Elastic precursor of the transformation from glycolipid-nanotube to -vesicle

By the combination of optical tweezer manipulation and digital video microscopy, the flexural rigidity of single glycolipid "nano" tubes has been measured below the transition temperature at which the lipid tubules are transformed into vesicles. Consequently, we have found a clear reduction of the rigidity obviously before the transition as temperature increasing. Further experiments of infrared spectroscopy (FT-IR) and differential scanning calorimetry (DSC) have suggested a microscopic change of the tube walls, synchronizing with the precursory softening of the nanotubes.Comment: 9 pages, 6 figure

Nonlinear Elasticity of the Sliding Columnar Phase

The sliding columnar phase is a new liquid-crystalline phase of matter composed of two-dimensional smectic lattices stacked one on top of the other. This phase is characterized by strong orientational but weak positional correlations between lattices in neighboring layers and a vanishing shear modulus for sliding lattices relative to each other. A simplified elasticity theory of the phase only allows intralayer fluctuations of the columns and has three important elastic constants: the compression, rotation, and bending moduli, $B$, $K_y$, and $K$. The rotationally invariant theory contains anharmonic terms that lead to long wavelength renormalizations of the elastic constants similar to the Grinstein-Pelcovits renormalization of the elastic constants in smectic liquid crystals. We calculate these renormalizations at the critical dimension $d=3$ and find that $K_y(q) \sim K^{1/2}(q) \sim B^{-1/3}(q) \sim (\ln(1/q))^{1/4}$, where $q$ is a wavenumber. The behavior of $B$, $K_y$, and $K$ in a model that includes fluctuations perpendicular to the layers is identical to that of the simple model with rigid layers. We use dimensional regularization rather than a hard-cutoff renormalization scheme because ambiguities arise in the one-loop integrals with a finite cutoff.Comment: This file contains 18 pages of double column text in REVTEX format and 6 postscript figure

Genetic Resistance to Malaria Is Associated With Greater Enhancement of Immunoglobulin (Ig)M Than IgG Responses to a Broad Array of Plasmodium falciparum Antigens

Background. People of the Fulani ethnic group are more resistant to malaria compared with genetically distinct ethnic groups, such as the Dogon people, in West Africa, and studies suggest that this resistance is mediated by enhanced antibody responses to Plasmodium falciparum antigens. However, prior studies measured antibody responses to <0.1% of P falciparum proteins, so whether the Fulani mount an enhanced and broadly reactive immunoglobulin (Ig)M and IgG response to P falciparum remains unknown. In general, little is known about the extent to which host genetics influence the overall antigen specificity of IgM and IgG responses to natural infections. Methods. In a cross-sectional study in Mali, we collected plasma from asymptomatic, age-matched Fulani (n = 24) and Dogon (n = 22) adults with or without concurrent P falciparum infection. We probed plasma against a protein microarray containing 1087 P falciparum antigens and compared IgM and IgG profiles by ethnicity. Results. We found that the breadth and magnitude of P falciparum-specific IgM and IgG responses were significantly higher in the malaria-resistant Fulani versus the malaria-susceptible Dogon, and, unexpectedly, P falciparum-specific IgM responses more strongly distinguished the 2 ethnic groups. Conclusions. These findings point to an underappreciated role for IgM in protection from malaria, and they suggest that host genetics may influence the antigen specificity of IgM and IgG responses to infection

A multiplexed immunoassay system based upon reciprocating centrifugal microfluidics

A novel, centrifugal disk-based micro-total analysis system (mu TAS) for low cost and high throughput semi-automated immunoassay processing was developed. A key innovation in the disposable immunoassay disk design is in a fluidic structure that enables very efficient micro-mixing based on a reciprocating mechanism in which centrifugal acceleration acting upon a liquid element first generates and stores pneumatic energy that is then released by a reduction of the centrifugal acceleration, resulting in a reversal of direction of flow of the liquid. Through an alternating sequence of high and low centrifugal acceleration, the system reciprocates the flow of liquid within the disk to maximize incubation/hybridization efficiency between antibodies and antigen macromolecules during the incubation/hybridization stage of the assay. The described reciprocating mechanism results in a reduction in processing time and reagent consumption by one order of magnitude.open121

Structural Properties of the Sliding Columnar Phase in Layered Liquid Crystalline Systems

Under appropriate conditions, mixtures of cationic and neutral lipids and DNA in water condense into complexes in which DNA strands form local 2D smectic lattices intercalated between lipid bilayer membranes in a lamellar stack. These lamellar DNA-cationic-lipid complexes can in principle exhibit a variety of equilibrium phases, including a columnar phase in which parallel DNA strands from a 2D lattice, a nematic lamellar phase in which DNA strands align along a common direction but exhibit no long-range positional order, and a possible new intermediate phase, the sliding columnar (SC) phase, characterized by a vanishing shear modulus for relative displacement of DNA lattices but a nonvanishing modulus for compressing these lattices. We develop a model capable of describing all phases and transitions among them and use it to calculate structural properties of the sliding columnar phase. We calculate displacement and density correlation functions and x-ray scattering intensities in this phase and show, in particular, that density correlations within a layer have an unusual $\exp(- {\rm const.} \ln^2 r)$ dependence on separation r. We investigate the stability of the SC phase with respect to shear couplings leading to the columnar phase and dislocation unbinding leading to the lamellar nematic phase. For models with interactions only between nearest neighbor planes, we conclude that the SC phase is not thermodynamically stable. Correlation functions in the nematic lamellar phase, however, exhibit SC behavior over a range of length scalesComment: 28 pages, 4 figure

Genomic Organization, Splice Variants and Expression of CGMl, a CD66-related Member of the Carcinoembryonic Antigen Gene Family

The tumor marker carcinoembryonic antigen (CEA) belongs to a family of proteins which are composed of one immunogiobulin variable domain and a varying number of immunoglobulin constant-like domains. Most of the membrane-bound members, which are anchored either by a glycosylphosphatidylinositol moiety or a transmembrane domain, have been shown to convey cell adhesion in vitro. Here we describe two splice variants of CGMI. a transmembrane member of the CEA family without immunoglobulin constant.like domains. CGM1a and CGM1c contain cytopiasmic domains of 71 and 31 amino acids, respectively, The cytoplasmic region of CGM1a is encoded by four exons (Cyt1-Cyt4). Differential splicing of the Cyt1 exon (53 bp)..

Identification of the Feline Humoral Immune Response to Bartonella henselae Infection by Protein Microarray

Background: Bartonella henselae is the zoonotic agent of cat scratch disease and causes potentially fatal infections in immunocompromised patients. Understanding the complex interactions between the host’s immune system and bacterial pathogens is central to the field of infectious diseases and to the development of effective diagnostics and vaccines. Methodology: We report the development of a microarray comprised of proteins expressed from 96 % (1433/1493) of the predicted ORFs encoded by the genome of the zoonotic pathogen Bartonella henselae. The array was probed with a collection of 62 uninfected, 62 infected, and 8 ‘‘specific-pathogen free’ ’ naïve cat sera, to profile the antibody repertoire elicited during natural Bartonella henselae infection. Conclusions: We found that 7.3 % of the B. henselae proteins on the microarray were seroreactive and that seroreactivity was not evenly distributed between predicted protein function or subcellular localization. Membrane proteins were significantly most likely to be seroreactive, although only 23 % of the membrane proteins were reactive. Conversely, we found that proteins involved in amino acid transport and metabolism were significantly underrepresented and did not contain any seroreactive antigens. Of all seroreactive antigens, 52 were differentially reactive with sera from infected cats, and 53 were equally reactive with sera from infected and uninfected cats. Thirteen of the seroreactive antigens were found to be differentially seroreactive between B. henselae type I and type II. Based on these results, we developed a classifier algorith

A Tale of Two Set Theories

We describe the relationship between two versions of Tarski-Grothendieck set theory: the first-order set theory of Mizar and the higher-order set theory of Egal. We show how certain higher-order terms and propositions in Egal have equivalent first-order presentations. We then prove Tarski's Axiom A (an axiom in Mizar) in Egal and construct a Grothendieck Universe operator (a primitive with axioms in Egal) in Mizar

Identification of Circulating Bacterial Antigens by In Vivo Microbial Antigen Discovery

Detection of microbial antigens in clinical samples can lead to rapid diagnosis of an infection and administration of appropriate therapeutics. A major barrier in diagnostics development is determining which of the potentially hundreds or thousands of antigens produced by a microbe are actually present in patient samples in detectable amounts against a background of innumerable host proteins. In this report, we describe a strategy, termed in vivo microbial antigen discovery (InMAD), that we used to identify circulating bacterial antigens. This technique starts with “InMAD serum,” which is filtered serum that has been harvested from BALB/c mice infected with a bacterial pathogen. The InMAD serum, which is free of whole bacterial cells, is used to immunize syngeneic BALB/c mice. The resulting “InMAD immune serum” contains antibodies specific for the soluble microbial antigens present in sera from the infected mice. The InMAD immune serum is then used to probe blots of bacterial lysates or bacterial proteome arrays. Bacterial antigens that are reactive with the InMAD immune serum are precisely the antigens to target in an antigen immunoassay. By employing InMAD, we identified multiple circulating antigens that are secreted or shed during infection using Burkholderia pseudomallei and Francisella tularensis as model organisms. Potential diagnostic targets identified by the InMAD approach included bacterial proteins, capsular polysaccharide, and lipopolysaccharide. The InMAD technique makes no assumptions other than immunogenicity and has the potential to be a broad discovery platform to identify diagnostic targets from microbial pathogens

Using detergent to enhance detection sensitivity of African trypanosomes in human CSF and blood by Loop-Mediated Isothermal Amplification (LAMP)

&lt;p&gt;&lt;b&gt;Background:&lt;/b&gt; The loop-mediated isothermal amplification (LAMP) assay, with its advantages of simplicity, rapidity and cost effectiveness, has evolved as one of the most sensitive and specific methods for the detection of a broad range of pathogenic microorganisms including African trypanosomes. While many LAMP-based assays are sufficiently sensitive to detect DNA well below the amount present in a single parasite, the detection limit of the assay is restricted by the number of parasites present in the volume of sample assayed; i.e. 1 per µL or 103 per mL. We hypothesized that clinical sensitivities that mimic analytical limits based on parasite DNA could be approached or even obtained by simply adding detergent to the samples prior to LAMP assay.&lt;/p&gt; &lt;p&gt;&lt;b&gt;Methodology/Principal Findings:&lt;/b&gt; For proof of principle we used two different LAMP assays capable of detecting 0.1 fg genomic DNA (0.001 parasite). The assay was tested on dilution series of intact bloodstream form Trypanosoma brucei rhodesiense in human cerebrospinal fluid (CSF) or blood with or without the addition of the detergent Triton X-100 and 60 min incubation at ambient temperature. With human CSF and in the absence of detergent, the LAMP detection limit for live intact parasites using 1 µL of CSF as the source of template was at best 103 parasites/mL. Remarkably, detergent enhanced LAMP assay reaches sensitivity about 100 to 1000-fold lower; i.e. 10 to 1 parasite/mL. Similar detergent-mediated increases in LAMP assay analytical sensitivity were also found using DNA extracted from filter paper cards containing blood pretreated with detergent before card spotting or blood samples spotted on detergent pretreated cards.&lt;/p&gt; &lt;p&gt;&lt;b&gt;Conclusions/Significance:&lt;/b&gt; This simple procedure for the enhanced detection of live African trypanosomes in biological fluids by LAMP paves the way for the adaptation of LAMP for the economical and sensitive diagnosis of other protozoan parasites and microorganisms that cause diseases that plague the developing world.&lt;/p&gt