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1.
Opt Express ; 32(2): 1305-1313, 2024 Jan 15.
Article in English | MEDLINE | ID: mdl-38297685

ABSTRACT

We present a study of homodyne measurements of two-mode, vacuum-seeded, quadrature-squeezed light generated by four-wave mixing in warm rubidium vapor. Our results reveal that the vacuum squeezing can extend down to measurement frequencies of less than 1 Hz, and the squeezing bandwidth, similar to the seeded intensity-difference squeezing measured in this system, reaches up to approximately 20 MHz for typical pump parameters. By dividing the squeezing bandwidth into smaller frequency bins, we show that different sideband frequencies represent independent sources of two-mode squeezing. These properties are useful for quantum sensing and quantum information processing applications. We also investigate the impact of group velocity delays on the correlations in the system, which allows us to optimize the useful spectrum.

2.
J Geophys Res Atmos ; 126(24): e2021JD035692, 2021 Dec 27.
Article in English | MEDLINE | ID: mdl-35865864

ABSTRACT

Accurate fire emissions inventories are crucial to predict the impacts of wildland fires on air quality and atmospheric composition. Two traditional approaches are widely used to calculate fire emissions: a satellite-based top-down approach and a fuels-based bottom-up approach. However, these methods often considerably disagree on the amount of particulate mass emitted from fires. Previously available observational datasets tended to be sparse, and lacked the statistics needed to resolve these methodological discrepancies. Here, we leverage the extensive and comprehensive airborne in situ and remote sensing measurements of smoke plumes from the recent Fire Influence on Regional to Global Environments and Air Quality (FIREX-AQ) campaign to statistically assess the skill of the two traditional approaches. We use detailed campaign observations to calculate and compare emission rates at an exceptionally high-resolution using three separate approaches: top-down, bottom-up, and a novel approach based entirely on integrated airborne in situ measurements. We then compute the daily average of these high-resolution estimates and compare with estimates from lower resolution, global top-down and bottom-up inventories. We uncover strong, linear relationships between all of the high-resolution emission rate estimates in aggregate, however no single approach is capable of capturing the emission characteristics of every fire. Global inventory emission rate estimates exhibited weaker correlations with the high-resolution approaches and displayed evidence of systematic bias. The disparity between the low-resolution global inventories and the high-resolution approaches is likely caused by high levels of uncertainty in essential variables used in bottom-up inventories and imperfect assumptions in top-down inventories.

3.
J Geophys Res Atmos ; 122(16): 8833-8851, 2017 Aug 27.
Article in English | MEDLINE | ID: mdl-33505826

ABSTRACT

Dry aerosol size distributions and scattering coefficients were measured on 10 flights in 32 clear-air regions adjacent to tropical storm anvils over the eastern Atlantic Ocean. Aerosol properties in these regions were compared with those from background air in the upper troposphere at least 40 km from clouds. Median values for aerosol scattering coefficient and particle number concentration >0.3 µm diameter were higher at the anvil edges than in background air, showing that convective clouds loft particles from the lower troposphere to the upper troposphere. These differences are statistically significant. The aerosol enhancement zones extended ~10-15 km horizontally and ~0.25 km vertically below anvil cloud edges but were not due to hygroscopic growth since particles were measured under dry conditions. Number concentrations of particles >0.3 µm diameter were enhanced more for the cases where Saharan dust layers were identified below the clouds with airborne lidar. Median number concentrations in this size range increased from ~100 l-1 in background air to ~400 l-1 adjacent to cloud edges with dust below, with larger enhancements for stronger storm systems. Integration with satellite cloud frequency data indicates that this transfer of large particles from low to high altitudes by convection has little impact on dust concentrations within the Saharan Air Layer itself. However, it can lead to substantial enhancement in large dust particles and, therefore, heterogeneous ice nuclei in the upper troposphere over the Atlantic. This may induce a cloud/aerosol feedback effect that could impact cloud properties in the region and downwind.

4.
Phys Rev Lett ; 114(24): 240401, 2015 Jun 19.
Article in English | MEDLINE | ID: mdl-26196968

ABSTRACT

Unitary transformations are the most general input-output maps available in closed quantum systems. Good control protocols have been developed for qubits, but questions remain about the use of optimal control theory to design unitary maps in high-dimensional Hilbert spaces, and about the feasibility of their robust implementation in the laboratory. Here we design and implement unitary maps in a 16-dimensional Hilbert space associated with the 6S(1/2) ground state of (133)Cs, achieving fidelities >0.98 with built-in robustness to static and dynamic perturbations. Our work has relevance for quantum information processing and provides a template for similar advances on other physical platforms.

5.
Phys Rev Lett ; 111(17): 170502, 2013 Oct 25.
Article in English | MEDLINE | ID: mdl-24206469

ABSTRACT

We implement arbitrary maps between pure states in the 16-dimensional Hilbert space associated with the ground electronic manifold of ^{133}Cs. This is accomplished by driving atoms with phase modulated radio-frequency and microwave fields, using modulation waveforms found via numerical optimization and designed to work robustly in the presence of imperfections. We evaluate the performance of a sample of randomly chosen state maps by randomized benchmarking, obtaining an average fidelity >99%. Our protocol advances state-of-the-art quantum control and has immediate applications in quantum metrology and tomography.

6.
J Acoust Soc Am ; 130(4): EL258-63, 2011 Oct.
Article in English | MEDLINE | ID: mdl-21974501

ABSTRACT

A nonlinear scatterer is simulated in the body of a sample and demonstrates a technique to locate and define the elastic nature of the scatterer. Using the principle of time reversal, elastic wave energy is focused at the interface between blocks of optical grade glass and aluminum. Focusing of energy at the interface creates nonlinear wave scattering that can be detected on the sample perimeter with time-reversal mirror elements. The nonlinearly generated scattered signal is bandpass filtered about the nonlinearly generated components, time reversed and broadcast from the same mirror elements, and the signal is focused at the scattering location on the interface.


Subject(s)
Acoustics , Nonlinear Dynamics , Signal Processing, Computer-Assisted , Sound , Acoustics/instrumentation , Aluminum , Computer Simulation , Elasticity , Equipment Design , Glass , Lasers , Motion , Scattering, Radiation , Sound Spectrography , Time Factors , Transducers , Vibration
7.
Nature ; 461(7265): 768-71, 2009 Oct 08.
Article in English | MEDLINE | ID: mdl-19812668

ABSTRACT

Chaotic behaviour is ubiquitous and plays an important part in most fields of science. In classical physics, chaos is characterized by hypersensitivity of the time evolution of a system to initial conditions. Quantum mechanics does not permit a similar definition owing in part to the uncertainty principle, and in part to the Schrödinger equation, which preserves the overlap between quantum states. This fundamental disconnect poses a challenge to quantum-classical correspondence, and has motivated a long-standing search for quantum signatures of classical chaos. Here we present the experimental realization of a common paradigm for quantum chaos-the quantum kicked top- and the observation directly in quantum phase space of dynamics that have a chaotic classical counterpart. Our system is based on the combined electronic and nuclear spin of a single atom and is therefore deep in the quantum regime; nevertheless, we find good correspondence between the quantum dynamics and classical phase space structures. Because chaos is inherently a dynamical phenomenon, special significance attaches to dynamical signatures such as sensitivity to perturbation or the generation of entropy and entanglement, for which only indirect evidence has been available. We observe clear differences in the sensitivity to perturbation in chaotic versus regular, non-chaotic regimes, and present experimental evidence for dynamical entanglement as a signature of chaos.

8.
Science ; 323(5920): 1443-8, 2009 Mar 13.
Article in English | MEDLINE | ID: mdl-19286547

ABSTRACT

Wind-driven upwelling in the ocean around Antarctica helps regulate the exchange of carbon dioxide (CO2) between the deep sea and the atmosphere, as well as the supply of dissolved silicon to the euphotic zone of the Southern Ocean. Diatom productivity south of the Antarctic Polar Front and the subsequent burial of biogenic opal in underlying sediments are limited by this silicon supply. We show that opal burial rates, and thus upwelling, were enhanced during the termination of the last ice age in each sector of the Southern Ocean. In the record with the greatest temporal resolution, we find evidence for two intervals of enhanced upwelling concurrent with the two intervals of rising atmospheric CO2 during deglaciation. These results directly link increased ventilation of deep water to the deglacial rise in atmospheric CO2.

9.
Int Angiol ; 25(4): 361-9, 2006 Dec.
Article in English | MEDLINE | ID: mdl-17164742

ABSTRACT

AIM: This pilot study provides preliminary information regarding safety and changes in exercise performance during treatment with ranolazine extended-release in patients with reproducible claudication during exercise treadmill testing (ETT). METHODS: We enrolled 45 patients with documented peripheral arterial disease, reproducible claudication on ETT, and ankle-brachial indices <0.85 at rest that decreased by at least 0.15% or 20% immediately postexercise. Randomized patients received double-blind treatment with either ranolazine 1 000 mg b.i.d. (n=22) or placebo (n=23) for 4 weeks. RESULTS: Compared with baseline, peak walking time (PWT) increased (mean+/-SEM) by 53+/-34 s with ranolazine (P=0.13) and by 41+/-33 s with placebo (P=0.22). Pain-free walking time during ETT increased by 62+/-18 s with ranolazine (P=0.002) and 36+/-18 s with placebo (P=0.045). Supplemental analyses, excluding patients with baseline exercise duration (16 min and (12 min, showed additional improvement with ranolazine on PWT. CONCLUSIONS: Ranolazine was well tolerated and these data provide a rationale for proceeding with a definitive trial.


Subject(s)
Acetanilides/therapeutic use , Intermittent Claudication/drug therapy , Piperazines/therapeutic use , Adult , Aged , Aged, 80 and over , Double-Blind Method , Female , Humans , Male , Middle Aged , Pilot Projects , Ranolazine
10.
Microb Pathog ; 41(1): 10-20, 2006 Jul.
Article in English | MEDLINE | ID: mdl-16725305

ABSTRACT

Bartonella henselae, the etiologic agent of cat scratch disease, bacillary angiomatosis and other clinical syndromes initiates infection through a trauma or wound to the skin suggesting involvement of extracellular matrix molecules. We have demonstrated in this study that B. henselae bound strongly fibronectin, collagen IX and X, but comparatively less laminin and collagen IV. B. henselae bound primarily the N- and C-terminal heparin (Hep-1 and Hep-2, respectively) and the gelatin-binding domains of fibronectin (Fn) but not the cell-binding domain. Binding to the Hep-binding domain was significantly inhibited by Hep suggesting common binding sites on the Fn molecule. Furthermore, glycosaminoglycans-mediated binding of B. henselae to soluble Fn showed that Hep but not dextran sulfate inhibited the bacterium binding to Fn. Unlike Fn, B. henselae bound strongly vitronectin only in the presence of Hep or dextran sulfate. Also, the binding of B. henselae to host cells could be inhibited by anti-B. henselae surface-reactive antibodies, the exogenous Fn or the anti-Fn polyclonal antibodies. Ligand blots, batch affinity purification and MALDI-TOF peptide fingerprinting identified B. henselae Pap31, Omp43 and Omp89 as the three major putative Fn-binding proteins (FnBPs) in B. henselae outer membrane proteins. We hypothesized that B. henselae wound associated infections involved interactions with extracellular matrix molecules. Taken together, the above data suggest that interactions between B. henselae and ECM molecules such as Fn may play an important role in the bacterium adherence to and invasion of host cells.


Subject(s)
Adhesins, Bacterial/metabolism , Bartonella henselae/metabolism , Extracellular Matrix Proteins/metabolism , Adhesins, Bacterial/isolation & purification , Bacterial Adhesion , Bartonella henselae/physiology , Cells, Cultured , Collagen/metabolism , Endothelial Cells/metabolism , Endothelium, Vascular , Fibronectins/metabolism , Gelatin/metabolism , Heparin/metabolism , Heparin/pharmacology , Humans , Protein Binding/drug effects , Protein Structure, Tertiary , Solubility
11.
Infect Immun ; 74(5): 2513-21, 2006 May.
Article in English | MEDLINE | ID: mdl-16622186

ABSTRACT

Bartonella henselae wound-associated infections suggest involvement of extracellular matrix molecules in adhesion and invasion. Pap31 was previously identified as a hemin-binding protein. Our recent studies suggest the protein is an adhesin that is recognized by the host's immune systems. In this study we examined the interactions of B. henselae Pap31 with fibronectin (Fn), heparin (Hep), and human umbilical vein endothelial cells (HUVECs). The cloned gene was expressed in Escherichia coli, and the purified Pap31 protein elicited strong antibody responses in mice and was reactive with rabbit anti-live B. henselae and mouse anti-Pap31 antibodies by Western blotting. Pap31 bound to immobilized Fn and to HUVECs in a dose-dependent manner and to Hep. Fn fragment-binding assays identified the Hep-1 and Hep-2 binding domains of human Fn and in particular the (12-13)FnIII repeat module as primary binding sites for this adhesin. Furthermore, Pap31 binding to the above Fn fragments could be inhibited by Hep, suggesting a common binding site involving the 13FnIII repeat module on the Hep-2 domain of Fn. Adherence of intact B. henselae to HUVECs was inhibited by increasing concentrations of anti-Pap31 antibodies. In addition, purified Pap31 coprecipitated effectively with Fn and anti-Fn antibodies. Taken together, these data suggest that Pap31 is an Fn-binding protein mediating the B. henselae-host interaction(s), and they implicate the 13FnIII repeat module as an important binding site for this adhesin on the Fn molecule. These interactions may be important initial steps leading to bacterial attachment and colonization that promote the establishment of B. henselae infections in vivo.


Subject(s)
Adhesins, Bacterial/metabolism , Bacterial Adhesion , Bartonella henselae/physiology , Fibronectins/metabolism , Amino Acid Sequence , Animals , Binding Sites , Cells, Cultured , Endothelial Cells/metabolism , Fibronectins/chemistry , Heparin/chemistry , Heparin/metabolism , Humans , Immunoprecipitation , Molecular Sequence Data , Rabbits , Repetitive Sequences, Amino Acid
12.
Mol Urol ; 5(1): 31-6, 2001.
Article in English | MEDLINE | ID: mdl-11689149

ABSTRACT

PURPOSE: The longitudinal colonization patterns by Escherichia coli of the vaginal introitus and urinary tract were investigated. MATERIALS AND METHODS: Cultures of the vaginal introitus and midstream urine were collected once a week for 12 consecutive weeks from five women with (patients) and five without (controls) a history of urinary tract infection (UTI). RESULTS: A total of 63 E. coli isolates was obtained from the 10 women, 26 from controls and 37 from patients. The bacterial counts of E. coli present in control individuals were uniformly low, < or = 200 E. coli/mL. The numbers in patients were higher and more variable, reaching > 10(5)/mL in urine and vaginal specimens. In 16 instances, E. coli was present in the urine and the vaginal introitus concurrently (matched isolates). Random amplified polymorphic DNA (RAPD) fingerprinting was used to characterize all matched E. coli isolates. Concurrent vaginal and urinary tract colonization was more common in the patient population, and usually, the same E. coli strain was present at both sites; only 15% of the matched isolates represented different strains. The RAPD fingerprinting was also carried out on selected isolates recovered from four patients and three control individuals over the 12-week study period. Colonization of the vaginal introitus and urinary tract in these individuals varied over time. Generally, however, a predominant E. coli strain was present in the vaginal milieu, urinary tract, or both, either continuously (for as long as 9 consecutive weeks in one patient) or intermittently. CONCLUSION: The results support the concept that the vaginal mucosa acts as reservoir of E. coli which may enter the urinary tract.


Subject(s)
Escherichia coli/isolation & purification , Urinary Tract/microbiology , Vagina/microbiology , Women's Health , Escherichia coli/genetics , Female , Humans , Longitudinal Studies , Mucous Membrane/microbiology , Random Amplified Polymorphic DNA Technique , Urinary Tract Infections/microbiology , Urinary Tract Infections/urine
13.
Blood ; 98(12): 3367-75, 2001 Dec 01.
Article in English | MEDLINE | ID: mdl-11719376

ABSTRACT

Graft-versus-host disease (GVHD) is a major cause of morbidity and mortality of allogeneic stem cell transplantation. Strategies to control GVHD while maintaining graft versus leukemia (GVL) include herpes simplex virus thymidine kinase (HSV-tk) gene transduction of donor T cells followed by treatment with ganciclovir (GCV). Alternatively, GVHD and GVL may be mediated by distinct processes. In this regard, whether cytokine polarization occurs and to what degrees various subsets of cytokine-producing T cells mediate GVHD or GVL has been an active area of research using cytokine or cytokine antibody infusion or genetically deficient mice. This study takes a different approach that allows simultaneous investigation into both the mechanisms underlying GVHD reactions and the efficacy of HSV-tk suicide gene-based T-cell deletion. A source of donor T cells, splenocytes from mice transgenic for HSV-tk controlled by elements of either the interleukin-2 (IL-2) or IL-4 promoters (IL-2-tk and IL-4-tk, respectively) was used, thus allowing investigation into the roles of T1 and T2 cells in ongoing GVHD reactions. To assess treatment rather than prevention of GVHD, GCV was started at peak disease. Remarkably, treatment at this late time point rescued mice from the clinical effects of GVHD caused by T cells expressing either transgene. Thus, both T1 and T2 cells play an important role in clinical GVHD in a minor histocompatibility antigen-mismatched setting. In addition, because clinical disease was reversible even at its maximum, these observations provide controlled evidence that this strategy of treating ongoing GVHD could be effective clinically.


Subject(s)
Disease Models, Animal , Graft vs Host Disease/immunology , Lymphocyte Subsets/immunology , T-Lymphocytes/immunology , Animals , Cell Death/genetics , Cytokines/biosynthesis , Cytokines/metabolism , Flow Cytometry , Ganciclovir/therapeutic use , Graft vs Host Disease/drug therapy , Graft vs Host Disease/pathology , Graft vs Leukemia Effect/immunology , Hematopoiesis , Hematopoietic Stem Cells/cytology , Hematopoietic Stem Cells/physiology , Interleukin-2/genetics , Interleukin-4/genetics , Male , Mice , Mice, Transgenic , Promoter Regions, Genetic , Simplexvirus/enzymology , Spleen/cytology , Thymidine Kinase/genetics , Thymus Gland/cytology , Weight Gain
14.
Arch Dermatol ; 137(8): 1105-6, 2001 Aug.
Article in English | MEDLINE | ID: mdl-11493113
15.
Microb Pathog ; 30(4): 221-8, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11312615

ABSTRACT

The cloning and sequencing of a gene from Rickettsia rickettsii which confers haemolytic activity on Escherichia coli strain TB1 is described. The open reading frame of the haemolysis-promoting gene, cjsT, is 1041 bp and encodes a putative protein with a molecular mass of 33 825 Da. CjsT has high sequence similarity to several bacterial proteases, particularly type IV signal peptidases. Cell lysates from an E. coli clone containing cjsT in pUC19 (pJON1) exhibited greater protease activity in functional assays than found in E. coli containing pUC19 alone. Disruption of the cjsT gene by insertional inactivation with a kanamycin cassette reduced both the protease and haemolytic activities conferred by cjsT. The protease inhibitors antipain and diisopropylfluorophosphate (DFP) both reduced the proteolytic activity of pJON1. The mechanism by which the R. rickettsii cjsT promotes haemolysis in E. coli remains unclear.


Subject(s)
Bacterial Proteins/genetics , Endopeptidases/genetics , Rickettsia rickettsii/enzymology , Amino Acid Sequence , Animals , Base Sequence , Chlorocebus aethiops , Cloning, Molecular , Endopeptidases/chemistry , Endopeptidases/metabolism , Escherichia coli/genetics , Genes, Bacterial , Molecular Sequence Data , Plasmids , Rickettsia rickettsii/genetics , Rickettsia rickettsii/pathogenicity , Vero Cells
16.
Int J Antimicrob Agents ; 17(4): 245-51, 2001 Apr.
Article in English | MEDLINE | ID: mdl-11295403

ABSTRACT

Urinary tract infections (UTIs) are the result of an interaction between bacterial virulence and host defense factors that compete to invade or protect the host, respectively. Research over the past 30 years has demonstrated that vaginal colonization with uropathogens precedes most UTIs. Receptivity of the vaginal mucosa for uropathogens is an essential initial step in vaginal mucosa colonization. When vaginal and buccal epithelial cells were collected from patients susceptible to reinfection and compared with such cells obtained from controls resistant to UTIs, the strains that caused cystitis adhered much more avidly to the epithelial cells from susceptible women. These genotypic traits for epithelial cell receptivity may be a major susceptibility factor in UTIs. The presence or absence of blood group determinants on the surface of uroepithelial cells may influence an individual's susceptibility to UTIs. The protective effect in women with the secretor phenotype may be due to fucosylated structures at the cell surface which decrease the availability of putative receptors for Escherichia coli. Susceptibility among women who do not secrete blood group antigens may be due to specific E. coli-binding glycolipids that are absent in women who secrete blood group antigens. Recent studies have shown that the vaginal fluid, which forms an interface between uropathogens and epithelial cells, also influences vaginal colonizations.


Subject(s)
Disease Susceptibility/microbiology , Escherichia coli/pathogenicity , Mouth Mucosa/microbiology , Urinary Tract Infections/microbiology , Vagina/microbiology , Bacterial Adhesion , Blood Group Antigens/analysis , Cystitis/blood , Cystitis/microbiology , Disease Susceptibility/blood , Epithelial Cells/metabolism , Epithelial Cells/microbiology , Female , Fucose/blood , Genetic Predisposition to Disease , Genotype , Glycolipids/blood , Glycosylation , Humans , Immunoglobulin A, Secretory/analysis , Mouth Mucosa/physiology , Phenotype , Receptors, Cell Surface/metabolism , Urinary Tract Infections/blood , Urinary Tract Infections/genetics , Vagina/physiology , Virulence
17.
J Biosci Bioeng ; 91(3): 305-10, 2001.
Article in English | MEDLINE | ID: mdl-16232994

ABSTRACT

For many applications avian antibody from egg yolk (IgY) offers advantages over the well-known mammalian antibodies. Different experimental techniques for the purification of IgY from chickens immunized with an alphagalactose-containing antigen (alphaGal-trisaccharide) were compared. These included ammonium sulfate precipitation, filtration with diatomaceous earth, treatment with deoxycholate, and thiophilic and affinity chromatography. Samples were tested for overall purity, protein and lipid content, and specific activity. Evaluated on the basis of these results and the simplicity of the process, the favored purification method is ammonium sulfate precipitation of diluted egg yolk directly followed by affinity chromatography. The high lipid content of IgY preparations is greatly reduced by either thiophilic or affinity chromatography. Affinity purification of ammonium sulfate precipitated material resulted in anti-alphaGal-trisaccharide IgY preparations with approximately 1% of the original protein content but approximately 100-fold higher specific activity for the alphaGal-trisaccharide epitope.

18.
Immunology ; 101(4): 467-73, 2000 Dec.
Article in English | MEDLINE | ID: mdl-11122450

ABSTRACT

Chicken antibodies (immunoglobulin Y; IgY) to the alpha Gal epitope (galactose alpha-1,3-galactose) bind to alpha Gal antigens of mouse and porcine tissues and endothelial cells in vitro and block human anti-alpha Gal antibody binding, complement activation and antibody-dependent cell-mediated lysis mechanisms. The activities and toxicity of anti-alpha Gal IgY have not been tested in vivo. In this study, we tested the effects of multiple injections of affinity-purified anti-alpha Gal IgY (AP-IgY) in both wild-type (WT) and alpha-1,3-galactosyltransferase knockout (Gal KO) mice. WT and Gal KO mice were injected once, twice, three, or four times intravenously (i.v.) with AP-IgY and killed at 1 hr or 24 hr. Mice displayed no toxicity to four injections of AP-IgY. Heart, lung, liver, kidney, spleen and pancreatic tissue were evaluated using immunohistochemical techniques for the presence of the alpha Gal epitope using the GSI-B4 lectin, and for bound IgY, as well as mouse IgM and IgG. The binding of AP-IgY antibodies to the endothelium of WT mouse tissues was essentially identical to the pattern of binding of the GSI-B4 lectin after injection of WT mice and death at 1 hr. WT mice killed 24 hr after i.v. injection of AP-IgY showed little remaining bound IgY in their endothelia, indicating that IgY is cleared over that time period. We also evaluated the blood drawn at the time of death for the presence of anti-alpha Gal IgY, anti-IgY IgM and anti-IgY IgG by enzyme-linked immunosorbent assay. Anti-alpha Gal IgY was almost undetectable in WT mouse sera at all injection and killing times. In contrast, Gal KO mouse sera showed increasing anti-alpha Gal IgY levels until 24 hr after the fourth injection, when anti-alpha Gal IgY levels were almost undetectable. Anti-IgY IgM and IgG levels in WT and Gal KO mouse sera showed a typical increase in anti-IgY IgM 24 hr after the second injection (3 days after the first injection) and an increase in anti-IgY IgG 24 hr after the third injection (5 days after the first injection). These results show that IgY binds to alpha Gal epitopes in the WT mice and is cleared sometime over a 24-hr time period and that IgY is an expected immunogen in mice eliciting a rather typical anti-IgY IgM and IgG response.


Subject(s)
Chickens/immunology , Disaccharides/immunology , Immunoglobulins/immunology , Animals , Antibodies, Anti-Idiotypic/blood , Disaccharides/genetics , Enzyme-Linked Immunosorbent Assay , Epitopes/metabolism , Immunoglobulin G/blood , Immunoglobulin M/blood , Immunoglobulins/blood , Immunoglobulins/metabolism , Mice , Mice, Knockout
19.
Infect Immun ; 68(10): 5970-8, 2000 Oct.
Article in English | MEDLINE | ID: mdl-10992509

ABSTRACT

Bacterial htrA genes are typically activated as part of the periplasmic stress response and are dependent on the extracytoplasmic sigma factor rpoE. A putative promoter region, P1, of the sigma(E)-type heat-inducible promoters has previously been identified upstream of the htrA gene of Bartonella henselae. Further analysis of the htrA mRNA by primer extension demonstrated that transcription initiates from P1 and a second region downstream of P1. This second promoter region, termed P2, had no sequence identity to sigma(E)-type heat-inducible promoters. Promoter regions were cloned individually and in tandem into pANT3 upstream of a promoterless version of the green fluorescent protein (GFP) gene (gfpmut3) and transformed into B. henselae by electroporation. The contiguous promoter region containing both P1 and P2 were necessary for the optimal transcriptional activation of the htrA gene. Promoter activity at 37 degrees C was distinctively higher than at 27 degrees C. However, thermal induction at 47 degrees C did not increase expression of gfpmut3. Invasion of human microvascular endothelial cells (HMEC-1) by B. henselae resulted in the formation of well-defined vacuoles containing clusters of bacteria exhibiting marked expression of gfpmut3 transcribed from the P1-P2 region. In addition, a moderate yet significant increase in the ratio of bacterial GFP to DNA was detected for intracellular bacteria compared to extracellular bacteria, indicating upregulation of htrA upon invasion of HMEC-1. The activation of specific genes in the intracellular environment may help us better understand the novel pathogenic mechanisms used by this bacterium.


Subject(s)
Bartonella henselae/genetics , Heat-Shock Proteins , Periplasmic Proteins , Serine Endopeptidases/genetics , Serine Endopeptidases/metabolism , Transcriptional Activation , Angiomatosis, Bacillary/microbiology , Bartonella henselae/metabolism , Bartonella henselae/pathogenicity , Base Sequence , Cell Line , Electroporation/methods , Endothelium, Vascular/cytology , Endothelium, Vascular/microbiology , Flow Cytometry , Green Fluorescent Proteins , Hot Temperature , Humans , Luminescent Proteins/genetics , Luminescent Proteins/metabolism , Microscopy, Fluorescence , Molecular Sequence Data , Promoter Regions, Genetic , Virulence
20.
Transplantation ; 70(5): 828-36, 2000 Sep 15.
Article in English | MEDLINE | ID: mdl-11003366

ABSTRACT

BACKGROUND: Acute vascular xenograft rejection (AVXR), also termed delayed xenograft rejection (DXR), occurs when hyperacute rejection (HAR) is prevented by strategies directed at xenoreactive natural antibodies and/or complement activation. We have hypothesized that AVXR/DXR is initiated in part by early components of the complement cascade, notably C1q. We have developed synthetic peptides (termed CBP2 and WY) that interfere with the interaction between C1q and antibody. METHODS: CBP2 and the WY-conjugates were used as inhibitors of immunoglobulin aggregate binding to solid phase C1q. Inhibition of complement activation by the peptides of the classical system was determined using lysis assays with sensitized sheep red blood cells or porcine aortic endothelial cells as targets and of the alternate complement pathway using guinea pig red blood cells as targets. Two transplant models were used to study the effects of administering peptides to recipients: rat heart transplant to presensitized mouse, and guinea heart transplant to PVG C6-deficient rats. RESULTS: CBP2 and WY-conjugates inhibited immunoglobulin aggregate binding to C1q. The peptides also inhibited human complement-mediated lysis of sensitized sheep red blood cells and porcine aortic endothelial cells in a dose-dependent manner and the WY-conjugates prevented activation of the alternate complement pathway as shown by inhibition of guinea pig red blood cells lysis with human serum. In addition, the use of the peptides and conjugates resulted in significant prolongation of xenograft survival. CONCLUSIONS: The CBP2 and WY peptides exhibit the functional activity of inhibition of complement activation. These peptides also prolong xenograft survival and thus provide reagents for the study of the importance of C1q and other complement components in transplant rejection mechanisms.


Subject(s)
Complement C1q/pharmacology , Immunoglobulins/pharmacology , Peptides/pharmacology , Transplantation, Heterologous , Animals , Complement C1q/antagonists & inhibitors , Cytotoxicity, Immunologic/drug effects , Drug Interactions , Graft Survival/drug effects , Heart Transplantation/immunology , Male , Mice , Mice, Inbred BALB C , Rats , Rats, Inbred Lew , Swine , Transplantation, Heterologous/immunology
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