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PubMed1986

Comparison of immunoblot analyses of spherule-endospore-phase extracellular protein and mycelial-phase antigen of Coccidioides immitis.

The extracellular proteins produced by Coccidioides immitis during growth of the spherule-endospore-phase and mycelial-phase antigen (coccidioidin) were studied by polyacrylamide gel electrophoresis followed by immunoblot analysis to detect specific serologic function. Filtrates obtained from 28- and 120-h growth of the spherule-endospore phase were compared with each other and with coccidioidin by using negative, immunoglobulin M (IgM) precipitin-positive, or complement fixation-positive pooled and single human sera followed by peroxidase-labeled anti-human IgA, IgE, IgG, or IgM (heavy chain specific) or peroxidase-labeled concanavalin A to detect the reaction. A total of 35 bands was seen in the stained gels. Different patterns were noted among the two spherule-endospore preparations and unheated and heated coccidioidin. At least 15 electrophoretically separate antigens were detected with positive serum ranging in approximate molecular weight (Mr) from 100,000 to 18,000. Most were clustered between 45 and 60 kilodaltons (kDa). Common bands were noted at 48 and 18 kDa. At least one band at 48 kDa was strongly reactive with complement fixation-positive serum demonstrated by reaction with anti-IgG and anti-IgE. In contrast, doublet bands in the 50- to 65-kDa area were highly reactive with IgM precipitin-positive serum detected by anti-IgM. IgM antibodies present in both positive sera reacted with a band at 46 kDa which was not reactive with IgG. Heating the antigens altered the reactivity of many of the antigens, including the 48-kDa band, but not the 46-kDa band.

Antigens, Fungal↗
PubMed1999

Influence of acute-phase parasite load on pathology, parasitism, and activation of the immune system at the late chronic phase of Chagas' disease.

To obtain low and high parasite loads in the acute phase of Chagas' disease, A/J mice were infected with 10(3) or 10(5) Trypanosoma cruzi trypomastigotes of the Y strain and treated on day 6 with benznidazol. One year later, chronically infected mice were screened for subpatent parasitemias, tissue pathology, and immune response. Mice infected with the high parasite inoculum showed higher levels of chronic parasitemias, heart and striated muscle inflammation, and activation of the immune system than did mice infected with the low inoculum. Concerning the activation of the immune system, the main findings for high-dose-infected mice were (i) increased numbers of splenocytes, with preferential expansion of CD8(+) and B220(-) CD5(-) cells, many of them bearing a macrophage phenotype; (ii) higher frequencies of B (B220(+)), CD4(+), and CD8(+) large lymphocytes; (iii) a shift of CD4(+) cells towards a CD45RBLow phenotype; (iv) increased frequencies of both CD45RBLow and CD45RBHigh large CD4(+) cells; (v) augmented numbers of total immunoglobulin (Ig)-secreting cells, with predominance of IgG2a-producing cells; and (vi) increased production of gamma interferon and interleukin 4. In addition, these mice presented lower IgM and higher IgG2a and IgG1 parasite-specific serum antibody levels. Our results indicate that the parasite load at the acute phase of T. cruzi infection influences the activation of the immune system and development of Chagas' disease pathology at the late chronic phase of the disease.

Acute Disease↗
PubMed1993

Axial filament formation in Bacillus subtilis: induction of nucleoids of increasing length after addition of chloramphenicol to exponential-phase cultures approaching stationary phase.

When chloramphenicol was added to a culture of Bacillus subtilis in early exponential growth, microscopic observation of cells stained by 4',6-diamidino-2-phenylindole showed nucleoids that had changed in appearance from irregular spheres and dumbbells to large, brightly stained spheres and ovals. In contrast, the addition of chloramphenicol to cultures in mid- and late exponential growth showed cells with elongated nucleoids whose frequency and length increased as the culture approached stationary phase. The kinetics of nucleoid elongation after the addition of chloramphenicol to exponential-phase cultures was complex. Immediately after treatment, the rate of nucleoid elongation was very rapid. The nucleoid then elongated steadily for about 4 min, after which the rate of elongation decreased considerably. Nucleoids of cells treated with 6-(p-hydroxyphenylazo)-uracil (an inhibitor of DNA synthesis) exhibited the immediate rapid elongation upon chloramphenicol treatment but not the subsequent changes. These observations suggest that axial filament formation during stationary phase (stage I of sporulation) in the absence of chloramphenicol results from changes in nucleoid structure that are initiated earlier, during exponential growth.

Bacillus subtilis↗
PubMed2006

Myocardial tissue phase mapping with cine phase-contrast mr imaging: regional wall motion analysis in healthy volunteers.

PURPOSE: To establish prospectively a database of normal three-dimensional systolic and diastolic endocardial and epicardial velocity values for all myocardial segments in healthy volunteers by using cine phase-contrast velocity magnetic resonance imaging, also called tissue phase mapping (TPM). MATERIALS AND METHODS: The study was approved by the institutional ethics committee and was conducted according to principles of the Declaration of Helsinki; each subject provided informed written consent. Ninety-six healthy volunteers (57 [59%] men, 39 [41%] women; mean age, 38 years +/- 12 [standard deviation]) underwent cardiac phase-contrast imaging with a black blood segmented k-space gradient-echo sequence for the analysis of three-dimensional myocardial velocity with high spatial resolution at 1.5 T on basal, midventricular, and apical short-axis views. Eighteen consecutive volunteers were imaged twice to determine interstudy reproducibility, and intra- and interobserver variability values were analyzed. Systolic and diastolic velocity curves were analyzed for peak velocity and time to peak velocity in the radial, circumferential, and longitudinal directions, as well as for torsion rate and longitudinal strain rate. Mixed-effects models with a random intercept for volunteers were used to test differences among the three ventricular sections and the transmural, endocardial, and epicardial parameters. RESULTS: TPM enabled reproducible assessment of myocardial velocity with small intra- and interobserver variability values. Systolic peak radial velocity was lowest at the apical level (P < .001); diastolic peak radial velocity was similar at all three myocardial levels (P = .73). As viewed from the apex, a relative counterclockwise rotation during systole was followed by a relative clockwise rotation of the apex against the base. Diastolic and systolic peak longitudinal velocity values decreased from base to apex (P < .001). A gradient between endocardium and epicardium was observed for radial velocity values, with greater endocardial velocity values (P < .001). CONCLUSION: TPM is a reproducible comprehensive modality for assessment of regional wall motion, and intra- and interobserver variability values are low.

Adult↗
PubMedDate not supplied

Effects of L-proline on phase I and phase II xenobiotic biotransformation capacities of rat and human hepatocytes in long-term collagen gel cultures.

L-Proline supplementation of the medium for collagen gel cultures of hepatocytes has been shown to improve albumin secretion. A study was made as to whether L-proline is also essential for the maintenance of xenobiotic biotransformation capacities in collagen gel sandwich and immobilisation cultures of rat and human hepatocytes. Key phase I (cytochrome P450-dependent monooxygenase [CYP)] and microsomal epoxide hydrase [mEH]) and phase II (glutathione S-transferase [GST]) biotransformation enzyme activities and the secretion of albumin in the culture medium were assessed in the absence and presence of L-proline. CYP and mEH activities were not affected by the addition of L-proline, whereas phase II alpha-Class GST activity of rat hepatocytes in collagen cultures was decreased. Species differences were demonstrated, as human hepatocytes showed a better maintenance of GST activities than their rat counterparts in the presence of L-proline. Albumin secretion, often considered to be a marker for differentiated cell function, does not parallel the biotransformation capacities of the hepatocytes in culture. Additional results demonstrated an L-proline-mediated enhancement of the proliferation rate of contaminating stellate cells in conventional monolayer culture. Transdifferentiation of stellate cells to proliferating myofibroblasts, along with an increased albumin secretion and collagen synthesis, are characteristic of fibrotic liver. Since the last two phenomena have been observed in L-proline-supplemented collagen gel cultures, it can be concluded that when stable collagen gel cultures of rat hepatocytes are needed for long-term pharmacotoxicological studies, it is preferable to use an L-proline-free culture medium. Further studies on medium optimisation are required for hepatocytes from species other than rat.

Adult↗
PubMed1992

Changes in the phase response curve of the circadian clock to a phase-shifting stimulus.

Experiments were conducted in hamsters to determine whether the phase response curve (PRC) to injections of the short-acting benzodiazepine triazolam is a fixed or a labile property of the circadian clock. The results indicated that (1) both the shape and the amplitude of the PRC to triazolam generated on the first day of transfer from a light-dark cycle (LD 14:10) to constant darkness (DD) (i.e., PRCLD) were different from those of the PRC generated after many days in DD (PRCDD); and (2) the phase-shifting effects of triazolam on the activity rhythms of hamsters transferred from LD 14:10 or 12:12 to DD changed dramatically within the first 8-9 days spent in DD. In an attempt to accelerate the resynchronization of the circadian clock of hamsters subjected to an 8-hr advance in the LD cycle, triazolam was given to the animals at a time selected on the basis of the characteristics of PRCLD. The activity rhythms of five of eight triazolam-treated animals were resynchronized to the new LD cycle within 2-4 days after the shift, whereas those of most of the control animals were resynchronized 21-29 days after the shift. These findings suggest that attempts to use pharmacological or nonpharmacological tools to phase-shift circadian clocks under entrained conditions should take into account information derived from PRCs generated at the time of transition from entrained to free-running conditions.

Animals↗
PubMed2000

Hydrophobicity parameters determined by reversed-phase liquid chromatography. Xiv. Application of a new hydrogen-accepting scale of monosubstituted pyrazines to analysis of the relationship between octanol-water partition coefficients and retention factors measured in different mobile phases.

We recently proposed a new H-accepting scale, SHA, for monosubstituted pyrazines, and demonstrated that this parameter works effectively in expressing the relationship between logP (P: 1-octanol/water partition coefficient) and logk' (k': retention factor derived from reversed phase liquid chromatography) with aqueous methanol solutions as the mobile phase, according to the equation: logk' = alogP+rhosigmaI+sSHA+const., where sigmaI represents the electronic substituent constant. In this work, we have extended the same treatment to analysis of logk' measured in mobile phases containing different organic modifiers such as 1-propanol, acetonitrile, and dioxane, and found that the above equation is still useful. By comparing the correlations obtained, it was confirmed that the parameter SHA could be universally utilized for representing the difference in H-bonding effects involved in different partitioning systems.

Chromatography, Liquid↗
PubMed2002

Enhanced protection against the migratory phase, but defective protection against the intestinal phase of Strongyloides venezuelensis infection in cotton rats, Sigmodon hispidus.

The protective capacity of the cotton rat, Sigmodon hispidus, against the migratory and intestinal phases of Strongyloides venezuelensis infection was examined. After subcutaneous infection with infective larvae (L(3)), adult worm recovery rates from male and female animals on Day 71 were only 0.10% and 0.06% of initial dose, respectively. To determine whether this enhanced protection was expressed during the migratory phase or the intestinal phase, larval recovery from the lungs of cotton rat was evaluated 3 days after subcutaneous L(3) infection. The larval recovery rate was only 0.5% of initial dose and about 40-fold lower than that from control mice. Protection in the intestine was also evaluated after intraduodenal implantation of adult worms. About 30% of implanted worms became established and worm burden remained constant until Day 28. Despite a high worm burden on Day 28, EPG was about 25-fold lower than the peak count. To evaluate expulsive capacity and monitor the cellular responses in the intestine of cotton rats, adult Nippostrongylus brasiliensis worms were implanted in addition to S. venezuelensis. Cotton rats were unable to expel adult S. venezuelensis worms, even after 21 days of observation. Although the number of mucosal mast cells increased significantly, the intraepithelial migration of mast cells was not observed. In contrast, N. brasiliensis was expelled by Day 6 in association with goblet cell hyperplasia. These results suggest that in cotton rats, the defective intestinal protection against adult S. venezuelensis worms results from dysfunction of mucosal mast cells.

Animals↗
PubMed2004

Phase noise and phase modulation in optical coherence tomography.

The signal in optical coherence tomography is often modulated either in phase or by use of the Doppler modulation generated by a depth-scanning mechanism. The effect of each type of modulation on the signal's amplitude is evaluated. The advantages of each type of modulation in terms of immunity to phase noise and penetration depth are discussed in relation to two envelope detection schemes, i.e., lock-in detection and rms-to-dc conversion. Phase noise due to drifts and demodulation instabilities causes distortion of the signal envelope and can be responsible in part for the speckle appearance of the image.

Animals↗
PubMed2006

Image reconstruction only by phase data in phase-shifting digital holography.

We describe data compression in phase-shifting digital holography. We demonstrate by experimentation that an image of a diffusely reflecting object can be reconstructed only by phase data of the derived complex amplitude. It is shown that reduction of the bit depth of the phase data does not seriously damage the image even down to 1 bit. We observe enhancement of halo in the image with low bit depths. This tendency is verified quantitatively by a one-dimensional simulation. Our procedure for smoothing the images that result from the data-compression methods is shown to be effective.

Journal Article↗
PubMed2006

Phase-shifting digital holography with a phase difference between orthogonal polarizations.

Phase-shifting digital holography with a phase difference between orthogonal polarizations is proposed. The use of orthogonal polarizations can make it possible to record two phase-shifted holograms simultaneously. By combining the holograms with the distributions of a reference wave and an object wave, the complex field of the object's wavefront can be obtained. Preliminary experimental results are shown to confirm the proposed method.

Journal Article↗
PubMed2000

Generalized algorithm for the unified analysis and simultaneous evaluation of geometrical spin-redirection phase and Pancharatnam phase in a complex interferometric system.

To permit unified analysis and simultaneous evaluation of geometrical spin-redirection phase and Pancharatnam phase, the conventional 2 x 2 Jones matrix calculation is generalized and a new scheme of 3 x 3 matrix calculation is proposed. With the proposed algorithm one can trace the polarization state changes and the geometric phase shifts caused by beam propagation along an arbitrary optical path that involves both reflection and refraction at surfaces with Fresnel shift and birefringence.

Algorithms↗
PubMed2004

Generalized phase-shifting interferometry with arbitrary unknown phase steps for diffraction objects.

A general method of extracting the arbitrary unknown and unequal phase steps in phase-shift interferometry from interferograms recorded on the diffraction field of an object and then reconstructing the object wave front digitally with our derived formulas is proposed. The phase steps are first calculated based on the statistical nature of the diffraction field and are further improved by an iterative approach. This method is simple, highly accurate, and usable for any frame number N (N > or = 3) and for both smooth and diffusing objects, as is verified by a series of computer simulations.

Journal Article↗
PubMed2006

Suppression of phase error in differential phase-shift keying data by amplitude regeneration.

A regenerator for differential phase-shift keying data, consisting of two Mach-Zehnder interferometers and two amplitude regenerators, is examined theoretically. The result, which shows that the phase error is suppressed if phase-preserving amplitude regenerators are used, provides a theoretical description of an already experimentally implemented regenerator.

Journal Article↗
PubMed1984

An evaluation of four different luminescence immunoassay systems: CELIA (chemiluminescent immunoassay), SPALT (solid-phase antigen luminescence technique), ILMA (immunoluminometric assay) and ILSA (immunoluminometric labelled second antibody). A critical study of macro solid phases for use in immunoassay systems, Part III.

The performance of different solid-phase luminescence immunoassays has been documented using four different assay concepts. These are CELIA (chemiluminescence immunoassay), SPALT (solid-phase antigen luminescence technique), ILMA (immunoluminometric assay) and ILSA (immunoluminometric labelled second-antibody assay). CELIA is analogous to a solid-phase radioimmunoassay and uses a labelled antigen, SPALT and ILSA use a labelled second (species-specific) antibody and ILMA a labelled substance-specific antibody, i.e. analogous to the immunoradiometric assay. Both bioluminescent and chemiluminescent labels have been used. Pyruvate kinase was used for bioluminescence and diazoluminol and N-(4-amino-butyl)-N-ethyl isoluminol hemisuccinamide for chemiluminescence. Relevant quality-control parameters and reference ranges have been given for the optimised assays. Assays described are: thyroxine, thyroxine binding globulin, cortisol, caeruloplasmin, ferritin and C-reactive protein. Luminescence immunoassays with coefficients of variation comparable with radioimmunoassay have been designed, values of under 5% being obtainable within the working range of the assay.

Animals↗
PubMedDate not supplied

Medium effects on zwitterionic-biradicaloid intermediates from two phenyl-alpha-oxoamides. Irradiations in fluid and solid protic media, neat solid phases, and the solid, smectic and isotropic phases of a completely saturated phosphonium salt at different temperatures.

The photochemical processes of two N,N-dialkyl phenyl-alpha-oxoamides, N,N-diisopropyl phenyl-alpha-oxoamide (1) and N,N-dibenzyl phenyl-alpha-oxoamide (2), are investigated at different temperatures in methanol and ethylene glycol (to probe the influences of H-bonding and viscosity), in the solid phase of D-sorbitol at room temperature (to compare with the results in the liquid alcohols and to assess the influence of a poorly organized "stiff" environment), in the neat solid phase (to probe the influence of well-ordered, "stiff" matrices), and in the solid, smectic A2 and isotropic phases of methyl-tris-tetradecylphosphonium tetrafluoroborate (1P14BF4) (to assess the ability of the intermediates to respond to subtle changes in the order and polarity of their local environments). From differences between the activation parameters for product pathways from irradiations in methanol and in 1P14BF4, we conclude that the zwitterionic pre-product intermediate from 1 is much more sensitive to the polarity, viscosity and order of its local environment than is the isomeric pre-product biradicaloid intermediate or either of the pre-product intermediates from 2. A very sensitive balance among the medium parameters, as well as internal steric and electronic factors of 1 and 2, controls the reactive pathways of the photochemically generated intermediates.

Journal Article↗
PubMed2002

Separation and control of the elution order of N-t-butyloxycarbonyl amino acids D/L isomers by reversed-phase HPLC using cyclodextrins as chiral selectors for the mobile phase.

The enantiomeric resolution of N-t-butyloxycarbonyl (N-t-Boc) amino acids D/L isomers by reversed-phase HPLC was investigated using cyclodextrins (CD's) as chiral selectors for the mobile phase. The use of a low pH (pH<4) for the mobile phase enabled the enantioseparation of N-t-Boc amino acids. The opposite elution order of D/L isomers was observed when hydroxypropyl-derivatized beta-CD was used instead of native beta-CD. A computer simulation of the enantioseparation showed that the ratio of the retention factors of the chiral selector and the sample determined the elution order and the resolution. When the retention factor of the chiral selector is smaller than that of the sample, an isomer having larger complex formation constant eluted faster. However, when the chiral selector had a larger retention factor than the sample, an opposite elution order of the isomers was obtained. The large difference in the retention factors between the chiral selector and the sample led to good enantiomeric separation.

Amino Acids↗