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At least 631 records · Page 35Linked to original sources
PubMed1999

New multicomponent capsules for immunoisolation.

A new generation of microcapsules based on the use of oligomers which participate in polyelectrolyte complexation reactions has been developed. These freeze-thaw stable capsules have been applied as a bioartificial pancreas and have resulted in normoglycemia for periods of six months in concordant xenotransplantations. The new chemistry permits the control of permeability and mechanical properties over a wide range and can be adapted both to microcapsule and hollow fiber geometries rendering it a robust tool for encapsulation in general. Methods, and metrics, for the characterization of the mechanical properties and permeability of microcapsules are presented.

Artificial Organs↗
PubMed2000

Broadband control of plate radiation using a piezoelectric, double-amplifier active-skin and structural acoustic sensing

The potential of a piezoelectric, double-amplifier active-skin with structural acoustic sensing (SAS) is demonstrated for the reduction of broadband acoustic radiation from a clamped, aluminum plate. The active-skin is a continuous covering of the vibrating portions of the plate with active, independently controllable piezoelectric, double-amplifier elements and is designed to affect control by altering the continuous structural radiation impedance rather than structural vibration. In simulation, acoustic models are sought for the primary and secondary sources that incorporate finite element methods. Simulation indicates that a total radiated power attenuation in excess of 10 dB may be achieved between 250 and 750 Hz with microphone error sensing, while under SAS the radiated power is reduced by nearly 8 dB in the same frequency range. In experiment, the adaptive feed forward filtered-x LMS (least mean square) algorithm, implemented on a Texas Instruments C40 DSP, was used in conjunction with the 6I6O control system. With microphone error sensing, 11.8-dB attenuation was achieved in the overall radiated power between 175 and 600 Hz, while inclusion of SAS resulted in a 7.3-dB overall power reduction in this frequency band.

Journal Article↗
PubMed1971

Irreversible effects on visible light on the retina: role of vitamin A.

Diffuse retinal irradiation by visible light produces in the rat the death of visual cells and pigment epithelium. Typically, cage illumination of 1500 lux from fluorescent light through a green filter leads to severe damage when continued for 40 hours. Vitamin A deficiency protects against this damage but experiments show that retinol released by light from rhodopsin is probably not the toxic agent. Protection against light damage depends on a long-range state of cell adaptation to light itself. The normal diurnal cycle of light and dark seems to be the essential factor in controlling visual cell viability and susceptibility.

Animals↗
PubMed1987

A rapid test for chitinase activity that uses 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide.

A total of 101 strains of bacteria from environmental and clinical sources, most of which were gram negative, were tested for chitobiase activity by using a filter paper spot test with 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide as the substrate. The results were compared with those obtained by a conventional plate method for chitinase activity by using colloidal chitin as the substrate. There was excellent agreement in the results for both methods. The filter paper spot test with 4-methylumbelliferyl-N-acetyl-beta-D-glucosaminide has the advantages of being rapid, simple to perform, and inexpensive. This method should be adaptable to a wider range of microorganisms, particularly those with unusual growth requirements.

Acetylglucosamine↗
PubMed1993

Conditional suicide system of Escherichia coli released into soil that uses the Bacillus subtilis sacB gene.

The sacB gene from Bacillus subtilis confers sucrose sensitivity upon gram-negative bacteria. The gene was investigated for use as a potential conditional suicide system for Escherichia coli released into soil. To ensure against the loss of the cell death function encoded under nonselective conditions, the nptI-sacR-B suicide cassette was inserted into the E. coli chromosome by using a circular nonreplicative integration vector. Stability studies yielded no loss of the suicide cassette in the integrated E. coli EL1026 strain. sacB induction in the absence of a selective pressure resulted in a lysis efficiency of up to 99.9%. The microcosm experiments confirmed the ability of the suicide cassette to limit the growth and reduce the survival of E. coli strains released into soil. Sucrose addition to sterile soil resulted in a 10(-3)-fold reduction of the final E. coli population density. sacB induction prevented the proliferation and triggered the rapid disappearance of E. coli from natural soil. Mutation to sucrose tolerance occurred at a frequency of 10(-5), making E. coli EL1026 a potential counterselectable donor strain for gene transfer studies. Specificity and potential adaptability to a wide range of gram-negative bacteria are additional conveniences of this conditional suicide system for the containment and counterselection of engineered microorganisms.

Bacillus subtilis↗
PubMed1994

Genetic Structure of Rhizobium etli biovar phaseoli Associated with Wild and Cultivated Bean Plants (Phaseolus vulgaris and Phaseolus coccineus) in Morelos, Mexico.

The genetic structure of Rhizobium etli biovar phaseoli was determined for five populations in three different locations in the state of Morelos, Mexico, by using starch gel electrophoresis for five to nine polymorphic loci. Two populations were sampled during two different years from nodules of cultivated and wild common bean plants (Phaseolus vulgaris). The three other populations were associated with wild runner beans (P. coccineus) and sampled during 1988. The Rhizobium populations differ genetically both among sites and among populations within the same site in different years, as shown by differences in allelic frequencies, genetic differentiation analysis, and differences in electrotypes. The total genetic diversity for the five populations during 1988 was H = 0.487; there were also high levels of genetic variation within each population. We found the highest linkage disequilibrium in a global analysis for all the populations. At a local scale, we also found significant linkage disequilibrium in two populations, although the distribution of the D' suggest some recombination at a local scale. The other three rhizobium populations exhibit low linkage disequilibrium. A cluster analysis (UPGMA) of pairwise genetic distances showed that bacteria isolated from most wild Phaseolus spp. are grouped by population, whereas those obtained from cultivated P. vulgaris are very heterogeneous. The analysis of the genetic structure of Rhizobium strains may allow the identification of strains that are naturally well adapted to a wide range of different environments, which may be useful for agricultural purposes or as a starting point for developing improved Rhizobium strains.

Journal Article↗
PubMed2004

Molecular evolution and mosaicism of leptospiral outer membrane proteins involves horizontal DNA transfer.

Leptospires belong to a genus of parasitic bacterial spirochetes that have adapted to a broad range of mammalian hosts. Mechanisms of leptospiral molecular evolution were explored by sequence analysis of four genes shared by 38 strains belonging to the core group of pathogenic Leptospira species: L. interrogans, L. kirschneri, L. noguchii, L. borgpetersenii, L. santarosai, and L. weilii. The 16S rRNA and lipL32 genes were highly conserved, and the lipL41 and ompL1 genes were significantly more variable. Synonymous substitutions are distributed throughout the ompL1 gene, whereas nonsynonymous substitutions are clustered in four variable regions encoding surface loops. While phylogenetic trees for the 16S, lipL32, and lipL41 genes were relatively stable, 8 of 38 (20%) ompL1 sequences had mosaic compositions consistent with horizontal transfer of DNA between related bacterial species. A novel Bayesian multiple change point model was used to identify the most likely sites of recombination and to determine the phylogenetic relatedness of the segments of the mosaic ompL1 genes. Segments of the mosaic ompL1 genes encoding two of the surface-exposed loops were likely acquired by horizontal transfer from a peregrine allele of unknown ancestry. Identification of the most likely sites of recombination with the Bayesian multiple change point model, an approach which has not previously been applied to prokaryotic gene sequence analysis, serves as a model for future studies of recombination in molecular evolution of genes.

Amino Acid Sequence↗
PubMed2026

A tiled amplicon protocol for culture-free whole-genome sequencing of M. tuberculosis from clinical specimens.

Whole-genome sequencing of Mycobacterium tuberculosis can be a valuable tool for TB surveillance and treatment, providing insights into transmission patterns and comprehensive drug susceptibility testing. However, the slow growth of M. tuberculosis means traditional culture-based sequencing methods can take weeks to return results, which has limited the widespread adoption of these techniques and limited their use in clinical decision-making. Tiled amplicon sequencing is a fast, reliable, and cost-effective method of whole-genome sequencing that can be done directly on clinical specimens and has been implemented at scale in academic and public health laboratories across the world; it was the cornerstone of SARS-CoV-2 sequencing and has been adapted for a wide range of viral pathogens. However, similar methods are not yet available for far larger bacterial genomes. Extending this approach to M. tuberculosis would significantly reduce the cost, labor, and turnaround time for whole-genome sequencing. We designed a tiled amplicon panel consisting of 5,128 primers that covers the entire M. tuberculosis genome, the largest tiled amplicon sequencing panel we are aware of to date. Applying our amplicon panels to clinical samples of sputum, we show the ability to recover whole-genome bacterial sequences without the need for culture. The resulting sequence data can be used to determine M. tuberculosis lineage and reliably identify markers of drug resistance. Using this approach in clinical settings could reduce the time needed for comprehensive drug susceptibility testing from weeks to days and enable genomic epidemiology to be performed at scale, even in resource-limited settings.IMPORTANCEWe have developed and tested an amplicon panel, TB-seq, for the priority pathogen Mycobacterium tuberculosis, demonstrating recovery of near-full genomes directly from patient sputum, including mixed and low-concentration samples. This approach significantly reduces the turnaround time for this slow-growing bacterium while maintaining high accuracy in detecting clinically relevant mutations, including those associated with drug resistance. Given the global burden of tuberculosis and the critical need for faster diagnostic solutions, we believe our method has the potential to improve clinical decision-making and public health strategies.

Mycobacterium tuberculosis↗
PubMed2005

por Variable-region typing by DNA probe hybridization is broadly applicable to epidemiologic studies of Neisseria gonorrhoeae.

The porin gene (porB) of Neisseria gonorrhoeae encodes the major outer membrane protein identified as PI or Por. To examine the utility of por variable-region (VR) typing, porB from 206 isolates was characterized by using oligonucleotide probes in a checkerboard hybridization assay that identifies the sequence types of five VRs of both PIA and PIB porB alleles. The strains represented temporally and geographically distinct isolates, isolates from a large cluster, epidemiologically linked partner isolates, and a collection of strains from disseminated gonococcal infections. By using rigorous epidemiologic criteria for transmission of infection between sex partners, por VR typing was more discriminatory than serovar typing in classifying isolates from both members of 43 epidemiologically linked pairs: 39 of 43 pairs were classified as coinciding by por VR typing compared to 43 of 43 by serovar determination (P = 0.058). porB sequence data confirmed the accuracy of the por VR method. Relationships between VR type and serovar typing monoclonal antibodies were observed for all six PIB and three of six PIA antibodies. por VR typing is a molecular tool that appears to have broad applicability. This method can be adapted to a wide range of technologies from simple hybridization to microarray and may allow for typing from noncultured clinical specimens.

Bacterial Typing Techniques↗
PubMed2003

A dual optic accommodating foldable intraocular lens.

AIM: To design an accommodating intraocular lens with extended accommodative range that can be adapted to current standard phacoemulsification and endocapsular implantation technique. METHOD: Ray tracing analysis and lens design; cadaver eye implantation. RESULTS: Ray tracing analysis indicated that axial movement of an exaggerated converging anterior optic linked by spring loaded haptics to a compensatory static diverging posterior optic produced greater change in conjugation power of the eye compared to axial movement of a single optic lens. A dual optic one piece foldable silicone lens was constructed and implanted via a 4 mm corneal incision into the capsular bag of two cadaver eyes. CONCLUSION: A dual optic intraocular lens design can increase the optical effect of a given displacement and suggests improvements for accommodating intraocular lenses.

Accommodation, Ocular↗
PubMed1999

Tumor necrosis factor receptor and Fas signaling mechanisms.

Four members of the tumor necrosis factor (TNF) ligand family, TNF-alpha, LT-alpha, LT-beta, and LIGHT, interact with four receptors of the TNF/nerve growth factor family, the p55 TNF receptor (CD120a), the p75 TNF receptor (CD120b), the lymphotoxin beta receptor (LT beta R), and herpes virus entry mediator (HVEM) to control a wide range of innate and adaptive immune response functions. Of these, the most thoroughly studied are cell death induction and regulation of the inflammatory process. Fas/Apo1 (CD95), a receptor of the TNF receptor family activated by a distinct ligand, induces death in cells through mechanisms shared with CD120a. The last four years have seen a proliferation in knowledge of the proteins participating in the signaling by the TNF system and CD95. The downstream signaling molecules identified so far--caspases, phospholipases, the three known mitogen activated protein (MAP) kinase pathways, and the NF-kappa B activation cascade--mediate the effects of other inducers as well. However, the molecules that initiate these signaling events, including the death domain- and TNF receptor associated factor (TRAF) domain-containing adapter proteins and the signaling enzymes associated with them, are largely unique to the TNF/nerve growth factor receptor family.

Animals↗
PubMed2001

The adipocyte: a model for integration of endocrine and metabolic signaling in energy metabolism regulation.

The ability to ensure continuous availability of energy despite highly variable supplies in the environment is a major determinant of the survival of all species. In higher organisms, including mammals, the capacity to efficiently store excess energy as triglycerides in adipocytes, from which stored energy could be rapidly released for use at other sites, was developed. To orchestrate the processes of energy storage and release, highly integrated systems operating on several physiological levels have evolved. The adipocyte is no longer considered a passive bystander, because fat cells actively secrete many members of the cytokine family, such as leptin, tumor necrosis factor-alpha, and interleukin-6, among other cytokine signals, which influence peripheral fuel storage, mobilization, and combustion, as well as energy homeostasis. The existence of a network of adipose tissue signaling pathways, arranged in a hierarchical fashion, constitutes a metabolic repertoire that enables the organism to adapt to a wide range of different metabolic challenges, such as starvation, stress, infection, and short periods of gross energy excess.

Adipocytes↗
PubMed1990

Rat hindlimb unloading: soleus histochemistry, ultrastructure, and electromyography.

Soleus muscle atrophy was induced by hindlimb unloading of male Sprague-Dawley rats (305 +/- 15 g) for 4, 7, and 10-14 days. Controls (291 +/- 14 g) were housed in vivarium cages. Soleus electromyogram (EMG) activity was recorded before and during tail suspension. Unloading caused progressive reduction in the muscle-to-body weight ratio. After 14 days, type I and IIa fibers decreased in area 63 and 47%, respectively. Subsarcolemmal mitochondria and myofibrils were degraded more rapidly than intermyofibrillar mitochondria and the cell membrane. After 10 days, 3% of the fibers exhibited segmental necrosis; affected fibers were all high-oxidative type IIa fibers. This suggested ischemic injury. By 13 days, 30% of the fibers possessed central corelike lesions involving primarily type I fibers. Video monitoring revealed abnormal plantar flexion of the hindfeet by 4 days; this posture shortened the soleus working range. Corelike lesions indicated adaptation to the shortened length. No morphological signs of denervation were detected. EMG activity shifted from tonic to phasic, and aggregate activity was 13% of normal after 7 days. These findings indicate that the atrophy and pathological changes result from unloaded contractions, reduced use, compromised blood flow, and shortened working length.

Adenosine Triphosphatases↗
PubMed2003

Processing of periodic whisker deflections by neurons in the ventroposterior medial and thalamic reticular nuclei.

Rats employ rhythmic whisker movements to sample information in their sensory environment. To study frequency tuning and filtering characteristics of thalamic circuitry, we recorded single-unit responses of ventroposterior medial (VPm) and thalamic reticular (Rt) neurons to 1- to 40-Hz sinusoidal and pulsatile whisker deflection in lightly narcotized rats. Neuronal entrainment was assessed by a measure of the relative modulation (RM) of firing at the stimulus frequency given by the first harmonic (F1) of the cycle time histogram divided by the mean firing rate (F0). VPm signaling of both sinusoidal and periodic pulsatile whisker movements improved gradually over 1-16 and was maximal at 20-40 Hz. By contrast, the RM of Rt responses increased over 1-8 Hz, but deteriorated progressively over the 12- to 40-Hz range. In Rt, response adaptation occurred at lower stimulus frequencies and to a greater extent than in VPm. Within a train of high-frequency stimuli, Rt responses progressively decremented, possibly due to the accumulation of inhibition, whereas those of VPm neurons augmented. Mean firing rates in Rt increased 42 spikes/s over 1-40 Hz, providing tonic (low RM) inhibition during high-frequency stimulation that may enhance VPm signal-to-noise ratios. Consistent with this view, VPm mean firing rates increased only 13 spikes/s over 1-40 Hz, and inter-deflection activity was suppressed to a greater extent than stimulus-evoked responses. Rt inhibition is likely to act in concert with actions of neuromodulators in optimizing thalamic temporal signaling of high-frequency whisker movements.

Action Potentials↗
PubMed2003

Neuroimaging of direction-selective mechanisms for second-order motion.

Psychophysical findings have revealed a functional segregation of processing for 1st-order motion (movement of luminance modulation) and 2nd-order motion (e.g., movement of contrast modulation). However neural correlates of this psychophysical distinction remain controversial. To test for a corresponding anatomical segregation, we conducted a new functional magnetic resonance imaging (fMRI) study to localize direction-selective cortical mechanisms for 1st- and 2nd-order motion stimuli, by measuring direction-contingent response changes induced by motion adaptation, with deliberate control of attention. The 2nd-order motion stimulus generated direction-selective adaptation in a wide range of visual cortical areas, including areas V1, V2, V3, VP, V3A, V4v, and MT+. Moreover, the pattern of activity was similar to that obtained with 1st-order motion stimuli. Contrary to expectations from psychophysics, these results suggest that in the human visual cortex, the direction of 2nd-order motion is represented as early as V1. In addition, we found no obvious anatomical segregation in the neural substrates for 1st- and 2nd-order motion processing that can be resolved using standard fMRI.

Analysis of Variance↗
PubMed2003

Interaction of mycoplasmas with host cells.

The mycoplasmas form a large group of prokaryotic microorganisms with over 190 species distinguished from ordinary bacteria by their small size, minute genome, and total lack of a cell wall. Owing to their limited biosynthetic capabilities, most mycoplasmas are parasites exhibiting strict host and tissue specificities. The aim of this review is to collate present knowledge on the strategies employed by mycoplasmas while interacting with their host eukaryotic cells. Prominant among these strategies is the adherence of mycoplasma to host cells, identifying the mycoplasmal adhesins as well as the mammalian membrane receptors; the invasion of mycoplasmas into host cells including studies on the role of mycoplasmal surface molecules and signaling mechanisms in the invasion; the fusion of mycoplasmas with host cells, a novel process that raises intriguing questions of how microinjection of mycoplasma components into eukaryotic cells subvert and damage the host cells. The observations of diverse interactions of mycoplasmas with cells of the immune system and their immunomodulatory effects and the discovery of genetic systems that enable mycoplasmas to rapidly change their surface antigenic composition have been important developments in mycoplasma research over the past decade, showing that mycoplasmas possess an impressive capability of maintaining a dynamic surface architecture that is antigenically and functionally versatile, contributing to the capability of the mycoplasmas to adapt to a large range of habitats and cause diseases that are often chronic in nature.

Animals↗
PubMed1995

Elevated concentrations of plasma lipids and apolipoproteins B, C-III, and E are associated with the progression of coronary artery disease in familial hypercholesterolemic swine.

We reported earlier that a complex familial hypercholesterolemia (c-FHC) phenotype characterized by elevated levels of total plasma cholesterol (TC) and apoB and reduced levels of HDL cholesterol (HDL-C) and apoA-I is associated with the development of spontaneous atherosclerotic lesions in swine. In this study, we investigated concentrations of plasma lipids and apolipoproteins B, C-III, and E in six parental animals of two cholesterol concentration phenotypes and their 32 offspring, which segregated into high, intermediate, and normal cholesterol phenotypes. Subsequently, we compared the extent of atherosclerotic lesion development in coronary arteries to the concentrations of plasma lipids and apolipoproteins in the parents and two offspring per family. Mean concentrations for the high (n = 23), intermediate (n = 13), and normal (n = 2) cholesterol level phenotypes at 4 months of age were TC, 316 +/- 62.2, 159 +/- 17.1, and 105 +/- 12 mg/dL; LDL cholesterol, 275 +/- 63.1, 113 +/- 16.4, and 67 +/- 18.4 mg/dL; HDL-C, 35 +/- 6.1, 41 +/- 5.7, and 33 +/- 6.4 mg/dL; triglycerides, 48 +/- 10.8, 39 +/- 8.0, and 29 +/- 5.7 mg/dL; apoB, 152 +/- 32.5, 80 +/- 7.2, and 48 +/- 5.7 mg/dL; apoC-III, 10 +/- 4.2, 8 +/- 1.7, and 3 +/- 0.1 mg/dL; and apoE, 17 +/- 3.4, 7 +/- 1.7, and 5 +/- 0.7 mg/dL, respectively. Histological analysis of the major coronary arteries from members of the three families showed considerable variation in the severity of lesions, ranging from foci of adaptive intimal thickening consisting of two to six layers of smooth muscle cells to advanced lesions containing necrotic cores, cholesterol clefts, calcification, and hemorrhage (type V). The most extensive lesions occurred only in animals of the high cholesterol phenotype (ie, c-FHC), in which the concentration of TC and apoB progressively increased after 4 months of age, apoC-III, apoE, and triglycerides increased or remained elevated, and HDL-C decreased, except for one animal. Data presented here show that the plasma cholesterol phenotypes in FHC animals are associated with levels of apolipoproteins B, C-III, and E and indicate that the increases in the studied parameters after 4 months of age correlate with the progression of coronary artery disease.

Animals↗
PubMed2006

Nanostructured bio-functional polymer brushes.

Structured poly(glycidyl methracrylate) (poly-GMA) brushes have been grafted onto flexible fluoro-polymer films using a radiation grafting process. The reactive epoxide of poly-GMA provides the basis for a versatile biofunctionalization of the grafted brushes. Structure definition by extreme ultraviolet (EUV) exposure allowed nanometer-scale resolution of periodic patterns. By variation of the exposure dose the height of the grafted structures can be adapted in a wide range. Derivatization of the grafted brushes included reaction with various amines with different side chains, hydrolysis of the epoxide to diols to increase protein resistance and introduction of ionic groups to yield poly-electrolytes. As an example for biofunctionalization, biotin was linked to the grafted brush and biofunctionality was demonstrated in a competitive biotin-streptavidin assay. In this article we also present a brief review of other approaches to obtain structured biofunctional polymer brushes.

Fluorocarbons↗