Development and characterization of mouse monoclonal antibodies reactive with chicken CD80

This study was carried out to develop and characterize mouse monoclonal antibodies (mAbs) against chicken CD80 (chCD80). A recombinant plasmid containing a chCD80/horse IgG4 fusion gene was constructed and expressed in CHO cells to produce recombinant chCD80/IgG4 protein. Chicken CD80 was purified from the chCD80/IgG4 fusion protein following enterokinase digestion, and used to immunize BALB/c mice, resulting in 158 hybridomas that produced mAbs against chCD80. Three mAbs with high binding specificity for recombinant chCD80/IgG4-transfected CHO cells were identified by flow cytometry, and one of these (#112) was selected for further characterization. Immunoprecipitation of CD80/IgG4-CHO cell extract, or lipopolysaccharide (LPS)-treated monocytes identified 35.0 kDa proteins. Immunohistochemical analysis revealed chCD80-expressing cells exclusively in the bursal follicles at the outer portion of the cortex, and throughout the red pulp and the outer boundary of the white pulp in the spleen. By immunofluorescence microscopy, chCD80 was observed on intestinal dendritic cells. LPS treatment of bursa or spleen monocytes for 24 or 48 h increased chCD80 expression. Finally, addition of chCD80 mAb to Con A-stimulated spleen cells inhibited the expression of major histocompatibility complex class II antigens and IL-2-driven proliferation of lymphoblast cells. In summary, these chCD80 mAbs will serve as valuable immunological reagents for basic and applied poultry immunology research.     

Morphology and electrical conductivity of PS/PMMA/SMMA blends filled with carbon black

An alternative strategy to reduce the percolation threshold of carbon black (CB) in polymer blends was investigated using random copolymer ternary blends of polystyrene (PS), poly(methyl methacrylate) (PMMA), and a styrene-methyl methacrylate random copolymer (SMMA). The target morphology was to selectively locate CB particles in the encapsulating layer of SMMA during melt mixing. The CB used in this study is BP-2000 from Cabot and has a strong selective affinity to PS. Even when the CB was premixed with SMMA, it moves to the PS phase during the melt mixing. However, we also observed the CB particles located at the interface between SMMA and PS phases. Through this study it is found that the interaction between polymers and CB particles is critical for selectively localizing CB particles in multi-component polymer blends. Although appropriate processing condition may retard the movement of CB particles to the polymer phase with affinity, it cannot prevent it completely and locate them to the SMMA phase, which is not thermodynamically favored. To locate CB particles in an encapsulating layer of ternary polymer blends, first of all, polymers forming it should have selective affinity to CB.     

Preparation of poly(vinyl acetate)/clay and poly(vinyl acetate)/poly(vinyl alcohol)/clay microspheres

Poly(vinyl acetate) (PVAc)/poly(vinyl alcohol)(PVA)/montmorillonite (MMT) clay nanocomposite microspheres with a core/shell structure have been developed via a suspension polymerization approach. In order to prepare the PVAc/MMT and PVAc/PVA/MMT nanocomposite microspheres, which are promising precursor of PVA/MMT nanocomposite microspheres, suspension polymerization of vinyl acetate with organophilic MMT and heterogeneous saponification were conducted. A quaternary ammonium salt, cetyltrimethylammonium bromide, was mixed with the MMT in the monomer phase prior to the suspension polymerization. The rate of conversion decreased with an increase in MMT concentration. The incorporation of MMT into the PVAc was verified by FT-IR spectroscopy. Organic vinyl acetate monomers were intercalated into the interlayer regions of organophilic clay hosts and followed by suspension polymerization. Partially saponified PVA/MMT nanocomposite microspheres with a core/shell structure were successfully prepared by heterogeneous saponification.     

Bioactive Compounds,Antioxidant and Binding Activities and Spear Yield of Asparagus officinalis L.

The aim of this investigation was to find a proper harvesting period and establishing fern number, which effects the spear yield, bioactive compounds and antioxidant activities of Asparagus officinalis L. Spears were harvested at 2, 4, and 6 weeks after sprouting. Control for comparison was used without harvest. Spears and total yield increased with prolonged spear harvest period. In harvest of 6 weeks long optimum spear yield was the highest and fern numbers were 5?~?8. Bioactive compounds (polyphenols, flavonoids, flavanols, tannins and ascorbic acid) and the levels of antioxidant activities by ferric-reducing/antioxidant power (FRAP) and cupric reducing antioxidant capacity (CUPRAC) assays in asparagus ethanol extracts significantly differed in the investigated samples and were the highest at 6 weeks harvest period (P?<?0.05). The first and the second segments from the tip significantly increased with the increase of catalase (CAT). It was interesting to investigate in vitro how human serum albumin (HSA) interacts with polyphenols extracted from investigated vegetables. Therefore the functional properties of asparagus were studied by the interaction of polyphenol ethanol extracts with HSA, using 3D- FL. In conclusion, antioxidant status (bioactive compounds, binding and antioxidant activities) improved with the harvesting period and the first segment from spear tip. Appropriate harvesting is effective for higher asparagus yield and its bioactivity.     

Echinochrome A Protects Mitochondrial Function in Cardiomyocytes against Cardiotoxic Drugs

Echinochrome A (Ech A) is a naphthoquinoid pigment from sea urchins that possesses antioxidant, antimicrobial, anti-inflammatory and chelating abilities. Although Ech A is the active substance in the ophthalmic and cardiac drug Histochrome^®, its underlying cardioprotective mechanisms are not well understood. In this study, we investigated the protective role of Ech A against toxic agents that induce death of rat cardiac myoblast H9c2 cells and isolated rat cardiomyocytes. We found that the cardiotoxic agents tert-Butyl hydroperoxide (tBHP, organic reactive oxygen species (ROS) inducer), sodium nitroprusside (SNP; anti-hypertension drug), and doxorubicin (anti-cancer drug) caused mitochondrial dysfunction such as increased ROS level and decreased mitochondrial membrane potential. Co-treatment with Ech A, however, prevented this decrease in membrane potential and increase in ROS level. Co-treatment of Ech A also reduced the effects of these cardiotoxic agents on mitochondrial oxidative phosphorylation and adenosine triphosphate level. These findings indicate the therapeutic potential of Ech A for reducing cardiotoxic agent-induced damage.     

Echinochrome A Treatment Alleviates Atopic Dermatitis-like Skin Lesions in NC/Nga Mice via IL-4 and IL-13 Suppression

Atopic dermatitis (AD) is a chronic inflammatory skin disease in which skin barrier dysfunction leads to dryness, pruritus, and erythematous lesions. AD is triggered by immune imbalance and oxidative stress. Echinochrome A (Ech A), a natural pigment isolated from sea urchins, exerts antioxidant and beneficial effects in various inflammatory disease models. In the present study, we tested whether Ech A treatment alleviated AD-like skin lesions. We examined the anti-inflammatory effect of Ech A on 2,4-dinitrochlorobenzene (DNCB)-induced AD-like lesions in an NC/Nga mouse model. AD-like skin symptoms were induced by treatment with 1% DNCB for 1 week and 0.4% DNCB for 5 weeks in NC/Nga mice. The results showed that Ech A alleviated AD clinical symptoms, such as edema, erythema, and dryness. Treatment with Ech A induced the recovery of epidermis skin lesions as observed histologically. Tewameter^® and Corneometer^® measurements indicated that Ech A treatment reduced transepidermal water loss and improved stratum corneum hydration, respectively. Ech A treatment also inhibited inflammatory-response-induced mast cell infiltration in AD-like skin lesions and suppressed the expression of proinflammatory cytokines, such as interferon-γ, interleukin-4, and interleukin-13. Collectively, these results suggest that Ech A may be beneficial for treating AD owing to its anti-inflammatory effects.     

Contributing ratio of the mechanical and the physicochemical actions to washing efficiency of mineral oil and protein soils

It was attempted to separate the detergency contributions from the mechanical actions and the physicochemical actions for two different types of soils. The mechanical washing efficiency was evaluated through the washing performed under the mechanical agitation, in the absence of detergent. The physicochemical washing efficiency was examined in the washing bath with detergent in the absence of mechanical agitation. Among the washing parameters, extended washing time and higher washing temperature were effective improving the mechanical and physicochemical washing efficiencies for mineral oil and carbon black. For blood soil, lower washing temperature and extended washing time enhanced the mechanical and physicochemical washing efficiencies. For the mineral oil, the presence of both mechanical and physicochemical actions led to the effective washing performance with synergistic effect. For the blood soil, mechanical actions were observed as more contributing than the physicochemical actions. The optimized washing program could be recommended as per the contributing ratios for the specific soil types, minimizing the energy consumption and detergent usage while achieving the washing performance.     

Synthesis and thermal properties of polyhydroxyamide copolymer and its derivatives

We synthesized a polyhydroxyamide (PHA) copolymer via low-temperature solution polymerization of 3,3'-dihydroxybenzidine with terephthaloyl chloride (80.0 mol%) and isophthaloyl chloride (20.0 mol%) in N,Ndimethylacetamide with the aid of LiCl. We prepared the PHA copolymer derivatives containing the fluorine-based substituents and investigated their solubility, cyclization behavior, and thermal properties using a differential scanning calorimeter (DSC), a thermogravimetric analyzer (TGA), and the simultaneous thermogravimetric analyzer coupled with a mass spectrometer (STA-MS). The chemical structures of the PHA copolymer and its derivatives, as well as the polybenzoxazoles (PBOs) obtained through thermal cyclization of the copolymer and derivatives, were determined by a fourier transform infrared (FT-IR) spectroscopic analysis. The PHA copolymer could be dissolved in organic solvents only with the aid of LiCl, while its derivatives were readily soluble in DMA_c and NMP without LiCl at room temperature. The DSC and TGA results demonstrated that the PHA copolymer derivatives could be converted to PBOs at a lower temperature than the PHA copolymer.     

Thermal properties of copolyetherester/silica nanocomposites

Thermal properties of copolyetherester/silica nanocomposites were examined by using DSC and TGA. The segmented block copolyetheresters with various hard segment structures and hard segment contents (HSC) were synthesized and their silica nanocomposite films were prepared by solution casting method. The nano-sized fumed silica particles were found to act as a nucleating agent of the copolyetheresters. The nanocomposites always showed reduced degree of supercooling or faster crystallization than the corresponding copolyetheresters. The nanocomposites also showed increased hard segment crystallinity except HSC 35 sample which had short hard segment length. In case of 2GT [poly(ethylene terephthalate)] copolyetheresters, which were not developed commercially because of their low crystallization rate, the hard segment crystallinity increased considerably. The copolyetherester/silica nanocomposites showed better thermal stability than copolyetheresters.     

A hybrid swarm population of Pinus densiflora × P. sylvestris inferred from sequence analysis of chloroplast DNA and morphological characters

To confirm a hybrid swarm population of Pinus densiflora × P. sylvestris in Jilin, China, we used needles and seeds from P. densiflora, P. sylvestris, and P. densiflora × P. sylvestris collected from natural stands or experimental stations to study whether shoot apex morphology of 4- year old seedlings can be correlated with the sequence of a chloroplast DNA simple sequence repeat marker (cpDNA SSRs). Total genomic DNA was extracted and subjected to sequence analysis of the pine cpDNA SSR marker Pt15169. Results show that morphological characters from 4-year old seedlings did not correlate with sequence variants of this marker. Marker haplotypes from all P. sylvestris trees had a CTAT element that was absent from all sampled P. densiflora trees. However, both haplotype classes involving this insertion/deletion element were found in a P. densiflora × P. sylvestris population and its seedling progeny. It was concluded that the P. densiflora × P. sylvestris accessions sampled from Jilin, China resulted from bi-directional crosses, as evidenced by both species’ cpDNA haplotypes within the hybrid swarm population.