Carbon dioxide is Mars' primary atmospheric constituent and is an active driver of Martian surface evolution. CO2 ice sublimation mechanisms have been proposed for a host of features that form in the contemporary Martian climate. However, there has been very little experimental work or quantitative modelling to test the validity of these hypotheses. Here we present the results of the first laboratory experiments undertaken to investigate if the interaction between sublimating CO2 ice blocks and a warm, porous, mobile regolith can generate features similar in morphology to those forming on Martian dunes today. We find that CO2 sublimation can mobilise grains to form (i) pits and (ii) furrows. We have documented new detached pits at the termini of linear gullies on Martian dunes. Based on their geomorphic similarity to the features observed in our laboratory experiments, and on scaling arguments, we propose a new hypothesis that detached pits are formed by the impact of granular jets generated by sublimating CO2. We also study the erosion patterns formed underneath a sublimating block of CO2 ice and demonstrate that these resemble furrow patterns on Mars, suggesting similar formation mechanisms.
Time of Flight secondary ion mass spectrometry (TOF-SIMS) has been used to explore the distribution of phospholipids in the plasma membrane of Tetrahymena pyriformis during cell division. The dividing cells were freeze dried prior to analysis followed by line scan and region of interest analysis at various stages of cell division. The results showed no signs of phospholipid domain formation at the junction between the dividing cells. Instead the results showed that the sample preparation technique had a great impact on one of the examined phospholipids, namely phosphatidylcholine (PC). Phosphatidylcholine and 2-aminoethylphosphonolipid (2-AEP) have therefore been evaluated in Tetrahymena cells that have been subjected to different sample preparation techniques: freeze drying ex situ, freeze fracture, and freeze fracture with partial or total freeze drying in situ. The result suggests that freeze-drying ex situ causes the celia to collapse and cover the plasma membrane.
In spray freeze drying (SFD) solutions are frozen by spraying into a very cold environment and subsequently dried by sublimation. In contrast to conventional freeze drying, spray freeze drying has the possibility to produce flowable lyophilizates which offers a variety of new pharmaceutical applications. Here, a drop jet nozzle is proposed as liquid dispenser that is able to produce droplets with a very narrow size distribution compared to standard methods. The drop jet nozzle is mounted in a spray tower designed to prevent direct contact of the product with the freezing medium. Various formulations have been tested containing lysozyme as model protein and stabilizers such as bovine serum albumin, polyvinylpyrrolidone or dextran in various concentrations and mannitol. Excellent free flowing and nearly monodispersed, porous particles are produced where particle properties can be controlled by formulation and process conditions. The particle diameter varied between 231±3μm and 310±10μm depending on the formulation composition. The lysozyme activity was >94±5% for all formulations exhibiting a full preservation of enzyme activity. This new method is very promising for the production of nearly monodisperse particulate lyophilizates in various therapeutic applications.
Polymeric micelles were studied as delivery carriers of diazepam, a practically insoluble drug in water, for rectal administration. The diazepam-loaded polymeric micelles were developed by using poloxamer 407 (P407), poloxamer 188, and D-α-tocopheryl poly(ethylene glycol) 1000 succinate (TPGS). Among the used polymers, TPGS resulted in polymeric micelles with good characteristics for encapsulation of diazepam which had the small particle size of 8-12 nm and narrow size distribution (PI 0.053-0.275). Additionally, 7.5% w/v of TPGS could entirely entrap the desired concentration of diazepam (5 mg/mL). To improve the physical stability upon lyophilization, an addition of P407 of 1% w/v prevented aggregation, increased physical stability, and maintained chemical stability of the lyophilized powders of diazepam-loaded polymeric micelles for 3 months storage at 4°C. The rate and amount of diazepam release from TPGS polymeric micelles mainly depended on the concentration of TPGS. The release data were fitted to Higuchi’s model suggesting that the drug release mechanism was controlled by Fickian diffusion. In conclusion, 10% w/v TPGS and 1% w/v P407 were the optimum formulation of lyophilized diazepam-loaded polymeric micelles.
As liquid liposomal formulations are prone to chemical degradation and aggregation, these formulations often require freeze drying (e.g. lyophilization) to achieve sufficient shelf-life. However, liposomal formulations may undergo oxidation during lyophilization and/or during prolonged storage. The goal of the current study was to characterize the degradation of 1, 2-dilinolenoyl-sn-glycero-3-phosphocholine (DLPC) during lyophilization, and to also probe the influence of metal contaminants in promoting the observed degradation. Aqueous sugar formulations containing DLPC (0.01 mg/ml) were lyophilized, and DLPC degradation was monitored using HPLC/UV and GC/MS methods. The effect of ferrous ion and sucrose concentration, as well as lyophilization stage promoting lipid degradation, was investigated. DLPC degradation increased with higher levels of ferrous ion. After lyophilization, 103.1% ± 1.1%, 66.9% ± 0.8%, and 28.7% ± 0.7% DLPC remained in the sucrose samples spiked with 0.0 ppm, 0.2 ppm and 1.0 ppm ferrous ion, respectively. Lipid degradation predominantly occurs during the freezing stage of lyophilization. Sugar concentration and buffer ionic strength also influence the extent of lipid degradation, and DLPC loss correlated with degradation product formation. We conclude that DLPC oxidation during the freezing stage of lyophilization dramatically compromises the stability of lipid-based formulations. In addition, we demonstrate that metal contaminants in sugars can become highly active when lyophilized in the presence of a reducing agent.
The aims of this study were to determine the stability of Podoviridae coliphage CA933P during lyophilization and storage in different media, and to establish similarities between the results obtained and those expected through mechanisms described for proteins stabilization during freeze-drying. PBS and SM buffer were assayed as lyophilization media. The effect of inorganic salts concentration as well as the addition of disaccharides on phage stability during freeze-drying and storage was also studied. The addition of low sucrose concentration (0.1 mol l(-1)) to SM buffer stabilized phage during freezing and drying steps of the lyophilization process, but higher sugar concentrations were detrimental to phage stability during freeze-drying. Sucrose stabilized phage during storage for at least 120 days. The lyoprotective effect of low concentrations of disaccharides during the drying step of the lyophilization of proteins as well as the stabilization of the freeze-dried product in time correlated with the results obtained for phage CA933P.
Micellar electrokinetic capillary chromatography with electrochemical detection has been used to quantify biogenic amines in freeze-dried Drosophila melanogaster brains. Freeze drying samples offers a way to preserve the biological sample while making dissection of these tiny samples easier and faster. Fly samples were extracted in cold acetone and dried in a rotary evaporator. Extraction and drying times were optimized in order to avoid contamination by red-pigment from the fly eyes and still have intact brain structures. Single freeze-dried fly-brain samples were found to produce representative electropherograms as a single hand-dissected brain sample. Utilizing the faster dissection time that freeze drying affords, the number of brains in a fixed homogenate volume can be increased to concentrate the sample. Thus, concentrated brain samples containing five or fifteen preserved brains were analyzed for their neurotransmitter content, and five analytes; dopamine N-acetyloctopamine, N-acetylserotonin, N-acetyltyramine, N-acetyldopamine were found to correspond well with previously reported values.
Sperm preservation is an important technique for maintaining valuable genetic resources in biomedical research and wildlife. In the mouse, the sperm cryopreservation method has been established and adopted by large-scale sperm preservation projects in cryobanks. Recently, a new sperm preservation method using freeze-drying has been studied in various mammals. Freeze-drying is the ultimate method by which sperm can be preserved long term in a refrigerator (4 °C). And it is possible to realize easy and safe transportation of sperm at an ambient temperature that requires neither liquid nitrogen nor dry ice. Furthermore, it has been demonstrated that the fertilizing ability of sperm cryopreserved or freeze-dried by the methods described in this chapter is well maintained during long-term preservation. This chapter introduces the latest protocols for cryopreservation and freeze-drying of mouse sperm, and the anticipated results of the fertilizing ability of these sperm preserved long-term.
Carbon dioxide is one of the most abundant species in cometary nuclei, but due to its high volatility CO2 ice is generally only found beneath the surface. We report the infrared spectroscopic identification of a CO2 ice-rich surface area, located in the Anhur region of comet 67P/Churyumov-Gerasimenko. Spectral modeling shows that about 0.1% of the 80×60 m area is CO2 ice. This exposed ice was observed a short time after exiting from local winter; following the increased illumination, the CO2 ice completely disappeared over about three weeks. We estimate the mass of the sublimated CO2 ice and the depth of the surface eroded layer. The presence of CO2 ice is interpreted as the result of the extreme seasonal changes induced by the rotation and orbit of the comet.
The equimolar C2 H2 -CO2 reaction has shown promise for carbon nanotube (CNT) production at low temperatures and on diverse functional substrate materials; however, the electron-pushing mechanism of this reaction is not well demonstrated. Here, the role of CO2 is explored experimentally and theoretically. In particular, 13 C labeling of CO2 demonstrates that CO2 is not an important C source in CNT growth by thermal catalytic chemical vapor deposition. Consistent with this experimental finding, the adsorption behaviors of C2 H2 and CO2 on a graphene-like lattice via density functional theory calculations reveal that the binding energies of C2 H2 are markedly higher than that of CO2 , suggesting the former is more likely to incorporate into CNT structure. Further, H-abstraction by CO2 from the active CNT growth edge would be favored, ultimately forming CO and H2 O. These results support that the commonly observed, promoting role of CO2 in CNT growth is due to a CO2 -assisted dehydrogenation mechanism.