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At least 325 records · Page 18Linked to original sources
PubMed2003

Chlamydia trachomatis type III secretion: evidence for a functional apparatus during early-cycle development.

The obligate intracellular bacterium Chlamydia trachomatis occupies a parasitophorous vacuole termed an inclusion. During its intracellular developmental cycle, C. trachomatis maintains this intracellular niche, presumably by expressing a type III secretion system, which deploys a set of host cell-interactive proteins including inclusion membrane-localized proteins termed Incs. Some Incs are expressed and secreted by 2 h (early cycle) after infection, whereas the expression of type III-specific genes is not detectable until 6-12 h (mid-cycle). To resolve this paradox, we investigated the presence of a type III apparatus on elementary bodies (EBs) that might function early in infection. We demonstrate the existence of the type III secretory apparatus by matrix-assisted laser desorption ionization time-of-flight (MALDI-TOF) and immunoblot analyses of purified EB extracts. Immunoblots using polyclonal antibodies specific for the core apparatus component CdsJ identified this protein in both EB and reticulate body (RB) extracts. Furthermore, CdsJ-specific signals were detected by immunoblot of whole infected-culture extracts and by indirect immunofluorescence of infected monolayers at times before the detection of cdsJ-specific message. Finally, expression of IncC, expressed by 2 h after infection during C. trachomatis infections, in Yersinia pseudotuberculosis resulted in its secretion via the Yersinia type III apparatus. Based on these data, we propose a model in which type III secretion pores are present on EBs and mediate secretion of early Incs and possible additional effectors. Mid-cycle expression of type III genes would then replenish secretion apparatus on vegetative RBs and serve as a source of secretion pores for subsequently formed EBs.

Bacterial Infections↗
PubMed2001

Cytometric analysis of an epitope-tagged transit peptide bound to the chloroplast translocation apparatus.

Chloroplast transit peptides are necessary and sufficient for the targeting and translocation of precursor proteins across the chloroplast envelope. However, the mechanism by which transit peptides engage the translocation apparatus has not been investigated. To analyse this interaction, we have developed a novel epitope-tagged transit peptide derived from the precursor of the small subunit of pea Rubisco. The recombinant transit peptide, His-S-SStp, contains a removable dual-epitope tag, His-S, at its N-terminus that permits both rapid purification via immobilized metal affinity chromatography and detection by blotting, flow cytometry and laser-scanning confocal microscopy. Unlike other chimeric precursors, which place the passenger protein C-terminal to the transit peptide, His-S-SStp bound to the translocation apparatus yet did not translocate across the chloroplast envelope. This early translocation intermediate allowed non-radioactive detection using fluorescent and chemiluminescent reporters. The physiological relevance of this interaction was confirmed by protein import competitions, sensitivity to pre- and post-import thermolysin treatment, photochemical cross-linking and organelle fractionation. The interaction was specific for the transit peptide since His-S alone did not engage the chloroplast translocation apparatus. Quantitation of the bound transit peptide was determined by flow cytometry, showing saturation of binding yet only slight ATP-dependence. The addition of GTP showed inhibition of the binding of His-S-SStp to the chloroplasts indicating an involvement of GTP in the formation of this early translocation intermediate. In addition, direct visualization of His-S-SStp and Toc75 by confocal microscopy revealed a patch-like labeling, suggesting a co-ordinate localization to discrete regions on the chloroplast envelope. These findings represent the first direct visualization of a transit peptide interacting with the chloroplast translocation apparatus. Furthermore, identification of a chloroplast-binding intermediate may provide a novel tool to dissect interactions between a transit peptide and the chloroplast translocation apparatus.

Amino Acid Sequence↗
PubMed2002

Asymmetry of the central apparatus defines the location of active microtubule sliding in Chlamydomonas flagella.

Regulation of ciliary and flagellar motility requires spatial control of dynein-driven microtubule sliding. However, the mechanism for regulating the location and symmetry of dynein activity is not understood. One hypothesis is that the asymmetrically organized central apparatus, through interactions with the radial spokes, transmits a signal to regulate dynein-driven microtubule sliding between subsets of doublet microtubules. Based on this model, we hypothesized that the orientation of the central apparatus defines positions of active microtubule sliding required to control bending in the axoneme. To test this, we induced microtubule sliding in axonemes isolated from wild-type and mutant Chlamydomonas cells, and then used electron microscopy to determine the orientation of the central apparatus. Transverse sections of wild-type axonemes revealed that the C1 microtubule is predominantly oriented toward the position of active microtubule sliding. In contrast, the central apparatus is randomly oriented in axonemes isolated from radial spoke deficient mutants. For outer arm dynein mutants, the C1 microtubule is oriented toward the position of active microtubule sliding in low calcium buffer, but is randomly oriented in high calcium buffer. These results provide evidence that the central apparatus defines the position of active microtubule sliding, and may regulate the size and shape of axonemal bends through interactions with the radial spokes. In addition, our results indicate that in high calcium conditions required to generate symmetric waveforms, the outer dynein arms are potential targets of the central pair-radial spoke control system.

Animals↗
PubMed2005

Spatial location and requirements for the assembly of the Agrobacterium tumefaciens type IV secretion apparatus.

Type IV secretion is used by pathogenic microorganisms to transfer effector macromolecules to eukaryotic target cells. The VirB/D4 apparatus of Agrobacterium tumefaciens transfers DNA and proteins to plant cells. We postulated that the cell pole is the site of assembly of the A. tumefaciens type IV apparatus. Using immunofluorescence microscopy, we now demonstrate that 10 of the VirB proteins localized primarily to one cell pole and a macromolecular VirB complex is assembled at the pole. Neither the assembly of the complex nor polar localization of a VirB protein requires ATP utilization by the VirB ATPases. The requirement of other VirB proteins for the polar localization of at least six VirB proteins indicates an essential role of protein-protein interaction in polar targeting. Four proteins (VirB3, VirB4, VirB8, and VirB11) could target themselves to a cell pole independent of a VirB protein. We provide evidence that VirB6-VirB10 are the structural components of the type IV apparatus. Using strains that express defined subsets of the virB genes, we demonstrate that VirB7-VirB10 are the minimum components sufficient for the assembly of a polar VirB complex. VirB6 associates with this complex to form the type IV secretion apparatus. VirB8 functions as the assembly factor and targets the apparatus to the cell pole.

Agrobacterium tumefaciens↗
PubMed1971

The synthesis of microtubule and other proteins of the oral apparatus in Tetrahymena pyriformis.

Several proteins, including microtubule proteins, have been isolated from the oral apparatus of the ciliate Tetrahymena. The synthesis of these proteins has been studied in relation to formation of this organelle system by the cell. Electron microscopy has shown that the isolated oral apparatus consists primarily of basal bodies, pellicular membranes, and a system of subpellicular microtubules and filaments. Cilia were removed during the isolation; therefore none of the proteins studied was from these structures. Evidence was obtained from the study of total oral apparatus protein which indicates that at least some of the proteins involved in formation of this organelle system may be synthesized and stored in the cytoplasm for use over long periods. This pattern of regulation was found for three individual proteins isolated from the oral apparatus fraction after extraction with a phenol-acetic acid solvent. A different pattern of regulation was found for microtubule proteins isolated from the oral apparatus of Tetrahymena. The data suggest that microtubule proteins, at least in logarithmically growing cells, are not stored in a cytoplasmic pool but are synthesized in the same cell cycle in which they are assembled into oral structures.

Animals↗
PubMed2001

New experimental apparatus for multimodal resonance imaging: initial EPRI and NMRI experimental results.

Electron paramagnetic resonance imaging (EPRI) is a recently developed imaging technique employed in the study of free radicals in living systems. A full understanding of many physiological and pathological processes involving free radicals has not yet been attempted. The reason for this is that whilst nuclear magnetic resonance imaging (NMRI) is able to generate very accurate images of soft tissues and organs, EPRI does not have this capability because of its sensitivity limitations and the large linewidths of paramagnetic probes. This work describes the development and optimization of a multimodal apparatus capable of performing both pulsed EPRI and NMRI experiments on the same sample. The instrument combines the possibilities offered by both techniques: the functional and biochemical information achieved with EPRI, and the high-resolution anatomical images generated by NMRI. At present, these experiments are performed by moving the sample from an EPRI spectrometer to an NMRI apparatus. Consequently, the acquisition times are very long and several problems arise in image reconstruction. On the other hand, a unique apparatus operating in the two modalities greatly reduces the acquisition times and makes it possible to relate accurately the observed distribution of electron spin density with the anatomical description of individual organs. The experiments are performed at 357 Gauss, corresponding to a resonance frequency of 1.52 MHz for NMR and 1 GHz for EPR. In the present work, a detailed description of the apparatus is reported, including the main magnet, the gradient assembly, the multimodal cavity and the transmitter and receiver systems. The preliminary experimental results obtained by this apparatus are presented.

Electron Spin Resonance Spectroscopy↗
PubMed1999

Mitral valve replacement with preservation of the subvalvular apparatus.

The introduction of the Starr-Edwards valve allowed complete replacement of diseased left-sided heart valves. With improved cardiopulmonary bypass, myocardial protection, and surgical techniques the mortality rate from aortic valve replacement decreased substantially, whereas the mortality rate from mitral valve replacement remained high, largely because of low cardiac output syndrome. Increasing use of mitral valve repair techniques resulted in a marked decrease in short-term and long-term morbidity and mortality when treating patients with mitral regurgitation. Some believed that this resulted from maintenance of the mitral annular papillary muscle continuity during mitral valve repair. Subsequent experimental and clinical studies have validated the positive short-term and long-term effects of maintaining the integrity of the mitral valve subvalvular apparatus. This article considers the history of the clinical use of preservation of the subvalvular apparatus, the physiologic studies examining this concept, and the clinical data available on its use. It also examines the following: 1) mitral stenosis versus mitral regurgitation and the preservation of the subvalvular apparatus; 2) whether the anterior, posterior, or both areas of the subvalvular apparatus should be preserved; and 3) the surgical techniques for the preservation of the subvalvular apparatus and valve implantation.

Animals↗
PubMed1998

Ultrahigh-vacuum reaction apparatus to study synchrotron-radiation-stimulated processes.

An ultrahigh-vacuum reaction apparatus to study synchrotron-radiation-stimulated processes has been constructed and placed on beamline 4B of the synchrotron radiation storage ring (UVSOR) at the Institute for Molecular Science. The apparatus is designed so that multiple synchrotron radiation processes such as etching and chemical vapour deposition can be carried out successively without breaking the high vacuum. It is equipped with IR reflection absorption spectroscopy (IRRAS) apparatus and reflective high-energy electron diffraction (RHEED) apparatus for in situ observations. The basic parameters of the apparatus including etching and deposition rates have been measured. IRRAS using buried metal layer substrates has been confirmed to be a very useful method of analyzing the reaction mechanisms of the synchrotron-radiation-stimulated processes.

Journal Article↗
PubMed1990

Immunohistochemical study of lysozyme in various benign sweat apparatus tumors.

We investigated the existence of lysozyme in various sweat apparatus tumors by adopting the avidin-biotin-peroxidase complex method. Positive reactions for lysozyme were found in four cases of apocrine cystadenoma, hidradenoma papilliferum, and an apocrine sweat apparatus benign tumor resembling "apocrine spiradenoma", all of which derive from apocrine sweat apparatus. On the other hand, in ten cases of syringoma, eccrine hidrocystoma, clear cell hidradenoma, eccrine spiradenoma, and eccrine poroma, which derive from eccrine sweat apparatus, no positive stainings for lysozyme were obtained. In four out of five cases of mixed tumor of the skin, the apocrine type exhibited positive results. Two cases of syringocystadenoma papilliferum were negative for lysozyme. The investigation of lysozyme in various sweat apparatus tumors is useful in determining the direction of differentiation in these tumors.

Adenoma, Sweat Gland↗
PubMed1995

Development of a new apparatus to observe microcirculation chronically in continuous flow blood pump research.

To observe microcirculation chronically is an important key to the successful evaluation of the continuous flow blood pump. In this study, we succeeded in developing a new apparatus by which microcirculation could be observed chronically without a microscope in a conscious animal. The apparatus utilizes a charge coupled device (CCD). A thin living tissue, such as mesentery, is put directly on a highly integrated CCD and is lit up through the tissue with a light-emitting diode (LED). The vascular nets in the tissue are projected onto the CCD like a contact photograph, which is then sent to a television screen and which can be used to analyze their motion and function. A 0.5-inch CCD having 250,000 pixels was used in this study. The cover glass of the CCD was removed, and a fiber optic plate was fixed onto the surface of the CCD for the tissue to be able to contact with the apparatus surface without clearance. The CCD as well as the LED were molded with epoxy resin for electrical insulation. The apparatus was 35 mm wide and 12 mm high with a micro stand for an LED, which can be easily implanted into an animal. The apparatus was implanted into a rabbit for 12 h. The configuration of arterioles and venules, tens of micrometers in diameter, and their motions in subcutaneous tissue could be observed.

Animals↗
PubMed1991

Experimental apparatus for selection of adherent microorganisms under stringent growth conditions.

A bioreactor apparatus is described for studying bacterial attachment. A cyclic, on-off, flow regime was imposed within the apparatus. Model calculations illustrate the utility of this flow pattern in the selection and maintenance of slow-growing, adherent organisms. The apparatus is believed to have general utility in testing bacterial attachment influenced by many types of experimental or environmental constraints, including variations in fluid dynamics, presence of toxic substances (metals or organics), nature of the substratum surface, concentrations of limiting nutrients, and competition between bacterial strains. As an example application, the apparatus was employed to test 14 bacterial strains for surface attachment in a nutrient-limited growth medium. The medium was developed, using the chemical equilibrium program MINEQL, for planned studies of biofilms in a solution with a chemically defined composition that permits calculation of trace metal speciation. The apparatus was used to select organisms with growth and attachment characteristics that could not be evaluated by conventional batch, or chemostat, culture conditions. When supplied with acetate, pyruvate, or succinate as a carbon and energy source, the gram-negative strains Pseudomonas cepacia 17616 and Zoogloea sp. WGO4 showed superior attachment characteristics to glass surfaces in the chemically defined medium but only moderate fluid-phase growth. The gram-positive Arthrobacter sp. strain 9G4D and gram-negative species P. pickettii and Zoogloea sp. WNJ8, when supplied with pyruvate as a carbon and energy source, were capable of superior growth in the fluid phase but formed only a low to moderate biofilm surface coverage.

Bacteria↗
PubMed2006

Apparatus for measuring rat body volume: a methodological proposition.

We propose a communicating-vessels system to measure body volume in live rats through water level detection by hydrostatic weighing. The reproducibility, accuracy, linearity, and reliability of this apparatus were evaluated in two tests using previously weighed water or six aluminum cylinders of known volume after proper system calibration. The applicability of this apparatus to measurement of live animals (Wistar rats) was tested in a transversal experiment with five rats, anesthetized and nonanesthetized. We took 18 measurements of the volume under each condition (anesthetized and nonanesthetized), totaling 90 measurements. The addition of water volumes (50-700 ml) produced a regression equation with a slope of 1.0006 +/- 0.0017, intercept of 0.75 +/- 0.81 (R(2) = 0.99999, standard error of estimate = 0.58 ml), and bias of approximately 1 ml. The differences between cylinders of known volumes and volumes calculated by the system were <0.4 ml. Mean volume errors were 0.01-0.07%. Among the live models, the difference between the volumes obtained for anesthetized and nonanesthetized rats was 0.31 +/- 2.34 (SD) ml (n = 90). These data showed that animal movement does not interfere with the volume measured by the proposed apparatus, and neither anesthesia nor fur shaving is needed for this procedure. Nevertheless, some effort should be taken to eliminate air bubbles trapped in the apparatus or the fur. The proposed apparatus for measuring rat body volume is inexpensive and may be useful for a range of scientific purposes.

Animals↗
PubMed1992

Functional construction of the sacroiliac ligamentous apparatus.

The sacroiliac ligamentous apparatus was examined as a part of a biomechanical pelvis-lower extremities system. The ligamentous apparatus of two pelves was freed, and the findings concerning the ligaments and their direction were drawn by a modular constructed, three-dimensional calculator model of the pelvic region. The ligamentous apparatus of the sacroiliac joint belongs to a functional system. Its task is to minimize every movement in this amphiarthrosis. The ligamentous apparatus shows an adaptation to strong or long-time-acting stresses. The junction between the os sacrum, pelvis and the ligamentous apparatus of the sacroiliac joint can be described as self-tightening. Local stresses are also reduced by the ligaments. A loosening in this system, which has to fix the os sacrum to the pelvic girdle, leads to a static insufficiency. The consequence is pain due to an irritation of the lumbosacral trunk. The exact description of the structure allows a representation according to the laws of similarity mechanics. With such a representation one can build up a computer-aided biomechanical model of the pelvis-lower extremities region. Examples for such a model are biomechanical finite-element models. By observing the laws of similarity mechanics (an exact description of geometric, physical and functional conditions) an efficient biomechanical model can be constructed that also takes into consideration the complex functional circumstances, in contrast to previous models. In order to construct such a model, one has to feed the findings of the examination into a data bank, which has to be demanded.

Biomechanical Phenomena↗
PubMed1994

Development of a thermoelectric cooling apparatus for high-voltage isoelectric focusing on a cellulose acetate membrane.

To develop an isoelectric focusing apparatus using a cellulose acetate membrane (Separax EF), we have designed a thermoelectric cooling isoelectric apparatus. This apparatus has two characteristics. Firstly, the cooling system was switched to a thermoelectric cooling system from an ice-cooling system. Secondly, the chamber lid of the electrophoretic apparatus was also devised so that samples could be applied without opening the chamber lid. With this apparatus we could perform the isoelectric focusing without worrying about room temperature and humidity in the laboratory. Applying 2000 V for an extra 5 min with our module cooling system, we achieved a much higher degree of resolution with three sheets of cellulose acetate membrane (Separax EF) overlaid for simultaneous electrophoresis. Thus, three types of information could be obtained from only one electrophoretic procedure.

Biocompatible Materials↗
PubMed1995

[Three simple maze tests employing mice housed in maze apparatuses].

This report deals with three different maze methods using spontaneous learning behavior. Forty-eight mice housed in an apparatus with a multiple maze mastered the maze task on the 5th day after the start of housing. Then they were divided into four groups, and two of the groups were treated with AF64A (3.5 nmol, i.c.v.) or trimethyltin (TMT, 3 mg/kg, p.o.), and the other two groups were treated with the vehicle as the respective control. Seven days after the treatment, their memory retrieval was tested. Subsequently, the same mice were housed in the apparatus with a T-maze. After the finish of the experiment using the T-maze, they were housed in the apparatus with an eight arm radial maze. The control groups mastered the T-maze task with a 3-sec delay on the 4th day after the start of housing and the radial maze task on the 10th day after the start of housing. Both the treatments lowered the performance in all maze tasks. These results show that the mice housed in the apparatus with maze learned to negotiate the maze spontaneously, and the apparatuses are useful for estimating memory in mice with little effort.

Animals↗
PubMed1998

New apparatus stimulating the region of tongue innervated by glossopharyngeal nerve and its application to monosodium glutamate (MSG) solution.

Some apparatuses stimulating the region of tongue innervated by chorda tympani nerve were well known and they were used popularly, but the apparatuses for glossopharyngeal nerve were not produced yet. In the study, the apparatus for the stimulations of glossopharyngeal nerve was trially produced. It was known that the substance of "Umami" was responded well in the tongue region innervated by glossopharyngeal nerve. The evoked potential for the stimulations of monosodium glutamate (MSG) solution which was known as substance of Umami was measured with the use of the trial stimulating apparatus. The evoked potential was detected by monopolar lead for the positions between Cz and A1 (left lobe) with the ground at Fpz. The evoked potential of MSG consisted of a positive peak wave with the latency at approximately 250 ms. The amplitudes increased and the latency decreased as the concentration of MSG increased. The evoked potential stimulating the tongue innervated by this glossopharyngeal nerve could be detected for the first time by the use of the trial apparatus.

Evoked Potentials↗
PubMed2000

[A study on application of multiple jointed contact-type measuring apparatus for dental use].

The measurement of oral maxillofacial region, including dentition and teeth, is important information for diagnosis and assessment of treatment in clinical dentistry. There are two kinds of the measuring systems, i.e., contact-type system and non-contact-type system. The latter mainly has been investigated in dentistry, because of its simplicity and quickness for measurement. However, since the objects in dentistry have complex shapes with curved surfaces and many dead angles, accuracy of values obtained by this system is insufficient for clinical use. Even though divided measurement is employed to overcome the problems, the increase of measuring element results in deterioration of the accuracy. In the present study, the accuracy of a contact-type measuring apparatus with 5 pivoting joints was investigated. For the cavity model, the depth and the width were measured. For the standardized dental cast which consists of three spheres, the diameter of three spheres and the distance of each sphere were measured. The values obtained by this apparatus were compared with those obtained by the precise 3D-measurement apparatus. The differences were insignificant in the cavity model. Significant difference was found in the standardized dental cast. However, the difference was very small, and so it is negligible in clinical use. By using this apparatus, the form of undercut and teeth was reconstructed completely. The results suggested that the multiple jointed contact-type measuring apparatus could be applied to the measurement in clinical dentistry.

Dental Equipment↗
PubMed1976

Halo-pelvic distraction apparatus. An analysis of one hundred and fifty consecutive patients.

One hundred and fifty consecutive patients on whom the halo-pelvic apparatus was used were analyzed. Sixty-six per cent had tuberculous kyphosis or paralytic scoliosis. The apparatus was of great value in holding and correcting spines with tuberculous kyphosis and the results were gratifying, even though the average correction of the kyphos was only 20 per cent. In paralytic scoliosis a 46 per cent correction was obtained, but presently Dwyer and Harrington instrumentation are used for the majority of such cases. The halo-pelvic apparatus was effective in holding and correcting severe congenital curves and kyphoscoliosis secondary to neurofibromatosis, especially when there were signs of cord compression. The complication rate was high early in the series, but has been lowered by strict patient selection and close adherence to specific guidelines in the application of the apparatus. The apparatus should be reserved for severe deformities when other means of correction or stabilization are inadequate.

Adolescent↗