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Background: During prostate stereotactic body radiation therapy (SBRT), prostate tumor translational motion may deteriorate the planned dose distribution. Most of the major advances in motion management to date have focused on correcting this one aspect of the tumor motion, translation. However, large prostate rotation up to 30° has been measured. As the technological innovation evolves toward delivering increasingly precise radiotherapy, it is important to quantify the clinical benefit of translational and rotational motion correction over translational motion correction alone. Purpose: The purpose of this work was to quantify the dosimetric impact of intrafractional dynamic rotation of the prostate measured with a six degrees-of-freedom tumor motion monitoring technology. Methods: The delivered dose was reconstructed including (a) translational and rotational motion and (b) only translational motion of the tumor for 32 prostate cancer patients recruited on a 5-fraction prostate SBRT clinical trial. Patients on the trial received 7.25 Gy in a treatment fraction. A 5 mm clinical target volume (CTV) to planning target volume (PTV) margin was applied in all directions except the posterior direction where a 3 mm expansion was used. Prostate intrafractional translational motion was managed using a gating strategy, and any translation above the gating threshold was corrected by applying an equivalent couch shift. The residual translational motion is denoted as (Formula presented.). Prostate intrafractional rotational motion (Formula presented.) was recorded but not corrected. The dose differences from the planned dose due to (Formula presented.) + (Formula presented.), ΔD((Formula presented.) + (Formula presented.)) and due to (Formula presented.) alone, ΔD((Formula presented.)), were then determined for CTV D98, PTV D95, bladder V6Gy, and rectum V6Gy. The residual dose error due to uncorrected rotation, (Formula presented.) was then quantified: (Formula presented.) = ΔD((Formula presented.) + (Formula presented.)) - ΔD((Formula presented.)). Results: Fractional data analysis shows that the dose differences from the plan (both ΔD((Formula presented.) + (Formula presented.)) and ΔD((Formula presented.))) for CTV D98 was less than 5% in all treatment fractions. ΔD((Formula presented.) + (Formula presented.)) was larger than 5% in one fraction for PTV D95, in one fraction for bladder V6Gy, and in five fractions for rectum V6Gy. Uncorrected rotation, (Formula presented.) induced residual dose error, (Formula presented.), resulted in less dose to CTV and PTV in 43% and 59% treatment fractions, respectively, and more dose to bladder and rectum in 51% and 53% treatment fractions, respectively. The cumulative dose over five fractions, ∑D((Formula presented.) + (Formula presented.)) and ∑D((Formula presented.)), was always within 5% of the planned dose for all four structures for every patient. Conclusions: The dosimetric impact of tumor rotation on a large prostate cancer patient cohort was quantified in this study. These results suggest that the standard 3–5 mm CTV-PTV margin was sufficient to account for the intrafraction prostate rotation observed for this cohort of patients, provided an appropriate gating threshold was applied to correct for translational motion. Residual dose errors due to uncorrected prostate rotation were small in magnitude, which may be corrected using different treatment adaptation strategies to further improve the dosimetric accuracy.

BACKGROUND: Hypofractionation in prostate radiotherapy is of increasing interest. Steep dose gradients and a large weight on each individual fraction emphasize the need for motion management. Real-time motion management techniques such as multileaf collimator (MLC) tracking or couch tracking typically adjust for translational motion while rotations remain uncompensated with unknown dosimetric impact. PURPOSE: The purpose of this study is to demonstrate and validate dynamic real-time rotation-including dose reconstruction during radiotherapy experiments with and without MLC and couch tracking. METHODS: Real-time dose reconstruction was performed using the in-house developed software DoseTracker. DoseTracker receives streamed target positions and accelerator parameters during treatment delivery and uses a pencil beam algorithm with water density assumption to reconstruct the dose in a moving target. DoseTracker's ability to reconstruct motion-induced dose errors in a dynamically rotating and translating target was investigated during three different scenarios: (1) no motion compensation and translational motion correction with (2) MLC tracking and (3) couch tracking. In each scenario, dose reconstruction was performed online and in real time during delivery of two dual-arc volumetric-modulated arc therapy prostate plans with a prescribed fraction dose of 7 Gy to the prostate and simultaneous intraprostatic lesion boosts with doses of at least 8 Gy, but up to 10 Gy as long as the organs at risk dose constraints were fulfilled. The plans were delivered to a pelvis phantom that replicated three patient-measured motion traces using a rotational insert with 21 layers of EBT3 film spaced 2.5 mm apart. DoseTracker repeatedly calculated the actual motion-including dose increment and the planned static dose increment since the last calculation in 84 500 points in the film stack. The experiments were performed with a TrueBeam accelerator with MLC and couch tracking based on electromagnetic transponders embedded in the film stack. The motion-induced dose error was quantified as the difference between the final cumulative dose with motion and without motion using the 2D 2%/2 mm γ-failure rate and the difference in dose to 95% of the clinical target volume (CTV ΔD95% ) and the gross target volume (GTV ΔD95% ) as well as the difference in dose to 0.1 cm3 of the urethra, bladder, and rectum (ΔD0.1CC ). The motion-induced errors were compared between dose reconstructions and film measurements. RESULTS: The dose was reconstructed in all calculation points at a mean frequency of 4.7 Hz. The root-mean-square difference between real-time reconstructed and film-measured motion-induced errors was 3.1%-points (γ-failure rate), 0.13 Gy (CTV ΔD95% ), 0.23 Gy (GTV ΔD95% ), 0.19 Gy (urethra ΔD0.1CC ), 0.09 Gy (bladder ΔD0.1CC ), and 0.07 Gy (rectum ΔD0.1CC ). CONCLUSIONS: In a series of phantom experiments, online real-time rotation-including dose reconstruction was performed for the first time. The calculated motion-induced errors agreed well with film measurements. The dose reconstruction provides a valuable tool for monitoring dose delivery and investigating the efficacy of advanced motion-compensation techniques in the presence of translational and rotational motion. ispartof: MEDICAL PHYSICS vol:49 issue:6 pages:3574-3584 ispartof: location:United States status: published

Human infection with parvovirus B19 causes a range of clinical manifestations, including benign erythema infectiosum in children, arthralgias in adults, aplastic crisis in patients with bone marrow failure, and potentially fatal congenital hydrops fetalis. Persistent parvovirus B19 infection is a rare disease presentation mostly seen in adult women or immunocompromised individuals. Treatment options include corticosteroids and intravenous immunoglobulin; however, viral clearance is difficult to obtain and rarely maintained. In this Grand Round, we report the case of a 43-year-old man with persistent parvovirus B19 infection and anaemia, who was refractory to standard treatment regimens, and whom we successfully treated with pegylated interferon alfa-2a. Initial treatment led to viral clearance and remission of anaemia, although secondary recurrence of virus required treatment extension. Despite extensive genetic and immunological evaluations, no underlying primary or secondary immunodeficiency was identified in the patient. We propose interferon alfa-2a as a treatment option for persistent parvovirus B19 infection and advocate long-term follow-up of patients and potentially repeated treatment.

Herpesvirus infections can lead to a number of severe clinical manifestations, particularly when involving the central nervous system (CNS), causing encephalitis and meningitis. However, understanding of the host factors conferring increased susceptibility to these diseases and their complications remains incomplete. Previous studies have uncovered defects in the innate Toll-like receptor 3 pathway and production of type I interferon (IFN-I) in children and adults that predispose them to herpes simplex encephalitis. More recently, there is accumulating evidence for an important role of IFN-independent cell-autonomous intrinsic mechanisms, including small nucleolar RNAs, RNA lariat metabolism, and autophagy, in restricting herpesvirus replication and conferring protection against CNS infection. The present review first describes clinical manifestations of HSV infection with a focus on neurological complications and then summarizes the host–pathogen interactions and innate immune pathways responsible for sensing herpesviruses and triggering antiviral responses and immunity. Next, we review the current landscape of inborn errors of immunity and the underlying genetic defects and disturbances of cellular immune pathways that confer increased susceptibility to HSV infection in CNS. Ultimately, we discuss some of the present outstanding unanswered questions relating to inborn errors of immunity and HSV CNS infection together with some perspectives and future directions for research in the pathogenesis of these severe diseases in humans.

Purpose: In radiotherapy, tumor motion may deteriorate the planned dose distribution. However, the dosimetric consequences of the motion are normally unknown for individual treatments. We here present a method for real-time motion-including tumor dose reconstruction and demonstrate its use for simulated stereotactic body radiotherapy (SBRT) of patients with liver cancer previously treated with Calypso-guided gating. Methods: Real-time motion-including dose reconstruction was performed using in-house developed software, DoseTracker, on offline replays of previous clinical treatments. The patient cohort consisted of fifteen patients previously treated in our clinic with three-fraction SBRT to the liver using conformal or IMRT plans. The tumor motion at treatment was monitored with implanted electromagnetic transponders. The dose reconstruction was performed for both the actual gated treatments and simulated nongated treatments using a 21 Hz data stream containing accelerator parameters and the recorded motion. The dose was reconstructed in the same calculation points within the planning target volume (PTV) as used by the treatment planning system (TPS). The reconstructed doses were compared with calculations performed in the TPS, in which the motion was modeled as a series of isocenter shifts. The comparison included point doses as a function of treatment time and the dose volume histogram (DVH) for the clinical target volume (CTV). The motion-induced reduction in the dose to 95% of the CTV, ∆D95%, and in the mean CTV dose, ΔDMean, was compared between DoseTracker and the TPS for each simulated fraction. DoseTracker currently assumes water density within the patient contour, so for comparison, the TPS calculations were performed with both CT density and water density. The calculation times were additionally analyzed. Results: Dose reconstruction was carried out for ninety SBRT sessions with calculation volumes ranging from 9.9 to 366.4 cm3 and median calculation times of 55–155 ms (equivalent to 18.2–6.5 Hz). Time-resolved trends of doses to a single calculation point in the patient were well replicated and dose differences between actual and planned calculations matched well. ΔDMean had a range of −0.1%–30.7%-points and was estimated by DoseTracker with a root-mean-square deviation (RMSD) to the TPS calculations of 0.43%–points (water density) and 0.79%-points (CT density). Similarly, ∆D95% had a range of 0.0%–35.2%-points and was estimated by DoseTracker with an RMSD of 0.80%-points (water density) and 1.33%-points (CT density). DoseTracker predicted losses in tumor dose coverage above 5%-points with high sensitivity (91.7%) and specificity (97.6%). Conclusions: Real-time dose reconstruction to moving tumors was demonstrated on offline replays of previous clinical treatments. DVHs of actually delivered dose are made available immediately after the end of treatment fractions. It shows promising results for liver SBRT with accurate estimation of CTV dose deteriorations caused by motion during treatment.

BACKGROUND Persons with dementia tend to be vulnerable to mobility challenges and hence face a greater risk of fall and subsequent fractures, morbidity, and mortality. Motion-based technologies (MBTs), also called sensor-based technologies or virtual reality, have the potential for assisting physical exercise and training as a part of a disease management and rehabilitation program, but little is known about its' use for people with dementia. OBJECTIVE The purpose of this pilot study was to investigate the feasibility and efficacy of MBT physical training at home for people with dementia. METHODS A 3-phase pilot study: (1) baseline start-up, (2) 15 weeks of group training at a local care center twice a week, and (3) 12 weeks of group training reduced to once a week, supplemented with individual MBT training twice a week at home. A total of 26 people with dementia from a municipality in Southern Denmark were eligible and agreed to participate in this study. Three withdrew from the study, leaving 23 participants for the final analysis. Feasibility was measured by the percentage of participants who trained with MBT at home, and their completion rate of total scheduled MBT sessions. Efficacy was evaluated by physical function, measured by Sit-to-Stand (STS), Timed-Up-and-Go (TUG), 6-minute Walk Test (6MW), and 10-meter Dual-task Walking Test (10MDW); cognitive function was measured by Mini-Mental State Examination (MMSE) and Neuropsychiatric Inventory-Questionnaire (NPI-Q); and European Quality of Life 5 dimensions questionnaire (EQOL5) was used for measuring quality of life. Descriptive statistics were applied accordingly. Wilcoxon signed-rank and rank-sum tests were applied to explore significant differences within and between the groups. RESULTS As much as 12 of 23 participants (52%) used the supplemental MBT training at home. Among them, 6 (50%) completed 75% or more scheduled sessions, 3 completed 25% or less, and 3 completed between 25% and 75% of scheduled sessions. For physical and cognitive function tests, supplementing with MBT training at home showed a tendency of overall stabilization of scores among the group of participants who actively trained with MBT; especially, the 10MDW test even showed a significant improvement from 9.2 to 7.1 seconds (P=.03). We found no positive effect on EQOL5 tests. CONCLUSIONS More than half of the study population with dementia used MBT training at home, and among them, half had an overall high adherence to the home training activity. Physical function tended to remain stable or even improved among high-adherence MBT individuals. We conclude that MBT training at home may be feasible for some individuals with dementia. Further research is warranted.

A design of a fully composite pylon with external grounding down-leads has been proposed for new-generation 400 kV transmission towers, able to save lines corridors and to reduce visual impact. This paper investigates and compares the backflashover performance of a composite pylon and two conventional metallic towers, which have been widely installed in Denmark. The transient models of overhead lines and all three towers were established respectively and the transient analysis was carried out in PSCAD. Monte Carlo method was used to estimate backflashover rate. The backflashover rate of composite pylon is 0.4526 cases per 100 km per year, which is in the same range, but slightly higher than that of metallic towers. The separated grounding down-leads of double circuits on composite pylon eliminates the danger of simultaneous backflashover of double circuits, which exists in transmission lines supported by metallic towers. After comparing the overvoltages to three phases of the three towers from backflashover, it is worthy considering that the installation of surge arresters at all three phases of composite pylon has a strong impact on the backflashover rather, but for the two metallic towers only the surge arresters at the upper phase has an impact.

EXECUTIVE SUMMARY Evaluations of improvements in long chronic-patient pathways must include both short- and long-term effects on patients; that is, effects on the full patient pathway. Otherwise, costs might be cut without considering the long-term effects and, consequently, the overall cost of the pathway could increase. Unfortunately, current methods of evaluation present several issues: (1) they do not provide valid insights regarding the effects of a given improvement effort until several years later, (2) they provide imprecise and biased results, and (3) the aggregated results are not useful for identifying and disseminating the best practices that lead to an improvement. In this article, the accelerated longitudinal design with decomposition of total costs (ALDD) method is applied to evaluate the effects of improvement efforts on inpatient utilization for long cardiac pathways at a Danish hospital. The results show that the ALDD method can deliver valid results much faster than traditional methods and can uncover hidden improvements in the local work processes of clinical teams. Application of the ALDD method at a hospital in Denmark identified a significant reduction (15.4%) in the mean total bed utilization per cardiac pathway and revealed that this reduction was caused by improvements in the work processes.

Purpose To clinically implement and characterize real-time motion-including tumor dose reconstruction during radiotherapy delivery. Methods Seven patients with 2–3 fiducial markers implanted near liver tumors received stereotactic body radiotherapy on a conventional linear accelerator. The 3D marker motion during a setup CBCT scan was determined online from the CBCT projections and used to generate a correlation model between tumor and external marker block motion. During treatment, the correlation model was updated by kV imaging every three seconds and used for real-time tumor localization. Using streamed accelerator parameters and tumor positions, in-house developed software, DoseTracker, calculated the dose to the moving tumor in real-time assuming water density in the patient. Post-treatment, the real-time tumor localization was validated by comparison with independent marker segmentations and 3D motion estimations. Dose reconstruction was validated by comparison with treatment planning system (TPS) calculations that modeled motion as isocenter shifts and used both actual CT densities and water densities. Results The real-time estimated tumor position had a mean 3D root-mean-square error of 1.7 mm (range: 0.9–2.6 mm). The motion-induced reduction in the minimum dose to 95% of the clinical target volume (CTV D95) per fraction was up to 12.3%-points. It was estimated in real-time by DoseTracker during patient treatment with a root-mean-square difference relative to the TPS of 1.3%-points (TPS CT) and 1.0%-points (TPS water). Conclusions The world’s first clinical real-time motion-including tumor dose reconstruction during radiotherapy was demonstrated. This marks an important milestone for real-time in-treatment quality assurance and paves the way for real-time dose-guided treatment adaptation.

Hepatocellular carcinoma (HCC) is the most common primary liver cancer, and the incidence of HCC is increasing. Recently, cancer immunotherapy has emerged as an efficient treatment against some cancers. Here we have used a mouse model of mutagen-induced HCC to explore the therapeutic usefulness of targeting the DNA-activated STING pathway in HCC. STING-deficient mice exhibited unaltered initial development of HCC, but had higher number of large tumors at late stages of disease. In the liver of STING-deficient HCC mice, we observed reduced levels of phospho-STAT1, autophagy, and cleaved caspase3. These responses were activated in the liver by treatment with a cyclic dinucleotide (CDN) STING agonist. Importantly, CDN treatment of mice after HCC development efficiently reduced tumor size. Initiation of CDN treatment at an even later stage of disease to allow HCC detection by MR scanning revealed that the majority of tumors regressed in response to CDN, but new tumors were also detected, which were unresponsive to CDN treatment. Overall, the modulation of the STING pathway affects the development of HCC, and holds promise for a use as a treatment of this disease, most likely in combination with other immunomodulatory treatments such as PD1 inhibitors or with standard of care.

This study validates a method of fast motion-including dose reconstruction for proton pencil beam scanning in the liver. The method utilizes a commercial treatment planning system (TPS) and calculates the delivered dose for any translational 3D target motion. Data from ten liver patients previously treated with photon radiotherapy with intrafraction tumour motion monitoring were used. The dose reconstruction method utilises an in-house developed program to incorporate beam's-eye-view tumour motion by shifting each spot in the opposite direction of the tumour and in-depth motion as beam energy changes for each spot. The doses are then calculated on a single CT phase in the TPS. Two aspects of the dose reconstruction were assessed: (1) The accuracy of reconstruction, by comparing dose reconstructions created using 4DCT motion with ground truth doses obtained by calculating phase specific doses in all 4DCT phases and summing up these partial doses. (2) The error caused by assuming 4DCT motion, by comparing reconstructions with 4DCT motion and actual tumour motion. The CTV homogeneity index (HI) and the root-mean-square (rms) dose error for all dose points receiving >70%, >80% and >90% of the prescribed dose were calculated. The dose reconstruction resulted in mean (range) absolute CTV HI errors of 1.0% (0.0-3.0)% and rms dose errors of 2.5% (1.0%-5.3%), 2.1% (0.9%-4.5%), and 1.8% (0.7%-3.7%) for >70%, >80% and >90% doses, respectively, when compared with the ground truth. The assumption of 4DCT motion resulted in mean (range) absolute CTV HI errors of 5.9% (0.0-15.0)% and rms dose errors of 6.3% (3.9%-12.6%), 5.9% (3.4%-12.5%), and 5.4% (2.6%-12.1%) for >70%, >80% and >90% doses, respectively. The investigated method allows tumour dose reconstruction with the actual tumour motion and results in significantly smaller dose errors than those caused by assuming that motion at treatment is identical to the 4DCT motion.

PURPOSE: In radiotherapy, dose deficits caused by tumor motion often far outweigh the discrepancies typically allowed in plan-specific quality assurance (QA). Yet, tumor motion is not usually included in present QA. We here present a novel method for online treatment verification by real-time motion-including 4D dose reconstruction and dose evaluation and demonstrate its use during stereotactic body radiotherapy (SBRT) delivery with and without MLC tracking.METHODS: Five volumetric modulated arc therapy (VMAT) plans were delivered with and without MLC tracking to a motion stage carrying a Delta4 dosimeter. The VMAT plans have previously been used for (non-tracking) liver SBRT with intra-treatment tumor motion recorded by kilovoltage intrafraction monitoring (KIM). The motion stage reproduced the KIM-measured tumor motions in 3D while optical monitoring guided the MLC tracking. Linac parameters and the target position were streamed to an in-house developed software program (DoseTracker) that performed real-time 4D dose reconstructions and 3%/3mm γ-evaluations of the reconstructed cumulative dose using a concurrently reconstructed planned dose without target motion as reference. Offline, the real-time reconstructed doses and γ-evaluations were validated against 4D dosimeter measurements performed during the experiments.RESULTS: In total, 181,120 dose reconstructions and 5,237 γ-evaluations were performed online and in real time with median computation times of 30 ms and 1.2 s, respectively. The mean (standard deviation) difference between reconstructed and measured doses was -1.2% (4.9%) for transient doses and -1.5% (3.9%) for cumulative doses. The root-mean-square deviation between reconstructed and measured motion-induced γ-fail rates was 2.0%-point. The mean (standard deviation) sensitivity and specificity of DoseTracker to predict γ-fail rates above a given threshold was 96.8% (3.5%) and 99.2% (0.4%), respectively, for clinically relevant thresholds between 1% and 30% γ-fail rate.CONCLUSIONS: Real-time delivery-specific QA during radiotherapy of moving targets was demonstrated for the first time. It allows supervision of treatment accuracy and action on treatment discrepancy within two seconds with high sensitivity and specificity. This article is protected by copyright. All rights reserved.

In nuclear medicine brain imaging, it is important to delineate regions of interest (ROIs) so that the outcome is both accurate and reproducible. The purpose of this study was to validate a new time-saving algorithm (DATquan) for accurate and reproducible quantification of the striatal dopamine transporter (DAT) with appropriate radioligands and SPECT and without the need for structural brain scanning. Methods: In a reconstructed DAT SPECT image, DATquan automatically calculated the ratio at steady state of specifically bound radioligand to nondisplaceable radioligand in tissue (BPND) within striatal ROIs that were delineated by use of a semiautomatic template-based alignment approach. DATquan was tested with 123 I-N-(3-iodoprop2E-enyl)-2-b-carbomethoxy-3b-(4-methylphenyl) SPECT images from 15 patients. In each image, ROIs were first manually delineated, and then corresponding BPND values were derived by an experienced physician. Afterward, 2 independent novice operators used DATquan to analyze the same 15 images. The resulting DATquan-derived BPND data were compared with the data retrieved by manual delineation to assess the accuracy and reproducibility of DATquan. Also, the operational aspects of DATquan were assessed on the basis of measurements of the mean running time of the algorithm as well as on the basis of quantification of the overlap of the DATquandelineated ROIs obtained by the 2 operators. Results: The mean algorithm running time was 3 min, and the operators’ striatal ROIs had a mean overlap of more than 82%. DATquanderived BPND values obtained by the 2 operators showed high agreement (the mean difference was 0.00 [SD, 0.05] in the striatum, 0.02 [SD, 0.26] in the putamen, and 0.03 [SD, 0.43] in the caudate nucleus). The interoperator variability was 2.2% (SD, 1.3%) in the striatum, 11.7% (SD, 9.9%) in the putamen, and 12.9% (SD, 4.0%) in the caudate nucleus. DATquan-derived BPND values showed high agreement with the values manually derived by the experienced delineator. Conclusion: DATquan is a freely available, accurate, and highly reproducible method for quantification of DAT binding in the brain by SPECT. Once implemented in clinics, DATquan will serve as a useful and time-saving tool.

Purpose A problem of current plan-specific quality assurance (QA) in radiotherapy is that it ignores organ motion, although motion may cause dose discrepancies much graver than the treatment delivery errors assessed by the QA procedure. Furthermore, the QA is not feasible for real-time adaptive treatments like MLC tracking, where the treatment machine behavior is determined on-the-fly as a response to tumor motion. To overcome these limitations, we have developed the software program DoseTracker that performs real-time motion-including dose reconstruction during treatment delivery. Here, we validate DoseTracker in experiments and simulated treatments. Methods DoseTracker performs real-time motion-including dose reconstruction, based on streamed linac parameters and target positions, using a pencil-beam algorithm. An arbitrary set of points can be chosen for the dose calculations and the calculations points can move independently as function of time. DoseTracker has been validated in phantom experiments and simulated patient treatments. The phantom experiments used a biplanar diode array on a programmable motion stage. DoseTracker performed online real-time reconstruction of the diode doses at 15 Hz and calculated the 3%/3 mm gamma failure rate (comparing motion doses with static doses) at 1 Hz during five VMAT liver SBRT treatments with and without MLC tracking. The gamma errors were retrospectively compared with diode measurements. In the treatment simulations, DoseTracker made (offline) real-time tumor dose reconstructions at 2–5 Hz in patient anatomy for four liver SBRT patients previously treated with motion monitoring by electromagnetic transponders. Simulations were performed with and without respiratory gating. The reduction in CTV D95 relative to the planned intent ( Δ D95) was retrospectively compared between DoseTracker and motion-including dose reconstructions performed in the treatment planning system (TPS) by isocenter shifts. DoseTracker currently assumes water densities so another set of TPS calculations were performed on water densities. Results Experiments yielded 2.0%-point gamma failure rate root-mean-square difference between DoseTracker and measurements. Simulated treatments yielded 1.2%-points (CT-densities) and 0.6%-point (water-densities) differences in Δ D95. Conclusions A program to reconstruct motion-induced dose errors was developed and tested in phantom studies and patients, yielding high accuracies, and allowing supervision of treatment correctness and action based on dose discrepancies. Planned improvements include CT-densities, rotations, etc.

Some Alternaria species are able to produce plant pathogenic as well as toxic metabolites. In both agriculture and the food industry it is important know if toxigenic Alternaria are present to rapidly employ the correct corrective actions. The purpose of this work was to establish a real-time PCR method, which can detect and quantify apple pathogenic and toxigenic Alternaria. An AM-toxin I primer set, which could recognize Alternaria DNA only, was designed by using primers complementary to the AM-toxin I gene. The method could detect small amounts of DNA (4 pg) and still obtain a large dynamic range (4 decades) without interference from apple material. Eight Alternaria isolates were analyzed for the presence of AM-toxin I gene and their production of secondary metabolites. Then analyses showed that all eight isolates contained the AM toxin gene and were able to produce the plant pathogenic tentoxin in addition to AM toxin I. The analyses also showed the production of tenuazonic acid, alternariols, Altenuene, altenusin and/or altertoxin I in pure culture. Analyses of inoculated apples showed that both the AM-toxin gene and alternariol monomethyl ether could be detected. Morphological analyses suggested that the eight Alternaria strains, though they all carried the AM toxin genes, probably belong to different but closely related un-described Alternaria taxa in the A. tenuissima species-group based on morphological and chemical differences.

Members of the RecQ helicase family are mutated in several human genomic instability syndromes, such as Werner and Bloom syndromes. The syndromes are characterized by premature ageing and cancer predisposition, respectively, and are therefore extensively used as model systems for studies of ageing and cancer. RecQ homologues are widely expressed enzymes, and genetic and biochemical investigations have pointed to their involvement in homologous recombinational DNA repair pathways. In the review we will focus on the implications of RecQ helicases for genome maintenance with specific emphasis on the homologues found in yeast.

Accumulation of the carcinogenic mycotoxin aflatoxin B1 has been reported from members of three different groups of Aspergilli (4) Aspergillus flavus, A. flavus var. parvisclerotigenus, A. parasiticus, A. toxicarius, A. nomius, A. pseudotamarii, A. zhaoqingensis, A. bombycis and from the ascomycete genus Petromyces (Aspergillus section Flavi), (2) Emericella astellata and E. venezuelensis from the ascomycete genus Emericella (Aspergillus section Nidulantes) and (3) Aspergillus ochraceoroseus from a new section proposed here: Aspergillus section Ochraceorosei. We here describe a new species, A. rambellii referable to Ochraceorosei, that accumulates very large amounts of sterigmatocystin, 3-O-methylsterigmatocystin and aflatoxin B1, but not any of the other known extrolites produced by members of Aspergillus section Flavi or Nidulantes. G type aflatoxins were only found in some of the species in Aspergillus section Flavi, while the B type aflatoxins are common in all three groups. Based on the cladistic analysis of nucleotide sequences of ITS1 and 2 and 5.8S, it appears that type G aflatoxin producers are paraphyletic and that section Ochraceorosei is a sister group to the sections Flavi, Circumdati and Cervini, with Emericella species being an outgroup to these sister groups. All aflatoxin producing members of section Flavi produce kojic acid and most species, except A. bombycis and A. pseudotamarii, produce aspergillic acid. Species in Flavi, that produce B type aflatoxins, but not G type aflatoxins, often produced cyclopiazonic acid. No strain was found which produce both G type aflatoxins and cyclopiazonic acid. It was confirmed that some strains of A. flavus var. columnaris produce aflatoxin B2, but this extrolite was not detected in the ex type strain of that variety. A. flavus var. parvisclerotigenus is raised to species level based on the specific combination of small sclerotia, profile of extrolites and rDNA sequence differences. A. zhaoqingensis is regarded as a synonym of A. nomius, while A. toxicarius resembles A. parasiticus but differs with at least three base pair differences. At least 10 Aspergillus species can be recognized which are able to biosynthesize aflatoxins, and they are placed in three very different clades.

In an 8-year study of the diversity and distribution of Penicillium commune contaminants in two different cheese dairies, swab and air samples were taken from the production plants, the processing environment and contaminated cheeses. A total of 321 Penicillium commune isolates were characterized using morphotypes (colony morphology and colours) and secondary metabolite profiles. Based on production of secondary metabolites the P. commune isolates were classified into 6 groups. The genetic diversity of the P. commune isolates was assessed using randomly amplified polymorphic DNA (RAPD) and amplified fragment length polymorphism (AFLP). For a sub-set of 272 P. commune isolates RAPD analysis generated 33 RAPD groups whereas AFLP profiling revealed 55 AFLP groups. This study conclusively showed that the discriminatory power of AFLP was high compared to RAPD and that AFLP fingerprinting matched morphotyping. P. commune isolates with identical profiles using all four typing techniques were interpreted as closely related isolates with a common origin and the distribution of these isolates in the processing environment indicated possible contamination points in the cheese dairies. The coating process and unpacking of cheeses with growth of P. commune seemed to cause the contamination problems. Several identical P. commune isolates remained present in the processing environment for more than 7 years in both dairies.

Eukaryotic DNA topoisomerase II is a dimeric nuclear enzyme essential for DNA metabolism and chromosome dynamics. It changes the topology of DNA by coupling binding and hydrolysis of two ATP molecules to the transport of one DNA duplex through a temporary break introduced in another. During this process the structurally and functionally complex enzyme passes through a cascade of conformational changes, which requires intra- and intersubunit communication. To study the importance of ATP binding and hydrolysis in relation to DNA strand transfer, we have purified and characterized a human topoisomerase II alpha heterodimer with only one ATP binding site. The heterodimer was able to relax supercoiled DNA, although less efficiently than the wild type enzyme. It furthermore possessed a functional N-terminal clamp and was sensitive to ICRF-187. This demonstrates that human topoisomerase II alpha can pass through all the conformations required for DNA strand passage and enzyme resetting with binding and hydrolysis of only one ATP. However, the heterodimer lacked the normal stimulatory effect of DNA on ATP binding and hydrolysis as well as the stimulatory effect of ATP on DNA cleavage. The results can be explained in a model, where efficient catalysis requires an extensive communication between the second ATP and the DNA segment to be cleaved.

We examined nine Aspergillus japonicus isolates and 10 Aspergillus aculeatus isolates by using molecular and biochemical markers, including DNA sequences of the ITS1-5.8S rRNA gene-ITS2 region, restriction fragment length polymorphisms (RFLP), and secondary-metabolite profiles. The DNA sequence of the internal transcribed spacers (ITS1 and ITS2) and the 5.8S rRNA gene could not be used to distinguish between A. japonicus and A. aculeatus but did show that these two taxa are more closely related to each other than to other species of black aspergilli. Aspergillus niger pyruvate kinase ( pki A) and pectin lyase A ( pel A) and Agaricus bisporus 28S rRNA genes, which were used as probes in the RFLP analysis, revealed clear polymorphism between these two taxa. The A. niger pki A and pel A probes placed six strains in an A. japonicus group and 12 isolates in an A. aculeatus group, which exhibited intraspecific variation when they were probed with the pel A gene. The secondary-metabolite profiles supported division of the isolates into the two species and differed from those of other black aspergilli. The strains classified as A. japonicus produced indole alkaloids and a polar metabolite, while the A. aculeatus isolates produced neoxaline, okaramins, paraherquamidelike compounds, and secalonic acid. A. aculeatus CBS 114.80 showed specific RFLP patterns for all loci examined. The secondary-metabolite profile of strain CBS 114.80 also differed from those of A. japonicus and A. aculeatus . Therefore, this strain probably represents a third taxon. This study provides unambiguous criteria for establishing the taxonomic positions of isolates of black aspergilli, which are important in relation to industrial use and legal protection of these organisms.

The genetic variability within 600 bp of DNA sequence from the ribosomal internal transcribed spacers (ITS 1 and ITS 2) and the 5.8 S rRNA gene was examined in 52 strains belonging to 29 terverticillate taxa in Penicillium subg. Penicillium from diverse sources and locations. The sequenced region is extremely conserved within the terverticillate penicillia, with only 29 positions differing in one or more taxa. The highest degree of ITS variability was seen among the species close to Penicillium roqueforti ( P. roqueforti, P. carneum and P. paneum ) whereas the ITS variability was very low between closely related taxa, e.g. among the taxa near P. aurantiogriseum. The relationships among the terverticillate penicillia, and related teleomorphs in Eupenicillium , were analysed from bootstrapped ITS sequence data sets using the neighbour-joining method. The terverticillate penicillia form a well supported clade with Eupenicillium crustaceum. Sequence analysis generally confirmed the overall taxonomy in Penicillium subg. Penicillium but relationships between all the terverticillate taxa could not be clearly established due to the low degree of ITS variability. Nevertheless, clades of Penicillium species sharing environmental characteristics did emerge, e.g. species growing on protein and lipid rich substrates (e.g. P. crustosum and P. commune ), species prevalent in dry habitats (e.g. P. chrysogenum and P. nalgiovense ), and those prevalent on carbohydrate rich substrates (e.g. P. aurantiogriseum and P. freii ).

Ichthyophonus hoferi Plehn and Mulsow, 1911 is thought to be one of the few pathogenic fungal infections of marine fish. The result of an attack is severe epizootics in herring stocks with drastic reduction in the population as a consequence. The exact phylogenetic position of the genusIchthyophonus is not known. In the present study, a combination of molecular data, ultrastructure and biochemical characters were utilized to investigate the phylogeny ofI.hoferi. The genomic DNA encoding the small subunit ribosomal RNA (18S rRNA) was amplified and sequenced. Comparisons with other eukaryotic 18S rRNA sequences indicate thatI. hoferi is not a member of the Fungi. In both the parsimony and the neighborjoining trees,I. hoferi is the sister taxon to the rosette agent. The clade encompassingI.hoferi and the rosette organism is the sister group to the chanoflagellate clade in the neighbor-joining tree, while in the parsimony tree theI. hoferi/rosette clade is equally distant to both the choanoflagellate and animal clades. Transmission electron microscopy showed thatI. hoferi has a defined cell wall, an endoplasm that consists of a fine granulated matrix with numerous ribosomes, several nuclei, vacuoles of varying density distributed throughout the cell, and mitochondria with tubular cristae. The cell wall ofI. hoferi contains chitin.

Regions of the genes encoding flagellin (flaA), the invasive associated protein (iap), listeriolysin O (hly) and 23S rRNA were sequenced for a range of Listeria monocytogenes isolates of different origin and serotypes. Several nucleotide sequence variations were found in the flaA, iap and hly genes. No differences were found for the rRNA genes, but our approach does not exclude the existence of differences between single copies of these genes. Based on the sequence differences, the L. monocytogenes strains can be divided into three distinct sequence types. Further, the presence of only a small number of sequence differences within each group indicates a strong degree of conservation within the groups. There was a complete correspondence among the groups of strains formed according to the analysis of the flaA, iap and hly genes, and the grouping correlates with serotype, pulsed field gel electrophoretic and multilocus enzyme electrophoretic data. Analysis of the region encoding the threonine-asparagine repeat units in the iap gene revealed some striking features. Sequence type 1 strains were found to have 16-17 repeats, sequence type 2 strains had 16-20 repeats whereas the two sequence type 3 strains analysed had only 11 repeats. Furthermore, within a 19 bp segment there was a 37% difference between the sequences of type 1 and 2 strains and that segment was absent in type 3 strains. Within the threonine-asparagine repeat region the nucleotide differences gave rise to four amino acid changes; however, all were changes among the three amino acids present in the repeat structure indicating a strong selective pressure on the composition of this region.

In the modern world, cities need to keep up with the demand for mobility, efficient infrastructure and environmental sustainability. The future smart cities use intelligent information and communication technologies to raise the quality of life. This includes computer vision as one of the main technologies. It can observe and analyse human activities from a distance in a noninvasive manner. Traditional computer vision utilises RGB cameras, but problems with this sensor include its light dependency, and the privacy issues that can be raised by people being observed. In this paper, we propose the use of thermal imaging in real-time smart city applications. Thermal cameras operate independently of light and measure the radiated infrared waves representing the temperature of the scene. In order to showcase the possibilities, we present five different applications which use thermal imaging only. These include both indoorand outdoor scenarios with the purposes of people detection, counting and tracking, as well as one application for traffic safety evaluation.

This paper presents a study investigating the potential use of human motion intensities as input for parametric urban design. Through a computer vision analysis of thermal images, motion intensity maps are generated and utilized as design drivers for urban design patterns; and, through a case study of a town square, human occupancy and motion intensities are used to generate situated flow topologies presenting new adaptive methods for urban design. These methods incorporate local flow as design drivers for canopy, pavement and furniture layout. The urban design solution may be configured due to various parameters such as security, comfort, navigation, efficiency, or aesthetics.

This paper illustrates responsive systems, which focus on the implementation of multi-objective adaptive design prototypes from sensored environments. The intention of the work is to investigate multi-objective criteria both as a material system and as a processing system by creating prototypes with structural integrity, where thermal energy flow through the prototype, to be understood as a membrane, can be controlled and the visual transparancy altered. The work shows performance based feedback systems and physical prototype models driven by information streaming, screening and application.

We have characterized a human topoisomerase IIalpha enzyme with a deletion of the conserved QTK loop, which extends from the transducer domain to the ATP-binding pocket in the GHKL domain. The loop has been suggested to play a role for interdomain communication in type II topoisomerases. The mutant enzyme performs only very low levels of strand passage, although it is able to cleave and ligate DNA as well as close the N-terminal clamp. Cleavage is nearly unaffected by ATP and ATP analogues relative to the wild-type enzyme. Although the enzyme is able to close the clamp, the clamp has altered characteristics, allowing trapping of DNA also in the absence of an ATP analogue. The enzyme furthermore retains intrinsic levels of ATPase activity, but the activity is not stimulated by DNA. Our observations demonstrate that the QTK loop is an important player for the interdomain communication in human topoisomerase IIalpha. First, the loop seems to play a role in keeping the N-terminal clamp in an open conformation when no nucleotide is present. Once the nucleotide binds, it facilitates clamp closure, although it is not essential for this event. The QTK loop, in contrast, is essential for the DNA-stimulated ATPase activity of human topoisomerase IIalpha.

DNA topoisomerase II is a multidomain homodimeric enzyme that changes DNA topology by coupling ATP hydrolysis to the transport of one DNA helix through a transient double-stranded break in another. The process requires dramatic conformational changes including closure of an ATP-operated clamp, which is comprised of two N-terminal domains from each protomer. The most N-terminal domain contains the ATP-binding site and is directly involved in clamp closure, undergoing dimerization upon ATP binding. The second domain, the transducer domain, forms the walls of the N-terminal clamp and connects the clamp to the enzyme core. Although structurally conserved, it is unclear whether the transducer domain is involved in clamp mechanism. We have purified and characterized a human topoisomerase II alpha enzyme with a two-amino acid insertion at position 408 in the transducer domain. The enzyme retains both ATPase and DNA cleavage activities. However, the insertion, which is situated far from the N-terminal dimerization area, severely disrupts the function of the N-terminal clamp. The clamp-deficient enzyme is catalytically inactive and lacks most aspects of interdomain communication. Surprisingly, it seems to have retained the intersubunit communication, allowing it to bind ATP cooperatively in the presence of DNA. The results show that even distal parts of the transducer domain are important for the dynamics of the N-terminal clamp and furthermore indicate that stable clamp closure is not required for cooperative binding of ATP.

In the present study, the use of aqueous polymer two-phase systems for separation of pathogenic bacteria from a complex food sample was investigated. Three different two-phase systems, a polyethylene glycol 3350/dextran T 500, a methoxy polyethylene glycol 5000/dextran T 500 and a polyethylene glycol 3350/hydroxypropyl starch system, were compared at pH 3 and pH 6 for their capacity to separate the pathogenic bacteria Listeria monocytogenes and Salmonella berta from a Cumberland sausage. In all three phase systems, the food particles partitioned to the lower phase. Best performance was obtained by the polymer combinations, polyethylene glycol 3350/dextran T 500 and polyethylene glycol 3350/hydroxypropyl starch. In these systems, Salmonella berta partitioned to the hydrophobic upper phase both at pH 3 and pH 6 with an average partitioning ratio of 80% and a recovery of 56%. Listeria monocytogenes partitioned to the upper phase at pH 3 only with an average partitioning ratio of 72% and a recovery of 45%. This method may become a valuable tool for separation of bacteria from complex food matrices.

Rapid identification of filamentous fungi is becoming increasingly important in food mycology both for monitoring the production process and for the identification of food spoilers. This paper describes the development and trial of two specifie PCR primer sets. A 336 bp fragment from species belonging to Penicillium subgenus Penicillium was amplified by the primers ITS 212d and ITS 549. The other primer set, ITS 183 and ITS 401 specifically identified two species. Penicillium roqueforti and P. carneum, both known as spoilers in the bread industry, by amplification of a 300 bp fragment. The future perspectives of PCR based identification of filamentous fungi in food are discussed.

The possible use of level-crossing spectroscopy to study properties of excited states generated by gas excitation of fast ionic beams has been investigated. With a 20-400 keV Be+ beam excited by means of helium gas, radiative lifetimes of 2D and 2F levels in Be II have been determined together with the fine-structure separation of the 5f 2F level in Be II and the hyperfine coupling constant for the 2s3d 1D level in Be I and the 2p 2p level in Be II.

Penicillium roqueforti is currently divided into two varieties, one used for cheese starter cultures, P. roqueforti var. roqueforti and one ubiquitous patulin-producing variety, P. roqueforti var. carneum. The ribosomal regions comprising the 5.8S gene and the internal transcribed spacers, ITS I and ITS II, have been analysed from 10 isolates belonging to each variety. The 10 P. roqueforti var. carneum isolates were separated into two groups of five on the basis of 12 base-pair differences in the ITS regions. One of the groups of P. roqueforti var. carneum in the following designated P. carneum differed from P. roqueforti var. roqueforti here designated P. roqueforti in just two positions, while the other group, here called P. paneum differed from P. roqueforti in 12 positions. Random Amplified Polymorphic DNA (RAPD) analysis substantiated these findings, and a comparison of secondary metabolites produced by the three groups showed that the P. roqueforti isolates all produce Penicillium Roqueforti (PR) toxin, marcfortines and fumigaclavine A, while the P. carneum isolates produce patulin, penitrem A and mycophenolic acid, as well as unidentified metabolites. P. paneum produces secondary metabolites in five chromophore families including the known mycotoxins patulin and botryodiploidin. On the basis of these findings it is proposed that P. roqueforti is reclassified into three species named P. roqueforti, P. carneum and P. paneum.

This work contributes to the growing body of work, conducted on the vicinities between well-being and biomedical treatments in health design. The article presents and discusses the design of the new delivery rooms at a Danish hospital in Hjørring, including the multi-sensory artwork: Nordjyske stemninger (Moods of Northern Jutland). The authors are both artists, architects, and researchers in this project, thus it is not the purpose of his article to report evaluation results. However, it is our intention to share and discuss contemporary healthcare design strategies and point to the importance of considering the interplay between cultural, social change, and environment in order to bridge the know-do gap in healing architecture. Based on our work we give a concrete example of a case aimed at re-introducing art in healthcare environments, supporting the caregiver, the laboring mother, and her companion in the existential and life-changing moment. The article includes descriptions of the design process including interviews, observations, and reflections. In this case, we want to argue that the gap between visions and implementation in evidence-based design and healing architecture, must be understood as a symptom of a deeper epistemological and philosophical challenge concerning the dichotomous and demarcating understanding of the relation between the human and its surroundings, obstructing ecological coherence and validity and silo stacking of results not utilizing the rich potentiality of interdisciplinarity synergies. As it is difficult to convey a bodily and sensory experience in only words and images, we hope that the reader will use their imagination while reading the descriptions of a situated experience throughout the article. The Ukrainian sculptor Alexander Archipenko described the cause and impulse of creative motivation as seeing the absence of a thing. With this lens we invite the reader behind the scenes in the creation of somesthetic design of the new delivery rooms, now being the background of more than 1.000 births a year in the Northern part of Jutland. The argument of this article uses artistic practice to explore a new potential healthcare practice, with overseen and neglected potentialities in a supportive somesthetic healthcare design. The article is structured in four parts: Healing environments, Somesthetic design framework, The sensory delivery room, and Reflection.