Crystallographic structure of an intact IgG1 monoclonal antibody.

The structure of an intact monoclonal antibody for phenobarbital, subclass IgG1, has been determined to 3.2 A resolution by X-ray crystallography. The molecule was visualized in a monoclinic unit cell having an entire immunoglobulin as the asymmetric unit.

Atomic force microscopy studies of living cells: visualization of motility, division, aggregation, transformation, and apoptosis.

Atomic force microscopy, in contact mode, has been used to image living mammalian cells in culture at both low and high resolutions. The method is shown practical for revealing cytoskeletal features beneath the cell membrane and their restructuring during a variety of cellular activities. Among the processes that we have visualized are locomotion, tissue formation, cell division, transformation by viruses, and cell death.

Polyamine metabolism in the thermotolerant mesophilic fungus Aspergillus fumigatus.

Biomass production by Aspergillus fumigatus was greatest at 40-45 degrees C and was associated with an increase in concentration of the diamine putrescine and activity of its biosynthetic enzyme ornithine decarboxylase. Concentrations of the other amines, cadaverine, spermidine and spermine were considerably lower than putrescine concentration and did not change significantly over the temperature range 20-50 degrees C. This is surprising in view of the greatly increased flux of label from ornithine through to spermidine at 45 and 50 degrees C, indicating an increased formation of this triamine.

Visualization of RNA crystal growth by atomic force microscopy.

The crystallization of transfer RNA (tRNA) was investigated using atomic force microscopy (AFM) over the temperature range from 4 to 16 degrees C, and this produced the first in situ AFM images of developing nucleic acid crystals. The growth of the (110) face of hexagonal yeast tRNAPhe crystals was observed to occur at steps on vicinal hillocks generated by multiple screw dislocation sources in the temperature range of 13.5-16 degrees C.

The crystallographic structure of the subtilisin protease from Penicillium cyclopium.

The major extracellular protease from the fungus Pencillium cyclopium was crystallized in the presence of p-phenylmethanesulfonyl fluoride (PMSF) and investigated by X-ray diffraction analysis. It was subsequently cloned and the amino acid sequence deduced from its cDNA. Although the sequence is only 49% identical to that of proteinase K of Tritirachium album, the three-dimensional structures of the two proteases are virtually identical.

Measurement of 160-fs, 248-nm pulses by two-photon fluorescence in fused-silica crystals.

Measurements of 160-fs, 248-nm ultrashort pulses are obtained through a two-photon fluorescence measurement based on the two-photon-induced color-center fluorescence in fused-silica crystals. The method proved to be reliable and advantageous in comparison with two-photon fluorescence techniques employing other materials, both solid state and gaseous.

Crystallization of xylanase from Erwinia chrysanthemi: influence of heat and polymeric substrate.

Xylanase from the bacterial plant pathogen Erwinia chrysanthemi (E.C. 3.

Molecular resolution imaging of macromolecular crystals by atomic force microscopy.

Atomic force microscopy (AFM) images at the molecular level have been obtained for a number of different protein and virus crystals. They can be utilized in some special cases to obtain information useful to crystal structure analyses by x-ray diffraction. In particular, questions of space group enantiomer, the packing of molecules within a unit cell, the number of molecules per asymmetric unit, and the dispositions of multiple molecules within the asymmetric unit may be resolved.

Menière's disease: differential diagnosis and treatment.

Menière's disease may be the underlying cause of dizziness, which is a common presenting complaint. This disease is differentiated from other causes of dizziness by its classic constellation of four symptoms; dizziness characterized as episodic spinning or whirling vertigo; fluctuating, low-frequency sensorineural hearing loss; tinnitus, and a sensation of fullness in the ear. Because these symptoms may or may not develop or occur simultaneously, a careful history and a complete physical examination are necessary for diagnosis.

Refined structure of an intact IgG2a monoclonal antibody.

The structure of an intact, anti-canine lymphoma monoclonal antibody (Mab231) was determined by molecular replacement and refined in a triclinic cell to an R-value of 20.9%, using synchrotron diffraction data from 2.8 to 20 A resolution.

The open conformation of a Pseudomonas lipase.

Background: . The interfacial activation of lipases results primarily from conformational changes in the enzymes which expose the active site and provide a hydrophobic surface for interaction with the lipid substrate. Comparison of the crystallization conditions used and the structures observed for a variety of lipases suggests that the enzyme conformation is dependent on solution conditions.

Crystallization of biological macromolecules from flash frozen samples on the Russian Space Station Mir.

One hundred eighty-three flash frozen, liquid-liquid diffusion and batch method protein and virus crystallization samples were launched aboard the Space Shuttle Discovery on June 27 (STS-71) and transferred to the Russian Space Station Mir on July 1, 1995. They were returned to earth November 20, 1995 (STS-74). Subsequent examination showed that of the 19 types of proteins and viruses investigated, 17 were crystallized during the period on Mir.

The important attributes of a nurse from the perspective of qualified and student nurses.

The studies described here set out to discover what student and qualified nurses understand to be the important attributes of a nurse. The first study was exploratory and used a repertory grid technique to ask 72 students, qualified nurses and nurse lecturers to compare themselves (as a nurse) with a number of other nurses and carers of their own choosing. The constructs that the participants used to make these comparisons were then categorized into attributes.

Crystal structure of turnip yellow mosaic virus.

The structure of turnip yellow mosaic virus (TYMV) has been solved to 3.2 A resolution and an R-value of 18.7%.

The crystal structure of peanut peroxidase.

Background: Peroxidases catalyze a wide variety of peroxide-dependent oxidations. Based on sequence alignments, heme peroxidases have been divided into three classes. Crystal structures are available for peroxidases of classes I and II, but until now no structure has been determined for class III, the classical extracellular plant peroxidases.

Crystal structure of an anti-anti-idiotype shows it to be self-complementary.

The structure of the Fab fragment of the mouse anti-anti-idiotypic monoclonal antibody (mAb) GH1002 was solved by X-ray crystallography. mAb GH1002 was elicited with the syngeneic anti-idiotype mAb MK2-23 which mimics the determinant defined by anti-human high molecular weight-melanoma associated antigen (HMW-MAA) mAb 763.74.

Incorporation of microcrystals by growing protein and virus crystals.

In the course of time-lapse video and atomic force microscopy (AFM) investigations of macromolecular crystal growth, we frequently observed the sedimentation of microcrystals and three-dimensional nuclei onto the surfaces of much larger, growing protein or virus crystals. This was followed by the direct incorporation over time of the smaller crystals into the bulk of the larger crystals. In some cases, clear indications were present that upon absorption of the small crystal onto the surface of the larger, there was proper alignment of the respective lattices, and consolidation proceeded without observable defect formation, i.

Structure of a ribonuclease B+d(pA)4 complex.

The structure of a tetragonal crystal of bovine pancreatic RNase B complexed with d(pA)(4) was determined by molecular replacement and difference Fourier methods. This crystal belongs to space group P4(1)2(1)2 and has unit-cell dimensions a = b = 44.5, c = 156.

Treating bulimia nervosa in primary care: a pilot study.

Objective: To develop and evaluate a simplified and condensed cognitive behavioral treatment for bulimia nervosa suitable for use in primary care.

Method: Once developed, the treatment was tested on a consecutive series of 11 patients.

Results: Six patients did well, 3 were nonresponders, and in the other 2 external events interfered with progress.

Structural comparison of the plant satellite viruses.

Detailed structures are now available for three plant satellite viruses, satellite tobacco necrosis virus (STNV), satellite tobacco mosaic virus (STMV), and satellite panicum mosaic virus (SPMV). It is, therefore, possible to compare the tertiary structure of viral protein subunits, their quaternary interactions, and the interactions of protein subunits with the RNA genome. This analysis indicates that, in spite of common function and preservation of a "jelly-roll" motif in the protein monomer, the three viruses are remarkably different.

Mechanisms of growth for protein and virus crystals.

The growth of six protein and virus crystals was investigated in situ using atomic force microscopy. Most of the crystals grew principally on steps generated by two-dimensional nucleation on surfaces though some grew by development of spiral dislocations. Apoferritin grew by a rarely encountered mechanism, normal growth, usually associated only with melt or vapour phase crystallization.

Determination of local refractive index for protein and virus crystals in solution by Mach-Zehnder interferometry.

To establish the importance of, and quantitatively evaluate, the macromolecular concentration gradients in the neighborhood of growing protein, virus, and nucleic acid crystals, a convenient, accurate, and nonintrusive method has been devised. This approach should prove particularly relevant in the rigorous comparison of crystals grown in a conventional laboratory setting with those grown in a microgravity environment. The method is based on precise determination of the local refractive index using Mach-Zehnder interferometry.