New insights into osmobiosis and chemobiosis in tardigrades.

Tardigrades are renowned for their ability to enter the extremotolerant state of latent life known as cryptobiosis. While it is widely accepted that cryptobiosis can be induced by freezing (cryobiosis) and by desiccation (anhydrobiosis), the latter involving formation of a so-called tun, the exact mechanisms underlying the state-as well as the significance of other cryptobiosis inducing factors-remain ambiguous. Here, we focus on osmotic and chemical stress tolerance in the marine tidal tardigrade Echiniscoides sigismundi. We show that E. sigismundi enters the tun state following exposure to saturated seawater and upon exposure to locality seawater containing the mitochondrial uncoupler DNP. The latter experiments provide evidence of osmobiosis and chemobiosis, i.e., cryptobiosis induced by high levels of osmolytes and toxicants, respectively. A small decrease in survival was observed following simultaneous exposure to DNP and saturated seawater indicating that the tardigrades may not be entirely ametabolic while in the osmobiotic tun. The tardigrades easily handle exposure to ultrapure water, but hypo-osmotic shock impairs tun formation and when exposed to ultrapure water the tardigrades do not tolerate DNP, indicating that tolerance towards dilute solutions involves energy-consuming processes. We discuss our data in relation to earlier and more contemporary studies on cryptobiosis and we argue that osmobiosis should be defined as a state of cryptobiosis induced by high external osmotic pressure. Our investigation supports the hypothesis that the mechanisms underlying osmobiosis and anhydrobiosis are overlapping and that osmobiosis likely represents the evolutionary forerunner of cryptobiosis forms that involve body water deprivation.

Thermotolerance experiments on active and desiccated states of Ramazzottius varieornatus emphasize that tardigrades are sensitive to high temperatures.

Global warming is already having harmful effects on habitats worldwide and it is therefore important to gain an understanding of how rising temperatures may affect extant animals. Here, we investigate the tolerance to high temperatures of Ramazzottius varieornatus, a tardigrade frequently found in transient freshwater habitats. Using logistic modelling on activity we evaluate the effect of 24 hour temperature exposures on active tardigrades, with or without a short acclimation period, compared to exposures of desiccated tardigrades. We estimate that the 50% mortality temperature for non-acclimated active tardigrades is 37.1 °C, with a small but significant increase to 37.6 °C following acclimation. Desiccated specimens tolerate much higher temperatures, with an estimated 50% mortality temperature of 82.7 °C following 1 hour exposures, but with a significant decrease to 63.1 °C following 24 hour exposures. Our results show that metabolically active tardigrades are vulnerable to high temperatures, yet acclimatization could provide a tolerance increase. Desiccated specimens show a much higher resilience-exposure-time is, however, a limiting factor giving tardigrades a restricted window of high temperature tolerance. Tardigrades are renowned for their ability to tolerate extreme conditions, but their endurance towards high temperatures clearly has an upper limit-high temperatures thus seem to be their Achilles heel.

Tolerance to Gamma Radiation in the Marine Heterotardigrade, Echiniscoides sigismundi.

Tardigrades belong to the most radiation tolerant animals on Earth, as documented by a number of studies using both low-LET and high-LET ionizing radiation. Previous studies have focused on semi-terrestrial species, which are also very tolerant to desiccation. The predominant view on the reason for the high radiation tolerance among these semi-terrestrial species is that it relies on molecular mechanisms that evolved as adaptations for surviving dehydration. In this study we report the first study on radiation tolerance in a marine tardigrade, Echiniscoides sigismundi. Adult specimens in the hydrated active state were exposed to doses of gamma radiation from 100 to 5000 Gy. The results showed little effect of radiation at 100 and 500 Gy but a clear decline in activity at 1000 Gy and higher. The highest dose survived was 4000 Gy, at which ca. 8% of the tardigrades were active 7 days after irradiation. LD50 in the first 7 days after irradiation was in the range of 1100-1600 Gy. Compared to previous studies on radiation tolerance in semi-terrestrial and limnic tardigrades, Echiniscoides sigismundi seems to have a lower tolerance. However, the species still fits into the category of tardigrades that have high tolerance to both desiccation and radiation, supporting the hypothesis that radiation tolerance is a by-product of adaptive mechanisms to survive desiccation. More studies on radiation tolerance in tardigrade species adapted to permanently wet conditions, both marine and freshwater, are needed to obtain a more comprehensive picture of the patterns of radiation tolerance.

Towards high-accuracy primary spectral radiometry from 400 K to 1300 K.

We describe the design, construction, calibration and use of a near-infrared thermodynamic radiation thermometer to measure blackbodies from 400 K to 1300 K. The motivation for this work is the pending redefinition of the kelvin and the need for direct, thermodynamic temperature measurements of the fixed-point blackbodies presently used in the realization of the temperature scale. The challenges of accurately measuring Planck radiances which vary greatly in radiance level and spectral shape are discussed. Methods to characterize the components used in the radiation thermometer design are described. The use of this radiation thermometer as a relative primary thermometer and the resulting residuals are shown. We describe radiometric calibration procedures for using the radiation thermometer as an absolute primary thermometer. Preliminary data showing the initial radiometric calibration steps are discussed.

Precise Measurement of Lunar Spectral Irradiance at Visible Wavelengths.

We report a measurement of lunar spectral irradiance with an uncertainty below 1 % from 420 nm to 1000 nm. This measurement uncertainty meets the stability requirement for many climate data records derived from satellite images, including those for vegetation, aerosols, and snow and ice albedo. It therefore opens the possibility of using the Moon as a calibration standard to bridge gaps in satellite coverage and validate atmospheric retrieval algorithms. Our measurement technique also yields detailed information about the atmosphere at the measurement site, suggesting that lunar observations are a possible solution for aerosol monitoring during the polar winter and can provide nighttime measurements to complement aerosol data collected with sun photometers. Our measurement, made with a novel apparatus, is an order of magnitude more accurate than the previous state-of-the-art and has continuous spectral coverage, removing the need to interpolate between filter passbands.

Stray light correction algorithm for multichannel hyperspectral spectrographs.

An algorithm is presented that corrects a multichannel fiber-coupled spectrograph for stray or scattered light within the system. The efficacy of the algorithm is evaluated based on a series of validation measurements of sources with different spectral distributions. This is the first application of a scattered-light correction algorithm to a multichannel hyperspectral spectrograph. The algorithm, based on characterization measurements using a tunable laser system, can be extended to correct for finite point-spread response in imaging systems.

The consultation as an interpretive dialogue about the child's health needs.

BACKGROUND: Though uniquely placed in the health care system, GPs only become aware of a small number of children with behavioural and emotional problems. Research evaluating the challenges and barriers in general practice for identifying children with problems is therefore important. OBJECTIVES: To identify and articulate GPs' experiences and perceptions of the GP-parent dialogue about children's health problems, in order to broaden our understanding of the challenges inherent to the dialogue. METHODS: The GPs' experiences and recollections were explored in a qualitative study comprising four focus group discussions and nine individual interviews. The focus of study was to explore GP consultations with children 0-5 years of age and their parent/s. RESULTS: Though expressing a family-focused approach to the child consultation, the GPs often did not succeed in making the consultation family focused. The analysis revealed that the GPs often were 'stuck' in the traditional role of expert and this made it difficult for them to explore the child's well-being. The consultation became more family focused when the GPs moved away from the role of expert. The GPs experienced that by sharing their uncertainty with parents, they often got more insight into the child's everyday life and family circumstances. CONCLUSION: The study indicates that through open reflective dialogue the GP is able to assess the child and strengthen mutual trust in the doctor-parent relationship to the benefit of children with special needs.

Best Practice Guidelines for Pre-Launch Characterization and Calibration of Instruments for Passive Optical Remote Sensing.

The pre-launch characterization and calibration of remote sensing instruments should be planned and carried out in conjunction with their design and development to meet the mission requirements. The onboard calibrators such as blackbodies and the sensors such as spectral radiometers should be characterized and calibrated using SI traceable standards. In the case of earth remote sensing, this allows inter-comparison and intercalibration of different sensors in space to create global time series of climate records of high accuracy where some inevitable data gaps can be easily bridged. The recommended best practice guidelines for this pre-launch effort is presented based on experience gained at National Institute of Standards and Technology (NIST), National Aeronautics and Space Administration (NASA) and National Oceanic and Atmospheric Administration (NOAA) programs over the past two decades. The currently available radiometric standards and calibration facilities at NIST serving the remote sensing community are described. Examples of best practice calibrations and intercomparisons to build SI (international System of Units) traceable uncertainty budget in the instrumentation used for preflight satellite sensor calibration and validation are presented.

Internal quantum efficiency modeling of silicon photodiodes.

Results are presented for modeling of the shape of the internal quantum efficiency (IQE) versus wavelength for silicon photodiodes in the 400 nm to 900 nm wavelength range. The IQE data are based on measurements of the external quantum efficiencies of three transmission optical trap detectors using an extensive set of laser wavelengths, along with the transmittance of the traps. We find that a simplified version of a previously reported IQE model fits the data with an accuracy of better than 0.01%. These results provide an important validation of the National Institute of Standards and Technology (NIST) spectral radiant power responsivity scale disseminated through the NIST Spectral Comparator Facility, as well as those scales disseminated by other National Metrology Institutes who have employed the same model.

Improved Near-Infrared Spectral Responsivity Scale.

A cryogenic radiometer-based system was constructed at the National Institute of Standards and Technology for absolute radiometric measurements to improve detector spectral power responsivity scales in the wavelength range from 900 nm to 1800 nm. In addition to the liquid-helium-cooled cryogenic radiometer, the system consists of a 100 W quartz-tungsten-halogen lamp light source and a 1 m single-grating monochromator for wavelength selection. The system was characterized and the uncertainty in spectral power responsivity measurements evaluated. A variety of photodetectors, including indium gallium arsenide photodiodes (InGaAs), germanium (Ge) photodiodes, and pyroelectric detectors, were subsequently calibrated. Over most of the spectral range, the spectral power responsivity of the photodetectors can be measured with a combined relative standard uncertainty of 0.4 % or less. This is more than a factor of two smaller than our previous capabilities, and represents a significant improvement in the near infrared (NIR) spectral power responsivity scale maintained at NIST. We discuss the characterization of the monochromator-based system and present results of photodetector spectral power responsivity calibrations.

Ultraviolet radiometry with synchrotron radiation and cryogenic radiometry.

The combination of a cryogenic radiometer and synchrotron radiation enables detector scale realization in spectral regions that are otherwise difficult to access. Cryogenic radiometry is the most accurate primary detector-based standard available to date, and synchrotron radiation gives a unique broadband and continuous spectrum that extends from x ray to far IR. We describe a new cryogenic radiometer-based UV radiometry facility at the Synchrotron Ultraviolet Radiation Facility II at the National Institute of Standards and Technology. The facility is designed to perform a variety of detector and optical materials characterizations. The facility combines a high-throughput, normal incidence monochromator with an absolute cryogenic radiometer optimized for UV measurements to provide absolute radiometric measurements in the spectral range from 125 nm to approximately 320 nm. We discuss results on photodetector characterizations, including absolute spectroradiometric calibration, spatial responsivity mapping, spectroreflectance, and internal quantum efficiency. In addition, such characterizations are used to study UV radiation damage in photodetectors that can shed light on the mechanism of the damage process. Examples are also given for UV optical materials characterization.

Surface mass spectrometry of biotinylated self-assembled monolayers.

Biotin and biotinylated self-assembled monolayers (SAMs) on gold have been investigated using time-of-flight secondary ion mass spectrometry, direct laser desorption, laser desorption with 193 nm photoionization of ion- and laser-desorbed species, and laser desorption with vacuum ultraviolet (VUV, 118 nm) photoionization. Our results indicate that direct laser desorption and laser desorption combined with 193 nm multiphoton ionization can detect a chromophoric molecule like biotin that is covalently bound to a SAM. However, secondary ion mass spectra were dominated by fragmentation, and ion desorption/193 nm photoionization detected no species related to biotin. The dominant features of the laser desorption/VUV mass spectra were neat and Au-complexed dimers of intact and fragmented biotinylated SAM molecules. Multiphoton and single-photon ionization of laser-desorbed neutrals from biotinylated SAMs both led to the production of ions useful for chemical analysis of the monolayer. Multiphoton ionization with ultraviolet radiation was experimentally less challenging but required a chromophore for ionization and resulted in significant fragmentation of the adsorbate. Single-photon ionization with VUV radiation was experimentally more challenging but did not require a chromophore and led to less fragmentation. X-ray photoelectron spectra indicated that the biotinylated SAM formed a disordered, 40-60 Å thick monolayer on Au. Additionally, projection photolithography with a Schwarzschild microscope was used to pattern the biotinylated SAM surface and laser desorption/photoionization was used to detect biotinylated adsorbates from the ∼10 µm sized pattern.

Microscopic laser desorption/postionization fourier transform mass spectrometry.

A microscopic laser desorption/postionization Fourier transform mass spectrometer (LD/FTMS) is described. The lateral resolution can be <1 µm with the inherent FTMS high mass resolution intact. Laser postionization allows a certain selectivity and an increase in sensitivity. This capability should allow materials characterization in a wide variety of cases. We demonstrate microscopic desorption and postionization for atoms and molecules.

Molecular analysis by ionization of laser-desorbed neutral species.

A powerful molecular surface analysis technique for the analysis of complex materials, such as polymer/additive systems, consists of laser desorption of surface molecules and subsequent ionization of these gas-phase molecules with resonant or nonresonant laser ionization. These molecular ions are subsequently detected by Fourier-transform mass spectrometry or time-of-flight mass spectrometry. We show that different wavelengths for the postionization step permit selectivity that provides important additional information on the chemical makeup of these complex materials. Near-UV wavelengthsselectively ionize aromatic polymer additives, far-UV wavelengths photoionize other nonaromatic species; and vacuum-UV wavelengths provide access to all the desorbed species. In addition to these applied results, we study many fundamental issues of laser desorption, such as desorption thresholds, velocity distributions, postionization wavelength selectivity, etc. The Fourier-transform mass spectrometer and time-of-flight mass spectrometer are discussed in detail.

[The patterns of anxiety and crying among normal Danish infants with and without colic]. / Uro- og graedemønster hos normale danske spaedbørn med og uden kolik.

On the basis of a descriptive, prospective investigation, the patterns of fussing and crying in normal Danish infants aged from 2-3 to 14 weeks are described. A total of 102 infants were included in the investigation and 85% completed the investigation. The parents carried out daily registration of the periods of fussing and crying. The great majority of the infants were very placid. The mean duration of fussing for the entire group was half an hour per week during the third week of life. This increased to 2 1/2 hours per week at the sixth week of life and then decreased gradually to 0 hours per week from the 12th week of life. The material was subdivided into quartiles. Infants in the first and second quartiles showed practically no fussing. Infants in the third quartile were fussy at the commencement of the period and became gradually more placid from the fifth week of life. Infants in the fourth quartile were fussy during the first 9-10 weeks of life, after which the fussing gradually diminished. In infants in the third and fourth quartiles, fussing was most marked in the evenings. Two pairs of parents considered that their infants had colic. Ten infants were defined as having infantile colic. In these infants, the patterns of fussing and crying showed a common pattern and this was significantly different from the remainder of the group. The basic variables for the 25% most placid infants and the ten infants presumed to have colic are compared.