Showing papers by "University of Virginia published in 1991"

Classification and characterization of heterotrophic microbial communities on the basis of patterns of community-level sole-carbon-source utilization.

Abstract: The BLOLOG redox technology based on tetrazolium dye reduction as an indicator of sole-carbon-source utilization was evaluated as a rapid, community-level method to characterize and classify heterotrophic microbial communities. Direct incubation of whole environmental samples (aquatic, soil, and rhizosphere) in BIOLOG plates containing 95 separate carbon sources produced community-dependent patterns of sole-carbon-source utilization. Principal-component analysis of color responses quantified from digitized images of plates revealed distinctive patterns among microbial habitats and spatial gradients within soil and estuarine sites. Correlation of the original carbon source variables to the principal components gives a functional basis to distinctions among communities. Intensive spatial and temporal analysis of microbial communities with this technique can produce ecologically relevant classifications of heterotrophic microbial communities.

Reduction by Granulocyte Colony-Stimulating Factor of Fever and Neutropenia Induced by Chemotherapy in Patients with Small-Cell Lung Cancer

Abstract: Background. Neutropenia and infection are major dose-limiting side effects of chemotherapy. Previous studies have suggested that recombinant methionyl granulocyte colony-stimulating factor (G-CSF) can reduce chemotherapy-related neutropenia in patients with cancer. We conducted a randomized clinical trial to test this hypothesis and the clinical implications. Methods. Patients with small-cell lung cancer were enrolled in a multicenter, randomized, double-blind, placebo-controlled trial of recombinant methionyl G-CSF to study the incidence of infection as manifested by fever with neutropenia (absolute neutrophil count, <1.0×l09 per liter, with a temperature ≥38.2°C) resulting from up to six cycles of chemotherapy with cyclophosphamide, doxorubicin, and etoposide. The patients were randomly assigned to receive either placebo or G-CSF, with treatment beginning on day 4 and continuing through day 17 of a 21 -day cycle. Results. The safety of the study treatment could be evaluated in 207 of the 211 pa...

Identification of the regulatory phosphorylation sites in pp42/mitogen-activated protein kinase (MAP kinase).

Abstract: Mitogen-activated protein kinase (MAP kinase) is a 42 kd serine/threonine protein kinase whose enzymatic activity requires phosphorylation of both tyrosyl and threonyl residues. As a step in elucidating the mechanism(s) for activation of this enzyme, we have determined the sites of regulatory phosphorylation. Following proteolytic digestion of 32P-labeled pp42/MAP kinase with trypsin, only a single phosphopeptide was detected by two-dimensional peptide mapping, and this peptide contained both phosphotyrosine and phosphothreonine. The amino acid sequence of the peptide, including the phosphorylation sites, was determined using a combination of Fourier transform mass spectrometry and collision-activated dissociation tandem mass spectrometry with electrospray ionization. The sequence for the pp42/MAP kinase tryptic phosphopeptide is similar (but not identical) to a sequence present in the ERK1- and KSS1-encoded kinases. The two phosphorylation sites are separated by only a single residue. The regulation of activity by dual phosphorylations at closely spaced threonyl and tyrosyl residues has a functional correlate in p34cdc2, and may be characteristic of a family of protein kinases regulating cell cycle transitions.

Celebrations of death : the anthropology of mortuary ritual

Abstract: This revised edition of a cross-cultural study of rituals surrounding death has become a standard text in anthropology, sociology and religion. Part of its fascination is that in understanding other people's death rituals we are able to gain a better understanding of our own. The authors refer to a wide variety of examples, from different continents and epochs. They compare the great tombs of the Berawan of Borneo and the pyramids of Egypt, as well as the dramas of medieval French royal funerals and the burial alive of the Dinka 'masters of the spear' in the Sudan, and other rituals which at first sight seem to have little in common. Many of these cases are anthropological classics, but the authors place them in an alternative context, so as to shape a novel synthesis on the anthropology of death ritual. A fresh introduction reviews theoretical developments in the field since the book first appeared in 1979.

Compact starbursts in ultraluminous infrared galaxies

Abstract: The 40 ultraluminous galaxies in the IRAS Bright Galaxy Sample of sources stronger than S = 5.24 Jy at lambda = 60 microns were mapped with approximately 0.25 arcsec resolution at 8.44 GHz. Twenty-five contain diffuse radio sources obeying the FIR-radio correlation; these are almost certainly starburst galaxies. Fourteen other galaxies have nearly blackbody FIR spectra with color temperatures between 60 and 80 K so their (unmeasured) FIR angular sizes must exceed approximately 0.25 arcsec, yet they contain compact (but usually resolved) radio sources smaller than this limit. The unique radio and FIR properties of these galaxies can be modeled by ultraluminous nuclear starbursts so dense that they 67 are optically thick to free-free absorption at about 1.49 GHz and dust absorption at about 25 microns. Only one galaxy (UGC 08058 = Mrk 231) is a dominated by a variable radio source too compact to be an ultraluminous starburst; it must be powered by a 'monster'.

Synthesis of water-soluble (aminoalkyl)camptothecin analogues: inhibition of topoisomerase I and antitumor activity.

Abstract: Water-soluble analogues of the antitumor alkaloid camptothecin (1) were prepared in which aminoalkyl groups were introduced into ring A or B. Most of the analogues were prepared by oxidation of camptothecin to 10-hydroxycamptothecin (2) followed by a Mannich reaction to give N-substituted 9-(aminomethyl)-10-hydroxycamptothecins (4-12) or by subsequent modification of Mannich product 4 (13, 15, 17, 19, 21). Others were obtained by modification of the hydroxyl group of 2 (25,26) or by total synthesis (35,42,43). These analogues, as well as some of their synthetic precursors, were evaluated for inhibition of topoisomerase I, cytotoxicity, and antitumor activity. Although there was not a quantitative correlation between these assays, compounds that inhibited topoisomerase I were also cytotoxic and demonstrated antitumor activity in vivo. Further evaluation of the most active water-soluble analogue led to the selection of 9-[(dimethylamino)methyl]-10-hydroxycamptothecin (4, SK&F 104864) for development as an antitumor agent. In addition to its water solubility, ease of synthesis from natural camptothecin, and high potency, 4 demonstrated broad-spectrum activity in preclinical tumor models and is currently undergoing Phase I clinical trials in cancer patients.

Microvascular architecture in a mammary carcinoma: branching patterns and vessel dimensions.

Abstract: The objective of this work was to introduce a tumor vessel classification scheme and to provide the first quantitative measurements of vessel branching patterns and the related vascular dimensions in a mammary carcinoma. Mammary adenocarcinoma R3230AC tumors, grown in the rat ovarian tissue-isolated tumor preparation, were infused with Batson's No. 17 polymer and maintained at an intravascular pressure of 50 mm Hg during polymerization. Maceration of the tumor in KOH allowed visualization of the vasculature. The vessel branching patterns, lengths, and diameters were measured in four tumors (4-5 g). A centrifugal ordering scheme was devised specifically to account for the unique features of tumor microvascular network topology. The arterial networks revealed two types of branching patterns. One type of arteriolar network exhibited decreasing vessel diameters and lengths with increasing branch order. In a second type of network, the diameter and length of the vessels displayed fluctuations in both variables at higher generations. Avascular and poorly vascularized regions with sparse capillary supply were present in the tumors, but analysis of several capillary networks in vascularized regions revealed a nonplanar meshwork of interconnected vessels. The meshworks were composed of vessels with a mean segment length of 67 microns, a mean diameter of 10 microns, and a mean intercapillary distance of 49 microns. Capillary path lengths ranged from 0.5 to 1.5 mm. Thus, tumor capillary diameter was greater than that in most normal tissues and, in the regions where capillary networks existed, intercapillary spacing was in the normal range. In the venous network, diameters decreased from 650 to 20 microns for the first to ninth order venules. Venule length decreased from 5 to 0.5 mm for first to fourth order but was fairly uniform (less than 500 microns) for higher orders. In conclusion, solid tumor vascular architecture, while exhibiting several features that are similar to those observed in normal tissues, has others that are not commonly seen in normal tissues. These features of the tumor microcirculation may lead to heterogeneous local hematocrits, oxygen tensions, and drug concentrations, thus reducing the efficacy of present day cancer therapies.

Dual defects in pulsatile growth hormone secretion and clearance subserve the hyposomatotropism of obesity in man.

Abstract: We have examined the mechanisms underlying reduced circulating GH concentrations in the obese human. Computer-assisted (deconvolution) analysis was used to determine endogenous GH secretory and clearance rates quantitatively from entire 24-h plasma GH concentration profiles. These analyses revealed that the half-life (t 1/2) of endogenous GH was significantly shorter in obese (11.7 +/- 1.6 min) than in normal weight subjects (15.5 +/- 0.81 min; P less than 0.01). The accelerated blood disposal rate of GH was not due to decreased circulating concentrations of GH-binding protein, since the latter were similar in obese (25 +/- 1.0%) and normal weight (24 +/- 2.3%) men. However, obese men had significantly fewer GH secretory bursts (3.2 +/- 0.53 vs. 9.7 +/- 0.67/day; P less than 0.01). Among the rare GH secretory bursts that occurred in obese subjects, there were significantly prolonged mean intersecretory burst intervals (282 +/- 65 vs. 131 +/- 11 min; P less than 0.05). The resultant daily GH production rate in obese men was reduced to one fourth that in normal weight individuals. Both GH secretion rate and burst frequency were negatively correlated with the degree of obesity (ponderal index). The decreases in GH burst frequency and half-life were specific, since GH secretory pulse amplitude (maximal rate of GH release), the mass of GH released per burst, and the duration of computer-resolved GH secretory bursts were not different in obese and normal weight men. We conclude that obese men harbor a double defect in GH dynamics involving both GH secretion and clearance, and that the severity of the GH secretory deficiency is proportionate to the degree of obesity.

The distribution of synapsin I and synaptophysin in hippocampal neurons developing in culture

Abstract: As a first step toward elucidating mechanisms involved in the sorting of synaptic vesicle proteins in neurons, we have used immunofluorescence microscopy to determine the distribution of two synaptic vesicle proteins, synapsin I and synaptophysin, in hippocampal neurons developing in culture. In mature cultures, synapsin I and synaptophysin immunoreactivity was concentrated in puncta that were restricted to sites where axons contacted neuronal cell bodies or dendrites. Electron-microscopic immunocytochemistry demonstrated that these puncta corresponded to vesicle-filled axonal varicosities that were exclusively presynaptic. At early stages of development, before cell-cell contact, both synapsin I and synaptophysin were preferentially localized in axons, where they were particularly concentrated in the distal axon and growth cone. In axons that did not contact other cells, immunostaining for these two proteins had a granular appearance, which persisted for at least 7 d, but focal accumulations of vesicles comparable to those seen at sites of synaptic contact were not observed. When neurons contacted one another, numerous puncta of synapsin I and synaptophysin formed within the first week in culture. Double-label immunofluorescence demonstrated that the two vesicle antigens were closely codistributed throughout these stages of development. These observations demonstrate that synaptic vesicle proteins assume a polarized distribution within nerve cells beginning early in development, as soon as the axon can be identified. In contrast, differences in microtubule polarity orientation that distinguish mature axons and dendrites, and that have been proposed to account for the selective sorting of some materials in nerve cells, first appear at a subsequent stage of development. The selective distribution of synaptic vesicle proteins to the axon occurs in isolated cells, independent of interactions with other cells. In contrast, the formation of large clusters of vesicles typical of presynaptic specializations requires contact with an appropriate postsynaptic target. Thus, in cultured hippocampal neurons, the localization of synaptic vesicles in presynaptic specializations is the result of sorting mechanisms intrinsic to individual neurons as well as to mechanisms mediated by cell-cell contact.

Circadian photoreception in the retinally degenerate mouse (rd/rd).

Abstract: We have examined the effects of light on circadian locomotor rhythms in retinally degenerate mice (C57BL/6J mice homozygous for the rd allele: rd/rd). The sensitivity of circadian photoreception in these mice was determined by varying the irradiance of a 15 min light pulse (515 nm) given at circadian time 16 and meauring the magnitude of the phase shift of the locomotor rhythm. Experiments were performed on animals 80 days of age. Despite the loss of visual photoreceptors in the rd/rd retina, animals showed circadian responses to light that were indistinguishable from mice with normal retinas (rd/+ and +/+). While no photoreceptor outersegments were identified in the retina of rd/rd animals (80–100 days of age), we did identify a small number of perikarya that were immunoreactive for cone opsins, and even fewer cells that contained rod opsin. Using HPLC, we demonstrated the presence and photoisomerization of the rhodopsin chromophore 11-cis retinaldehyde. The rd/rd retinas contained about 2% of 11-cis retinaldehyde found in +/+ retinas. We have yet to determine whether the opsin immunoreactive perikarya or some other unidentified cell type mediate circadian light detection in the rd/rd retina.

Geometrically necessary, incidental and subgrain boundaries

Abstract: During the deformation of polycrystals, the grains break up into domains within which the selection of operative slip systems differs. The domains then subdivide into “cell blocks”. Locally, each group of cell blocks comes near to fulfilling the Taylor criterion when taken collectively, but the number of active glide systems in any one cell block is fewer than predicted. The boundaries between cell blocks and/or domains accommodate the lattice misorientations which result from glide on the different slip system combinations. They are therefore named “geometrically necessary bpundaries”. They, like all boundaries capable of accommodating variable lattice misorientations, are composed of dislocations. Microscopically, such boundaries appear as “dense dislocation walls” and “microbands”. Geometrically necessary boundaries are distinguished from ordinary dislocation cell boundaries by the absence of a change of glide systems across the latter. In materials deforming with a cell structure, ordinary dislocation cell boundaries as well as traditional “deformation bands” arise from the mutual trapping of dislocations into low-energy configurations. Such cell boundaries or walls are therefore named “incidental dislocation boundaries”. The misorientation across incidental boundaries is typically much smaller than for geometrically necessary boundaries. A further distinction is their respective on the flow stress. The average spacing of dislocation cell walls is inverse proportional to the flow stress whereas geometrically necessary boundaries obey the Hall-Petch relationship. Since they tend to occur more frequently the incidental boundaries typically control the flow stress. At increasing strain the angles between dislocation cells increase and different slip system combinations can operate in neighbouring cells. Cell walls are then no longer incidental boundaries but geometrically necessary boundaries. Such boundaries are termed “subgrain boundaries”.

Thin film deposition and microelectronic and optoelectronic device fabrication and characterization in monocrystalline alpha and beta silicon carbide

Abstract: The deposition of silicon carbide thin films and the associated technologies of impurity incorporation, etching, surface chemistry, and electrical contacts for fabrication of solid-state devices capable of operation at temperatures to 925 K are addressed. The results of several research programs in the United States, Japan and the Soviet Union, and the remaining challenges related to the development of silicon carbide for microelectronics are presented and discussed. It is concluded that the combination of alpha -SiC on alpha -SiC appears especially viable for device fabrication. In addition, considerable progress in the understanding of the surface science, ohmic and Schottky contacts, and dry etching have recently been made. The combination of these advances has allowed continual improvement in Schottky diode p-n junction, MESFET, MOSFET, HBT, and LED devices. >

Inhibition of proteolysis protects hippocampal neurons from ischemia.

Abstract: Intense proteolysis of cytoskeletal proteins occurs in brain within minutes of transient ischemia, possibly because of the activation of calcium-sensitive proteases (calpains). This proteolytic event precedes overt signs of neuronal degeneration, is most pronounced in regions of selective neuronal vulnerability, and could have significant consequences for the integrity of cellular function. The present studies demonstrate that (i) the early phase of enhanced proteolysis is a direct response to hypoxia rather than other actions of ischemia, (ii) it is possible to pharmacologically inhibit the in vivo proteolytic response to ischemia, (iii) inhibition of proteolysis is associated with a marked reduction in the extent of neuronal death, and (iv) protected neurons exhibit normal-appearing electrophysiological responses and retain their capacity for expressing long-term potentiation, a form of physiological plasticity thought to be involved in memory function. These observations indicate that calcium-activated proteolysis is an important component of the post-ischemic neurodegenerative response and that targeting this response may be a viable therapeutic strategy for preserving both the structure and function of vulnerable neurons.

Magnetic properties of free cobalt clusters

Abstract: We report in this paper results of Stern-Gerlach experiments for all the systems we have studied to date. Three distinct behaviors are observed. Cobalt exhibits superparamagnetic behavior, with a magnetic moment per atom of 2.29 ± 0.08µ B for N = 115. The rare earths gadolinium and terbium exhibit superparamagnetic behavior for certain clusters, while other clusters exhibit behavior consistent with their moment being locked to the crystal lattice. In both cases, the moments are considerably reduced from the bulk values. The third behavior is no measurable deflection at all, as demonstrated by vanadium, palladium, chromium, and aluminum.

Laparoscopic cholecystectomy. Treatment of choice for symptomatic cholelithiasis.

Abstract: Our initial experience with laparoscopic cholecystectomy (LC) for symptomatic cholelithiasis has involved 152 patients. Patient age ranged from 17 to 83 years; most were female (78%). Their average weight was 170 pounds (range, 75 to 365 lbs.). Twenty-two per cent had a single gallstone, while 9% had two to three stones and 64% had more than three stones. Exclusion criteria initially included upper abdominal scarring, severe acute cholecystitis, choledocholithiasis, and inability to tolerate general anesthesia. The first two of these are now only relative contraindications with increased experience. Thirteen of the one hundred fifty-two procedures (8.5%) required conversion to an open operation. Average time of operation was 138 minutes. Intraoperative cholangiography was attempted in 78% of cases and was completed successfully in 66% of those attempted. There have been no deaths. The complication rate has been low: 4% major, 0% life-threatening, and 7.2% minor complications. Postoperative analgesic requirements are remarkably low: 36% of patients required no narcotics after leaving the recovery room. Eighty-seven per cent of patients successfully undergoing LC were discharged by the first postoperative day. Most patients resumed normal activities within 1 week after discharge. Laparoscopic cholecystectomy offers the majority of patients with symptomatic cholelithiasis an improved treatment option, resulting in significantly less postoperative pain, hospitalization, and recuperation time.

Recombinant human interleukin-1 induces meningitis and blood-brain barrier injury in the rat. Characterization and comparison with tumor necrosis factor.

Abstract: The diversity of infectious agents capable of inducing meningitis and blood-brain barrier (BBB) injury suggests the potential for a common host mediator. The inflammatory polypeptides, IL-1 and TNF, were tested in an experimental rat model as candidate mediators for induction of meningitis and BBB injury. Intracisternal challenge of rIL-1 beta into rats induced neutrophil emigration into cerebrospinal fluid (CSF) and significantly increased BBB permeability to systemically administered 125I-BSA as early as 3 h later (P less than 0.05). This injury was reversible, dose dependent and significantly inhibited by prior induction of systemic neutropenia (via intraperitoneal cyclophosphamide) or preincubation of the rIL-1 beta inoculum (50 U) with an IgG monoclonal antibody to rIL-1 beta. Similar kinetics and reversibility of CSF inflammation and BSA permeability were observed using equivalent dose inocula of rIL-1 alpha. rTNF-alpha was less effective as an independent inducer of meningitis or BBB injury over an inoculum range of 10(1) U (0.0016 micrograms/kg)-10(6) U (160 micrograms/kg) when injected intracisternally, but inoculum combinations of low concentrations of rTNF alpha (10(3) U) and rIL-1 beta (0.0005-5.0 U) were synergistic in inducing both meningitis and BBB permeability to systemic 125I-BSA. These data suggest that in situ generation of interleukin-1 within CSF (with or without TNF) is capable of mediating both meningeal inflammation and BBB injury seen in various central nervous system infections.

Nerve growth factor in the urinary bladder of the adult regulates neuronal form and function.

Abstract: Urethral obstruction produces increased voiding frequency (0.7 +/- 0.06 to 1.1 +/- 0.08 h-1) and hypertrophy of the urinary bladder (89 +/- 1.7 to 708 +/- 40 mg) with profound increments in the dimensions of afferent (4, 6) and efferent neurons (299 +/- 4.7 to 573 +/- 8.6 microns2) supplying this organ in the rat. We discovered that hypertrophied bladders of rat and human contain significantly more nerve growth factor (NGF) per milligram wet weight, protein, and DNA than normal bladders. The temporal correlation between NGF content, neuronal hypertrophy, and bladder weight was consistent with a role for this growth factor in the neurotrophic effects associated with obstruction. Autoimmunity to NGF abolished the hypertrophy of NGF-sensitive bladder neurons in the pelvic ganglion after obstruction. Relief of urethral obstruction reduced bladder size (349 +/- 78 mg), but neuronal hypertrophy (460.2 +/- 10.2 microns2) and elevated NGF levels were only partially reversed. Bladder hypertrophy (133 +/- 4.3 mg) induced by osmotic diuresis slightly increased ganglion cell area (365.2 +/- 6.1 microns2) and only doubled NGF content of the bladder. These findings provide important new evidence that parenchymal cells in the hypertrophied bladder can synthesize NGF and possibly other molecular messengers that act to alter the size and function of neurons in adult animals and man.

Cholesterol-induced fluid-phase immiscibility in membranes.

Abstract: The fluid-phase behavior of binary mixtures of cholesterol with phosphatidylcholines is investigated using magnetic resonance methods. Phospholipid biradicals provide the electron spin resonance spectroscopic resolution of two immiscible fluid phases in the dipalmitoylphosphatidylcholine-cholesterol system. Isotropic chemical shifts of the phospholipid carbonyl carbons in binary mixtures with cholesterol measured using solid-state high-resolution nuclear magnetic resonance methods furnish evidence for a putative hydrogen bond between the 3 beta-hydroxyl of cholesterol and the sn-2 carbonyl of the phospholipid. The location in the bilayer of cholesterol in the two fluid phases is determined by measuring spin label-enhanced spin-lattice relaxation rates of the 13C nuclei of both the phospholipid and cholesterol molecules. These results suggest, in a time-averaged sense, that in the cholesterol-poor fluid phase the cholesterol molecule essentially spans the bilayer, whereas in the cholesterol-rich fluid phase the molecule is present in both monolayers of the bilayer.

Physical and chemical factors influencing transport of microorganisms through porous media.

Abstract: Resting-cell suspensions of bacteria isolated from groundwater were added as a pulse to the tops of columns of clean quartz sand. An artificial groundwater solution (AGW) was pumped through the columns, and bacterial breakthrough curves were established and compared to test the effects of ionic strength of the AGW, cell size (by using strains of similar cell surface hydrophobicity but different size), mineral grain size, and presence of heterogeneities within the porous media on transport of the bacteria. The proportion of cells recovered in the effluent ranged from nearly 90% for AGW of a higher ionic strength (I = 0.0089 versus 0.00089 m), small cells (0.75-micron-diameter spheres versus 0.75 by 1.8-micron rods), and coarse-grained sand (1.0 versus 0.33 mm) to less than 1% for AGW of lower ionic strength, large cells, and fine-grained sand. Differences in the widths of peaks (an indicator of dispersion) were significant only for the cell size treatment. For treatments containing heterogeneities (a vein of coarse sand in the center of a bed of fine sand), doubly peaked breakthrough curves were obtained. The first peak represents movement of bacteria through the transmissive coarse-grained vein. The second peak is thought to be dominated by cells which have moved (due to dispersion) from the fine-grained matrix to the coarse-grained vein near the top of the column and thus had been retarded, but not retained, by the column. Strength of effects tests indicated that grain size was the most important factor controlling transport of bacteria over the range of values tested for all of the factors examined.(ABSTRACT TRUNCATED AT 250 WORDS)

Information architecture: in search of efficient flexibility

Abstract: This article addresses how information systems architecture can be used to support organizations in the 1990s-organizations that face the dual challenge of "speed and flexibility" and "low cost and efficiency." At the heart of this challenge is the basic notion that information systems have been anything but flexible in the past and that, for many firms, information systems are more disablers of flexibility than enablers. The article discusses two architectural solutions to this problem: "the high road and the low road," and the benefits and pitfalls of teach. We conclude that neither solution will succeed on its own and that firms need to combine elements of both to meet the challenges of the 1990s. This article is based on some of the things we have learned through research, case writing, and consulting wile working with a variety of organizations over the past three years. These experience shave illustrated the importance of and the struggle with IS architecture for today's global competitors. The content is intended to help guide, provoke, stimulate, and entertain others who believe that the integration of information technology with organizational strategy and structure os of paramount concern to senior managers.