High-resolution magic-angle spinning H nuclear magnetic resonance studies of lipid dispersions using spherical glass ampoules

A very useful high-resolution magic-angle spinning (MAS) ¹H NMR method for studying lipid dispersions is presented. The sample can be loaded into the spherical glass ampoule very easily, and a spinning speed of more than 10 kHz can be achieved without the problems of sample leakage or water loss. The line width at half height for the HDO peak is less than 1.5 Hz, and the method can be implemented by anyone who has access to a solid-state MAS NMR spectrometer. By using the spherical glass ampoule method we found two water peaks, which could be ascribed to bulk water outside of the multilamellar liposome (peak at high frequency) and interlamellar water (peak at low frequency), in both POPC (1-palmitoyl-2-oleoyl-sn-glycero-3-phosphocholine) and DOPC (1,2-dioleoyl-sn-glycero-3-phosphocholine) liposomes. These are the first two examples of lipids without exchangeable protons that exhibit two distinct water resonances. The respective ¹H spin-lattice relaxation times (T₁) were measured, yielding values twice as long for bulk water as compared with interlamellar water. Both the chemical shift and spin relaxation results demonstrate the ability of MAS ¹H NMR to rapidly monitor changes in physical properties that accompany water interactions with zwitterionic phosphatidylcholines.     

On the measurement of carbon dioxide uptake with a glass electrode

1. It has been shown that the glass electrode method as previously described by the author for measurements of transient rates in photosynthesis is free of systematic errors. 2. Justification has been given for the use of the steady state-gas flow method for measuring rates of assimilation under conditions in which the solution is known not to be seriously depleted of CO₂. 3. The decline of photosynthetic rates under light saturation at CO₂ pressures less than several tenths per cent of an atmosphere has been shown to be a real phenomenon. It has been suggested that there may be a real discrepancy between this finding and those of some other investigators due to a difference in the previous history of the algal suspensions.     

On the active streaming experiment of actomyosin solutions in glass microcapillaries.

The active-streaming experiments of Oplatka et al. (Oplatka, A. and Tirosh, R. (1973) Biochim. Biophys. Acta 305, 684-688 and Oplatka, A. Gadasi, H., Tirosh, R. Lamed, Y., Muhlrad, A. and Liron, N. (1974) J. Mechanochem. Cell Motil. 2, 295-306) with actomyosin solution in a glass microcapillary is reexamined under various conditions with several kinds of reference material. It is found that vigorous streaming took place in the actomyosin solution as reported by Oplatka et al. However, streaming which is indistinguishable from that observed in the actomyosin solution in the presence of actomyosin ATPase activity also occurred, even when the ATPase activity was blocked. The streaming also cannot be confirmed as being active when using acto-heavy meromyosin solution. There is a possibility that the streaming experiment provides interesting information on the microscopic state of solutions which is not directly related to the chemo-mechanical conversion.     

Amplification of spontaneous emission of neon-like argon in a fast gas-filled capillary discharge

The evolution of the CAPEX facility and its basic diagnostics are described. The experiments carried out in the last modification of this facility accomplished with the demonstration of amplified spontaneous emission of neon-like argon (Ar⁸⁺) at the wavelength 46.88 nm. The first version of the facility, CAPEX1, operated with a plastic capillary and had a short high-power passive prepulse and an imperfect gas-filling system. In the second version, CAPEX2, a ceramic capillary was used, the prepulse amplitude was lowered, and the gas-filling system was improved. In the third, most successful version, CAPEX3, the capillary bending was reduced, a longer external prepulse was used, and the gas-filling system was further optimized. For each version, results of X-ray measurements are presented and interpreted.     

Hard shell gas-filled contrast enhancement particles for colour Doppler ultrasound imaging of tumors

Hollow hard shell particles of 200 nm and 2 micron diameter with a 10 nm thick porous silica shell have been synthesized using polystyrene templates and a sol-gel process. The template ensures than the hollow particles are monodispersed, while the charged silica surface ensures that they remain suspended in solution for weeks. When filled with perfluorocarbon gas, the particles behave as an efficient contrast agent for colour Doppler ultrasound imaging in human breast tissue. The silica shell provides unique properties compared to conventional soft shell particles employed as ultrasound contrast agents: uniform size control, strong adsorption to tissue and cells immobilizing particles at the tissue injection site, a long imaging lifetime, and a silica surface that can be easily modified with biotargeting ligands or small molecules to adjust the surface charge and polarity.     

Meiotic recombination: sealing the partnership at the junction

Crossovers ensure proper chromosome segregation in meiosis. A heterodimer of MutS proteins, hMSH4-hMSH5, has recently been found to interact with recombination intermediates in a manner that suggests a mechanism for directing meiotic DNA double strand break repair towards a crossover pathway.     

A new method to detect the presence of continuous gas-filled pathways for oxygen diffusion in legume nodules

A new method is presented which evaluates the contribution ofgas-filled and water-filled pathways across the barrier whichrestricts O₂ diffusion into infected cells of nodules. SinceO₂ will move through air about 104 times faster than throughwater any continuous gas-filled pathways which traverse thecortex would form a major route for O₂ transport. However, microscopicevidence for the existence of such direct connections is ambiguous. On theoretical grounds, O₂ should diffuse through a He atmosphereabout 3.7 times faster than through air or Ar, but O₂ flux acrossa liquid barrier should be unaffected by changes in the backgroundmixing gas. Thus, if O₂ influx to the infected cells is increasedwhen the ambient gas phase is changed from air to He/21% O₂this is evidence for a continuous pathway of gas-filled poresacross the whole width of the cortical barrier. This theoreticalapproach was validated by measuring the rates of diffusion ofO₂ through milli-pore filters in background atmospheres of eitherair, Ar or He. These membranes were used dry, to simulate the‘open pore’ model for nodule diffusion, or wettedwith water or gum Arabic (which is similar to the glycoproteinsassociated with the barrier) to simulate the ‘closed pore’situation. Knowing the diffusion constants for the gas- and water-filledpathways that may be involved in the nodule barrier, we thenevaluated the contribution of these components to the totaldiffusion resistance by examining the effect of Ar/O₂ and He/O₂gas mixtures on H₂ production and respiration of nodules. Theresults indicate that, in unstressed soyabean nodules, abouthalf of the O₂ flux to infected cells is via inter-connectedgas-filled pores, which ‘close’ to produce a liquid-filledbarrier as the diffusion resistance increases in response tostress. In unstressed lupin nodules all of the O₂ flux crossesa liquid-filled barrier. Key words: Oxygen diffusion, nodules, nitrogen fixation     

Quantifying the cleanliness of glass capillaries

I used capillary rise methods to investigate the lumenal surface properties of quartz (fused silica, Amersil T-08), borosilicate (Corning 7800), and high-lead glass (Corning 0010) capillaries commonly used to make patch pipets. I calculated the capillary rise and contact angle for water and methanol from weight measurements. The capillary rise was compared with the theoretical maximum value calculated by assuming each fluid perfectly wetted the lumenal surface of the glass (i.e., zero contact angle, which reflects the absence of surface contamination). For borosilicate, high-lead, and quartz capillaries, the rise for water was substantially less than the theoretical maximum rise. Exposure of the borosilicate, lead, and quartz capillaries to several cleaning methods resulted in substantially better—but not perfect—agreement between the theoretical maximum rise and calculated capillary rise. By contrast, the capillary rise for methanol was almost identical in untreated and cleaned capillaries, but less than its theoretical maximum rise. The residual discrepancy between the observed and theoretical rise for water could not be improved on by trying a variety of cleaning procedures, but some cleaning methods were superior to others. The water solubility of the surface contaminants, deduced from the effectiveness of repeated rinsing, was different for each of the three types of capillaries examined: Corning 7800>quartz>Corning 0010. A surface film was also detected in quartz tubing with an internal filament. I conclude that these borosilicate, quartz, and high-lead glass capillaries have a film on the lumenal surface, which can be removed using appropriate cleaning methods. The surface contaminants may be unique to each type of capillary and may also be hydrophobic. Two simple methods are presented to quantitate the cleanliness of glass capillary tubing commonly used to make pipets for studies of biological membranes. It is not known if the surface film is of importance in electrophysiological studies of biological membranes.     

Cortex shatters the glass ceiling

Recreating developmental structures in vitro has been a primary challenge for stem cell biologists. Recent studies in Cell Stem Cell (Eiraku et al., 2008) and Nature (Gaspard et al., 2008) demonstrate that embryonic stem cells can recapitulate early cortical development, enabling them to generate specific cortical subtypes.