Collinear Laser Spectroscopy of 2

Transition frequencies and fine-structure splittings of the 2 ^{3}S_{1}→2 ^{3}P_{J} transitions in helium-like ^{12}C^{4+} were measured by collinear laser spectroscopy on a 1-ppb level. Accuracy is increased by more than 3 orders of magnitude with respect to previous measurements, enabling tests of recent nonrelativistic (NR) QED calculations including terms up to mα^{7}. Deviations between the theoretical and experimental values are within theoretical uncertainties and are ascribed to mα^{8} and higher-order contributions in the series expansion of the NR QED calculations. Finally, prospects for an all-optical charge radius determination of light isotopes are evaluated.

Nuclear Charge Radius of

Collinear laser spectroscopy was performed on the isomer of the aluminium isotope ^{26m}Al. The measured isotope shift to ^{27}Al in the 3s^{2}3p ^{2}P_{3/2}^{○}→3s^{2}4s ^{2}S_{1/2} atomic transition enabled the first experimental determination of the nuclear charge radius of ^{26m}Al, resulting in R_{c}=3.130(15) fm. This differs by 4.5 standard deviations from the extrapolated value used to calculate the isospin-symmetry breaking corrections in the superallowed ß decay of ^{26m}Al. Its corrected Ft value, important for the estimation of V_{ud} in the Cabibbo-Kobayashi-Maskawa matrix, is thus shifted by 1 standard deviation to 3071.4(1.0) s.

Non-invasive evaluation of human mucosal structures by multiphoton laser scanning tomography in vitro.

BACKGROUND: Mucous membranes may be affected by various diseases and the diagnostic accessibility is limited. Multiphoton laser tomography (MPT) is a useful tool for in vivo evaluation of superficial skin structures and might also be useful for the imaging of mucosa. OBJECTIVES: In order to investigate the suitability of MPT for the evaluation of mucous membranes, tissue samples of different donors and anatomical localizations have been imaged. METHODS: Human mucosa samples from the urinary bladder, palatine tonsil and ocular conjunctiva were investigated by MPT and subsequently compared with conventional histology. RESULTS: Horizontal images of the epithelium and the underlying connective tissue were obtained using the autofluorescence and second harmonics generation signals by MPT. The stratification of multilayered epithelium was consistently reproduced. Also inflammatory changes like lymphocytic infiltrates and widened intercellular spaces were imaged in some cases. CONCLUSION: Mucous tissue samples were obtained and MPT imaging was successfully performed. In the direct comparison with histology of the same tissue samples, the characteristic morphology of the epithelia was found. Further studies are necessary in order to compare images of healthy and diseased mucosal epithelium and the use of MPT for its evaluation in vivo.

Experienced and inexperienced observers achieved relatively high within-observer agreement on video mobility scoring of dairy cows.

Assessment of lameness prevalence and severity requires visual evaluation of thelocomotion of a cow. Welfare schemes including locomotion assessments are increasingly being adopted, and more farmers and their veterinarians might implement a locomotion-scoring routine together. However, high within-observer agreement is a prerequisite for obtaining valid mobility scorings, and within-observer agreement cannot be estimated in a barn, because the gait of cows is dynamic and may change between 2 occasions. The objective of this study was to estimate the within-observer agreement according to the observers' educational background and experience with cattle, based on video recordings with very diverse types of gait. Groups of farmers, bovine veterinarians, first- and fourth-year veterinary students, researchers, and cattle-inexperienced sensory assessors evaluated mobility using a 5-point mobility score system developed specifically for walking cows (n=102 observers). The evaluation sessions were similar for all groups, lasted 75 min, and were organized as follows: introduction, test A, short training session, break, and test B. In total, video recordings of 22 cows were displayed twice in a random order (11 cows in each test × 2 replicates). Data were analyzed applying kappa coefficient, logistic regression, and testing for random effects of observers. The crude estimates of 95% confidence interval for weighted kappa in test A and B ranged, respectively, from 0.76 to 0.80 and 0.70 to 0.75. When adjusting for the fixed effects of video sample and gait scoring preferences, the probability of assigning the same mobility score twice to the same cow varied from 55% (sensory assessors) to 72% (fourth-year veterinary students). The random effect of the individual observers was negligible. That is, in general observers could categorize the mobility characteristics of cows quite well. Observers who preferred to assess the attributes back arch or the overall mobility score (based on uneven gait) had the highest agreement, respectively, 69 or 68%. The training session seemed insufficient to improve agreement. Nonetheless, even novice observers were able to achieve perfect agreement up to 60% of the 22 scorings with merely the experience obtained during the study (introduction and training session). The relatively small differences between groups, together with a high agreement, demonstrate that the new system is easy to follow compared with previously described scoring systems. The mobility score achieves sufficiently high within-observer repeatability to allow between-observer agreement estimates, which are reliable compared with other more-complex scoring systems. Consequently, the new scoring scale seems feasible for on-farm applications as a tool to monitor mobility within and between cows, for communication between farmers and veterinarians with diverse educational background, and for lamenessbenchmarking of herds.

COMPliance and Arthralgia in Clinical Therapy: the COMPACT trial, assessing the incidence of arthralgia, and compliance within the first year of adjuvant anastrozole therapy.

BACKGROUND: This prospective study evaluated the relationship between arthralgia and compliance during the first year of adjuvant anastrozole therapy in postmenopausal women with hormone receptor-positive early breast cancer. PATIENTS AND METHODS: COMPliance and Arthralgia in Clinical Therapy (COMPACT) was an open-label, multicenter, noninterventional study conducted in Germany. Patients had started adjuvant anastrozole 3-6 months before the study start. The primary end points were arthralgia, compliance, and the relationship between compliance and arthralgia, assessed at specific time points. RESULTS: Overall, 1916 patients received upfront anastrozole. Mean arthralgia scores were increased from baseline at each visit up to 9 months. Compliance with anastrozole therapy gradually decreased over time from baseline to 9 months (P<0.001). At 9 months, investigators estimated that >95% of patients were compliant versus patient reports of <70%. There was a significant association between arthralgia mean scores and noncompliance at 6 months (P<0.0001), 9 months (P<0.0001), and overall (P<0.0001). Over time, new events or impairment of existing arthralgias were reported in 14% (3 months), 11% (6 months), and 9% (9 months) of patients. CONCLUSION: Arthralgia is important in the clinical management of women with early breast cancer and may contribute to noncompliance and clinical outcomes. CLINICALTRIALSGOV IDENTIFIER: NCT00857012.

[Google translate is not sufficient to overcome language barriers in neonatal medicine]. / Erste Erfahrungen mit Google Translate in der Neonatologie.

Language barriers hinder the interaction with patients and relatives. The use of language services increases knowledge, satisfaction and the use of medical care and thus improves patient's clinical outcome. The recommended use of professional interpreters (PI) is not always feasible. We tested an online translation tool as an alternative for PI for the transla-tion of standardized sentences from a neonatal doctor-/nurse-relative-interview.Translation of 20 sentences from a German neonatal intensive care unit parent information brochure to English, Portuguese and Arabic, using Google Translate (GT). Assessment of accuracy concerning grammar and content, in a second step simplification of all incorrect sentences, translation by GT and critical re-assessment and evaluation.An average of 42% of the sentences was correctly translated concerning grammar and content. The proportion of incorrectly translated sentences varied between 45-70%. By simpli-fication another 23% were translated correctly.Translations by GT were often incorrect in content and grammar. We suppose that the design of GT, which is a statistical translation engine, might be an explanation for this phenomenon. Presently, GT cannot guarantee unambiguous translations and cannot substitute PIs, only in particular circumstances, the use of GT or similar engines may be justified. For future use of electronic translation services, we suggest to compile a catalogue of sentences containing central information, which can be translated into defined foreign languages without misinterpretation or loss of information.

[Molecular pathology of the lungs. New perspectives by next generation sequencing]. / Molekularpathologie der Lunge. Perspektiven durch neue Sequenzierformen.

Lung cancer is one of the most frequent malignancies in the western world. Its frequent association with a wide spectrum of mutations in genes encoding various signal transducers that are often linked to therapy response, emphasizes the obvious need for improved, fast and highly efficient approaches in molecular pathology. Comprehensive analyses of the mutation status of progression and therapy relevant genes can be performed by the novel sequencing forms named next generation sequencing (NGS) providing extremely high capacities for ultra-deep sequence analyses. The 454 pyrosequencing method, the sequencing by synthesis and the semiconductor sequencing platform are now available for parallel sequencing approaches of multitudinous target genes linked to multiple tumor DNA applications. The "one molecule, one clone, one read" principle by the NGS approaches supplies not only information on allele frequencies and mutation rates but also has the advantage of a very sensitive detection of low frequency variants.

Safety assessment by multiphoton fluorescence/second harmonic generation/hyper-Rayleigh scattering tomography of ZnO nanoparticles used in cosmetic products.

Zinc oxide nanoparticles (ZnO NPs) are commonly used as UV filters in commercial sunscreen products. Their penetration into the skin is intensively discussed in the literature. In the present in vivo study, penetration of ZnO NPs (30 nm in size) into human skin was investigated by multiphoton tomography. Based on the non-linear effects of a second harmonic generation and hyper-Rayleigh scattering, the distribution of ZnO NPs in the horny layers of the epidermis, as well as the furrows, wrinkles and orifice of the hair follicles was analyzed. This method permitted distinguishing between the particulate and dissolved forms of Zn. A detection limit of 0.08 fg/µm(3) was estimated. Taking advantage of this sensitivity, it was clearly shown that ZnO NPs penetrate only into the outermost layers of stratum corneum, furrows and into the orifices of the hair follicles and do not reach the viable epidermis.

[How to handle unkown fluids in fluid-preserved collections : Cloudy prospects?] / Zum Umgang mit Präparaten in unbekannten Konservierungslösungen in Feuchtpräparatesammlungen : Trübe Aussichten?

BACKGROUND: Fluid collections are gaining more importance for research and teaching, but they are facing preservation problems. In the case of historical collections, the methods of fixation and preservation are poorly documented. The liquid used is unknown. In order to ensure the preservation of such collections, it is essential to have available a preservation liquid that is compatible with the most common historical liquids and techniques. MATERIALS AND METHODS: A universal liquid based on historical recipes was developed for such problematic preparations. RESULTS: The use of distilled water, glycerin and ethanol (80%) in a ratio of 10:6:1 offers a good alternative that is harmless to the health of the user. It can be used for colour-preserving conserved preparations and for pure ethanol and formaldehyde preparations and is recommended as a universal solution for preparations in unknown preservation liquids.

Keratinocyte morphology of human skin evaluated by in vivo multiphoton laser tomography.

BACKGROUND: Multiphoton tomography (MPT) is a novel non-invasive imaging method in dermatology allowing the depiction of the epidermis with sub-cellular resolution. Here, we present a descriptive characterization of unaffected human epidermis, morphometric data on human keratinocytes and some epidermal parameters in vivo and a morphological characterization of keratinocyte changes in actinic keratoses. METHODS: In a clinical setting, 57 volunteers of different age groups were examined using MPT. RESULTS: The morphological appearance of keratinocytes showed polygonal cells in the horny layer, a granular cytoplasm in the stratum granulosum, smaller prickle cells in the stratum spinosum and hyperpigmented small round basal cells. Actinic keratoses presented remarkable differences including widened inter-cellular spaces, heterogeneity in cellular fluorescence and shape as well as an increased ratio of nuclear to cellular size. Finally, the thickness of the epidermis was significantly increased in actinic keratoses compared with the control. CONCLUSION: In vivo MPT provides high-resolution images allowing the identification and quantification of cellular morphometric parameters. First observations of morphology and morphometry of actinic keratoses are reported.

Two-photon microscopy of deep intravital tissues and its merits in clinical research.

Multiphoton excitation laser scanning microscopy, relying on the simultaneous absorption of two or more photons by a molecule, is one of the most exciting recent developments in biomedical imaging. Thanks to its superior imaging capability of deeper tissue penetration and efficient light detection, this system becomes more and more an inspiring tool for intravital bulk tissue imaging. Two-photon excitation microscopy including 2-photon fluorescence and second harmonic generated signal microscopy is the most common multiphoton microscopic application. In the present review we take diverse ocular tissues as intravital samples to demonstrate the advantages of this approach. Experiments with registration of intracellular 2-photon fluorescence and extracellular collagen second harmonic generated signal microscopy in native ocular tissues are focused. Data show that the in-tandem combination of 2-photon fluorescence and second harmonic generated signal microscopy as two-modality microscopy allows for in situ co-localization imaging of various microstructural components in the whole-mount deep intravital tissues. New applications and recent developments of this high technology in clinical studies such as 2-photon-controlled drug release, in vivo drug screening and administration in skin and kidney, as well as its uses in tumourous tissues such as melanoma and glioma, in diseased lung, brain and heart are additionally reviewed. Intrinsic emission two-modal 2-photon microscopy/tomography, acting as an efficient and sensitive non-injurious imaging approach featured by high contrast and subcellular spatial resolution, has been proved to be a promising tool for intravital deep tissue imaging and clinical studies. Given the level of its performance, we believe that the non-linear optical imaging technique has tremendous potentials to find more applications in biomedical fundamental and clinical research in the near future.

Clinical study on the effects of a cosmetic product on dermal extracellular matrix components using a high-resolution multiphoton tomograph.

BACKGROUND/PURPOSE: The aim of this study was to demonstrate the effects of selected plant extracts in a cosmetic cream on the dermal network components after a 3-month treatment using an in vivo multiphoton tomographic device. METHODS: Twenty-four Caucasian women aged between 45 and 65 applied randomly a cosmetic emulsion B containing active ingredients (soy and jasmine) twice a day on one arm and its vehicle A (without active ingredients) on the other arm during 3 months. Measurements were performed on the internal side of the forearm before starting the treatment (T0), after 4 week (T4) and 12 weeks (T12) treatment. Measurements consisted of a multi-layers acquisitions using a multiphoton tomograph with subcellular resolution. Optical sections (about 6 microm thick) were recorded from 0 to about 200 microm using two different wavelengths: 760 and 820 nm. To compare the series of images and obtain an objective quantification of the signal of second harmonic generation (SHG) and autofluorescence, the method used consisted of taking the integrated brightness of an image (same rectangular area for all images) as a measure of the signal. Following this step, a ratio between brightness of images from the area treated with cream A or B and brightness of untreated area was calculated and used as an assessment of treatment efficacy. The parameter used for statistical analysis (variance analysis) is the difference before and after 12 weeks of treatment by either cream A or B of the signal ratios calculated in the upper dermis (118-130 microm) and those from a deeper region of the upper dermis (165-178 microm). RESULTS: Signals (autofluorescence+SHG) of extracellular matrix do not change significantly with time (weeks 0, 4 and 12) when cream A (vehicle with no active ingredient) is applied. Treatment with cream B results in an enhancement in the signal level of extracellular matrix at week 12. The comparison of signals, in both areas (118-130 microm and 160-178 microm), show an higher increase in the deeper region than in the more superficial one for product B while we do not notice any change with product A. CONCLUSION: The multiphoton tomograph provided excellent high-resolution images, which describe clearly the different skin layers, single cells and extracellular matrix components in all the 24 volunteers. Statistic analyses reveal a real effect for product B with selected plant extracts, known to increase collagen synthesis. Changes observed are characteristics of modifications in dermal collagen and elastin content. To our knowledge, it is the first time that it was possible to demonstrate in vivo the effect of a cosmetic product on the superficial dermal layer, in a non-invasive and non-destructive process, i.e. without cutting the skin.

Fulminant herpes simplex virus hepatic failure in pregnancy requiring liver transplantation.

Herpes simplex virus hepatitis is a rare but potentially fatal condition that usually affects the immunocompromised, including pregnant women. This case report details the course of fulminant hepatic failure in a woman at 31 weeks gestation resulting in emergent delivery of the fetus and liver transplant in the mother.

[Multiphoton microscopy and in vivo tomography in dermatologic imaging]. / Multiphotonenmikroskopie und In-vivo-Multiphotonentomographie in der dermatologischen Bildgebung.

Multiphoton microscopy (MPM) and in vivo multiphoton tomography (MPT) are non-invasive examination techniques that allow for the evaluation of cellular as well as extra-cellular structures by working at a subcellular resolution level. These techniques are thus appropriate not only for clinical diagnostics but also for scientific issues in basic and applied research. MPM and MPT are based on the stimulation of biogenic fluorophores by two or more long-wave, low-energy photons and the evocation of second harmonic generation (SHG). Thus, the evaluation quality of cell clusters and tissues is similar to histological sections. At the same time the dermal fiber network can be assessed. MPT was developed further for the application in non-invasive in vivo diagnostics of skin diseases. This review presents the capabilities of multiphoton-based diagnostics in the evaluation of transcutaneous metabolism. In addition, the multiphoton techniques employed for the evaluation of physiologic and pathologic changes of the dermal fiber network as well as in the diagnosis of dermal and epidermal disorders by visual biopsy. Besides the morphological classification of benign and malignant skin tumors or allergic or inflammatory skin lesions, the techniques also allow for recording metabolic processes.

A new Collinear Apparatus for Laser Spectroscopy and Applied Science (COALA).

We present a new collinear laser spectroscopy setup that has been designed to overcome systematic uncertainty limits arising from high-voltage and frequency measurements, beam superposition, and collisions with residual gas that are present in other installations utilizing this technique. The applied methods and experimental realizations are described, including an active stabilization of the ion-source potential, new types of ion sources that have not been used for collinear laser spectroscopy so far, dedicated installations for pump-and-probe measurements, and a versatile laser system referenced to a frequency comb. The advanced setup enables us to routinely determine transition frequencies, which was so far demonstrated only for a few cases and with lower accuracy at other facilities. It has also been designed to perform accurate high-voltage measurements for metrological applications. Demonstration and performance measurements were carried out with Ca+ and In+ ions.

Rigid and high-numerical-aperture two-photon fluorescence endoscope.

We present a rigid miniaturized optical system block fiber-optic two-photon endoscope based on a compact two-axis piezo scanner system and a miniature high (0.65) NA GRIN lens objective. The optical system is scanned as a whole by a piezo scanner allowing always an on-axis beam irradiation of the optical system. A photonic crystal fiber is used for excitation and ultrashort laser pulses can be delivered with typical power up to 100 mW at 800 nm. Two-photon fluorescence signal is collected by the use of a multimode fiber. Lateral resolution values for the system were experimentally measured to be 0.67 microm vertically and 1.08 microm horizontally. Axial resolution was found to be 5.8 microm. The endoscope is highly flexible and controllable in terms of time acquisition, resolution, and magnification. Fluorescence images were acquired over a 420 microm x 420 microm field of view. Results presented here demonstrate the ability of the system to resolve subcellular details and the potential of the technology for in vivo applications.

[Femtosecond laser ablation and scanning microscopy of the internal retinal limiting membrane: an experimental study].

The investigation was undertaken to study whether femtosecond laser ablation and microscopy might be used in the internal retinal borderline membrane. Ablation of internal limiting membrane preparations removed using or not using indocyanine green was made by a low-energy femtosecond laser. Examination of the preparations by laser and electron microscopy revealed precision laser cuts of the internal retinal borderline membrane. The use of indocyanine green during laser ablation reduced laser irradiation parameters as compared to the dye not being applied. Low-energy femtosecond lasers enable precision contactless ablation of the internal borderline membrane to be carried out without collateral damage to the adjacent tissue. The parameters of laser impulses, particularly low ones used in the ablation of indocyanine green-stained preparations, prove the photosensitizing effect of the dye.

Nonlinear optical endoscope based on a compact two axes piezo scanner and a miniature objective lens.

We report on a nonlinear optical endoscope that adopts a hollow core photonic crystal fiber for single-mode illumination delivery and a multimode one for signal collection. Femtosecond laser pulses up to 100 mW can be delivered at a centered wavelength of 800 nm. The two-photon fluorescence response of our system is shown to have axial and lateral resolutions of 5.8 microm and 0.6 microm respectively. Fluorescence detection was obtained at different wavelengths between 790 and 840 nm which could allow SHG detection for example. The maximal field-of-view of the acquired images is 420 microm x 420 microm. Detection efficiency is greater by using an avalanche photodiode in comparison to a photo multiplier tube. Results presented here demonstrate the ability of the system to resolve cellular details and the potential of the device for future in vivo imaging diagnosis.

[Guideline for the Early Detection of Breast Cancer in Germany 2008. Recommendations from the short version]. / Stufe-3-Leitlinie Brustkrebs-Früherkennung in Deutschland 2008 : Auszug aus der Kurzfassung: Handlungsempfehlungen.

The updated 2008 German Guideline for Early Detection of Breast Cancer provides evidence-based and consensus-based recommendations of the knowledge gained by the German Society for Surgery and the German Society of Plastic, Aesthetic, and Reconstructive Surgeons together with 29 professional societies, associations, and nonmedical organizations. The guideline is meant to assist physicians, healthy women, and patients in medical decisions with recommendations regarding the diagnostic chain in early detection of breast cancer. In addition to these recommendations, the guideline also includes descriptions of quality assurance for resources, procedures, outcomes, and evaluation using a set of quality indicators. It updates the previous version from 2003. The guideline's recommendations are presented. They are described in detail in the full publication (in German) Geburtsh Frauenh 2008; 68:251-261. The long version of the Guideline, methods report, and evidence report are available on the internet at www.awmf-leitlinien.de (reg. no. 077/001) with free access.

Clinical two-photon microendoscopy.

Two-photon medical imaging has found its way into dermatology as an excellent method for noninvasive skin cancer detection without need of contrast agents as well as for in situ drug screening of topically-applied cosmetical and pharmaceutical components. There is an increasing demand to apply the multiphoton technology also for deep-tissue skin imaging as well as for intracorporal imaging. We report on the first clinical use of multiphoton endoscopes, in particular of a miniaturized rigid two-photon GRIN lens endoscope. The microendoscope was attached to the multiphoton tomograph DermaInspect and employed to detect the extracellular matrix proteins collagen and elastin in the human dermis of volunteers and patients with ulcera by in vivo second harmonic generation and in vivo two-photon autofluorescence.