Molluscan pleiotropic FADD involved in innate immune signaling and induces apoptosis.

Fas-associated protein with death domain (FADD) was initially identified as a crucial adaptor protein in the apoptotic pathway mediated by death receptor (DR). Subsequently, many studies have confirmed that FADD plays a vital role in innate immunity and inflammatory responses in animals. However, the function of this pleiotropic molecule in mollusk species has not been well explored. In this study, we successfully verified the gene sequence of FADD in the Zhikong scallop (Chlamys farreri) and designated it as CfFADD. The CfFADD protein contains a conserved death effector and death domains. Phylogenetic analysis showed that CfFADD is a novel addition to the molluscan FADD family with a close evolutionary relationship with molluscan FADD subfamily proteins. CfFADD mRNA expression in various scallop tissues was significantly induced by challenge with pathogen-associated molecular patterns (lipopolysaccharide, peptidoglycan, and poly(I:C)), suggesting its role in innate immunity in scallops. Co-immunoprecipitation showed that CfFADD interacted with the scallop DR (tumor necrosis factor receptor) and a signaling molecule involved in the Toll-like receptor pathway (interleukin-1 receptor-associated kinase), confirming that CfFADD may be involved in DR-mediated apoptosis and innate immune signaling pathways. Further studies showed that CfFADD interacted with CfCaspase-8 and activated caspase-3. HEK293T cells exhibited distinct apoptotic features after transfection with a CfFADD-expression plasmid, suggesting a functional DR-FADD-caspase apoptotic pathway in scallops. Overexpression of CfFADD led to a significant dose-dependent activation of interferon ß and nuclear factor-κB reporter genes, demonstrating the key role of CfFADD in innate immunity. In summary, our research has confirmed the critical roles of CfFADD in innate immunity and apoptosis and provides valuable information for developing comparative immunology theories.

Molecular cloning and functional study of a Rel homologous gene in sea urchin Strongylocentrotus intermedius.

NF-κB (Nuclear factor-kappa B) family proteins are versatile transcription factors that play crucial regulatory roles in cell development, growth, apoptosis, inflammation, and immune response. However, there is limited research on the function of these key genes in echinoderms. In this study, an NF-κB family gene (SiRel) was identified in sea urchin Strongylocentrotus intermedius. The gene has an open reading frame length of 1809 bp and encodes for 602 amino acids. Domain prediction results revealed that the N-terminal of SiRel protein encodes a conserved Rel homology domain (RHD), including the RHD-DNA binding domain and the RHD-dimerization domain. Multiple sequence comparison results showed that the protein sequences of these two domains were conserved. Phylogenetic analysis indicated that SiRel clustered with Strongylocentrotus purpuratus p65 protein and Rel protein of other echinoderms. Results from quantitative real-time PCR demonstrated detectable SiRel mRNA expression in all tested sea urchin tissues, with the highest expression level found in the gills. And SiRel mRNA expression levels were significantly induced after LPS (Lipopolysaccharide) and poly(I:C) (Polyinosinic:polycytidylic acid) stimulation. In addition, SiRel protein expression can be found in cytoplasm and nucleus of HEK293T cells. Co-immunoprecipitation results showed that SiRel could interact with sea urchin IκB (Inhibitor of NF-κB) protein. Western blotting and dual-luciferase reporter gene assay results indicated that overexpression of SiRel in HEK293T cells could impact the phosphorylation levels of JNK (c-Jun N-terminal kinase) and Erk1/2 (Extracellular signal-regulated kinases1/2) and activate interleukin-6 (IL-6), activating protein 1 (AP-1), interferon (IFN)α/ß/γ, and signal transducer and activator of transcription 3 (STAT3) reporter genes in HEK293T cells. In conclusion, this study reveals that SiRel plays an important role in the innate immune response of sea urchins and enriches our understanding of comparative immunology theory.

The Chemokine CXCL7 is Correlated with LDH-A and Predicts the Prognosis of Patients with Colorectal Cancer.

OBJECTIVE: The aims of this study were to evaluate the correlation between chemokine (C-X-C) ligand 7 (CXCL7) expression and glycolysis and to explore the prognostic significance of CXCL7 in colorectal cancer (CRC). METHODS: The expression of CXCL7 and lactate dehydrogenase A (LDH-A) was measured by immunohistochemistry in tissue from 158 CRC patients. Patients were divided into high expression and low expression groups based on receiver operating characteristic curves and a cut-off value. The correlation between CXCL7 and LDH-A expression was evaluated. The overall survival (OS) times of CRC patients were explored. The risk factors related to prognosis were assessed. RESULTS: Significantly higher expression of CXCL7 and LDH-A was detected in CRC tissue than in non-CRC tissue, and was associated with N stage and tumor-node-metastasis (TNM) stage. CXCL7 expression was strongly correlated with LDH-A expression in CRC tissue. High expression of CXCL7 was validated as an independent risk factor for OS. CONCLUSION: Increased expression of CXCL7 was positively correlated with LDH-A expression and was an independent risk factor for CRC prognosis.

Fluorinated TiO2 Hollow Spheres for Detecting Formaldehyde under UV Irradiation.

The fluorinated titanium dioxide (F-TiO2) hollow spheres with varying F to Ti molar ratios were prepared by a simple one-step hydrothermal method followed by thermal processing. The diameter of the F-TiO2-0.3 hollow spheres with a nominal ratio of F:Ti = 0.3:1 was about 200-400 nm. Compared with the sensor based on pristine TiO2 sensing materials, the F-TiO2-0.3 sensor displayed an enhanced sensing performance toward gaseous formaldehyde (HCHO) vapor at room temperature under ultraviolet (UV) light irradiation. The F-TiO2-0.3 sensor demonstrated an approximately 18-fold enhanced response (1.56) compared to the pristine TiO2 sensor (0.085). The response and recovery times of the F-TiO2-0.3 sensor to 10 ppm HCHO were about 56 s and 64 s, respectively, and a limit-of-detection value of 0.5 ppm HCHO was estimated. The F-TiO2-0.3 sensor also demonstrated good repeatability and selectivity to HCHO gas under UV light irradiation. The outstanding HCHO gas-sensing properties of the F-TiO2-0.3 sensor were related to the following factors: the excitation effect caused by the UV light facilitated surface chemical reactions with analyte gas species; the hollow sphere structure provided sufficient active sites; and the surface fluoride (≡Ti-F) created additional chemisorption sites on the surface of the TiO2 material.

UV-Activated Au Modified TiO2/In2O3 Hollow Nanospheres for Formaldehyde Detection at Room Temperature.

Au modified TiO2/In2O3 hollow nanospheres were synthesized by the hydrolysis method using the carbon nanospheres as a sacrificial template. Compared to pure In2O3, pure TiO2, and TiO2/In2O3 based sensors, the Au/TiO2/In2O3 nanosphere-based chemiresistive-type sensor exhibited excellent sensing performances to formaldehyde at room temperature under ultraviolet light (UV-LED) activation. The response of the Au/TiO2/In2O3 nanocomposite-based sensor to 1 ppm formaldehyde was about 5.6, which is higher than that of In2O3 (1.6), TiO2 (2.1), and TiO2/In2O3 (3.8). The response time and recovery time of the Au/TiO2/In2O3 nanocomposite sensor were 18 s and 42 s, respectively. The detectable formaldehyde concentration could go down as low as 60 ppb. In situ diffuse reflectance Fourier transform infrared spectroscopy (DRIFTS) was used to analyze the chemical reactions on the surface of the sensor activated by UV light. The improvement in the sensing properties of the Au/TiO2/In2O3 nanocomposites could be attributed to the nanoheterojunctions and electronic/chemical sensitization of the Au nanoparticles.

UV-Enhanced Formaldehyde Sensor Using Hollow In2O3@TiO2 Double-Layer Nanospheres at Room Temperature.

Hollow In2O3@TiO2 double-layer nanospheres were prepared via a facile water bath method using the sacrifice template of carbon nanospheres. It is shown that the size of the In2O3/TiO2 nanocomposites is 150-250 nm, the thickness of the In2O3 shell is about 10 nm, and the thickness of the TiO2 shell is about 15 nm. The sensing performances of the synthesized In2O3/TiO2 nanocomposites-based chemiresistive-type sensor to formaldehyde (HCHO) gas under UV light activation at room temperature have been studied. Compared to the pure In2O3- and pure TiO2-based sensors, the In2O3/TiO2 nanocomposite sensor exhibits much better sensing performances to formaldehyde. The response of the In2O3/TiO2 nanocomposite-based sensor to 1 ppm formaldehyde is about 3.8, and the response time and recovery time are 28 and 50 s, respectively. The detectable formaldehyde concentration can reach as low as 0.06 ppm. The role of the formed In2O3/TiO2 heterojunctions and the involved chemical reactions activated by UV light have been investigated by AC impedance spectroscopy and the in situ diffuse reflectance Fourier transform infrared spectroscopy. The improvement of the sensing properties of In2O3/TiO2 nanocomposites could be attributed to the nanoheterojunctions between the two components and the "combined photocatalytic effects" of UV-light-emitting diode irradiation. Density functional theory calculations demonstrated that introducing heterojunctions could improve the adsorption energy and charge transfer between formaldehyde and sensing materials.

Scallop RIG-I-like receptor 1 responses to polyinosinic:polycytidylic acid challenge and its interactions with the mitochondrial antiviral signaling protein.

Retinoic acid-inducible gene I (RIG-I)-like receptors (RLRs) are a class of pattern recognition receptors located in the cytoplasm that play a key role in antiviral innate immunity in animals. However, few studies have been conducted on the function of RLR proteins in invertebrates. In this study, the complete coding sequence of the RLR gene of the Zhikong scallop, Chlamys farreri, was obtained and named CfRLR1 with an aim to study the response of CfRLR1 to polyinosinic:polycytidylic acid [poly (I:C)] stimulation and the interaction between the CfRLR1 and C. farreri mitochondrial antiviral signaling (MAVS) protein. Sequence analysis revealed that CfRLR1 encodes 1161 amino acids, and the encoded protein covers two tandem caspase activation and recruitment domains (CARDs), a helicase domain, and a C-terminal regulatory domain. Phylogenetic analysis revealed that CfRLR1 belongs to the RLR family of mollusks. Quantitative real-time polymerase chain reaction showed that CfRLR1 mRNA was expressed in all tested tissues, with its highest expression observed in feet and gill tissues. Furthermore, CfRLR1 expression in the gill tissues was significantly induced after the poly (I:C) challenge. Finally, the results of co-immunoprecipitation and yeast two-hybrid assays revealed that CfRLR1 can bind to the CfMAVS protein via CARD-CARD interactions. Overall, our results elucidate the immune function of invertebrate RLR proteins and provide valuable information on viral disease control for scallop farming.

Ambient ultraviolet B radiation induced valve behavioral acclimation of Pacific oyster which resulted from the different response strategies of smooth and striated adductor muscles.

Light not only conveys image-forming vision but also has an impact on various physiological functions. In particular, ultraviolet B (UVB) radiation has the closest relationship with living organisms. For Pacific oysters (Crassostrea gigas), alteration of valve behavior is one of the most important ways responding to ambient UVB. In the present study, the response of adult C. gigas to sunlight (especially UVB) was evaluated by monitoring valve activity and further elucidated at the physiological and metabolomic levels. After exposure, the valve activity of C. gigas demonstrated flexible acclimation to the ambient conditions. The potential adjustment of osmoregulation and oxidative stress might be related to ambient UVB radiation. Mycosporine-like amino acids might contribute to the protection of C. gigas against UVB, while precursors of ß-alanine and degradation products of 5-hydroxytryptamine might adjust the contraction of the adductor muscles. The different responses of the adductor muscles (smooth and striated) were manifested in signal transduction and metabolisms of energy and nucleotide. This study not only indicated the correlation between the valve behavioral changes in oysters and light radiation, especially UVB, but illustrated the acclimation strategies of oysters to ambient light (UVB) environment.

Role of nickel dopant on gas response and selectivity of electrospun indium oxide nanotubes.

It has been demonstrated that the incorporation of Ni into metal oxide-based gas sensors often enhances the sensing performance by increasing the catalytic and heterojunction effects. However, it remains unclear how these two effects work either individually or synergistically in gas sensing. Herein, a series of Ni-doped In2O3 nanotubes (NIO NTs) with different doping concentrations were synthesized through a traditional electrospinning technique. The as-prepared NIO NTs were uniform, with length of micron scale, an average diameter of approximately 70 nm, and a tube wall thickness of approximately 10 nm. Following their incorporation into gas sensors, the NIO NTs often showed improved sensing properties (including excellent response and selectivity) for ethanol vapor compared to the pristine In2O3 NTs. Specifically, at 100 ppm ethanol and 220 °C, the response of NIO-7 NTs (7 mol% Ni) was approximately four-fold higher than that of pristine In2O3 NTs (49.74 vs. 13.39). The gas sensing test results indicated that the improved sensing performance was due to the formation of a heterojunction between In2O3 and NiO, as well as to the catalysis effects of Ni3+ ions. Additionally, simulation results indicated that the improved gas selectivity could be due to the Ni doping-induced change in surface adsorption energies of the tested gases.

Oyster Versatile IKKα/ßs Are Involved in Toll-Like Receptor and RIG-I-Like Receptor Signaling for Innate Immune Response.

IκB kinases (IKKs) play critical roles in innate immunity through signal-induced activation of the key transcription factors nuclear factor-κB (NF-κB) and interferon regulatory factors (IRFs). However, studies of invertebrate IKK functions remain scarce. In this study, we performed phylogenetic analysis of IKKs and IKK-related kinases encoded in the Pacific oyster genome. We then cloned and characterized the oyster IKKα/ß-2 gene. We found that oyster IKKα/ß-2, a homolog of human IKKα/IKKß, responded to challenge with lipopolysaccharide (LPS), peptidoglycan (PGN), and polyinosinic-polycytidylic acid [poly(I:C)]. As a versatile immune molecule, IKKα/ß-2 activated the promoters of NF-κB, TNFα, and IFNß, as well as IFN-stimulated response element (ISRE)-containing promoters, initiating an antibacterial or antiviral immune state in mammalian cells. Importantly, together with the cloned oyster IKKα/ß-1, we investigated the signal transduction pathways mediated by these two IKKα/ß proteins. Our results showed that IKKα/ß-1 and IKKα/ß-2 could interact with the oyster TNF receptor-associated factor 6 (TRAF6) and that IKKα/ß-2 could also bind to the oyster myeloid differentiation factor 88 (MyD88) protein directly, suggesting that oyster IKKα/ßs participate in both RIG-I-like receptor (RLR) and Toll-like receptor (TLR) signaling for the reception of upstream immune signals. The fact that IKKα/ß-1 and IKKα/ß-2 formed homodimers by interacting with themselves and heterodimers by interacting with each other, along with the fact that both oyster IKKα/ß proteins interacted with NEMO protein, indicates that oyster IKKα/ßs and the scaffold protein NEMO form an IKK complex, which may be a key step in phosphorylating IκB proteins and activating NF-κB. Moreover, we found that oyster IKKα/ßs could interact with IRF8, and this may be related to the IKK-mediated activation of ISRE promotors and their involvement in the oyster "interferon (IFN)-like" antiviral pathway. Moreover, the expression of oyster IKKα/ß-1 and IKKα/ß-2 may induce the phosphorylation of IκB proteins to activate NF-κB. These results reveal the immune function of oyster IKKα/ß-2 and establish the existence of mollusk TLR and RLR signaling mediated by IKKα/ß proteins for the first time. Our findings should be helpful in deciphering the immune mechanisms of invertebrates and understanding the development of the vertebrate innate immunity network.

Characterization of the IRF2 proteins isolated from the deep-sea mussel Bathymodiolus platifrons and the shallow-water mussel Modiolus modiolus.

Interferon regulatory factors (IRFs) are transcription factors that play important roles in immune defense, stress response, hematopoietic differentiation, and cell apoptosis. IRFs of invertebrate organisms and their functions remain largely unexplored. In the present study, for the first time new IRFs (BpIRF2 and MmIRF2) were identified in the deep-sea mussel Bathymodiolus platifrons and the shallow-water mussel Modiolus modiolus. The open reading frame of BpIRF2 and MmIRF2 encoded putative proteins of 354 and 348 amino acids, respectively. Comparison and phylogenetic analysis revealed that both IRF2 proteins were new identified invertebrate IRF molecular. As transcriptional factors, both BpIRF2 and MmIRF2 could activate the interferon-stimulated response element-containing promoter and BpIRF2 could interact with itself. Moreover, both BpIRF2 and MmIRF2 were localized to the cytoplasm and nucleus. Collectively, these results demonstrated that IRF2 proteins might be crucial in the innate immunity of deep-sea and shallow-water mussels.

Molecular Characterization and Functional Analysis of a Putative Octopamine/Tyramine Receptor during the Developmental Stages of the Pacific Oyster, Crassostrea gigas.

Octopamine (OA) and its precursor, tyramine (TA), participate in invertebrate development such as growth, maturation, and reproduction by activating their corresponding G protein-coupled receptors (GPCRs). Although OA was first discovered in mollusks (octopus), subsequent studies on OA, TA and related receptors have primarily been conducted in Ecdysozoa, especially in insects. Accordingly, only limited reports on OA/TA receptors in mollusks are available and their physiological roles remain unclear. Here, a full-length cDNA encoding a putative 524 amino acid OA/TA receptor (CgGPR1) was isolated from the Pacific oyster Crassostrea gigas. CgGPR1 was most closely related to the Lymnaea stagnalis OA receptor OAR2 in sequence. Phylogenetic analysis showed that CgGPR1 belongs to a poorly studied subfamily of invertebrate OA/TA receptors. The spatio-temporal expression of CgGPR1 in C. gigas larvae was examined by quantitative real-time PCR and Western blot analysis. CgGPR1 was expressed during all developmental stages of C. gigas with higher levels at mid-developmental stages, indicating its potential role in embryogenesis and tissue differentiation. Immunoreactive fluorescence of CgGPR1 was mainly observed in the velum, foot, gill and mantle of C. gigas larvae. CgGPR1 transcripts were detected in all the tested organs of adult C. gigas, with highest level in the mantle. Pharmacological analysis showed that cAMP and Ca2+ concentrations remained unchanged in HEK293 cells expressing CgGPR1 upon addition of OA, TA or related amines, suggesting that CgGPR1 modulates other unknown molecules rather than cAMP and Ca2+. Our study sheds light on CgGPR1 function in oysters.

Molecular characterization and functional analysis of tumor necrosis factor receptor-associated factor 2 in the Pacific oyster.

Tumor necrosis factor receptor (TNFR)-associated factors (TRAFs) are a family of crucial adaptors, playing vital roles in mediating signal transduction in immune signaling pathways, including RIG-I-like receptor (RLR) signaling pathway. In the present study, a new TRAF family member (CgTRAF2) was identified in the Pacific oyster, Crassostrea gigas. Comparison and phylogenetic analysis revealed that CgTRAF2 could be a new member of the invertebrate TRAF2 family. Quantitative real-time PCR revealed that CgTRAF2 mRNA was highly expressed in the digestive gland, gills, and hemocytes, and it was significantly up-regulated after Vibrio alginolyticus and ostreid herpesvirus 1 (OsHV-1) challenge. The CgTRAF2 mRNA expression profile in different developmental stages of oyster larvae suggested that CgTRAF2 could function in early larval development. CgTRAF2 mRNA expression pattern, after the silence of CgMAVS (Mitochondrial Antiviral Signaling) -like, indicated that CgTRAF2 might function downstream of CgMAVS-like. Moreover, the subcellular localization analysis revealed that CgTRAF2 was localized in cytoplasm, and it may play predominately important roles in signal transduction. Collectively, these results demonstrated that CgTRAF2 might play important roles in the innate immunity and larval development of the Pacific oyster.

Cloning and characterization of a novel caspase-8-like gene in Crassostrea gigas.

Cysteine-dependent aspartate-directed proteases, or caspases, play key roles in apoptosis and immune defense. In this study, we cloned the first caspase-8-like gene (CgCaspase8-2) identified in the pacific oyster, Crassostrea gigas. The 2572-bp cDNA encodes a putative protein of 714 amino acids that contains two tandem death effector domains (DEDs) at the N-terminal, and P20 and P10 domains at the C-terminal. The conserved pentapeptide motif QACQG was also identified in the deduced CgCaspase8-2 protein. Phylogenetic analysis indicated that CgCaspase8-2 was clustered with initiator caspases in the invertebrate subgroup, but the similarity between CgCaspase8-2 and other invertebrate caspase-8s was low. CgCaspase8-2 protein was localized in the cytoplasm, and over-expression of CgCaspase8-2 in HEK293T cells induced cell death, suggesting a role in apoptosis. Quantitative real-time PCR results demonstrated that CgCaspase8-2 was widely expressed in various tissues and developmental stages, with the highest CgCaspase8-2 expression levels detected in hemolymph and the blastula stage. Furthermore, CgCaspase8-2 transcripts showed no change in response to a bacterial challenge but exhibited notable up-regulation post-poly (I:C) challenge, suggesting that CgCaspase8-2 is specifically involved in immune responses against viruses. In summary, CgCaspase8-2 is involved in both apoptotic and immune function.

Alternative splicing and immune response of Crassostrea gigas tumor necrosis factor receptor-associated factor 3.

Diverse alternative splicing isoforms play an important role in immune diversity and specificity. Their role in molluscan host-defense is however poorly understood. We characterized two alternative isoforms of tumor necrosis factor receptor-associated factor 3 (TRAF3) in the Pacific oyster, Crassostrea gigas, which were named CgTRAF3-S and CgTRAF3-L. An intron was retained in CgTRAF3-L, introducing a premature termination codon. Comparison and phylogenetic analysis revealed that CgTRAF3 shared a higher identity with other species, suggesting the conservation of the two gene transcripts. Quantitative real-time PCR was performed and the expression levels of CgTRAF3 isoforms were found to be significantly changed after Vibrio anguillarum and ostreid herpesvirus 1 challenges. These two isoforms represented contrary trends, indicating that CgTRAF3-L might function as a negative regulator of CgTRAF3-S. We also investigated the expression level of the transcripts of the two CgTRAF3 isoforms, following the silence of C. gigas mitochondrial anti-viral signaling protein like gene (CgMAVS-like). We concluded that CgTRAF3 might be involved in a MAVS-mediated immune signaling pathway. This study suggests that CgTRAF3 may be a response to bacterial and viral stimulation and that the two isoforms may be involved in immune response pathways. It is also possible that the two alternative splicing isoforms could be inter-coordinated and may promote survival of these oysters under immune stress conditions.

[A pivotal role of cortactin, a CTTN encoding protein, in endocytosis of human colon cancer].

OBJECTIVE: To explore the role of cortactin in endocytosis of colon cancer cells and clarify the significance of its over-expression in colon cancer tissues. METHODS: Immunohistochemistry and Western blot were employed to detect the expression of cortactin in benign and malignant tissues and cells. Cell endocytosis was examined in cancer cells after siRNA treatment and DNA transfection with plasmid encoding cortactin wild type and domain deletion mutants. RESULTS: Cortactin was over-expressed in colon cancer tissues than in adjacent normal tissues. The expression rate was 77.5% in cancer tissues and 47.5% in normal tissues (P < 0.05). The value of transferrin uptake was 0.61 ± 0.02 in siRNA treated cancer cells and 1.01 ± 0.16 in the control cells (P < 0.05). Intact molecule and sufficient level of cortactin was required for an optimal endocytosis of cancer cells. Cortactin was involved in coated-vesicle transportation in cells. CONCLUSION: Endocytosis in colon cancer cells is dependent on an intact expression of CTTN. An over-expression of cortactin facilitates the signaling in invasion and metastasis related to endocytosis.

Zirconia nanoparticles-coated column for the capillary electrochromatographic separation of iron-binding- and phosphorylated-proteins.

A ZrO(2) nanoparticles (ZrO(2)NPs)-coated column was prepared through a sol-gel process using zirconium(iv) oxychloride, which reacted with silanol groups of the fused-silica capillary. The condensation reaction was carried out at 350 °C for 8 h. Electroosmotic flow (EOF) measurements and scanning electron microscopy (SEM) images were used to characterize the ZrO(2)NPs fabricated on the inner wall of the capillary. Below the pI value (pH 5-6), cathodic EOF elucidated that the phosphate buffer adsorbs tightly on the zirconia surface, resulting in a negatively charged surface. In this work, iron-binding proteins, phosphorylated proteins and glycoproteins were selected as the model compounds. The effects of pH, concentration, buffer type and the organic modifier were studied to optimize the separation efficiency. Iron-binding proteins exhibited a retention time for myoglobin (Mb) < hemoglobin (Hb), which corresponded to the binding constants for ZrO(2)NPs. The α- and ß-subunit of Hb could be separated in borate buffer (20 mM, pH 9.0) with MeOH (20%, v/v). Greater affinity of α-casein and bovine serum albumin (BSA) for the stationary phase as the pH decreased was found by comparison with that of conalbumin (ConA) and transferrin (Tf). Interestingly, 14 peaks for glycoisoforms of ovalbumin (OVA) were observed using borate buffer (40 mM, pH 9.0). The established method was also applied to the determination of analytes in the egg whites of chicken and duck eggs.

Novel stationary phase for complexation gas chromatography originating from ionic liquid and metallomesogen.

A metallomesogen of a polycatenar oxazoline copper(II) complex, [Cu(S-C(12))(2)], that exhibited a columnar mesophase with a helical organization was prepared and employed as the stationary phase for the GC separation with polycyclic aromatic hydrocarbons (PAHs) as model compounds. For introducing the mesogen into the capillary column, an ionic liquid (BeMIM-TfO) was used as the vehicle. The results of thermal analyses and UV-vis spectroscopy indicated that some beneficial interactions occurred between the metallomesogen and the ionic liquid. Various parameters affecting the separation efficiency were studied. Different ratios of BeMIM-TfO and Cu(S-C(12))(2) (1:0, 1:1, 1:2 and 1:3 (w/w)) were tested for the separation of the PAHs. As the amount of Cu(S-C(12))(2) was increased, complete separation could be achieved. The stationary phase with the ratio of 1:1 provided the most satisfactory result having average theoretical plate number of 5.2 x 10(3)plates/m. With an optimized temperature program, 11 PAH mixtures were completely separated within 27 min. The interaction between PAH and these fascinating and interesting stationary phases was discussed.