Detection of magnetomotive ultrasound signals from human tissue.

Rectal cancer is a common cancer, with presently a 5-year survival of 67 %. Treatment is based on tumor stage, but current staging methods, such as magnetic resonance imaging (MRI) or ultrasound, are limited in the ability to correctly stage the disease. Magnetomotive ultrasound is a developing modality that has a potential to improve rectal cancer staging. Magnetic nanoparticles are set in motion by an external magnetic field, and the resulting motion signature is detected by ultrasound. Here, we report on magnetomotive images of magnetic nanoparticles in human tissue, using a prototype system where a rotating permanent magnet provides the varying magnetic field, and an ultrasound transducer array encircling the magnet, detects the induced motion. Prior to surgery, a patient with a low rectal tumor was injected at three sites close to the tumor with magnetic nanoparticles. Postsurgical magnetomotive ultrasound scanning revealed the three injection sites, with no obvious artefactual signals. A phantom study showed detection of nanoparticles beyond 40 mm, where 30 mm is the expected maximum distance to mesorectal lymph nodes. Magnetomotive ultrasound image of iron oxide nanoparticles in human tissue. Prior to surgery a patient was injected with nanoparticles, and the excised tissue specimen was imaged with a prototype magnetomotive ultrasound system. The three colored areas overlaid on the standard B-mode greyscale image, correspond to the three injection sites.

A novel mouse model of radiation-induced cancer survivorship diseases of the gut.

A deeper understanding of the radiation-induced pathophysiological processes that develop in the gut is imperative to prevent, alleviate, or eliminate cancer survivorship diseases after radiotherapy to the pelvic area. Most rodent models of high-dose gastrointestinal radiation injury are limited by high mortality. We therefore established a model that allows for the delivering of radiation in fractions at high doses while maintaining long-term survival. Adult male C57/BL6 mice were exposed to small-field irradiation, restricted to 1.5 cm of the colorectum using a linear accelerator. Each mouse received 6 or 8 Gy, two times daily in 12-h intervals in two, three, or four fractions. Acute cell death was examined at 4.5 h postirradiation and histological changes at 6 wk postirradiation. Another group was given four fractions of 8 Gy and followed over time for development of visible symptoms. Irradiation caused immediate cell death, mainly limited to the colorectum. At 6 wk postirradiation, several crypts displayed signs of radiation-induced degeneration. The degenerating crypts were seen alongside crypts that appeared perfectly healthy. Crypt survival was reduced after the fourth fraction regardless of dose, whereas the number of macrophages increased. Angiogenesis was induced, likely as a compensatory mechanism for hypoxia. Four months postirradiation, mice began to show radiation-induced symptoms, and histological examination revealed an extensive crypt loss and fibrosis. Our model is uniquely suitable for studying the long-term trajectory and underlying mechanisms of radiation-induced gastrointestinal injury.NEW & NOTEWORTHY A novel mouse model for studying the long-term trajectory of radiation-induced gut injury. The method allows for the use of high doses and multiple fractions, with minor impact on animal health for at least 3 mo. Crypt loss and a slow progression of fibrosis is observed. Crypt degeneration is a process restricted to isolated crypts. Crypt degeneration is presented as a convenient proxy endpoint for long-term radiation-induced gut injury.

Determination of intrinsic energy dependence of point-like inorganic scintillation detector in brachytherapy.

BACKGROUND: Inorganic scintillation detectors (ISDs) are promising for in vivo dosimetry in brachytherapy (BT). ISDs have fast response, providing time resolved dose rate information, and high sensitivity, attributed to high atomic numbers. However, the conversion of the detector signal to absorbed dose-to-water is highly dependent on the energy spectrum of the incident radiation. This dependence is comprised of absorbed dose energy dependence, obtainable with Monte Carlo (MC) simulation, and the absorbed dose-to-signal conversion efficiency or intrinsic energy dependence requiring measurements. Studies have indicated negligible intrinsic energy dependence of ZnSe:O-based ISDs in Ir-192 BT. A full characterization has not been performed earlier. PURPOSE: This study characterizes the intrinsic energy dependence of ZnSe:O-based ISDs for kV X-ray radiation qualities, with energies relevant for BT. METHODS: Three point-like ISDs made from fiber-coupled cuboid ZnSe:O-based scintillators were calibrated at the Swedish National Metrology Laboratory for ionizing radiation. The calibration was done in terms of air kerma free-in-air, K air ${K}_{{\mathrm{air}}}$ , in 13 X-ray radiation qualities, Q $Q$ , from 25 to 300 kVp (CCRI 25-250 kV and ISO 4037 N-series), and in terms of absorbed dose to water, D w ${D}_{\mathrm{w}}$ , in a Co-60 beam, Q 0 ${Q}_0$ . The mean absorbed dose to the ISDs, relative to K air ${K}_{{\mathrm{air}}}$ and D w ${D}_{\mathrm{w}}$ , were obtained with the MC code TOPAS (Geant4) using X-ray spectra obtained with SpekPy software and laboratory filtration data and a generic Co-60 source. The intrinsic energy dependence was determined as a function of effective photon energy, E e f f ${E}_{eff}$ , (relative to Co-60). The angular dependence of the ISD signal was measured in a 25 kVp (0.20 mm Al HVL) and 135 kVp beam (0.48 mm Cu HVL), by rotating the ISDs 180° around the fiber's longitudinal axis (perpendicular to the beam). A full 360° was not performed due to setup limitations. The impact of detector design was quantified with MC simulation. RESULTS: Above 30 keV E e f f ${E}_{eff}$ the intrinsic energy dependence varied with less than 5 ± 4% from unity for all detectors (with the uncertainty expressed as the mean of all expanded measurement uncertainties for individual E e f f ${E}_{eff}$ above 30 keV, k = 2). Below 30 keV, it decreased with up to 17% and inter-detector variations of 13% were observed, likely due to differences in detector geometry not captured by the simulations using nominal geometry. In the 25 kVp radiation quality, the ISD signal varied with 24% over a ∼45° rotation. For 135 kVp, the corresponding variation was below 3%. Assuming a 0.05 mm thicker layer of reflective paint around the sensitive volume changed the absorbed dose with 6.3% at the lowest E e f f ${E}_{eff}$ , and with less than 2% at higher energies. CONCLUSION: The study suggests that the ISDs have an intrinsic energy dependence relative to Co-60 lower than 5 ± 4% in radiation qualities with E e f f ${E}_{eff}\ $ > 30 keV. Therefore, they could in principle be calibrated in a Co-60 beam quality and transferred to such radiation qualities with correction factors determined only by the absorbed dose energy dependence obtained from MC simulations. This encourages exploration of the ISDs' applications in intensity modulated BT with Yb-169 or other novel intermediate energy isotopes.

Meaning of working for older nurses and nursing assistants in Sweden: A qualitative study.

Meaningful work is related to the motivation to continue to work in older ages and later retirement. This qualitative study addresses calls for further research on the meaning of working for older workers using the Interpretative Phenomenological Analysis approach to explore in-depth the dimensions underlying the subjective experience of meaningful work among 27 nurses and nursing assistants aged 55-75 years. The findings show that work was perceived as a primary source of: (1) personal identity (2) purpose and contribution, (3) competence and accomplishment, (4) social contacts and belongingness, (5) activity, routines and purposeful use of time, and (6) economic security and freedom. These qualitative findings may be applied in interventions aiming to encourage extended working lives in key welfare occupations, which are facing significant staff shortages.

Cooperation between viral interferon regulatory factor 4 and RTA to activate a subset of Kaposi's sarcoma-associated herpesvirus lytic promoters.

The four Kaposi's sarcoma-associated herpesvirus (KSHV)-encoded interferon (IFN) regulatory factor homologues (vIRF1 to vIRF4) are used to counter innate immune defenses and suppress p53. The vIRF genes are arranged in tandem but differ in function and expression. In KSHV-infected effusion lymphoma lines, K10.5/vIRF3 and K11/vIRF2 mRNAs are readily detected during latency, whereas K9/vIRF1 and K10/vIRF4 mRNAs are upregulated during reactivation. Here we show that the K10/vIRF4 promoter responds to the lytic switch protein RTA in KSHV-infected cells but is essentially unresponsive in uninfected cells. Coexpression of RTA with vIRF4 is sufficient to restore regulation, a property not shared by other vIRFs. The K9/vIRF1 promoter behaves similarly, and production of infectious virus is enhanced by the presence of vIRF4. Synergy requires the DNA-binding domain (DBD) and C-terminal IRF homology regions of vIRF4. Mutations of arginine residues within the putative DNA recognition helix of vIRF4 or the invariant cysteines of the adjacent CxxC motif abolish cooperation with RTA, in the latter case by preventing self-association. The oligomerization and transactivation functions of RTA are also essential for synergy. The K10/vIRF4 promoter contains two transcription start sites (TSSs), and a 105-bp fragment containing the proximal promoter is responsive to vIRF4/RTA. Binding of a cellular factor(s) to this fragment is altered when both viral proteins are present, suggesting a possible mechanism for transcriptional synergy. Reliance on coregulators encoded by either the host or viral genome provides an elegant strategy for expanding the regulatory potential of a master regulator, such as RTA.

Computational prediction of drug solubility in lipid based formulation excipients.

PURPOSE: To investigate if drug solubility in pharmaceutical excipients used in lipid based formulations (LBFs) can be predicted from physicochemical properties. METHODS: Solubility was measured for 30 structurally diverse drug molecules in soybean oil (SBO, long-chain triglyceride; TGLC), Captex355 (medium-chain triglyceride; TGMC), polysorbate 80 (PS80; surfactant) and PEG400 co-solvent and used as responses during PLS model development. Melting point and calculated molecular descriptors were used as variables and the PLS models were validated with test sets and permutation tests. RESULTS: Solvation capacity of SBO and Captex355 was equal on a mol per mol scale (R (2) = 0.98). A strong correlation was also found between PS80 and PEG400 (R (2) = 0.85), identifying the significant contribution of the ethoxylation for the solvation capacity of PS80. In silico models based on calculated descriptors were successfully developed for drug solubility in SBO (R (2) = 0.81, Q (2) = 0.76) and Captex355 (R (2) = 0.84, Q (2) = 0.80). However, solubility in PS80 and PEG400 were not possible to quantitatively predict from molecular structure. CONCLUSION: Solubility measured in one excipient can be used to predict solubility in another, herein exemplified with TGMC versus TGLC, and PS80 versus PEG400. We also show, for the first time, that solubility in TGMC and TGLC can be predicted from rapidly calculated molecular descriptors.

Osseointegration of titanium with an antimicrobial nanostructured noble metal coating.

Nanometer scale surface features on implants and prostheses can potentially be used to enhance osseointegration and may also add further functionalities, such as infection resistance, to the implant. In this study, a nanostructured noble metal coating consisting of palladium, gold and silver, never previously used in bone applications, was applied to machined titanium screws to evaluate osseointegration after 6 and 12 weeks in rabbit tibiae and femurs. Infection resistance was confirmed by in vitro adhesion test. A qualitatively and quantitatively similar in vivo bone response was observed for the coated and uncoated control screws, using histology, histomorphometry and electron microscopy. The bone-implant interface analysis revealed an extensive bone formation and direct bone-implant contact. These results demonstrate that the nanostructured noble metal coating with antimicrobial properties promotes osseointegration and may therefore be used to add extra implant functionality in the form of increased resistance to infection without the use of antibiotics. FROM THE CLINICAL EDITOR: The authors of this paper demonstrate that nanostructured noble metal coating of implants and prostheses used in orthopedic procedures promotes osseointegration and may be used to add extra implant functionality in the form of increased resistance to infection without the use of antibiotics.

Management of paediatric traumatic brain injury in Sweden: a national cross-sectional survey.

BACKGROUND: Previous studies have shown variations in management routines for children with traumatic brain injury (TBI) in Sweden. It is unknown if this management has changed after the publication of the Scandinavian Neurotrauma Committee guidelines in 2016 (SNC16). Also, knowledge of current practice routines may guide development of an efficient implementation strategy for the guidelines. The aim of this study is therefore to describe current management routines in paediatric TBI on a hospital/organizational level in Sweden. Secondary aims are to analyse differences in management over time, to assess the current dissemination status of the SNC16 guideline and to analyse possible variations between hospitals. METHODS: This is a sequential, cross-sectional, structured survey in five sections, covering initial management routines for paediatric TBI in Sweden. Respondents, with profound knowledge of local management routines and recommendations, were identified for all Swedish hospitals with an emergency department managing children (age 0-17 year) via phone/mail before distribution of the survey. Responses were collected via an on-line survey system during June 2020-March 2021. Data are presented as descriptive statistics and comparisons were made using Fisher exact test, when applicable. RESULTS: 71 of the 76 identified hospitals managed patients with TBI of all ages and 66 responded (response rate 93%). 56 of these managed children and were selected for further analysis. 76% (42/55) of hospitals have an established guideline to aid in clinical decision making. Children with TBI are predominately managed by inexperienced doctors (84%; 47/56), primarily from non-paediatric specialities (75%; 42/56). Most hospitals (75%; 42/56) have the possibility to admit and observe children with TBI of varying degrees and almost all centres have complete access to neuroradiology (96%; 54/56). In larger hospitals, it was more common for nurses to discharge patients without doctor assessment when compared to smaller hospitals (6/9 vs. 9/47; p < 0.001). Presence of established guidelines (14/51 vs. 42/55; p < 0.001) and written observation routines (16/51 vs. 29/42; p < 0.001) in hospitals have increased significantly since 2006. CONCLUSIONS: TBI management routines for children in Sweden still vary, with some differences occurring over time. Use of established guidelines, written observation routines and information for patients/guardians have all improved. These results form a baseline for current management and may also aid in guideline implementation.

Approaching retirement: A qualitative study of older nursing assistants' experiences of work in residential care and late-career planning.

In many countries, eldercare workers are approaching retirement. To remain attractive to older and experienced workers, organizations need to understand how employees nearing retirement think about and experience their work situation. This qualitative study investigated how older nursing assistants within residential care for older people experienced aging at work, their psychosocial work environment, and their late-career planning. Semi-structured interviews with eight nursing assistants (aged 55-61 years) in Sweden were analyzed using an Interpretative Phenomenological Analysis approach. The results show that the psychosocial work environment was perceived as stressful and considered a long-term health risk, and that (future) health and work ability were key factors determining nursing assistants' late-career planning. Moreover, personal resources and social support from colleagues seemed protective against job demands. Aspects considered in late-career planning also included personal finances and meaningfulness of work. While these findings may be sample-specific, they still provide insights into the experiences of an important occupational group. This means that the findings can be useful for organizations aiming at promoting successful and sustainable aging-in-workplace and encouraging extended working lives.

Postnatal hearing loss: a study of children who passed neonatal TEOAE hearing screening bilaterally.

BACKGROUND: Universal newborn hearing screening (UNHS) contributes to the early diagnosis of hearing loss. However, not all permanent pediatric hearing impairments can be identified by UNHS. AIMS/OBJECTIVE: To investigate children who have successfully passed the UNHS, but have later-onset hearing loss at an early stage. METHODS: UNHS of children, was reviewed retrospectively from databases at Karolinska University Hospital, Sweden. Gender, age, the reason for contact, the first contact and the most recent audiogram, the hearing diagnosis, the degree of hearing loss when they were enrolled in hearing rehabilitation, and the hearing aids they used were analysed. RESULTS: 63 children who had passed the UNHS at birth and were diagnosed with a hearing impairment at a later stage were included in the study. The average age was 3.3 and 3.9 years old when the children were diagnosed and were finally enrolled in the hearing habilitation, respectively. The reasons for diagnostic evaluation of a suspected hearing loss at present study are preschool hearing tests at the Child Health Care Centres, parents suspect, and/or delayed speech and language development. CONCLUSIONS AND SIGNIFICANCE: Our findings suggest that a passed UNHS does not exclude a future delayed onset of hearing loss, particularly in children with risk factors.

Wide-scale use of Notch signaling factor CSL/RBP-Jkappa in RTA-mediated activation of Kaposi's sarcoma-associated herpesvirus lytic genes.

For Kaposi's sarcoma-associated herpesvirus (KSHV; also called human herpesvirus 8 [HHV8]), the switch from latency to active lytic replication requires RTA, the product of open reading frame 50 (ORF50). RTA activates transcription from nearly 40 early and delayed-early viral promoters, mainly through interactions with cellular DNA binding proteins, such as CSL/RBP-Jkappa, Oct-1, C/EBPalpha, and c-Jun. Reliance on cellular coregulators may allow KSHV to adjust its lytic program to suit different cellular contexts or interpret signals from the outside. CSL is a key component of the Notch signaling pathway and is targeted by several viruses. A search with known CSL binding sequences from cellular genes found at least 260 matches in the KSHV genome, many from regions containing known or suspected lytic promoters. Analysis of clustered sites located immediately upstream of ORF70 (thymidylate synthase), ORF19 (tegument protein), and ORF47 (glycoprotein L) uncovered RTA-responsive promoters that were validated using mRNAs isolated from KSHV-infected cells undergoing lytic reactivation. Notably, ORF19 behaves as a true late gene, indicating that RTA regulates all three phases of the lytic program. For each new promoter, the response to RTA was dependent on CSL, and 5 of the 10 candidate sites were shown to bind CSL in vitro. Analysis of individual sites highlighted the importance of a cytosine residue flanking the core CSL binding sequence. These findings broaden the role for CSL in coordinating the KSHV lytic gene expression program and help to define a signature motif for functional CSL sites within the viral genome.

The latency-associated nuclear antigen interacts with MeCP2 and nucleosomes through separate domains.

Kaposi's sarcoma-associated herpesvirus (KSHV)-infected cells express the latency-associated nuclear antigen (LANA) involved in the regulation of host and viral gene expression and maintenance of the KSHV latent episome. Performance of these diverse functions involves a 7-amino-acid chromatin-binding motif (CBM) situated at the amino terminus of LANA that is capable of binding directly to nucleosomes. LANA interacts with additional chromatin components, including methyl-CpG-binding protein 2 (MeCP2). Here, we show that the carboxy-terminal DNA-binding/dimerization domain of LANA provides the principal interaction with MeCP2 but that this association is modulated by the CBM. Both domains are required for LANA to colocalize with MeCP2 at chromocenters, regions of extensive pericentric heterochromatin that can be imaged by fluorescence microscopy. Within MeCP2, the methyl-CpG-binding domain (MBD) is the primary determinant for chromatin localization and acts together with the adjacent repression domains (the transcription repression domain [TRD] and the corepressor-interacting domain [CRID]) to redirect LANA to chromocenters. MeCP2 facilitates repression by LANA bound to the KSHV terminal repeats, a function that requires the MeCP2 C terminus in addition to the MBD and CRID/TRD. LANA and MeCP2 can also cooperate to stimulate transcription of the human E2F1 promoter, which lacks a LANA DNA-binding sequence, but this function requires both the N and C termini of LANA. The ability of LANA to establish multivalent interactions with histones and chromatin-binding proteins such as MeCP2 would enable LANA to direct regulatory complexes to specific chromosomal sites and thereby achieve stable reprogramming of cellular gene expression in latently infected cells.

Investigation of a synthetic diamond detector response in kilovoltage photon beams.

PURPOSE: An important characteristic of radiation dosimetry detectors is their energy response which consists of absorbed-dose and intrinsic energy responses. The former can be characterized using Monte Carlo (MC) simulations, whereas the latter (i.e., detector signal per absorbed dose to detector) is extracted from experimental data. Such a characterization is especially relevant when detectors are used in nonrelative measurements at a beam quality that differs from the calibration beam quality. Having in mind the possible application of synthetic diamond detectors (microDiamond PTW 60019, Freiburg, Germany) for nonrelative dosimetry of low-energy brachytherapy (BT) beams, we determined their intrinsic and absorbed-dose energy responses in 25-250 kV beams relative to a 60 Co beam, which is usually the reference beam quality for detector calibration in radiotherapy. MATERIAL AND METHODS: Three microDiamond detectors and, for comparison, two silicon diodes (PTW 60017) were calibrated in terms of air-kerma free in air in six x-ray beam qualities (from 25 to 250 kV) and in terms of absorbed dose to water in a 60 Co beam at the national metrology laboratory in Sweden. The PENELOPE/penEasy MC radiation transport code was used to calculate the absorbed-dose energy response of the detectors (modeled based on blueprints) relative to air and water depending on calibration conditions. The MC results were used to extract the relative intrinsic energy response of the detectors from the overall energy response. Measurements using an independent setup with a single ophthalmic BEBIG I25.S16 125 I BT seed (effective photon energy of 28 keV) were used as a qualitative check of the extracted intrinsic energy response correction factors. Additionally, the impact of the thickness of the active volume as well as the presence of extra-cameral components on the absorbed-dose energy response of a microDiamond detector was studied using MC simulations. RESULTS: The relative intrinsic energy response of the microDiamond detectors was higher by a factor of 2 in 25 and 50 kV beams compared to the 60 Co beam. The variation in the relative intrinsic energy response of silicon diodes was within 10% over the investigated photon energy range. The use of relative intrinsic energy response correction factors improved the agreement among the absorbed dose to water values determined using microDiamond detectors and silicon diodes, as well as with the TG-43 formalism-based calculations for the 125 I seed. MC study of microDiamond detector design features provided a possible explanation for inter-detector response variation at low-energy photon beams by differences in the effective thickness of the active volume. CONCLUSIONS: MicroDiamond detectors had a non-negligible variation in the relative intrinsic energy response (factor of 2) which was comparable to that in the absorbed-dose energy response relative to water at low-energy photon beams. Silicon diodes, in contrast, had an absorbed-dose energy dependence on photon energy that varied by a factor of 6, whereas the intrinsic energy dependence on beam quality was within 10%. It is important to decouple these two responses for a full characterization of detector energy response especially when the user and reference beam qualities differ significantly, and MC alone is not enough.

Long-term mucosal injury and repair in a murine model of pelvic radiotherapy.

Chronic intestinal injury after pelvic radiotherapy affects countless cancer survivors worldwide. A comprehensive understanding of the long-term injury dynamics is prevented in available animal models. With linear accelerators that are used to treat cancer in patients, we irradiated a small volume encompassing the colorectum in mice with four fractions of 8 Gy per fraction. We then determined the long-term dynamics of mucosal injury, repair, and the duration of inflammation. We show that crypt fission, not cell proliferation, is the main long-term mechanism for rescuing crypt density after irradiation, and provides a potentially wide window for clinical interventions. Persisting macrophage aggregations indicate a chronic mucosal inflammation. A better understanding as to how crypt fission is triggered and why it fails to repair fully the mucosa may help restore bowel health after pelvic radiotherapy. Moreover, anti-inflammatory interventions, even if implemented long after completed radiotherapy, could promote bowel health in pelvic cancer survivors.

Out-of-field doses from secondary radiation produced in proton therapy and the associated risk of radiation-induced cancer from a brain tumor treatment.

PURPOSE: To determine out-of-field doses produced in proton pencil beam scanning (PBS) therapy using Monte Carlo simulations and to estimate the associated risk of radiation-induced second cancer from a brain tumor treatment. METHODS: Simulations of out-of-field absorbed doses were performed with MCNP6 and benchmarked against measurements with tissue-equivalent proportional counters (TEPC) for three irradiation setups: two irradiations of a water phantom using proton energies of 78-147 MeV and 177-223 MeV, and one brain tumor irradiation of a whole-body phantom. Out-of-field absorbed and equivalent doses to organs in a whole-body phantom following a brain tumor treatment were subsequently simulated and used to estimate the risk of radiation-induced cancer. Additionally, the contribution of absorbed dose originating from radiation produced in the nozzle was calculated from simulations. RESULTS: Out-of-field absorbed doses to the TEPC ranged from 0.4 to 135 µGy/Gy. The average deviation between simulations and measurements of the water phantom irradiations was about 17%. The absorbed dose contribution from radiation produced in the nozzle ranged between 0 and 70% of the total dose; the contribution was however small in absolute terms. The absorbed and equivalent doses to the organs ranged between 0.2 and 60 µGy/Gy and 0.5-151 µSv/Gy. The estimated lifetime risk of radiation-induced second cancer was approximately 0.01%. CONCLUSIONS: The agreement of out-of-field absorbed doses between measurements and simulations was good given the sources of uncertainties. Calculations of out-of-field organ doses following a brain tumor treatment indicated that proton PBS therapy of brain tumors is associated with a low risk of radiation-induced cancer.

IAEA-SSDL bilateral comparisons for diagnostic level air kerma measurement standards.

Comparisons of national standard of air kerma for conventional and mammographic diagnostic X-ray radiation qualities were conducted by the IAEA. Eleven secondary standards dosimetry laboratories provided calibration data for Exradin A3 and Radcal RC6M transfer ionization chambers circulated. Each comparison result expressed as the ratio of the participant and IAEA calibration coefficient were within the acceptance limit of ±2.5%. From the 67 results of 11 participants and 10 available beam qualities, the comparison result was within its standard uncertainty in 63 cases, and within the expanded (k = 2) uncertainty in four cases. Detailed calibration uncertainty budgets from participant laboratories are presented. The relative standard calibration uncertainty of each participant was in the range of 0.5-1.3%. These results indicate that the calibration related uncertainty component is reasonable low for a clinical measurement. In addition to the calibration coefficient, other corrections should be applied for clinical measurement to achieve the recommended accuracy.

Distinct NF-kappaB regulation by shear stress through Ras-dependent IkappaBalpha oscillations: real-time analysis of flow-mediated activation in live cells.

NF-kappaB, a transcription factor central to inflammatory regulation during development of atherosclerosis, is activated by soluble mediators and through biomechanical inputs such as flow-mediated shear- stress. To investigate the molecular mechanisms underlying shear stress mediated signal transduction in vascular cells we have developed a system that applies flow-mediated shear stress in a controlled manner, while inserted in a confocal microscope. In combination with GFP-based methods, this allows continuous monitoring of flow induced signal transduction in live cells and in real time. Flow-mediated shear stress, induced using the system, caused a successive increase in NF-kappaB-regulated gene activation. Experiments assessing the mechanisms underlying the NF-kappaB induced activity showed time and flow rate dependent effects on the inhibitor, IkappaBalpha, involving nuclear translocation characterized by a biphasic or cyclic pattern. The effect was observed in both endothelial- and smooth muscle cells, demonstrated to impact noncomplexed IkappaBalpha, and to involve mechanisms distinct from those mediating cytokine signals. In contrast, effects on the NF-kappaB subunit relA were similar to those observed during cytokine stimulation. Further experiments showed the flow induced inter-compartmental transport of IkappaBalpha to be regulated through the Ras GTP-ase, demonstrating a pronounced reduction in the effects following blocking of Ras activity. These studies show that flow-mediated shear stress, regulated by the Ras GTP-ase, uses distinct mechanisms of NF-kappaB control at the molecular level. The oscillatory pattern, reflecting inter-compartmental translocation of IkappaBetaalpha, is likely to have fundamental impact on pathway regulation and on development of shear stress-induced distinct vascular cell phenotypes.

Monte Carlo simulations related to gas-based optical diagnosis of human sinusitis.

We investigate the feasibility of using diode laser gas spectroscopy for sinusitis diagnostics. We simulate light propagation using the Monte Carlo concept, as implemented by the Advanced Systems Analysis Program (ASAP) software. Simulations and experimental data are compared for a model based on two scattering bodies representing human tissue, with an air gap in-between representing the sinus cavity. Simulations are also performed to investigate the detection geometries used in the experiments, as well as the influence of the optical properties of the scattering bodies. Finally, we explore the possibility of performing imaging measurements of the sinuses. Results suggest that a diagnostic technique complementary to already existing ones could be developed.

Gas monitoring in human sinuses using tunable diode laser spectroscopy.

We demonstrate a novel nonintrusive technique based on tunable diode laser absorption spectroscopy to investigate human sinuses in vivo. The technique relies on the fact that free gases have spectral imprints that are about 10.000 times sharper than spectral structures of the surrounding tissue. Two gases are detected; molecular oxygen at 760 nm and water vapor at 935 nm. Light is launched fiber optically into the tissue in close proximity to the particular maxillary sinus under study. When investigating the frontal sinuses, the fiber is positioned onto the caudal part of the frontal bone. Multiply scattered light in both cases is detected externally by a handheld probe. Molecular oxygen is detected in the maxillary sinuses on 11 volunteers, of which one had constantly recurring sinus problems. Significant oxygen absorption imprint differences can be observed between different volunteers and also left-right asymmetries. Water vapor can also be detected, and by normalizing the oxygen signal on the water vapor signal, the sinus oxygen concentration can be assessed. Gas exchange between the sinuses and the nasal cavity is also successfully demonstrated by flushing nitrogen through the nostril. Advantages over current ventilation assessment methods using ionizing radiation are pointed out.

Fluorescence monitoring of a topically applied liposomal Temoporfin formulation and photodynamic therapy of nonpigmented skin malignancies.

Meso-tetra(hydroxyphenyl)chlorin (mTHPC) (INN: Temoporfin) is a potent photodynamically active substance in clinical use today. Usually, the substance is given systemically and a known drawback with this administration route is a prolonged skin light sensitization. For the first time to our knowledge, a liposomal Temoporfin gel formulation for topical application was studied in connection with photodynamic therapy (PDT) of nonpigmented skin malignancies in humans. Intervals of 4 hr between drug administration and light irradiation were used. Sensitizer distribution within tumor and surrounding normal skin was investigated by means of point monitoring and imaging fluorescence spectroscopy before, during, and after PDT, showing high tumor selectivity. Furthermore, the bleaching of Temoporfin was studied during the PDT procedure by monitoring the fluorescence following excitation by using a therapeutic light. A 30-35% light-induced photometabolization was shown. No pain occurred during or after treatment. It was also observed that the treated area did not show any swollen tissue or reddening, as is often seen in PDT using topical delta-aminolevulinic acid. On controlling the patients one week after treatment, healing progress was observed in several patients and no complications were registered.