its pivotal role, the enzyme has poor kinetics and substrate specificity. Structurally, Rubisco exists in different forms ranging from dimers of large subunits to hexa-decamers of large and small subunits. These structurally distinct Rubiscos utilize complex machinery for biogenesis including a multiverse of chaperones. It is our aim to devise a simple in vitro method to investigate the folding and assembly requirements for different forms of Rubisco. An understanding of in vitro Rubisco reconstitution will provide critical information on additional components required for successful Rubisco re-engineering in in vivo systems, knowledge of which will help in producing platforms for recombinant expression of heterologous Rubisco systems. To this end we have adopted the continuous spectrophotometric Rubisco activity assay to quantitatively evaluate folding and assembly kinetics in addition to specific chaperone requirements of different Rubisco systems. Chaperonin GroEL/GroES and Mg-ATP are necessary and sufficient for both dimeric and hexameric form II Rubisco, consisting only of large subunits, to fold and assemble from denatured subunits. It was observed that the dimeric Rhodospirillum rubrum form II Rubisco had a more rapid maturation rate compared to the hexameric Acidithiobacillus ferrooxidans form II Rubisco likely due to the requirement for higher order oligomerization of the hexamer. Interestingly, in contrast to the well-studied chaperone dependent form I cyanobacterial Rubisco, the assembly of the hexadecameric proteobacterial form I Acidithiobacillus ferrooxidans Rubisco occurs independently of ancillary proteins. Instead, it requires only the small subunits to assemble into the functional hexa-decamers following successful folding by GroEL/GroES. In conclusion, our method will permit systematic in vitro screening and determination of conditions that allow comparative assessment of folding and assembly of all forms of Rubiscos. Simultaneously, we are aiming at constructing Rubisco engineering platforms using a cyanobacterial system, a well-established model organism for photosynthetic research. We have generated cyanobacterial strains (termed cyano-red) using Synechocystis sp. PCC 6803 in which the form IC red-type Rubisco from Rhodobacter sphaeroides along with its activase is expressed, folded and assembled properly. Further biochemical and physiological characterization of this cyano-red strain will provide us with novel insights on the biogenesis of different Rubiscos using this cyanobacterial platform. Doctor of Philosophy