Molecular cloning and expression analysis of pig CD81

CD81, also known as TAPA-1 (target of antiproliferative antibody 1), is a member of the tetraspanin family of proteins and a component of the B cell co-receptor complex. Several studies have shown that CD81 plays significant roles in a variety of immune responses, including activation of B cells and T cells. In this study, we cloned pig Cd81 cDNA using RT-PCR coupled with rapid amplification of cDNA ends (RACE)-PCR and determined the complete cDNA sequence of pig Cd81. Pig Cd81 cDNA contains an open reading frame (711 bp) encoding 236 amino acids. The identity of pig CD81 with those of human, cattle, rat, and mouse are 90.30%, 92.26%, 86.22%, and 86.22%, respectively. Alignment of the CD81 amino acid sequence with those of mammalian species showed that the large extracellular loop (LEL) is the most divergent, whereas other domains are largely conserved. Pig Cd81 mRNA was detected by RT-PCR in a broad range of tissues, including lymphoid tissues as well as nonlymphoid tissues, indicated variety of cellular functions of CD81 in most pig tissues. Flow cytometry analyses demonstrated that human CD81 antibody recognizes a pig CD81 on the cell surface. Further, immunohistochemistry analysis using human CD81 antibody on pig spleen was revealed that CD81 expression is widely diffused in spleen tissue. Future study will be focused on defining the functional role of CD81 during the course of pig infectious diseases.     

Glycol Chitosan-Based Fluorescent Theranostic Nanoagents for Cancer Therapy

Theranostics is an integrated nanosystem that combines therapeutics with diagnostics in attempt to develop new personalized treatments with enhanced therapeutic efficacy and safety. As a promising therapeutic paradigm with cutting-edge technologies, theranostic agents are able to simultaneously deliver therapeutic drugs and diagnostic imaging agents and also monitor the response to therapy. Polymeric nanosystems have been intensively explored for biomedical applications to diagnose and treat various cancers. In recent years, glycol chitosan-based nanoagents have been developed as dual-purpose materials for simultaneous diagnosis and therapy. They have shown great potential in cancer therapies, such as chemotherapeutics and nucleic acid and photodynamic therapies. In this review, we summarize the recent progress and potential applications of glycol chitosan-based fluorescent theranostic nanoagents for cancer treatments and discuss their possible underlying mechanisms.     

Long-Term Fertilization Effects on Rice Productivity and Nutrient Efficiency in Korean Paddy

ABSTRACT

Long-term fertilization tests evaluated rice (Oryza sativa) productivity in relation to application of nitrogen (N)-phosphorus (P)-potassium (K) (120-34.9-66.7 kg ha^{? 1}, respectively) during 1967–1972 and N-P-K (150-43.7-83.3 kg ha^{? 1}, respectively) during 1973–2000. The comparison treatments (NP, PK, and NK) and the control (not fertilized) were selected for calculating nutrient efficiency. Rice grain yield increased at a 17.78 kg ha^{? 1} yr^{? 1} in the control, mainly due to development of improved cultivars. Phosphorus management was found to be important for indigenous fertility and rice productivity in this paddy soil. Yield increased significantly with P fertilization. Without N fertilization (PK), rice productivity increased 56.85 kg ha^{? 1} yr^{? 1} from 62% of NPK at the initial stage to 74% after passing 34 years, which might be affected by increasing biological N fixation with P accumulation in soil. In NK treatment, rice yield increased at a relatively low rate (37.82 kg hr^{? 1} yr^{? 1}) from the same rice productivity with that of NPK in 1967 to 91% after 34 years. In comparison, yield increased at a high rate (62.82 kg hr^{? 1} yr^{? 1}) without K fertilization (NP) from ca. 90% of NPK and might exceed the yield of NPK after 64 years of long-term fertilization. Therefore, K fertilization level might be readjusted after long-term fertilizing in paddy soil.     

Development of an enzyme-linked immunosorbent assay for the pyrethroid cypermethrin

A competitive enzyme-linked immunosorbent assay (ELISA) for the detection of cypermethrin was developed. Two haptens, the trans- and cis-isomers of 3-[(+/-)-cyano-3-(2,2-dichloroethenyl)-2,2-dimethylcyclopropanecarbonyloxy]methyl]phenoxyacetic acid, were conjugated with thyroglobulin as immunogens. Four antisera were generated and screened against six different coating antigens. The assay that was the most sensitive for cypermethrin was optimized and characterized. The IC(50) for cypermethrin was 13.5 +/- 4.3 microg/L, and the lower detection limit (LDL) was 1.3 +/- 0.5 microg/L. This ELISA had relatively low cross-reactivities with other major pyrethroids, such as deltamethrin, phenothrin, resmethrin, fluvalinate, and permethrin. Methanol was found to be the best organic cosolvent for this ELISA, with an optimal sensitivity observed at a concentration of 40% (v/v). The assay parameters were unchanged at pH values between 5.0 and 8.0, whereas higher ionic strengths strongly suppressed the absorbances. To increase the sensitivity of the overall method, a C(18) sorbent-based solid-phase extraction was applied to various domestic and environmental water samples. The water samples, fortified with cypermethrin, were analyzed according to this method. Good recoveries and correlation with spike levels were observed.     

Giant piezoelectricity on Si for hyperactive MEMS

Microelectromechanical systems (MEMS) incorporating active piezoelectric layers offer integrated actuation, sensing, and transduction. The broad implementation of such active MEMS has long been constrained by the inability to integrate materials with giant piezoelectric response, such as Pb(Mg(1/3)Nb(2/3))O(3)-PbTiO(3) (PMN-PT). We synthesized high-quality PMN-PT epitaxial thin films on vicinal (001) Si wafers with the use of an epitaxial (001) SrTiO(3) template layer with superior piezoelectric coefficients (e(31,f) = -27 ± 3 coulombs per square meter) and figures of merit for piezoelectric energy-harvesting systems. We have incorporated these heterostructures into microcantilevers that are actuated with extremely low drive voltage due to thin-film piezoelectric properties that rival bulk PMN-PT single crystals. These epitaxial heterostructures exhibit very large electromechanical coupling for ultrasound medical imaging, microfluidic control, mechanical sensing, and energy harvesting.     

Application of electrospun polyurethane web to breathable water-proof fabrics

Electrospun web may possibly be widely applied to protective garments or specialty textiles due to its high level of protection as well as comfort. Of particular interest in this study is to develop waterproof-breathable fabric by applying electrospun web of polyurethane directly onto the substrate fabric. The optimal electrospinning condition was examined with regards to the concentration, applied voltage and tip-to-collector distance. Solvent-electospinning of polyurethane was performed at the optimum condition, using N,N-dimethylacetamide as solvent. The thickness of 0.02 mm of electrospun web was applied onto the polyester/nylon blended fabric. For comparison, the polyester/nylon fabrics were coated with 0.02 mm thickness of polyurethane resin membranes adopting four different conditions. The electrospun PU web/fabric was compared to resin coated fabrics in terms of water-proof and breathable properties. The electrospun web applied fabric showed higher air permeability, vapor transmission, and thermal insulation properties than resin coated fabrics, which can be translated as greater comfort sensation of electrospun applied fabrics. However, water resistance value of electrospun web applied fabric did not reach that of resin coated fabrics.     

Interaction Between Sorghum Protein Extraction and Precipitation Conditions on Yield,Purity, and Composition of Purified Protein Fractions

Sorghum proteins have the potential to be used as a bio‐industrial renewable resource for applications such as biodegradable films and packaging. This project was designed to evaluate the effect of interactions between sorghum protein extraction and precipitation conditions on the yield, purity, and composition of sorghum protein fractions. Proteins were extracted with 70% ethanol under nonreducing conditions, with ultrasound, or under reducing conditions using either sodium metabisulfite or glutathione as the reducing agent. Several conditions were used to isolate the extracted proteins through precipitation, including lowering ethanol concentrations alone or in combination with lowering to pH 2.5, or by adding 1M NaCl to the extract. Combinations of these conditions were also tested. All precipitation conditions effectively precipitated proteins and lowering the pH and adding 1M NaCl to the extracts enhanced precipitation in some cases. However, the conditions that precipitated the maxium amount of protein or highest purity of protein varied according to how the proteins were initially extracted. Precipitated proteins were characterized by RP‐HPLC, SEC, HPCE, and SDS‐PAGE to compare the protein fractions composition. Nonreduced and sonicated samples had a much wider M_w distribution than reduced extracts. Thus, extraction and precipitation conditions influenced the isolated proteins yield, purity, and composition. Because the extraction and purification processes influenced the composition, purity, and biochemical properties, it may be possible to prepare protein fractions with unique functionalities for specific end‐uses.     

Papaya seed represents a rich source of biologically active isothiocyanate

In the present study, papaya (Carica papaya) seed and edible pulp were carefully separated and then the contents of benzyl isothiocyanate and the corresponding glucosinolate (benzyl glucosinolate, glucotropaeolin) quantified in each part. The papaya seed with myrosinase inactivation contained >1 mmol of benzyl glucosinolate in 100 g of fresh seed. This content is equivalent to that of Karami daikon (the hottest Japanese white radish) or that of cress. The papaya seed extract also showed a very high activity of myrosinase and, without myrosinase inactivation, produced 460 micromol of benzyl isothiocyanate in 100 g of seed. In contrast, papaya pulp contained an undetectable amount of benzyl glucosinolate and showed no significant myrosinase activity. The n-hexane extract of the papaya seed homogenate was highly effective in inhibiting superoxide generation and apoptosis induction in HL-60 cells, the activities of which are comparable to those of authentic benzyl isothiocyanate.     

Synthesis of zeolite as ordered multicrystal arrays

Zeolites are crystalline nanoporous aluminosilicates widely used in industry. In order for zeolites to find applications as innovative materials, they need to be organized into large two- and three-dimensional (2D and 3D) arrays. We report that uniformly aligned polyurethane films can serve as templates for the synthesis of uniformly aligned 2D and possibly 3D arrays of silicalite-1 crystals, in which the orientations of the crystals are controlled by the nature of the polymers. We propose that the supramolecularly organized organic-inorganic composites that consist of the hydrolyzed organic products and the seed crystals are responsible for this phenomenon.     

Microsatellite-based analysis of genetic diversity in 91 commercial Brassica oleracea L. cultivars belonging to six varietal groups

Brassica oleracea L. includes various types of important vegetables that show extremely diverse phenotypes. To elucidate the genetic diversity and relationships among commercial cultivars derived by different companies throughout the world, we characterized the diversity and genetic structure of 91 commercial B. oleracea cultivars belonging to six varietal groups, including cabbage, broccoli, cauliflower, kohlrabi, kale and kai-lan. We used 69 polymorphic microsatellite markers showing a total of 359 alleles with an average number of 5.20 alleles per locus. Polymorphism information content (PIC) values ranged from 0.06 to 0.73, with an average of 0.40. Among the six varietal groups, kohlrabi cultivars exhibited the highest heterozygosity level, whereas kale cultivars showed the lowest. Based on genetic similarity values, an UPGMA clustering dendrogram and a two-dimensional scale diagram (PCoA) were generated to analyze genetic diversity. The cultivars were clearly separated into six different clusters with a tendency to cluster into varietal groups. Model-based structure analysis revealed six genetic groups, in which cabbage cultivars were divided into two subgroups that were differentiated by their head shape, whereas cauliflower and kai-lan cultivars clustered together into a single group. Furthermore, we identified 18 SSR markers showing 27 unique alleles specific to only one cultivar that can be used to discriminate 22 cultivars from the others. Our phylogenetic and population structure analysis presents new insights into the genetic structure and relationships among 91 B. oleracea cultivars and provides valuable information for breeding of B. oleracea species. In addition, we demonstrate the utility of SSR markers as a powerful tool for discriminating between the cultivars. The SSR markers described herein will also be helpful for Distinctness, Uniformity and Stability (DUS) test of new cultivars.