Using chiral tactoids as optical probes to study the aggregation behavior of chromonics.

Tactoids are nuclei of an orientationally ordered nematic phase that emerge upon cooling the isotropic phase. In addition to providing a natural setting for exploring chromonics under confinement, we show that tactoids can also serve as optical probes to delineate the role of temperature and concentration in the aggregation behavior of chromonics. For high concentrations, we observe the commonly reported elongated bipolar tactoids. As the concentration is lowered, breaking of achiral symmetry in the director configuration is observed with a predominance of twisted bipolar tactoids. On further reduction of concentration, a remarkable transformation of the director configuration occurs, wherein it conforms to a unique splay-minimizing configuration. Based on a simple model, we arrive at an interesting result that lower concentrations have longer aggregates at the same reduced temperature. Hence, the splay deformation that scales linearly with the aggregate length becomes prohibitive for lower concentrations and is relieved via twist and bend deformations in this unique configuration. Raman scattering measurements of the order parameters independently verify the trend in aggregate lengths and provide a physical picture of the nematic-biphasic transition.

Curvature-Induced Twist in Homeotropic Nematic Tori.

We confine a nematic liquid crystal with homeotropic anchoring to stable toroidal droplets and study how geometry affects the equilibrium director configuration. In contrast to the case of cylindrical confinement, we find that the equilibrium state is chiral-a twisted and escaped radial director configuration. Furthermore, we find that the magnitude of the twist distortion increases as the ratio of the ring radius to the tube radius decreases; we confirm this with computer simulations of optically polarized microscopy textures. In addition, numerical calculations also indicate that the local geometry indeed affects the magnitude of the twist distortion. We further confirm this curvature-induced twisting using bent cylindrical capillaries.

Clinical significance of overexpression of NRG1 and its receptors, HER3 and HER4, in gastric cancer patients.

BACKGROUND: Neuregulin 1 (NRG1), a ligand for human epidermal growth factor (HER) 3 and HER4, can activates cell signaling pathways to promote carcinogenesis and metastasis. METHODS: To investigate the clinicopathologic significance of NRG1 and its receptors, immunohistochemistry was performed for NRG1, HER3, and HER4 in 502 consecutive gastric cancers (GCs). Furthermore, HER2, microsatellite instability (MSI), and Epstein-Barr virus (EBV) status were investigated. NRG1 gene copy number (GCN) was determined by dual-color fluorescence in situ hybridization (FISH) in 388 available GCs. RESULTS: NRG1 overexpression was observed in 141 (28.1%) GCs and closely correlated with HER3 (P = 0.034) and HER4 (P < 0.001) expression. NRG1 overexpression was significantly associated with aggressive features, including infiltrative tumor growth, lymphovascular, and neural invasion, high pathologic stage, and poor prognosis (all P < 0.05), but not associated with EBV, MSI, or HER2 status. Multivariate analysis identified NRG1 overexpression as an independent prognostic factor for survival (P = 0.040). HER3 and HER4 expressions were observed in 157 (31.3%) and 277 (55.2%), respectively. In contrast to NRG1, expression of these proteins was not associated with survival. NRG1 GCN gain (GCN ≥ 2.5) was detected in 14.7% patients, including two cases of amplification, and was moderately correlated with NRG1 overexpression (κ, 0.459; P < 0.001). CONCLUSIONS: Although our results indicate a lack of prognostic significance of HER3 and HER4 overexpression in GC, overexpression of their ligand, NRG1, was associated with aggressive clinical features and represented an independent unfavorable prognostic factor. Therefore, NRG1 is a potential prognostic and therapeutic biomarker in GC patients.

Enhanced Alignment of Water-Soluble Polythiophene Using Cellulose Nanocrystals as a Liquid Crystal Template.

Cellulose nanocrystals (CNCs) are bioderived, rodlike particles that form a chiral nematic liquid crystal (LC) in water. In this work, CNCs were used to induce long-range order in a semiconducting polymer, poly[3-(potassium-4-butanoate) thiophene-2,5-diyl] (PPBT). When mixed with CNCs, it was found that PPBT was incorporated into the liquid crystal "template" to form ordered structures with highly birefringent domains, as observed under polarized light. We show that the π-π interactions between polymer chains, which contribute considerably to the energetics of the semiconducting system, are directly influenced by the presence and packing of the liquid crystal phase. Upon increasing the concentration of CNCs from the isotropic to chiral nematic regime, we observe a bathochromic shift in the UV-vis spectra and the emergence of the 0-0 vibrational peak, suggesting enhanced π-π stacking leading to chain coplanarization. Furthermore, the chiral nature of the PPBT/CNC mixture was evidenced by a negative peak in circular dichroism (CD) spectroscopy, promoting the notion that the polymer chains followed the helicoidal twist of the chiral nematic liquid crystal host. At high temperatures, the peak height ratios and overall intensities of the UV-vis and CD spectra associated with PPBT decreased as the chiral nematic pitch grew larger in size.

Macroscopic alignment of chromonic liquid crystals using patterned substrates.

We demonstrate an efficient technique to align lyotropic chromonic liquid crystals (LCLCs) using secondary sputtering lithography (SSL). Monodomains of LCLCs prepared using SSL maintained their stable alignment for days. A generalization of Berreman's theory was employed to determine the anchoring strength of LCLCs on tessellated surface patterns. The anchoring energy initially increases with the amplitude (A) of the grooves and excellent alignment of LCLCs was observed when the amplitude of the grooves is equal to half its wavelength (λ). We also note that the anchoring energy levels off above qA∼ 3 (where q = 2π/λ), which suggests that increasing qA beyond a certain value does not provide any further advantage for the alignment of LCLCs. This finding provides a useful optimization criterion for the fabrication of the patterned cells to achieve stable monodomain alignment of LCLCs. Our analysis also explains why good alignment of LCLCs has been a difficult task.

Disclination elastica model of loop collision and growth in confined nematic liquid crystals.

Theory and modeling are used to characterize disclination loop-loop interactions in nematic liquid crystals under capillary confinement with strong homeotropic anchoring. This defect process arises when a mesogen in the isotropic phase is quenched into the stable nematic state. The texture evolution starts with +1/2 disclination loops that merge into a single loop through a process that involves collision, pinching and relaxation. The process is characterized with a combined Rouse-Frank model that incorporates the tension and bending elasticity of disclinations and the rotational viscosity of nematics. The Frank model of disclinations follows the Euler elastica model, whose non-periodic solution, known as Poleni's curve, is shown to locally describe the loop-loop collision and to shed light on why loop-loop merging results in a disclination intersection angle of approximately 60°. Additional Poleni invariants demonstrate how tension and bending pinch the two loops into a single +1/2 disclination ring. The Rouse model of disclination relaxation yields a Cahn-Hilliard equation whose time constant combines the confinement, tension/bending stiffness ratio and disclination diffusivity. Based on predictions made using this three stage process, a practical procedure is proposed to find viscoelastic parameters from defect geometry and defect dynamics. These findings contribute to the evolving understanding of textural transformations in nematic liquid crystals under confinement using the disclination elastica methodology.

Mechanisms and shape predictions of nematic disclination branching under conical confinement.

Liquid crystals (LCs) are self-organizing anisotropic viscoelastic soft materials that flow like viscous liquids and display anisotropies like crystals. When a nematic liquid crystal is confined to a capillary tube with strong anchoring conditions, disclination defects of higher (+1) and lower (+1/2) topological charges can coexist, connected through a defect branch point. The shape of the +1/2 disclination lines emanating from the branch point are functions of confinement and bulk elasticity. Previous work shows that nematic liquid crystals under cylindrical confinement display a radial (one +1 line)-to-planar polar (two +1/2 lines) defect texture transition through the nucleation and uniform motion of a disclination branch point. Here we present analysis, scaling and modeling based on a non-linear non-local nematic elastic equation that shows that a branch point also can be generated from disclinations in a liquid crystal confined to different conical geometries with homeotropic anchoring conditions. The cone aperture increases the bending stiffness but decreases the curvature of the disclination. These competing effects lead to a decrease in the total disclination curvature, increase in elastic energy and volume of the branching region. The results are summarized into power laws and integrated into a shape/energy diagram that reveals the effects of confinement and its gradient (cone angle) on disclination shape selection. These new findings are useful to assess the Frank elasticity of new nematic liquid crystals and to predict novel defect structures in complex confinement, including biological microfluidics and mesophase fiber spinning.

Identification and characterization of proteins isolated from microvesicles derived from human lung cancer pleural effusions.

Microvesicles (MVs, also known as exosomes, ectosomes, microparticles) are released by various cancer cells, including lung, colorectal, and prostate carcinoma cells. MVs released from tumor cells and other sources accumulate in the circulation and in pleural effusion. Although recent studies have shown that MVs play multiple roles in tumor progression, the potential pathological roles of MV in pleural effusion, and their protein composition, are still unknown. In this study, we report the first global proteomic analysis of highly purified MVs derived from human nonsmall cell lung cancer (NSCLC) pleural effusion. Using nano-LC-MS/MS following 1D SDS-PAGE separation, we identified a total of 912 MV proteins with high confidence. Three independent experiments on three patients showed that MV proteins from PE were distinct from MV obtained from other malignancies. Bioinformatics analyses of the MS data identified pathologically relevant proteins and potential diagnostic makers for NSCLC, including lung-enriched surface antigens and proteins related to epidermal growth factor receptor signaling. These findings provide new insight into the diverse functions of MVs in cancer progression and will aid in the development of novel diagnostic tools for NSCLC.

Proteomic analysis of microvesicles derived from human colorectal cancer ascites.

The presence of malignant ascites in the peritoneal cavity is a poor prognostic indicator of low survival rate. Various cancer cells, including those of colorectal cancer (CRC), release microvesicles (exosomes) into surrounding tissues and peripheral circulation including malignant ascites. Although recent progress has revealed that microvesicles play multiple roles in tumor progression, the protein composition and the pathological function of malignant ascites-derived microvesicles are still unknown. Here, we report the first global proteomic analyses of highly purified microvesicles derived from human CRC ascites. With 1-D SDS-PAGE and nano-LC-MS/MS analyses, we identified a total of 846 microvesicular proteins from ascites of three CRC patients with high confidence; 384 proteins were identified in at least two patients. We identified proteins that might function in tumor progression via disruption of epithelial polarity, migration, invasion, tumor growth, immune modulation, and angiogenesis. Furthermore, we identified several potential diagnostic markers of CRC including colon-specific surface antigens. Our proteomic analyses will help to elucidate diverse functions of microvesicles in cancer progression and will aid in the development of novel diagnostic tools for CRC.

Raman scattering study of phase biaxiality in a thermotropic bent-core nematic liquid crystal.

Polarized Raman spectroscopy was used to investigate the development of orientational order and the degree of phase biaxiality in a bent-core mesogenic system. The values of the uniaxial order parameters and , and biaxial order parameters , , and , and their evolution with temperature were determined. The temperature dependence of almost all order parameters reveals a second order transition from the uniaxial to biaxial nematic phase with increasing to ∼0.22 before a first order transition to the smectic-C phase, upon cooling.

Gram-positive bacteria produce membrane vesicles: proteomics-based characterization of Staphylococcus aureus-derived membrane vesicles.

Although archaea, Gram-negative bacteria, and mammalian cells constitutively secrete membrane vesicles (MVs) as a mechanism for cell-free intercellular communication, this cellular process has been overlooked in Gram-positive bacteria. Here, we found for the first time that Gram-positive bacteria naturally produce MVs into the extracellular milieu. Further characterizations showed that the density and size of Staphylococcus aureus-derived MVs are both similar to those of Gram-negative bacteria. With a proteomics approach, we identified with high confidence a total of 90 protein components of S. aureus-derived MVs. In the group of identified proteins, the highly enriched extracellular proteins suggested that a specific sorting mechanism for vesicular proteins exists. We also identified proteins that facilitate the transfer of proteins to other bacteria, as well to eliminate competing organisms, antibiotic resistance, pathological functions in systemic infections, and MV biogenesis. Taken together, these observations suggest that the secretion of MVs is an evolutionally conserved, universal process that occurs from simple organisms to complex multicellular organisms. This information will help us not only to elucidate the biogenesis and functions of MVs, but also to develop therapeutic tools for vaccines, diagnosis, and antibiotics effective against pathogenic strains of Gram-positive bacteria.

Imaging optical scattering of butterfly wing scales with a microscope.

A new optical method is proposed to investigate the reflectance of structurally coloured objects, such as Morpho butterfly wing scales and cholesteric liquid crystals. Using a reflected-light microscope and a digital single-lens reflex (DSLR) camera, we have successfully measured the two-dimensional reflection pattern of individual wing scales of Morpho butterflies. We demonstrate that this method enables us to measure the bidirectional reflectance distribution function (BRDF). The scattering image observed in the back focal plane of the objective is projected onto the camera sensor by inserting a Bertrand lens in the optical path of the microscope. With monochromatic light illumination, we quantify the angle-dependent reflectance spectra from the wing scales of Morpho rhetenor by retrieving the raw signal from the digital camera sensor. We also demonstrate that the polarization-dependent reflection of individual wing scales is readily observed using this method, using the individual wing scales of Morpho cypris. In an effort to show the generality of the method, we used a chiral nematic fluid to illustrate the angle-dependent reflectance as seen by this method.

Polypeptide Composite Particle-Assisted Organization of π-Conjugated Polymers into Highly Crystalline "Coffee Stains".

We demonstrate that homopolypeptides covalently tethered to anisotropically shaped silica particles induce crystalline ordering of representative semiconducting polymers. Films drop-cast from chloroform dispersions of poly(γ-stearyl-l-glutamate) (PSLG) composite particles and poly(3-hexythiophene) (P3HT) led to highly ordered crystalline structures of P3HT. Hydrophobic-hydrophobic interactions between the alkyl side chains of P3HT and PSLG were the main driving force for P3HT chain ordering into the crystalline assemblies. It was found that the orientation of rigid P3HT fibrils on the substrate adopted the directionality of the evaporating front. Regardless of the PSLG-coated particle dimensions used, the drop-cast films displayed patterns that were shaped by the coffee ring and Marangoni effects. PSLG-coated particles of high axial ratio (4.2) were more efficient in enhancing the electronic performance of P3HT than low axial ratio (2.6) homologues. Devices fabricated from the ordered assemblies displayed improved charge-carrier transport performance when compared to devices fabricated from P3HT alone. These results suggest that PSLG can favorably mediate the organization of semiconducting polymers.

Orientational Order of a Lyotropic Chromonic Liquid Crystal Measured by Polarized Raman Spectroscopy.

Lyotropic chromonic liquid crystals are distinct from thermotropic nematics from a fundamental standpoint as the structure of the aggregating columns is a function of both the temperature and concentration. We report on the thermal evolution of orientational order parameters, both the second (=scalar) (⟨P200⟩ (=S)) and fourth (⟨P400⟩) order, of sunset yellow FCF aqueous solutions, measured using polarized Raman spectroscopy for different concentrations. The order parameter increases with the concentration, and their values are high in comparison with those of thermotropic liquid crystals. On the basis of Raman spectroscopy, we provide the strongest evidence yet that the hydrozone tautomer of SSY is the predominant form in aqueous solutions in the isotropic, nematic, and columnar phases, as well as what we believe to be the first measurements of (⟨P400⟩) for this system.

Spontaneous emergence of chirality in achiral lyotropic chromonic liquid crystals confined to cylinders.

The presumed ground state of a nematic fluid confined in a cylindrical geometry with planar anchoring corresponds to that of an axial configuration, wherein the director, free of deformations, is along the long axis of the cylinder. However, upon confinement of lyotropic chromonic liquid crystals in cylindrical geometries, here we uncover a surprising ground state corresponding to a doubly twisted director configuration. The stability of this ground state, which involves significant director deformations, can be rationalized by the saddle-splay contribution to the free energy. We show that sufficient anisotropy in the elastic constants drives the transition from a deformation-free ground state to a doubly twisted structure, and results in spontaneous symmetry breaking with equal probability for either handedness. Enabled by the twist angle measurements of the spontaneous twist, we determine the saddle-splay elastic constant for chromonic liquid crystals for the first time.

Enhanced mobility and effective control of threshold voltage in P3HT-based field-effect transistors via inclusion of oligothiophenes.

Improved organic field-effect transistor (OFET) performance through a polymer-oligomer semiconductor blend approach is demonstrated. Incorporation of 2,5-bis(3-dodecylthiophen-2-yl)thieno[3,2-b]thiophene (BTTT) into poly(3-hexylthiophene) (P3HT) thin films leads to approximately a 5-fold increase in charge carrier mobility, a 10-fold increase in current on-off ratio, and concomitantly, a decreased threshold voltage to as low as 1.7 V in comparison to single component thin films. The blend approach required no pre- and/or post treatments, and processing was conducted under ambient conditions. The correlation of crystallinity, surface morphology and photophysical properties of the blend thin films was systematically investigated via X-ray diffraction, atomic force microscopy and optical absorption measurements respectively, as a function of blend composition. The dependence of thin-film morphology on the blend composition is illustrated for the P3HT:BTTT system. The blend approach provides an alternative avenue to combine the advantageous properties of conjugated polymers and oligomers for optimized semiconductor performance.

Cloning and characterization of a new cysteine proteinase secreted by Paragonimus westermani adult worms.

The cysteine proteinases of Paragonimus westermani are known to play important roles in invasion and pathogenesis to hosts and in immune modulation and nutrient uptake. In this study, we have cloned a new cysteine proteinase of P. westermani, PwCP2, from adult worms and tested its diagnostic usefulness. The PwCP2 gene had an open reading frame of 816 base pairs and a conserved catalytic triad of cysteine, histidine, and asparagine residues. The mature form of recombinant PwCP2 (rPwCP2) lacking a proregion was overexpressed in Escherichia coli and used to produce antiserum. Western blot and immunohistochemical analyses using this antiserum showed that PwCP2 was expressed as a mature form, 24-kD product in a crude extract and in the excretory-secretory product of P. westermani, and was localized mainly in the intestinal epithelium of the adult worm. Western blot analysis using the rPwCP2 showed not only high sensitivity (90%) and specificity (100%) to sera from patients with paragonimiasis westermani, but also no cross-reactivity with sera from patients with clonorchiasis, sparganosis, or cysticercosis. Furthermore, an enzyme-linked immunosorbent assay using rPwCP2 exhibited a sensitivity of 93% and a specificity of 93% with sera of rats infected with P. westermani metacercariae. These results suggest that the excretory-secretory PwCP2 can be used for the diagnosis of paragonimiasis.

Theoretical predictions of disclination loop growth for nematic liquid crystals under capillary confinement.

The combination of low elasticity modulus, anisotropy, and responsiveness to external fields drives the rich variety of experimentally observed pattern formation in nematic liquid crystals under capillary confinement. External fields of interest in technology and fundamental physics are flow fields, electromagnetic fields, and surface fields due to confinement. In this paper we present theoretical and simulation studies of the pattern formation of nematic liquid crystal disclination loops under capillary confinement including branching processes from a m=+1 disclination line to two m=+1/2 disclination curves that describe the postnucleation and growth regime of the textural transformation from radial to planar polar textures. The early postnucleation and growth of emerging disclination loops in cylindrical capillaries are characterized using analytical and computational methods based on the nematic elastica that takes into account line tension and line bending stiffness. Using subdiffusive growth and constant loop anisotropy, we found that the solution to the nematic elastica is a cusped elliptical geometry characterized by exponential curvature variations. The scaling laws that govern the loop growth reflect the tension to bending elasticity balance and reveal that the loop dilation rate depends on the curvature and normal velocity of the disclination. The line energy growth is accommodated by the decrease in branch-point curvature. These findings contribute to the evolving understanding of textural transformations in nematic liquid crystals under confinement using the nematic elastic methodology.

Performance characteristics of nested polymerase chain reaction vs real-time polymerase chain reaction methods for detecting Mycobacterium tuberculosis complex in paraffin-embedded human tissues.

OBJECTIVES: Nucleic acid amplification tests on formalin-fixed, paraffin-embedded (FFPE) tissue specimens enable Mycobacterium tuberculosis complex (MTB) detection and rapid tuberculosis diagnosis in the absence of microbiologic culture tests. We aimed to evaluate the efficacy of different polymerase chain reaction (PCR) methods for detecting Mycobacterium species in FFPE tissues. METHODS: We examined 110 FFPE specimens (56 nonmycobacterial cases, 32 MTB, and 22 nontuberculous mycobacteria [NTM] determined by acid-fast bacilli [AFB] culture) to assess five PCR methods: nested PCR (N-PCR) (Seeplex MTB Nested ACE Detection; Seegene, Seoul, South Korea), an in-house real-time PCR (RT-PCR) method, and three commercial RT-PCR methods (AccuPower MTB RT-PCR [Bioneer, Seoul, Korea], artus M tuberculosis TM PCR [Qiagen, Hilden, Germany], and AdvanSure tuberculosis/NTM RT-PCR [LG Life Sciences, Seoul, Korea]). RESULTS: The results of N-PCR, in-house RT-PCR, and AdvanSure RT-PCR correlated well with AFB culture results (concordance rates, 94.3%, 87.5%, and 89.5%, respectively). The sensitivity of N-PCR (87.5%) was higher than that of the RT-PCR methods, although these differences were not statistically significant between N-PCR and the in-house and AdvanSure RT-PCR methods (68.8% and 80.0%, respectively). All the PCR methods had high specificities, ranging from 98.2% to 100%. Only two NTM cases were detected by AdvanSure RT-PCR, implying a very low sensitivity. CONCLUSIONS: Well-designed RT-PCR and N-PCR can effectively identify MTB in FFPE specimens.

Rapid, sensitive, and specific detection of Mycobacterium tuberculosis complex by real-time PCR on paraffin-embedded human tissues.

The detection of Mycobacterium tuberculosis complex (MTB) in clinical specimens is important for diagnosing and caring for patients in whom tuberculosis is clinically suspected. We collected 129 FFPE specimens, including 56 nontuberculosis cases, 63 MTB cases, and 10 nontuberculous mycobacteria (NTM) cases determined by acid-fast bacilli (AFB) culture. We performed AFB staining; nested MTB PCR, targeting the IS6110 gene; and real-time MTB PCR, targeting the senX3-regX3 intergenic region in the 129 FFPE specimens. The sensitivity and specificity of AFB staining were 37.0% and 98.2%, respectively, using AFB culture results as the reference standard. The sensitivity and specificity of detecting MTB were 68.3% and 98.5%, respectively, by nested PCR; and 74.6% and 98.5% by real-time PCR, respectively. Among the 129 specimens, four were positive by AFB staining but negative by nested or real-time PCR. NTM grew in all four of these cases by AFB culture. AFB density in FFPE tissue sections significantly correlated with MTB DNA load. Thus, real-time PCR is a useful diagnostic tool for rapid and sensitive MTB detection in FFPE specimens, whereas NTM should be included in differential diagnoses of cases positive by AFB staining but negative by PCR.