The first dual-phase xenon TPC equipped with silicon photomultipliers and characterisation with 37 Ar.

For the first time, a small dual-phase (liquid/gas) xenon time projection chamber was equipped with a top array of silicon photomultipliers for light and charge readout. Here we describe the instrument in detail, as well as the data processing and the event position reconstruction algorithms. We obtain a spatial resolution of ∼ 1.5 mm in the horizontal plane. To characterise the detector performance, we show calibration data with internal 83 m Kr and 37 Ar sources, and we detail the production of the latter as well as its introduction into the system. We finally compare the observed light and charge yields down to electronic recoil energies of 2.82 keV to predictions based on NEST v2.0.

Nondestructive application of laser-induced fluorescence spectroscopy for quantitative analyses of phenolic compounds in strawberry fruits (Fragaria x ananassa).

Laser-induced fluorescence spectroscopy (LIFS) was nondestructively applied on strawberries (EX = 337 nm, EM = 400-820 nm) to test the feasibility of quantitatively determining native phenolic compounds in strawberries. Eighteen phenolic compounds were identified in fruit skin by UV and MS spectroscopy and quantitatively determined by use of rp-HPLC for separation and diode-array or chemical reaction detection. Partial least-squares calibration models were built for single phenolic compounds by means of nondestructively recorded fluorescence spectra in the blue-green wavelength range using different data preprocessing methods. The direct orthogonal signal correction resulted in r (2) = 0.99 and rmsep < 8% for p-coumaroyl-glucose, and r (2) = 0.99 and rmsep < 24% for cinnamoyl-glucose. In comparison, the correction of the fluorescence spectral data with simultaneously recorded reflectance spectra did not further improve the calibration models. Results show the potential of LIFS for a rapid and nondestructive assessment of contents of p-coumaroyl-glucose and cinnamoyl-glucose in strawberry fruits.

Fluorimetric detection of protoporphyrins as an indicator for quality monitoring of fresh intact pork meat.

In fresh meat production fast and non-destructive quality monitoring along the distribution chain is a key aspect to guaranteeing high quality and safe products for consumption. The applicability of fluorescence spectroscopy using protoporphyrins as indicators for meat ageing was investigated. Porcine musculus longissimus dorsi (MLD) was stored in slices over 20 days at 5 and 12°C and measured every day with an excitation of 420nm and an emission range of 550-750nm. Additionally, pH, drip loss and colour were examined to assess possible correlations. The obtained spectra of the MLD showed an increase in three peaks at 592, 638 and 705nm which could be reconstructed using the spectra of standard solutions of protoporphyrin IX (PP) and zinc protoporphyrin IX (ZnPP) or magnesium protoporphyrin (MgPP), respectively. Using principal component analysis (PCA) on the fluorescence spectral data, the meat slices stored at 5°C showed differences in the fluorescence signal after the 10th day and 5th day when stored at 12°C. An interrelationship between the additional analyses and the fluorescence intensities on these relevant days could not be established. In conclusion, the increase of ZnPP fluorescence due to temperature related changes of physiological meat properties is capable of serving as a quality indicator with regards to inadequate conditioning (e.g. during transportation and/or storage) of pork meat.

Saccharomyces cerevisiae secretes and correctly processes human interferon hybrid proteins containing yeast invertase signal peptides.

Synthetic oligonucleotides coding for the yeast invertase secretion signal peptide were fused to the gene for the mature form of human interferon (huIFN-alpha 2). Two plasmids (E3 and F2) were constructed. E3 contained the invertase signal codons in a reading frame with the mature huIFN-alpha 2 gene. F2 had a deletion of the codon for alanine at amino acid residue-5 in the invertase signal and an addition of a methionine codon located between the coding sequences for the invertase signal and mature huIFN-alpha 2. Both hybrid genes were located adjacent to the promoter from the 3-phosphoglycerate kinase gene on the multicopy yeast expression plasmid, YEp1PT. Yeast transformants containing these plasmids produced somewhat more IFN than did the same expression plasmid containing the IFN gene with its human secretion signal sequence. HuIFN-alpha 2, purified from the medium of yeast cells containing E3, was found to be processed at the correct site. The huIFN-alpha 2 made by plasmid F2 was found to be completely processed at the junction between the invertase signal (a variant) and the methionine of methionine-huIFN-alpha 2. These results strongly suggested that the invertase signal (or its variant) attached to huIFN was efficiently recognized by the presumed signal recognition particle and was cleaved by the signal peptidase in the yeast cells. These results also suggested that amino acid changes on the right side of the cleavage site did not necessarily prevent cleavage or secretion.

3D surface accuracy of CAD generated skull defect contour.

The creation of a satisfactory cosmetic outcome in the repair of cranial defects relies on manual skill. However, computer aided design is gaining acceptance in the creation of custom cranial implants. The purpose of this work is to demonstrate the accuracy of a CAD generated skull defect contours using 3D difference maps. 3D multi-slice CT scanning was carried out on a life size plastic skull. Surface models were generated of the original skull and of temporofrontal and parietal defects. Surface contours were interpolated towards the centre of the defect from the edges where it was blended. The CAD contour deviation ranged from 0.0 mm to 2.0 mm with 80% of the total defect area less than 0.66 mm as measured by difference maps. CAD techniques can be used to produce contours for the repair of cranial defects with minimum deviation from the original skull contour. This enables accurate design and production of cranial implants.

"Virtual unwrapping" of a mummified hand.

The purpose of this work is to demonstrate the feasibility of medical virtual reality technologies in the investigation of a mummified hand. The Ulster Museum obtained the mummy hand, which originated from Thebes, without any identifying information. The mummified hand was investigated using conventional X-ray and 3D multi-slice Computed Tomography (CT). Imaging revealed a range of fractures of the wrist, metacarpals and phalanges whilst 3D CT demonstrated internal structures using volume rendering. The absence of any features of bone healing at the fracture sites would imply that they occurred just prior to death or in the mummified state possibly during excavation. Conventional X-ray imaging indicated that the hand, although small, was likely to have originated from an adult. Medical imaging and virtual reality display will enable us to produce a rapid prototyped model using fused deposition technology. Therefore, further paleopathological research can be performed on the replica without the need to handle the original specimen.

Calcium ion-flux across phosphatidylcholine membranes mediated by ionophore A23187.

The antibiotic A23187 carries Ca2+ across Müller-Rudin membranes made from 1,2-dierucoyl-sn-glycero-3-phosphocholine and n-decane. The conductance of the membranes is not increased by the Ca2+-transport. The flux depends linearly on Ca2+ concentration and ionophore concentration (above pH 6). It increases with increasing pH, approximately by a factor of 4-5 between pH 6 and pH 8. Maximal Ca2+-fluxes of about 10(-10) mol-cm-2-s-1 were found. A counter transport of H+ could not be detected. The complex formation between A23187 and Ca2+ in egg phosphotidylcholine vesicles was studied spectroscopically. The results are consistent with the formation of a 2:1 complex. Optical absorption measurements on single phophatidylcholine membranes were used to calculate the concentration of membrane-bound ionophore A23187.

Properties of bilayer membranes in the presence of dipicrylamine. A comparative study by optical absorption and electrical relaxation measurements.

In order to test the question if a pool of lipophilic ions may exist in black lipid membranes which cannot be detected by electrical relaxation measurements we have performed simultaneously measurements of the optical absorption of a lipophilic ion. The absorbance of membrane-bound dipicrylamine at 410 nm was measured with a sensitive spectrophotometer which can detect absorbance changes larger than or equal to 4-10(-5). A minimal concentration of about 6-10(11) dipicrylamine ions per cm2 of the membrane could be detected with this instrument. The dipicrylamine concentration in the membrane obtained with the optical method Ntopt is compared with the concentrations Ntel obtained from simultaneous electrical relaxation meausurements. Ntopt and Ntel agreed at low dipicrylamine concentrations (10(-8)--10(-7) M in the aqueous phase) and showed saturation at higher concentrations (up to 5-10(-6) M). In the saturation range Ntopt was maximally four times higher than Ntel. The significance of this difference is discussed together with general aspects of the saturation phenomenon.

CAD generated mold for preoperative implant fabrication in cranioplasty.

Intraoperative fabrication of acrylic cranial implants may be difficult and will increase operation time. In addition forming implants directly on the defect, intracranial tissues are exposed to heat of polymerization and residual monomer, that occurs, when autopolymerizing methyl methacrylate is used intraoperatively. Furthermore the cosmetical result may be unacceptable. Preoperatively formed acrylic implants may reduce these disadvantages compared to conventional techniques in cranioplasty. We will present methods for preoperative fabrication of cranial implants for a cadaver specimen. Implants were fabricated using a Rapid prototyping (RP) models of the skull built by Fused Deposition Modeling (FDM). In addition a mold of the defect was generated by CAD techniques, that can serve as a template for implant design.

Stereotactic radiotherapy of extracranial targets: CT-simulation and accuracy of treatment in the stereotactic body frame.

BACKGROUND AND PURPOSE: Evaluation of set-up accuracy and analysis of target reproducibility in the stereotactic body frame (SBF), designed by Blomgren and Lax from Karolinska Hospital, Stockholm. Different types of targets were analyzed for the risk of target deviation. The correlation of target deviation to bony structures was analyzed to evaluate the value of bones as reference structures for isocenter verification. MATERIALS AND METHODS: Thirty patients with 32 targets were treated in the SBF for primary or metastatic peripheral lung cancer, liver metastases, abdominal and pelvic tumor recurrences or bone metastases. Set-up accuracy and target mobility were evaluated by CT-simulation and port films. The contours of the target at isocenter level, bony structures and body outline were compared by matching the CT-slices for treatment planning and simulation using the stereotactic coordinates of the SBF as external reference system. The matching procedure was performed by using a 3D treatment planning program. RESULTS: Set-up accuracy represented by bony structures revealed standard deviations (SD) of 3.5 mm in longitudinal, 2.2 mm in anterior-posterior and 3.9 mm in lateral directions. Target reproducibility showed a SD of 4.4 mm in longitudinal, 3.4 mm ap and 3.3 mm in lateral direction prior to correction. Correlation of target deviation to bones ranged from 33% (soft tissue targets) to 100% (bones). CONCLUSION: A security margin of 5 mm for PTV definition is sufficient, if CT simulation is performed prior to each treatment to correct larger target deviations or set-up errors. Isocenter verification relative to bony structures is only safe for bony targets but not for soft tissue targets.

Intra-arterial cerebral thrombolysis for acute ischemic stroke in a community hospital.

BACKGROUND AND PURPOSE: Advances in thrombolytic therapy, brain imaging, and neurointerventional techniques provide new therapeutic options for acute stroke. Intra-arterial thrombolysis has proved to be a potent therapeutic tool. To show that this procedure can be performed in community hospitals, we describe our experience with a group of 11 patients treated for middle cerebral artery occlusions. METHODS: Twenty-two patients seen during a period of 1 year with clinical findings of acute major-vessel stroke met screening criteria and were evaluated under an institutional review board-approved protocol. After CT scanning, 17 of those patients met strict criteria, gave informed consent, and underwent angiography. Eleven patients had M1 and M2 middle cerebral artery occlusions and received local thrombolytic therapy with urokinase. Recanalization efficacy, complications, and outcome data were compiled. RESULTS: The average score on the National Institutes of Health Stroke Scale was 22.2 at the onset of treatment and 12.5 after therapy, with 91% of patients showing neurologic improvement. Complete (TIMI 3) recanalization occurred in 73% of cases and partial recanalization (TIMI 2) in 18%. At the 90-day follow-up evaluation, 56% of patients had good outcomes (modified Rankin score, 0 to 1). One intracranial hemorrhage occurred. CONCLUSION: Intra-arterial thrombolysis can be performed in a community hospital by radiologists with interventional and neuroradiologic skills given appropriate institutional preparation.

The in vitro colony assay: a predictor of clinical outcome.

PURPOSE: To determine the predictive power of an in vitro colony assay on the clinical normal-tissue complication rate. MATERIAL AND METHODS: Primary skin fibroblasts from 88 individuals were generated from the skin biopsies of patients who received a standardized radiotherapy. Tissue was cultured for three to six passages, irradiated with doses between 1 and 8 Gy under defined conditions, seeded and finally the colonies were stained and counted after 10-14 days. The survival curves were fitted by the L-Q model and the SF2, alpha/beta and plating efficiency were calculated. RESULTS: The parameters SF2 and plating efficiency were stable throughout the 4-year test period. Intra-individual differences between repeated experiments were significantly lower than inter-individual test results. For the observed acute skin and late normal-tissue reactions other than skin the in vitro parameter SF2 correlated significantly (p<0.005). For late skin reactions this correlation was not found. DISCUSSION: In contrast to other publications, a clear correlation was found between the in vitro test results and clinically observed early reactions. The lack of correlation for late skin reactions suggests that the combination of intrinsic radiation sensitivity and exogenous factors may alter the clinically observed reaction of certain tissues to a different extent.

In vitro radiosensitivity measured in lymphocytes and fibroblasts by colony formation and comet assay: comparison with clinical acute reactions to radiotherapy in breast cancer patients.

PURPOSE: To compare colony-forming and comet assays on fibroblasts and lymphocytes of 32 breast cancer patients irradiated after breast-conserving operations and to correlate the results with acute clinical radiation reactions in the skin. MATERIAL AND METHODS: Skin fibroblasts were isolated and cultivated before radiotherapy and lymphocytes were drawn prior to the first and directly after the final external irradiation. The colony-forming assay was performed with fibroblasts and the comet assay with lymphocytes and fibroblasts of breast cancer patients according to standard protocols. The clinical radiation reactions of the patients were graded according to the RTOG system. RESULTS: No significant correlation (p =0.09) was detected between clinical acute skin reactions and the in vitro clonogenic data in fibroblasts. Results of the comet assay in lymphocytes, however, showed a significant correlation (p <0.05) with the clinical data when patients were divided into two groups with average and elevated acute reactions. Apart from initial damage, fibroblasts did not show significant differences between the two patient groups. Repeated comet assays in lymphocytes of the same patient drawn before treatment and before and after external radiotherapy demonstrated good reproducibility of the test and no significant impact of preceding radiation treatment. There was a good correlation (r =0.65) between the comet assay results in fibroblasts and lymphocytes of the same individual. CONCLUSIONS: In this cohort of patients, a significant correlation between the in vitro results of the comet assay in lymphocytes and clinical acute reactions was detected. The results of the comet assay and of fibroblast colony formation did not correlate with in vitro radiosensitivity.

Phylogeny and diversity of Achromatium oxaliferum.

Achromatium oxaliferum was first described in 1893 by Schewiakoff as an unusually large bacterium living in freshwater sediments. Up to now no pure culture is available. Physical enrichments of achromatia collected from the acidic Lake Fuchskuhle, which houses a peculiar, smaller variety, and the neutral Lake Stechlin were investigated by the cultivation-independent rRNA approach. PCR in combination with cloning and sequencing was used for the retrieval of 24 partial and 4 nearly full-length 16S rRNA sequences that formed two distinct phylogenetic clusters. Fluorescence-in-situ-hybridization (FISH) with four 16S rRNA-targeted oligonucleotide probes unambiguously assigned the different sequences to either regular, large A. oxaliferum cells or to the smaller Lake Fuchskuhle population, tentatively named "A. minus". The two Achromatium sp. 16S rRNA sequence clusters form a stable deep branch in the gamma subclass of the class Proteobacteria. The closest cultivated relatives are Chromatium vinosum, Rhabdochromatium marinum and Ectothiorhodospira halophila with 16S rRNA similarities of 86.2 to 90.5%. Profound differences in the population structure of achromatia were revealed in the two lakes by FISH. In one sample from Lake Stechlin three genotypes could be visualized, and 49% of the cells were assigned to A. oxaliferum clone AST01, 28% to Achromatium sp. genotype AFK192/AFK433 and 23% to Achromatium sp. genotype AFK192/AST433. In contrast, a morphologically and phylogenetically homogeneous population of "A. minus". was present in Lake Fuchskuhle.

Recent developments in cornea transplantation (1997-1999).

Between 1997 and 1999 a steady increase in cornea donations was achieved, but the number of transplantations remained stable because many grafts did not pass quality control. Intermediate organ culture of entire bulbi was examined as a possible solution to reduce post-mortem times and increase suitability for transplantation.

Evaluation of seizure observation and documentation.

This study evaluated seizure documentation after participants observed videotaped seizures to determine how their background (educational level, employment position, years of experience, practice frequency, and familiarity with epilepsy and seizures) and attributes of the seizures themselves affected their skill. Observer variables did not show significant differences when mean seizure rating scores of 348 documented seizures for 58 participants were compared. Combinations of variables were significant; certified nursing assistants (CNAs) with clinical seizure experience had significantly higher mean seizure rating scores than those without experience (p < .01). There were significant differences in the mean percentile scores for each seizure observed (p < .001). Seizures with excess motor activity had the highest scores. Most common observations for any seizure observed were location and description of movement. Participants had significantly higher scores when observing a seizure a second time (p < .001). Differences in education, employment position, and years of employment are not limiting factors in performing seizure observation and documentation for persons who have been trained and certified in the skill. Opportunities to periodically view seizures improved seizure observation and documentation. Because motor movement observations predominate seizure documents, seizure observation instruction should not only emphasize key observational details but also highlight less frequent observations, such as responsiveness, that may play a key role in seizure classification. Using videotaped seizures and a scoring tool are useful for initial training and certification as well as periodic retraining to maintain quality in the skill.

Anatomical accuracy in medical 3D modeling.

In complex surgery, medical modeling has become an accepted tool for diagnosis, simulation and the planning of surgical interventions [1]. However, the question concerning the accuracy of the model, i.e. the equivalence between the model itself on the one hand and the original anatomical situation on the other hand, remains unanswererd in the current literature.

Medical biomodelling in surgical applications: results of a multicentric European validation of 466 cases.

The study started in September 1999 and ended in April 2002. It is based on a questionnaire [www.phidias.org] assessing case-related questions due to the application of stereolithographic models. Each questionnaire contains over 50 items. These variables take into account diagnosis, indications and benefits of stereolithographic models with view on different steps of the surgical procedures: preoperative planning, intraoperative application and overall outcome after surgical intervervention. These questionnaires were completed by the surgeons who performed operation. Over the time course of our multicentric study (30 months), we evaluated 466 cases. The study population consists of n=231 male and n= 235 female patients. 54 surgeons from 9 European countries were involved. There are main groups of diagnosis that related to the use of a model. Most models were used in maxillofacial surgery. The operative planning may help to determine the resection line of tumor and optimize reconstructive procedures. Correction of large calvarian defects can be simulated and implants can be produced preoperatively. Overall in 58 % of all cases a time- saving effect was reported. The study strongly suggests, that medical modeling has utility in surgical specialities, especially in the craniofacial and maxillofacial area, however increasingly in the orthopedic field. Due to our results, medical modeling optimizes the preoperative surgical planning. Surgeons are enabeled to perform realistic and interactive simulations. The fabrication of implants, its design and fit on the model, allow to reduce operation time and in consequence risk and cost of operation. In addition, the understanging of volumetric data is improved, especially if medical models are combined with standart imaging modalities. Finally, surgeons are able to improve communication between their patientents and colleagues.

Temperature elevation during simulated polymethylmethacrylate (PMMA) cranioplasty in a cadaver model.

The aims of this study were to: (i) obtain temperature measurements during in vitro polymerisation of polymethylmethacrylate (PMMA) disks of a range of thicknesses; and (ii) obtain tissue temperature measurements at various locations within a skull defect during a simulated PMMA cranioplasty procedure using a cadaver. In vitro, higher temperatures were recorded with increasing PMMA thickness. During the simulated cranioplasty, the maximum temperature was observed inside the PMMA sample, with nearby tissues being exposed to temperatures of greater than 50 degrees C over prolonged periods. There is conflicting information in the literature concerning the sensitivity of brain tissue and bone to elevated temperatures. Preoperatively fabricated PMMA cranioplasty prostheses are recommended.