Characterization of crystals of a cytochrome oxidase (nitrite reductase) from Pseudomonas aeruginosa by x-ray diffraction and electron microscopy

Cytochrome oxidase from Pseudomonas aeruginosa has been crystallized from 2 m-ammonium sulfate. The crystals occur principally as thin diamond-shaped plates of space group P2₁2₁2 with unit cell dimensions of 92 Å × 115 Å × 76 Å. Determination of the density of glutaraldehyde-fixed, water-equilibrated crystals (1.167 g/cm³), coupled with the unit cell volume (804,000 ų), indicates that there is one subunit (~63,000 Mr) per asymmetric unit. X-ray diffraction data which were limited to 12 Å resolution due to small crystal size were obtained for the hk0 and 0kl zones using precession photography. Amplitude and phase data for the hk0, 0kl, and h0l zones were obtained from computer-based Fourier analysis of appropriate micrographs recorded from negatively stained microplates and thin sections of larger crystals using minimal beam electron microscopy. For crystals embedded in the presence of tannic acid it was possible to achieve 20 Å resolution which is comparable to the resolution achieved with negative staining of thin crystalline arrays. In addition, unstained electron diffraction on glutaraldehyde-fixed, glucose-stabilized plates was recorded to a resolution of 9 Å. The three-dimensional packing of the cytochrome oxidase dimer in the unit cell has been deduced from computer reconstructed images of the three principal projections along the crystallographic axes. The cytochrome oxidase dimer is located in the unit cell with the dimer axis coincident with a crystallographic 2-fold axis; thus within the resolution of the present data in projection (9 Å) the two subunits are identical, in agreement with biochemical evidence. The crystals have been prepared with the enzyme in the fully oxidized state and upon reduction a progressive cracking of the crystals is observed, possibly due to a conformational change dependent on the oxidation state of the heme iron.     

Macrophage function in tumor-bearing mice: dissociation of phagocytic and chemotactic responsiveness

Infusion of CBA mice with lymphoid cells from the H-2 compatible but Mls-antigen incompatible C3H × CBA hybrid results in a specifically reduced capacity of the recipients lymphocytes to react in the MLC against C3H-cells. Although this reduction is immunologically specific the results of this investigation have shown that such mice exhibit a strongly reduced capacity to produce humoral antibodies against heterologous erythrocytes and a T-cell independent antigen (PVP).     

Predatory behaviour of lutzia on Culex fatigans

The carnivorous mosquito Lutzia (= Culex) raptor devours 20 to 50 larvae (third instar) of Culex fatigans in a day. The predatory capacity of L. raptor is not influenced by changes in volume of water, but significantly influenced by changes in prey density. With increasing prey density, the percentage of prey killed and left unconsumed increases. The duration required to subdue and consume a single larva is 15 min for L. raptor previously deprived of food for 3 to 24 hrs; the handling duration of prey increases to 20 min for the predator previously deprived food for 1 hr. The duration increases with increasing prey size; L. raptor requires 1, 6, 62 or 113 min to handle a single II, III, IV or mini pupa of Culex fatigans weighing 0.2,1.2,4.2 and 4.0 mg respectively.     

Simultaneous achievement of accurate CT number and image quality improvement for myocardial perfusion CT at 320-MDCT volume scanning

PurposeTo investigate differences in image-to-image variations between full- and half-scan reconstruction on myocardial CT perfusion (CTP) study.MethodsUsing a cardiac phantom we performed ECG-gated myocardial CTP on a second-generation 320-multidetector CT volume scanner. The heart rate was set at 60 bpm; once per second for a total of 24 s were performed. CT images were acquired at 80- and 120 kVp and subjected to full- and half-scan reconstruction. On images acquired at the same slice level we then measured image-to-image variations, coefficients of variance (CV), and image noise.ResultsThe image-to-image variations with full- and half-scan reconstruction were 1.3 HU vs. 27.2 HU at 80 kVp (p < 0.001) and 0.70 HU vs. 9.3 HU at 120 kVp (p < 0.001) even though the mean HU value was almost the same for both reconstruction methods. The CV of 80- and 120-kVp images of the left ventricular cavity decreased by 0.16% and 0.17%, respectively, with full-scan reconstruction; with half-scan reconstruction it decreased by 3.34% and 2.30%, respectively. Compared with half-scan reconstruction, the image noise was reduced by 27.2% at 80 kVp and by 28.0% at 120 kVp with full-scan reconstruction.ConclusionMyocardial CTP with full-scan reconstruction substantially decreased image-to-image variations and provided accurate CT attenuation.     

On the Estimation of the Population Mean with Known Coefficient of Variation

For the estimation of population mean a class of estimators has been proposed when the coefficient of variation is known and its efficiency is compared with the usual unbiased estimator and the estimators suggested by various researchers. The properties of the proposed class of estimators have been also discussed in the case of normal population.     

The molecular size of glycans liberated by hydrazinolysis from semliki forest virus proteins

The glycans of well characterized, [6-³H]galactose-labelled glycopeptides, GC-4 from bovine IgG₁ as well as GP-V-2 and GP-V-5 from α₁-acid glycoprotein, were liberated by hydrazinolysis. Molecular weights close to the expected values were observed by gel filtration. Desialated glycans of Semliki Forest virus proteins were likewise liberated by hydrazinolysis and subjected to gel filtration. Metabolically labelled [1-³H]galactose-oligosaccharides of the mixed viral proteins revealed an apparent molecular weight of 1800. The bi-antennary glycan liberated from the reference glycopeptide GC-4 was of 1750 daltons. A mixture of [2-³H]mannose-labelled E₁-and E₂-proteins of the virus contained L-type glycans of 1800 daltons (formerly called A-type), and M-type glycans of 1200 daltons (formerly called B-type). A fraction of the E₃-glycans isolated by affinity chromatography on Concanavalin A-Sepharose showed an average molecular weight of 2150, a value intermediate between the three- and four-antennary glycans liberated from the reference glycopeptides GP-V-5 and GP-V-2. The rest of the E₃-glycans were of 1850 daltons, a value close to the bi-antennary GC-4 glycan. We suggest that the comparatively large size of the E₃-glycans and the exposed position of E₃-proteins on the viral surface may be interrelated.