Electronmicroscopic investigations on the role of vesicle-like bodies in inner ear maculae for fish otolith growth.

The presence, morphology and possible origin of vesicle-like bodies (VBs) within the inner ear otolithic membrane of developmental stages of cichlid fish Oreochromis mossambicus and adult swordtail fish Xiphophorus helleri was analysed by means of transmission and scanning electron microscopy (TEM and SEM, respectively) employing various fixation procedures. The VBs are believed to be involved in the formation of the otolith (or statolith in birds and mammals) regarding the supply of the otolith's organic material. Increasing the osmolarity of the fixation medium decreased the number of VBs seen. Decalcification ended up in a complete disappearance of the VBs. Whilst a fixation with glutaraldehyde followed by OSO4 fixation yielded numerous VBs, only few of them were observed when the tissue was fixed with glutaraldehyde and OSO4 simultaneously. Therefore, the results strongly suggest that the VBs are fixative (i.e., glutaraldehyde) induced artifacts, so-called blisters. With this, the supply of an oto- or statolith's organic material remains obscure. Possibly, it is provided by secretion from the supporting cells as has been hypothesized earlier.     

Neuronal feedback between brain and inner ear for growth of otoliths in fish.

Previous investigations revealed that fish inner ear otolith growth (concerning otolith size and calcium-incorporation) depends on the amplitude and the direction of gravity, suggesting the existence of a (negative) feedback mechanism. In search for the regulating unit, the vestibular nerve was unilaterally transected in neonate swordtail fish (Xiphophorus helleri) which were subsequently incubated in the calcium-tracer alizarin-complexone. Calcium incorporation ceased on the transected head sides, indicating that calcium uptake is neurally regulated.     

Efficacy of an ototoxic aminoglycoside (gentamicin) on the differentiation of the inner ear of cichlid fish.

Previous investigations revealed that the growth of fish inner ear otoliths depends on the amplitude and the direction of gravity, thus suggesting the existence of a (negative) feedback mechanism. In the course of these experiments, it was shown that altered gravity both affected otolith size (and thus the provision of the proteinacious matrix) as well as the incorporation of calcium. It is hitherto unknown, as of whether sensory hair cells are involved either in the regulation of otolith growth or in the provision of otolithic material (such as protein or inorganic components) or even both. The ototoxic aminoglycoside gentamicin (GM) damages hair cells in many vertebrates (and is therefore used for the treatment of Meniere's disease in humans). The present study was thus designed to determine as of whether vestibular sensory cells are needed for otolith growth by applying GM in order to induce a (functionally relevant) loss of these cells. Developing cichlid fish Oreochromis mossambicus were therefore immersed in 120 mg/l GM for 10 or 21 days. At the beginning and at the end of the experimental periods, the fish were incubated in the calcium-tracer alizarin complexone (AC). After the experiment, otoliths were dissected and the area grown during GM-exposure (i.e., the area enclosed by the two AC labellings) was determined planimetrically. The results showed that incubating the animals in a GM-solution had no effect on otolith growth, but the development of otolith asymmetry was affected. Ultrastructural examinations of the sensory hair cells revealed that they had obviously not been affected by GM-treatment (no degenerative morphological features observed). Overall, the present results suggest that hair cells are not affected by GM concerning their possible role in (general) otolith growth, but that these cells indeed might have transitionally been impaired by GM resulting in a decreased capacity of regulating otolith symmetry.     

Calcium-tracers disclose the site of biomineralization in inner ear otoliths of fish.

Since changing gravity (concerning direction and amplitude) strongly affects inner ear otolith growth and otolithic calcium incorporation in developing fish, it was the aim of the present study to locate the site of mineralization in order to gain cues and insights into the provenance of the otoliths inorganic compounds. Therefore, larval cichlid fish (Oreochromis mossambicus) were incubated in the calcium-tracer alizarin complexone (AC; red fluorescence). After maintenance in aquarium water for various periods (1, 2, 3, 6, 9 and 12 h; 1, 2, 3, 5, 6, 7, 15, 29, 36 and 87 d), the animals were incubated in the calcium-tracer calcein (CAL; green fluorescence). AC thus labeled calcium being incorporated at the beginning of the experiment and would subsequently accompany calcium in the course of a possible dislocation, whereas CAL visualized calcium being deposited right at the end of the test. Subsequently, the otoliths were analyzed using a laser scanning microscope and it was shown that the initial site of calcium incorporation was located directly adjacent to the sensory epithelium and the otolithic membrane. Later, calcium deposits were also found on further regions of the otoliths' surface area, where they had been shifted to in the course of dislocation. This finding strongly indicates that the sensory epithelium plays a prominent role in otolithic biomineralization, which is in full agreement with an own electron microscopical study [ELGRA News 23 (2003) 63].     

Histochemical localisation of carbonic anhydrase in the inner ear of developing cichlid fish,Oreochromis mossambicus

Inner ear otolith growth in terms of mineralisation mainly depends on the enzyme carbonic anhydrase (CAH). CAH is located in specialised, mitochondria-rich macular cells (ionocytes), which are involved in the endolymphatic ion exchange, and the enzyme is responsible for the provision of the pH-value necessary for otolithic calcium carbonate deposition.     

Calcium gradients in the fish inner ear sensory epithelium and otolithic membrane visualized by energy filtering transmission electron microscopy (EFTEM).

Inner ear otolith formation in fish is supposed to be performed by the molecular release of proteinacious precursor material from the sensory epithelia, followed by an undirected and diffuse precipitation of calcium carbonate (which is mainly responsible for the functionally important weight of otoliths). The pathway of calcium into the endolymph, however, still remains obscure. Therefore, the presence of calcium within the utricle of larval cichlid fish Oreochromis mossambicus was analyzed by means of energy filtering transmission electron microscopy (EFTEM). Electron spectroscopic imaging (ESI) and electron energy loss spectra (EELS) revealed discrete calcium precipitations, which were especially numerous in the proximal endolymph as compared to the distal endolymph. A decreasing proximo-distal gradient was also present within the proximal endolymph between the sensory epithelium and the otolith. Further calcium particles covered the peripheral proteinacious layer of the otolith. They were especially pronounced at the proximal surface of the otolith. Other calcium precipitates were found to be accumulated at the macular junctions. These results strongly suggest that the apical region of the macular epithelium is involved in the release of calcium and that calcium supply of the otoliths takes place in the proximal endolymph.     

The relationship between ionospheric total electron content (TEC) over East Asia and the tropospheric circulation around the Qinghai-Tibet Plateau obtained with a partial correlation method

This paper reports a study on the relationship between ionospheric total electron content (TEC) over East Asia and the tropospheric circulation around the Qinghai-Tibet Plateau. Ionospheric TEC over East Asia are obtained from 25 observatories during 1996–2004. By applying a partial correlation method which can eliminate the influences of solar and geomagnetic activities, we find no significant correlation between TEC and the Asian zonal circulation index (Iz), but find a positive correlation between the day-to-day variability of TEC and Iz. We suggest that the positive correlation is closely related with the topography of the Qinghai-Tibet Plateau. The dynamical effect on airflow of the plateau can generate vortexes, and the vortexes may continuously excite internal gravity waves (IGWs) which transmit up to the ionosphere and cause regional wave disturbances. This study gives evidence for the dynamical mechanism of ionosphere–troposphere coupling and shows the importance of the Qinghai-Tibet Plateau in the ionosphere–troposphere coupling over East Asia.     

Ballooning perturbations in the inner magnetosphere of the Earth: Spectrum,stability and eigenmode analysis

We investigate the generation of ballooning perturbations in the inner magnetosphere of the Earth in the dipole model of the geomagnetic field taking into account ionospheric boundary conditions. The ionosphere is considered as a thin layer with finite conductivity. The eigenmode spectrum is discrete and consists of Alfvén, slow magnetosonic, flute and incompressible modes. Their interaction depends on ionospheric conductivity. The decay rate is small in noon and night sectors and large in dawn and dusk sectors. The lowest stability threshold α/γ ≈ 4.25 is determined by flute modes.     

Comparisons between primrose lake (54°N, 110°W) ROCOB winds (20–60 km) and Saskatoon (52°N, 107°W) M. F. radar winds (60–110 km): 1978–1982

The development of the new CIRA will require the combination of winds from many sources, e.g. rockets (ROCOB) up to ～60 km, and radar winds ～60–110 km. Difficulties are that such rocket data have larger errors at 60–65 km, and tidal effects may become significant. Radar data for 60–80 km may also have tidal contamination, due to ? 16h of data per day: from 80–110 km tidal corrections are usually reliable.Comparisons are made between the unique Saskatoon MF radar set, which is continuous from mid 1978–1983, and the ROCOB data from Primrose Lake, which is only 340 km northwest. While the agreement is satisfactory, special care is required when matching the two regions: particular problems are the low rocket sampling rate, and the unexpectedly large amplitude of the diurnal tide. Important differences from the zonal winds of CIRA-72 emerge, especially in winter months. Meridional cross-sections differ from previous data models in the extent of the summer equatorward flow.     

Development of space-fertilized eggs and formation of primordial germ cells in the embryos of Medaka fish.

In the second International Microgravity Laboratory (IML-2) mission in 1994, four small Japanese killifish (Medaka, Oryzias latipes) made a space travel of 15 days aboard a space shuttle. These four adult Medaka fish successfully mated in space for the first time among vertebrate animals. Moreover, the eggs they laid developed normally, at least in their external appearance, hatching as fry (baby fish) in space. Fish mated and laid eggs every day during the first week. Near the end of the mission most of the eggs had a well-developed body with two pigmented eyes. In total, 43 eggs were laid (detected), out of which 8 fry hatched in space, as truly 'space-originated' babies. A further 30 fry hatched within 3 days after landing. This is the normal hatching rate, compared with the ground-based data. Among the 8 space-originated fry, four were killed for histological sections, and germ cells at the gonadal region were counted for each fry. Their numbers were in the range of the germ cells of the normal control fry (ground-kept samples). Thus, as embryos developed normally in their external appearance, inside the embryos the formation of primordial germ cells took place normally in space, and their migration to the genital ridges was not hindered by microgravity. The two of the remaining space-originated fry have grown up and been creating their offspring in the laboratory. This proved that the primordial germ cells formed in space were also normal from a functional point of view. The four space-travelled adult fish re-started mating and laying eggs on the 7th day after landing and continued to do so every day afterward. Fertilization rate and hatchability of these eggs were as high as the eggs laid by the laboratory-kept fish. This fact implies that in gametogenesis of adult fish, there are no specific stages of germ cells extremely susceptible to microgravity.     

Probability of cell hits in selected organs and tissues by high-LET particles at the ISS orbit.

The fluence of high-LET particles (HLP) with LET infinity H2O greater than 15 keV micrometers-1 in selected organs and tissues were measured with plastic nuclear track detectors using a life-size human phantom on the 9th Shuttle-Mir Mission (STS-91). The planar-track fluence of HLP during the 9.8-day mission ranged from 1.9 x 10(3) n cm-2 (bladder) to 5.1 x 10(3) n cm-2 (brain) by a factor of 2.7. Based on these data, a probability of HLP hits to a matured cell of each organ or tissue was roughly estimated for a 90-day ISS mission. In the calculation, all cells were assumed to be spheres with a geometric cross-sectional area of 500 micrometers2 and the cell-hit frequency from isotropic space radiation can be described by the Poisson-distribution function. As results, the probability of one or more than 1 hit to a single cell by HLP for 90 days ranged from 17% to 38%; that of two or more than 2 hits was estimated to be 1.3-8.2%.     

Mean winds of the mesosphere and lower thermosphere (60–110 km): A global distribution from radar systems (MF,Meteor, VHF)

During the last decade a large number of radars (～12) have been developed, which have produced substantial quantities of tidally-corrected mean winds data. The distribution of the radars is not global, but many areas are well covered: the Americas with Poker Flat (65°N), Saskatoon (52°N), Durham (43°N), Atlanta (34°N), Puerto Rico (18°N); Europe with Kiruna (68°), Garchy (47°N) and Monpazier (44°N); and Oceania with Christchurch (44°S), Adelaide (35°S), Townsville (20°S), and Kyoto (35°N). Zonal and meridional wind height-time cross-sections from $\text{60}\text{80}$ km (MF/Meteor Radar) to ～110 km have been prepared for the last 5–6 years. They are compared with cross-sections from CIRA-72 for zonal winds, and Groves (1969) for meridional winds.It is shown that while CIRA-72 is still a useful model for many purposes, significant differences exist between it and the new radar data. The latter demonstrate important seasonal, latitudinal, longitudinal and hemispheric variations. The new meridional cross-sections are of great value. The common features with Groves (1969) are the equatorward cells in summer near 85 km; however their strength (～10 ms^?1) and size are less. Systematic and somewhat different variations emerge at higher (?52°N) and middle (35–44°) latitudes.     

The 16-day waves in the mesosphere and lower thermosphere over Wuhan (30.6°N, 114.5°E) and Adelaide (35°S, 138°E)

Winds from a meteor radar at Wuhan (30.6°N, 114.5°E) and a MF radar at Adelaide (35°S, 138°E) are used to study the 16-day waves in the mesosphere and lower thermosphere (MLT). The height range is 78–98 km at Wuhan and 70–98 km at Adelaide. By comparison, it is found that the zonal components at both sites are generally larger than the meridional ones, and eastward motion of the zonal background winds is favorable for the 16-day waves penetration to the MLT region. The zonal maximum amplitude appears in the autumn (September–October) around 86–98 km at Wuhan and in the winter months and early spring (July–October) around 72–82 km at Adelaide. Differences are found in wave amplitudes and time of appearance between the two years of 2002 and 2003. In 2003, the intensity of the wave amplitudes is relatively smaller than that for 2002 at both sites. The summer 16-day waves are comparatively weaker at Adelaide in both years, but stronger in 2002 at Wuhan near the mesopause and the lower thermosphere (86–98 km). The strong summer waves at Wuhan may come from the winter southern hemisphere.     

Role of shielding in modulating the effects of solar particle events: Monte Carlo calculation of absorbed dose and DNA complex lesions in different organs.

Distributions of absorbed dose and DNA clustered damage yields in various organs and tissues following the October 1989 solar particle event (SPE) were calculated by coupling the FLUKA Monte Carlo transport code with two anthropomorphic phantoms (a mathematical model and a voxel model), with the main aim of quantifying the role of the shielding features in modulating organ doses. The phantoms, which were assumed to be in deep space, were inserted into a shielding box of variable thickness and material and were irradiated with the proton spectra of the October 1989 event. Average numbers of DNA lesions per cell in different organs were calculated by adopting a technique already tested in previous works, consisting of integrating into "condensed-history" Monte Carlo transport codes--such as FLUKA--yields of radiobiological damage, either calculated with "event-by-event" track structure simulations, or taken from experimental works available in the literature. More specifically, the yields of "Complex Lesions" (or "CL", defined and calculated as a clustered DNA damage in a previous work) per unit dose and DNA mass (CL Gy-1 Da-1) due to the various beam components, including those derived from nuclear interactions with the shielding and the human body, were integrated in FLUKA. This provided spatial distributions of CL/cell yields in different organs, as well as distributions of absorbed doses. The contributions of primary protons and secondary hadrons were calculated separately, and the simulations were repeated for values of Al shielding thickness ranging between 1 and 20 g/cm2. Slight differences were found between the two phantom types. Skin and eye lenses were found to receive larger doses with respect to internal organs; however, shielding was more effective for skin and lenses. Secondary particles arising from nuclear interactions were found to have a minor role, although their relative contribution was found to be larger for the Complex Lesions than for the absorbed dose, due to their higher LET and thus higher biological effectiveness.     

Relationships between soil properties of the abandoned fields and spectral data derived from the advanced spaceborne thermal emission and reflection radiometer (ASTER)

The structural transformation of Polish agriculture after 1989 has been leading to significant changes in land use. As a result a large part of formerly ploughed fields lies abandoned and they occur across considerable variability in soil typological units. Accurate information about soil conditions within the abandoned fields facilitates proper management in the new socio-economic situation. Such information can be collected using satellite images since the structure and condition of the vegetation growing on the abandoned fields reflects soil properties. The objective of this study is to evaluate the relationships between physical and chemical attributes of soil within the abandoned fields and spectral reflectance patterns recorded by ASTER sensors onboard Terra Satellite. Soil samples were collected at five abandoned fields which have not been ploughed since 2000 and analyzed in the laboratory to determine their physical and chemical properties. Nine ASTER nearly cloud-free pictures were used for this study in order to derive the remote-sensing attributes of the abandoned fields. In order to evaluate the relationships between soil fertility and remotely sensed data, partial least-squares (PLS) and a multiple linear regression (MLR) analysis between these two datasets were carried out. In the regression analysis, only soil TEB (total exchangeable bases) stock in the whole profile displayed the highest correlation with remotely sensed data acquired in April and May and the best predictors were NDVI and LSWI vegetation indices.     

Effects of spaceflight (STS-87) on tropisms and plastid positioning in protonemata of the moss Ceratodon purpureus.

Apical cells of moss protonemata represent a single-celled system that perceives and reacts to light (positive and negative phototropism) and to gravity (negative gravitropism). Phototropism completely overrides gravitropism when apical cells are laterally irradiated with relatively high red light intensities, but below a defined light intensity threshold gravitropism competes with the phototropic reaction. A 16 day-long exposure to microgravity conditions demonstrated that gravitropism is allowed when protonemata are laterally illuminated with light intensities below 140 nmol m-2s-1. Protonemata that were grown in darkness in microgravity expressed an endogenous tendency to grow in arcs so that the overall culture morphology resembled a clockwise spiral. However this phenomenon only was observed in cultures that had reached a critical age and/or size. Organelle positioning in dark-grown apical cells was significantly altered in microgravity. Gravisensing most likely involves the sedimentation of starch-filled amyloplasts in a well-defined area of the tip cell. Amyloplasts that at 1-g are sedimented were clustered at the apical part of the sedimentation zone in microgravity. Clustering observed in microgravity or during clino-rotation significantly differs from sedimentation-induced plastid aggregations after inversion of tip cells at 1-g.     

A preliminary study of the single crest phenomenon in total electron content (TEC) in the equatorial anomaly region around 120°E longitude between 1999 and 2012

The diurnal variations in total electron content (TEC) in the equatorial ionisation anomaly (EIA) region are not always represented by two crests on both sides of the magnetic equator. Sometimes, only an obvious single crest is evident at equatorial and low latitudes. In this paper, we focus on analysis of the morphological features of the single crest phenomenon in TEC around 120°E longitude during geomagnetic quiet days (Kp < 4₋). The variations in TEC are also compared with morphological parameters (foF2 and hmF2) derived from the International Reference Ionosphere extended to Plasmasphere (IRI–Plas) model. Our results show that the single crest phenomenon occurs mainly on days with extremely low solar activity, while the corresponding F2 layer critical frequency showed obvious asymmetry, or even only a single peak.     

Accumulation of stress protein 72 (HSP72) in muscle and spleen of goldfish taken into space.

Using Western blot analysis, here, we report the levels of HSP72 in several organs from goldfish which were taken into space on the NASA space shuttle. A remarkable accumulation of HSP72 was detected in muscle and spleen of those fish taken into space as compared with controls. These results suggested that the HSP72 induction is a kind of stress response at the molecular level introduced by the space environment consisting of microgravity and/or cosmic radiation as stressors.