A New Versatile System for Freeze-Substitution, Freeze-Drying and Low Temperature Embedding of Biological Specimens

A universal system for freeze-substitution (FS), freeze-drying (FD) and low temperature embedding (LTE) has been developed, suited to perform standardized procedures of cryoprocessing biological and medical specimens as well as systematic studies of dehydration and embedding at various low and high temperatures. In a 35 I Dewar vessel with 110 mm neck diameter an aluminum tube is mounted to the bottom of the liquid nitrogen (LN2x) reservoir and extends to the lower part of the cylindrical neck. At its top an aluminum plate serves as a contact surface for either the FS chamber or the FD chamber. FS and subsequent LTE are carried out in an environment of dry cold nitrogen gas provided by evaporating nitrogen from the dewar. Different capsules and moulds may be used for cryodehydration and LTE. FD of bulk specimens or cryosections takes place in an absolutely clean vacuum provided by a cryosorption pump integrated in the FD apparatus. Most of the Hp molecules from the frozen specimen are trapped by large cold surfaces inside the drying chamber. Due to the low LN2 consumption during FS or FD (3-4 l LN2/day) both procedures may be carried out for 8-10 days without refilling the dewar. A few representative results show that well frozen biological material is stabilized by prolonged FS or FD at temperatures of about -80°C without use of chemical fixatives like OsO4 in the substitution medium during FS or by OsO4 vapor fixation after FD

A protosolar nebula origin for the ices agglomerated by Comet 67P/Churyumov-Gerasimenko

The nature of the icy material accreted by comets during their formation in the outer regions of the protosolar nebula is a major open question in planetary science. Some scenarios of comet formation predict that these bodies agglomerated from crystalline ices condensed in the protosolar nebula. Concurrently, alternative scenarios suggest that comets accreted amorphous ice originating from the interstellar cloud or from the very distant regions of the protosolar nebula. On the basis of existing laboratory and modeling data, we find that the N$_2$/CO and Ar/CO ratios measured in the coma of the Jupiter family comet 67P/Churyumov-Gerasimenko by the ROSINA instrument aboard the European Space Agency's Rosetta spacecraft match those predicted for gases trapped in clathrates. If these measurements are representative of the bulk N$_2$/CO and Ar/CO ratios in 67P/Churyumov-Gerasimenko, it implies that the ices accreted by the comet formed in the nebula and do not originate from the interstellar medium, supporting the idea that the building blocks of outer solar system bodies have been formed from clathrates and possibly from pure crystalline ices. Moreover, because 67P/Churyumov-Gerasimenko is impoverished in Ar and N$_2$, the volatile enrichments observed in Jupiter's atmosphere cannot be explained solely via the accretion of building blocks with similar compositions and require an additional delivery source. A potential source may be the accretion of gas from the nebula that has been progressively enriched in heavy elements due to photoevaporation.Comment: The Astrophysical Journal Letters, in pres

Vitamin D receptor distribution in intestines of domesticated sheep Ovis ammon f. aries

The biologically active form of vitamin D, i.e., 1,25-dihydroxycholecalciferol or calcitriol, plays an important role in bone metabolism and calcium homeostasis, which is often disturbed at the onset of lactation in high milk-yielding domestic ruminants. Gene transcription is modulated via vitamin D receptors, but nongenomic effects of vitamin D via membrane receptors have also been described. In the intestines, vitamin D promotes calcium absorption via vitamin D receptors. Vitamin D receptors are of clinical relevance, but have not been systematically assessed within all segments of the intestine in any species. Thus, we present for the first time an immunohistochemical study of the distribution patterns of the vitamin D receptor protein in sheep, which may be the basis for present and future investigations on mineral homeostasis in domestic ruminants. Tissue probes of the intestines were collected from five lambs and five nonlactating and nonpregnant dams, fixed in formalin, embedded in paraffin, and used for the assessment of vitamin D receptor protein. Nuclear vitamin D receptor immunoreaction was scored semiquantitatively and exhibited a segment-specific distribution pattern. Goblet cells always were devoid of any vitamin D receptor immunoreaction. Surface epithelial cells and enterocytes of the crypt openings generally demonstrated only a weak immunoreaction. Basally and/or intermediately located crypt epithelial cells exhibited stronger immunoreactions in duodenum, jejunum, and colon descendens. This basal/intermediate to superficial gradient was most pronounced in the duodenum and less evident in jejunum and colon descendens and not observed in ileum and cecum. There were no age-dependent variations in vitamin D receptor protein expression. Results demonstrate that intestinal vitamin D receptor distribution patterns are segment-specific and strongest immunoreactions correlate with highest intestinal calcium absorptive activities, as reported in literature. Strong expression of vitamin D receptors within the lower half of crypts also suggests a role for calcitriol in epithelial differentiation and cellular homeostasis

The Rosetta Ion and Electron Sensor (IES)measurement of the development of pickup ions from comet 67P/Churyumov-Gerasimenko

The Rosetta Ion and Electron Sensor (IES) has been measuring solar wind ions intermittently since exiting from hibernation in May 2014. On 19 August, when Rosetta was ~80 km from the comet 67P/Churyumov-Gerasimenko, which was ~3.5 AU from the Sun, IES began to see ions at its lowest energy range, ~4–10 eV. We identify these as ions created from neutral species emitted by the comet nucleus, photoionized by solar UV radiation in the neighborhood of the Rosetta spacecraft (S/C), and attracted by the small negative potential of the S/C resulting from the population of thermal electrons. Later, IES began to see higher-energy ions that we identify as having been picked up and accelerated by the solar wind. IES continues to measure changes in the solar wind and the development of the pickup ion structure

Messung von Herzfrequenzen und Kotcortisolmetaboliten bei Pferden am Zürcher Sechseläuten

The Zurich’s Sechseläuten is a traditional festival of the Zurich guilds, in which around 500 horses take part. After a parade through the old town of Zurich, the riders gather at a big square to canter around a burning woodpile topped with an exploding effigy (the «Böögg»). The level of stress experienced by the horses partaking in this event is subjected to increasing scrutiny. The aim of this study was to evaluate the stress load of the horses participating in the Sechseläuten procession more objectively by measuring heart rate and faecal cortisol metabolites. Twenty-three horse-rider pairs were voluntarily recruited from the guilds for participation in the study. For comparison purposes, three sections of the procession were simulated in terms of gait and distance travelled in a normal riding environment during a test day and observed during the Sechseläuten itself 16 days later: Counter march Bahnhofstrasse, parade Limmatquai and parade around the burning «Böögg». All horses were equipped with a heart rate measuring system with GPS tracking (Polar Team Pro®, Polar Electro Oy, Finland). Faecal samples for measuring cortisol metabolites were taken rectally immediately before and 24 hours after the events. Thirteen of the 23 horses were sedated during the Sechseläuten (Acepromazine, 0,1–0,2 mg/kg p.o.). The differences between the observation periods and groups (sedated/unsedated) were tested by analysis of variance (P<0,05). Walking and cantering speeds in the three sections did not differ between the test day and the Sechseläuten itself. Compared to the test day, the average heart rate was increased in all sections of the real event. However, neither the initial cortisol metabolite value nor the increase after the events showed a significant difference between the test day and the Sechseläuten. The average heart rate of the sedated group did not differ from the unsedated group during the walking sections but was elevated during canter around the Böögg. The increase in heart rate at the Sechseläuten indicated a stress load, but together with the unchanged cortisol responses, this can be classified as moderate. The influence of a sedation were inconclusive and should be further investigated

Immunohistochemical demonstration of cyclooxygenase-2 (COX-2) and prostaglandin receptors EP2 and FP expression in the bovine intercaruncular uterine wall around term

During parturition, uterine-derived prostaglandins (PG) play an outstanding role regarding the functional elimination of the corpus luteum and the promotion of uterine contraction. The rate-limiting enzyme cyclooxygenase-2 (COX-2), highly regulated in a cell-type and localization specific manner throughout pregnancy, is involved in uterine prostanoid production. Prostaglandins exert their effects via G-protein-coupled receptors. Distribution and cellular localization of these receptors are decisive factors for prostaglandin-mediated actions. Since both COX-2 and PG receptors have only been assessed during pregnancy in the cow, these parameters were localized immunohistochemically near term to evaluate their specific role at parturition. Thus, during two periods, segments of the intercaruncular uterine wall were collected from cows at slaughter being eight and nine months pregnant, from cattle during caesarean section, and after spontaneous calving. Results reveal that COX-2 was mainly localized in the cytoplasm of surface epithelial cells with a high expression in animals with induced parturition. The enzyme could also be found in lower concentrations within the glandular epithelium without any effect of gestational time or labour. In contrast to relaxant prostaglandin E receptor type 2 (EP2), not showing any change in all tissue layers observed, contractile prostaglandin F(2alpha) receptor (FP) was modulated during the peripartal period revealing a peak expression in animals with induced parturition. FP was localized in surface and glandular epithelial cells as well as in endometrial stroma and myometrial smooth muscle cells. Our study indicates that labour and induction of parturition may have an effect on amounts of immunohistochemically detectable COX-2 and FP. EP2 remains rather unchanged during the peripartal period. COX-2 and FP thus contribute via changes in amount and distribution to mechanisms associated with parturition

Organics in comet 67P – a first comparative analysis of mass spectra from ROSINA–DFMS, COSAC and Ptolemy

The ESA Rosetta spacecraft followed comet 67P at a close distance for more than 2 yr. In addition, it deployed the lander Philae on to the surface of the comet. The (surface) composition of the comet is of great interest to understand the origin and evolution of comets. By combining measurements made on the comet itself and in the coma, we probe the nature of this surface material and compare it to remote sensing observations. We compare data from the double focusing mass spectrometer (DFMS) of the ROSINA experiment on ESA's Rosetta mission and previously published data from the two mass spectrometers COSAC (COmetary Sampling And Composition) and Ptolemy on the lander. The mass spectra of all three instruments show very similar patterns of mainly CHO-bearing molecules that sublimate at temperatures of 275 K. The DFMS data also show a great variety of CH-, CHN-, CHS-, CHO2- and CHNO-bearing saturated and unsaturated species. Methyl isocyanate, propanal and glycol aldehyde suggested by the earlier analysis of the measured COSAC spectrum could not be confirmed. The presence of polyoxymethylene in the Ptolemy spectrum was found to be unlikely. However, the signature of the aromatic compound toluene was identified in DFMS and Ptolemy data. Comparison with remote sensing instruments confirms the complex nature of the organics on the surface of 67P, which is much more diverse than anticipated

Influence of spacecraft outgassing on the exploration of tenuous atmospheres with in situ mass spectrometry

Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/94765/1/jgra20796.pd

First detection of a diamagnetic cavity at comet 67P/Churyumov-Gerasimenko

International audienceContext: The Rosetta magnetometer RPC-MAG has been exploring the plasma environment of comet 67P/Churyumov-Gerasimenko since August 2014. The first months were dominated by low-frequency waves which evolved into more complex features. However, at the end of July 2015, close to perihelion, the magnetometer detected a region that did not contain any magnetic field at all.Aims: These signatures match the appearance of a diamagnetic cavity as was observed at comet 1P/Halley in 1986. The cavity here is more extended than previously predicted by models and features unusual magnetic field configurations, which need to be explainedMethods: The onboard magnetometer data were analyzed in detail and used to estimate the outgassing rate. A minimum variance analysis was used to determine boundary normals.Results. Our analysis of the data acquired by the Rosetta Plasma Consortium instrumentation confirms the existence of a diamagnetic cavity. The size is larger than predicted by simulations, however. One possible explanation are instabilities that are propagating along the cavity boundary and possibly a low magnetic pressure in the solar wind. This conclusion is supported by a change in sign of the Sun-pointing component of the magnetic field. Evidence also indicates that the cavity boundary is moving with variable velocities ranging from 230−500 m/s