Comparisons of the low-resolution structures of ornithine decarboxylase by electron microscopy and X-ray crystallography: The utility of methylamine tungstate stain and butvar support film in the study of macromolecules by transmission electron microscopy

The structure of ornithine decarboxylase (M_r ≈? 1.04 × 10⁶) from Lactobacillus 30a was investigated by electron microscopy and x-ray crystallography. Electron micrographs showed the structure to be well preserved in methylamine tungstate stain. The molecules interacted little with the Butvar support film, yielding three unique projections: a hexagonal ring (front view) and two rod-shaped projections (edge views). Stereo pairs revealed a novel feature of the Butvar film in that some molecules were suspended in the stain in random orientations. Consequently, the relatedness of the hexagonal ring and the rod-shaped particles could be demonstrated since some particle shapes interconverted when the stage was tilted ± 45°. The two edge views were related by a 30° rotation about the sixfold axis. Image averaging of the three primary views suggested a dodecamer (point group symmetry 622) composed of two hexameric rings, apparently in an eclipsed configuration. To investigate the structural organization of the complex, the dissociation of the enzyme was studied by electron microscopy. The dissociation process involved the initial breakage of the ring followed by separation of dimers from the ring (one subunit from each of the two hexamers). Thus, the dodecamer forms as a hexamer of dimers rather than a dimer of hexamers. These structural studies were confirmed and extended by x-ray crystallographic analysis. A 4.0-Å resolution electron density map revealed two hexameric rings, consisting of six closely associated dimers, tilted approximately 10° with respect to the molecular twofold axis. Electron density projections of the three primary views of the molecule derived from the x-ray data corresponded closely to those obtained from image averaging of the electron microscopy data, thereby establishing in a novel way the reliability of the electron microscopy studies. Methylamine tungstate stain and Butvar support film therefore offer unique advantages for investigating protein structures by electron microscopy.     

Enhancing the tuning range of a single resonant band long period grating while maintaining the resonant peak depth by using an optimized high index indium tin oxide overlay

We report laboratory test results of a long period grating (LPG) that can maintain a constant resonant peak depth over an enhanced tuning range when it is coated with an indium tin oxide (ITO) electrode that has optimized thickness and refractive index. The authors have experimentally demonstrated a LPG coated with ITO that can be tuned in excess of 200 nm with an ambient refractive index change of less than 0.01. To the best of the authors' knowledge, this is the highest sensitivity reported for a LPG to date. In addition to the tuning performance, the resonant peak remains within 1 dB of its maximum depth for at least 100 nm of the tuning range.     

Fabrication and performance of low loss optical components made byion exchange in glass

Passive optical components made by photolithography with ion exchange in glass have been developed. The technology used to fabricate these components is described, and their performance characterization is discussed. It is shown that this technique produces highly uniform low-loss devices     

Evolution of cooperativity in hemoglobins: what can invertebrate hemoglobins tell us?

While vertebrate hemoglobins typically are tetrameric and show highly regulated and cooperative ligand binding, little is known of the evolution of these properties. We are studying the structural and functional properties of the hemoglobins from Caudina arenicola, an echinoderm. The echinoderms are in the lineage most closely related to the vertebrates to express hemoglobin. C. arenicola has three sets of red cells, in the water vascular system, the coelomic cavity, and in an intestinal vein. Each of these expresses a distinct array of globins. The hemoglobins are cooperative and exhibit unusual ligand-linked associative properties, being dimeric when oxygenated and forming tetramers and higher aggregates on deoxygenation. The major coelomic hemoglobins have been subjected to a detailed examination by a combination of ligand binding analyses and protein and DNA sequencing, as well as X-ray crystallography. Two typical globin introns were identified, along with a unique intron that bisects an N-terminal extension of the globin from the remainder of the gene. X-ray crystallographic analysis shows that the subunit interfaces of C. arenicola hemoglobins differ radically from those of vertebrate hemoglobins and indeed from some other invertebrate hemoglobins, but closely resemble the packing arrangements found in a clam hemoglobin (Scapharca). However, the residues implicated in cooperativity in these two types of hemoglobins differ substantially.     

Effects of brain death on myocardial function and ischemic tolerance of potential donor hearts

BACKGROUND: An increasing number of experimental and clinical studies reports hemodynamic instability in the donor organism after brain death. However, the relative importance of brain death-related cardiac dysfunction on posttransplantation cardiac function and the reversibility of the observed changes remain controversial. In this study a load-independent analysis of cardiac function after brain death was performed. Special interest was focused on a possible interactive influence of brain death and cardiac preservation on postischemic cardiac function. METHODS: In 12 anesthetized dogs, brain death was induced by inflation of a subdural balloon; 12 sham-operated animals served as control subjects. After a 2-hour observation in situ, the hearts were explanted and perfused parabiotically either immediately or after hypothermic ischemic preservation (4 hours, 4 degrees C). Heart rate, cardiac output, left ventricular pressure, the maximum of left ventricular pressure development and aortic pressure were measured in situ. In addition, the slope of the end-systolic pressure-volume relationship, coronary blood flow, and myocardial oxygen consumption were estimated in the cross-circulated hearts. RESULTS: In spite of a brain death-associated hemodynamic deterioration in situ (expressed as low mean aortic pressure and significant decrease of maximal dP/dt), myocardial function was similar to control after explantation, if assessed ex vivo. Furthermore, after hypothermic ischemic preservation and reperfusion, complete functional recovery of control and brain-dead hearts could be observed. CONCLUSIONS: These data indicate that hemodynamic instability after brain death may rather reflect altered loading conditions than irreversible myocardial damage or primary cardiac dysfunction. Furthermore, there is no evidence for a brain death-related impairment of ischemic tolerance.     

Explanation of launch condition choice for GRIN multimode fiberattenuation and bandwidth measurements

In order to predict the attenuation and bandwidth of a graded index (GRIN) multimode fiber system accurately, the fiber composing the system is best characterized by a restricted launch attenuation measurement and an over-filled-launch (OFL) bandwidth measurement of the fibers. The rationale for these launches is provided in terms of the power evolution in the fiber and its impact on system performance, and the basic physics of GRIN multimode fiber. Experimental results are shown that demonstrate the results     

Remote sensing of the Indo-Pacific region: ocean colour,sea level,winds and sea surface temperatures

The 1997–1998 ENSO (El Niño-Southern Oscillation) was not only the largest event of the century but also the most comprehensively observed. Satellite data were employed for ocean colour, sea level, winds, sea surface temperature (SST), and outgoing longwave radiation (OLR) were used to describe the response of the surface marine ecosystem associated with the ENSO event. Some of the large-scale anomalies in ocean colour include elevated biological activity to the north of the Equator in the Pacific coincident with lower sea levels associated with the classic ENSO-horseshoe pattern ecosystem response to the anomalous upwelling in the eastern Indian Ocean caused by the 1997–1998 dipole event, and the dramatic eastward propagating feature in the Equatorial Pacific in response to the La Niña dynamics. Ocean general circulation model (OGCM) experiments show that capturing the high-frequency wind changes is crucial for simulating the La Niña and the coupled biological–physical model (OBGCM) runs clearly show that higher frequency winds are also important for capturing the mean upwelling and nutrient supply into the euphotic zone. Thus, the QuickSCAT winds are expected to play a major role in ecosystem modelling in the future. This study shows the utility of satellite data for understanding not only ocean circulation but also the coupled ecosystem variability. Morcover, it is also shown that spatio-temporal resolution of the satellite winds will directly affect the accuracy of oceanic and ecosystem simulations.     

Vascularization in Bioartificial Parenchymal Tissue: Bioink and Bioprinting Strategies

Among advanced therapy medicinal products, tissue-engineered products have the potential to address the current critical shortage of donor organs and provide future alternative options in organ replacement therapy. The clinically available tissue-engineered products comprise bradytrophic tissue such as skin, cornea, and cartilage. A sufficient macro- and microvascular network to support the viability and function of effector cells has been identified as one of the main challenges in developing bioartificial parenchymal tissue. Three-dimensional bioprinting is an emerging technology that might overcome this challenge by precise spatial bioink deposition for the generation of a predefined architecture. Bioinks are printing substrates that may contain cells, matrix compounds, and signaling molecules within support materials such as hydrogels. Bioinks can provide cues to promote vascularization, including proangiogenic signaling molecules and cocultured cells. Both of these strategies are reported to enhance vascularization. We review pre-, intra-, and postprinting strategies such as bioink composition, bioprinting platforms, and material deposition strategies for building vascularized tissue. In addition, bioconvergence approaches such as computer simulation and artificial intelligence can support current experimental designs. Imaging-derived vascular trees can serve as blueprints. While acknowledging that a lack of structured evidence inhibits further meta-analysis, this review discusses an end-to-end process for the fabrication of vascularized, parenchymal tissue.     

Development of system specification for laser-optimized 50-/spl mu/m multimode fiber for multigigabit short-wavelength LANs

This paper presents the scientific arguments used in the specification development process by the Telecommunications Industry Association (TIA) Working Group FO-2.2.1 to develop the new multimode fiber and vertical-cavity surface-emitting laser specifications for high-speed application in data communications. Numerous engineering and commercial tradeoffs are described. The specification minimizes the link failure rate and overall link cost through utilization of communication-theory-based modeling and experimental verification. This was balanced against the reality of manufacturing costs attempting to maximize the yield of individual link components. The specific application used as an example has 50-/spl mu/m graded-index multimode fiber operating at 10 Gb/s (e.g., 10 Gb/s Ethernet and fiber channel). The link performance is determined by the interaction of the fiber intermodal dispersion measured by the differential modal delay, and the transceiver launch distribution into the multimode fiber measured by encircled flux. A theoretically based model and the simulation approach that were used to simulate 40 000 links are described. The information from these simulations was used to determine the specification limits. In addition, sensitivity to the specification limits was evaluated. The experimental results of a round robin conducted by the TIA are presented, which confirm that the modeled performance would yield the expected results in actual practice.     

Measurements for enhanced bandwidth performance over 62.5-/spl mu/m multimode fiber in short-wavelength local area networks

The Telecommunications Industry Association (TIA) FO-2.2.1 Working Group on the modal dependence of bandwidth has conducted industrywide interlaboratory comparisons on measurements aimed at improving the bandwidth performance of short-wavelength, laser-based, multimode-fiber local area networks (LANs). Measurements of both transceiver encircled flux and fiber restricted-mode-launch bandwidth can together successfully predict an enhanced system performance, provided that the proper limiting criteria are selected. System performance is determined by a measurement of effective bandwidth and/or intersymbol interference. Recommendations for source and fiber selection criteria come from a risk analysis based on an extensive multilaboratory comparison involving 95 fibers and 69 laser transceivers. For this paper, enhanced system performance is defined as a performance that allows operation at a data rate of at least one gigabit per second over a 500-m length of 62.5/125-/spl mu/m graded-index glass fiber.