Cryo-container

Medical Gases can be produced in bulk and delivered in special customized packages (compressed gas cylinders of different sizes, compressed gas bimdles, as refrigerated liquid) in mobile cryo-containers or in stationary cryo-tanks including evaporator. In addition, a large number of gas mixtures are individually produced by medical institutes according to specific orders and, due to lacking authorizations, delivered as specific pharmaceutical. 

Fig. 10 (a) Chemical structure of PEG-6-PCL copolymer, (b) CLSM image of PEG-6-PCL polymersomes containing membrane-encapsulated Nile Red (2 mol%) and aqueous entrapped Calcein dyes. Scale bar 5 pm. (c) Cryo-TEM image of PEG-6-PCL polymersomes. Scale bar 100 nm. Reprinted from [228] with permission... 

The more recently developed cryo-TEM technique has started to be used with increasing frequency for block copolymer micelle characterization in aqueous solution, as illustrated by the reports of Esselink and coworkers [49], Lam et al. [50], and Talmon et al. [51]. It has the advantage that it allows for direct observation of micelles in a glassy water phase and accordingly determines the characteristic dimensions of both the core and swollen corona provided that a sufficient electronic contrast is observed between these two domains. Very recent studies on core-shell structure in block copolymer micelles as visualized by the cryo-TEM technique have been reported by Talmon et al. [52] and Forster and coworkers [53]. In a very recent investigation, cryo-TEM was used to characterize aqueous micelles from metallosupramolecular copolymers (see Sect. 7.5 for further details) containing PS and PEO blocks. The results were compared to the covalent PS-PEO counterpart [54]. Figure 5 shows a typical cryo-TEM picture of both types of micelles. 

Fig. 20 Cryo-TEM image of isolated micelles and segmented rods made from miktoarm ABC block copolymers containing PEO, PB and perfluorinated polyether blocks (scale bar is 50 nm). A schematic representation of the stacking of block copolymer chains in segmented worms is also shown. Reprinted with permission from [284], Copyright (2004) American Association for the Advancement of Science...

The structure determination proceeded in steps starting with separate structural analyses of the component protein VP7. Information from a 22 A resolution cryo-EM reconstruction of the viral core (Grimes et al., 1997) was then combined with the high-resolution atomic structure for the VP7(T13) trimer (Grimes et al., 1995) to yield a model providing phase information to a higher resolution than the EM reconstruction alone. Infectious BTV-1 crystals (unit cell dimensions a = 796 A,b = 822 A, and c = 753 A), containing half a particle in the asymmetric unit. 

Accurate selection of translation termination factors to ribosomes containing a stop codon in the A-site is less well understood. A picture is only beginning to emerge as the bacterial 708 ribosome and class I release factor RF2 atomic models have recently been fitted into cryo-EM stmctures. Via multiple interactions RF2 connects the ribosomal decoding site with the PTC and functionally mimics a tRNA molecule in the A-site. In the complex RF2 is close to helices 18, 44, and 31 of the 168 rRNA, small subunit ribosomal protein 812, helices 69, 71, 89, and 92 of the 238 rRNA, the L7/L12 stalk, and protein LI 1 of the large subunit (Arkov et al. 2000 Klaholz et al. 2003 Rawat et al. 2003). The L7/L12 stalk inter-... 

Visualization of the bacterial RF3 on the ribosome by cryo-EM revealed that RE3 could be bound in two different conformational states associated with significant conformational changes within the ribosome (Klaholz et al. 2004). In state 1 ribosomes contain a tRNA in the P-site and RE3 is loosely associated. In state 2 the tRNA has moved to the E-site and RF3 is more tightly attached to the ribosomes (Klaholz et al. 2004). How eRF3 structurally influences eREl and the eukaryotic ribosome awaits future investigation. 

Figure 10.10 Transmission electron micrograph of ferritin entrapped in POPC liposomes (palmitoyloleoylphosphatidylcholine). Cryo-TEM micrographs of (a) ferritin-containing POPC liposomes prepared using the reverse-phase evaporation method, followed by a sizing down by extrusion through polycarbonate membranes with 100 nm pore diameters ([POPC] = 6.1 mM) and (b) the vesicle suspension obtained after addition of oleate to pre-formed POPC liposomes ([POPC] = 3 mM, [oleic acid - - oleate] = 3 mM). (Adapted from Berclaz et al, 2001a, b.)...

Figure 10.12 Number-weighted size distributions as obtained by cryo-TEM (adapted from Berclaz et al, 2001a, b). (a) Distribution for the pre-formed POPC vesicles ([POPC] = 1.9 mM). (b) Distribution for the vesicle suspension obtained upon addition of oleate to pre-formed ferritin-containing POPC vesicles ([POPC] = 0.2 mM [oleic acid -I- oleate] = 5 mM). Empty ( ) and ferritin-containing ( ) vesicles are represented individually in the histogram, (c) Direct comparison of the number-weighted size distribution of the pre-formed POPC vesicles, which contained at least one ferritin molecule ( ) with the number-weighted size distribution of the ferritin-containing vesicles obtained after oleate addition to pre-formed POPC vesicles ( ). Note that the total of all ferritin-containing vesicles was set to 100%.

Percutaneous penetration of 7V-nitrosodiethanolamine was measured using cryo-preserved human trunk skin and three vehicle formulations (isopropyl myristate, sunscreen cream or a 10% shampoo) containing 7V-nitroso[ C]diethanolamine. The absorption rate of a low dermal dose (10 ixg/cm ) of 7V-nitrosodiethanolamine was a linear function of the concentration (0.06, 0.2 or 0.6 Xg/ xL) applied to the skin. The peak rates for the isopropyl m uistate and shampoo vehicles were seen within five hours and for the sunscreen somewhat later. Total 48-h absorption ranged from 35 to 65% of the dose and was formulation-dependent (isopropyl m uistate > shampoo > sunscreen). A total absorption of 4-6 x JcaE was estimated to equate to an applied N-nitrosodiethanolamine dose of 10 x%lcaE. When applied as a large infinite dose (0.5 mg/cm ), total 7V-nitrosodiethanolamine absorption (4-35% of the applied dose) followed a different rank order (shampoo > isopropyl m uistate > sunscreen), probably due to the barrier-damaging properties of the vehicles. The permeability coefficient for isopropyl myristate was 3.5 X 10 cm/h (Franz etal., 1993). 

Figure 29-4 Structure of 23S-28S ribosomal RNAs. (A) The three-dimensional structure of RNA from the 50S subunit of ribosomes of Halocirculci marismortui. Both the 5S RNA and the six structural domains of the 23S RNA are labeled. Also shown is the backbone structure of protein LI. From Ban et al.17 Courtesy of Thomas A. Steitz. (B) The corresponding structure of the 23S RNA from Thermus thermophilus. Courtesy of Yusupov et al.33a (C) Simplified drawing of the secondary structure of E. coli 23S RNA showing the six domains. The peptidyltransferase loop (see also Fig. 29-14) is labeled. This diagram is customarily presented in two halves, which are here connected by dashed lines. Stem-loop 1, which contains both residues 1 and 2000, is often shown in both halves but here only once. From Merryman et al.78 Similar diagrams for Haloarcula marismortui17 and for the mouse79 reveal a largely conserved structure with nearly identical active sites. (D) Cryo-electron microscopic (Cryo-EM) reconstruction of a 50S subunit of a modified E. coli ribosome. The RNA has been modified genetically to have an...

Cryo 1°C freezing containers (cat. no. 5100 Nalgene, Nalge Europe Ltd, Rotherwas, Hereford, UK). 

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