Sesame oleosin and prepro-2S albumin expressed as a fusion polypeptide in transgenic rice were split, processed and separately assembled into oil bodies and protein bodies

A recombinant polypeptide containing the precursor protein of a sesame storage protein, 2S albumin, fused to the C-terminus of a sesame oleosin was expressed in transgenic rice seeds under the control of a rice glutelin promoter. The recombinant polypeptide of 32 kDa, equivalent to the resultant molecular mass of sesame oleosin (15 kDa) and prepro-2S albumin (17 kDa), was detected in the endoplasmic reticulum fraction of maturing transgenic rice seeds, but not in the purified oil bodies or the soluble extract of transgenic seeds. However, sesame oleosin presumably fused with a 2 kDa C-terminal appendix originating from the signal sequence of prepro-2S albumin, was found in the purified oil bodies, and mature sesame 2S albumin apparently processed into two subunits (9 and 4 kDa) linked by disulfide bonds was detected in extracts of transgenic seeds. Immunogold labeling revealed that the sesame oleosin and 2S albumin were separately located in oil bodies and protein bodies of embryo cells of transgenic rice seeds. While sesame 2S albumin was also detected in protein bodies of endosperm cells of transgenic seeds, the co-expressed sesame oleosin, probably degraded due to the lack of oil bodies in this tissue, and was not detected. The results provide a new technique for introducing two recombinant polypeptides separately into rice oil bodies and protein bodies from one expression construct.     

Aroma finishing of PET fabrics with PVAc nanoparticles containing lavender oil

A durable aroma finishing for PET fabric was carried out by adopting poly(vinyl acetate) (PVAc) nanoparticles containing lavender oil (LO) in core. Relatively small size of PVAc nanoparticles (ca. 244 nm of mean particle diameter) was expected to resist the frictional destruction of the particles, which is frequently observed in cases of microcapsules. PVAc nanoparticles containing LO in core were prepared by emulsification-diffusion method and their application as an aroma releasing agent for PET fabrics was assessed through the observation of releasing profiles of LO in ethanol for experimental acceleration. Melamine-formaldehyde (MF) microcapsules containing LO were also prepared and treated on fabrics for comparison. PVAc nanoparticles treated on PET fabric showed higher initial releasing amount, which was ascribed to the enhanced surface area. After 2 days of releasing, PET fabric treated with PVAc nanoparticles showed slower and more stable releasing profile and reached about 12 ppm of cumulative release after 16 days, which was under two thirds of that with MF microcapsules. PVAc nanoparticles can be used as an agent for durable aroma finishing of PET fabrics.     

Compressional behavior of carbon nanotube reinforced mesophase pitch-based carbon fibers

The tensile-recoil compressional behavior of the carbon nanotube reinforced mesophase pitch (MP)-based composite carbon fibers (CNT-re-MP CFs) was investigated by using Instron and SEM. The CNT-re-MP CFs exhibited improved, or at least equivalent, compressive strength as compared with commercial MP-based carbon fibers. Particularly, when CNT of 0.1 wt% was reinforced, the ratios of recoil compressive strengths to tensile strength of CNT-re-MPCFs were much higher (the difference is at least 10% or higher) than those for the commercial counterparts and even than those for PAN-based commercial carbon fibers. FESEM micrographs showed somewhat different fractography from that of a typical shear failure as the CNT content increased.     

Acid-sensitivity and physical properties of polymethylmethacrylate and polyurethane films containing polymeric styryl dye

Solution-cast films of polymethylmethacrylate (PMMA) and polyurethane (PU) containing polymeric styryl dye up to 5% by weight were prepared to investigate their acid-sensitivity and mechanical and thermal properties. Original red samples due to styryl dye turned out yellow very rapidly as they were exposed to acid vapor of hydrochloric acid or p-toluenesulfonic acid. According to UV/VIS spectroscopic measurements, characteristic peak intensities near 430 nm and 520 nm increased or decreased relatively with exposed amounts of acid, respectively. Both PMMA and PU samples showed uniform color distribution due to a good miscibility between polymer and dye which can be evidenced by measurements of glass transition temperature. No significant difference in acid-sensitivity was found between PU and PMMA except relative absorbance. However, dependence of their mechanical properties on dye content was somewhat different with PU or PMMA. In case of PMMA, modulus and breaking stress increased up to about 50% with increasing dye content, whereas those of PU samples showed only slight increase. It was ascribed to whether the matrix polymer was in the glassy or rubbery state.     

A Comparative Study of Non-contact Infrared and Digital Rectal Thermometer Measurements of Body Temperature in the Horse

Equine body temperature taken with a non-contact infrared thermometer (NCIT) does not correlate well with the equine body temperature taken with a digital rectal thermometer (DRT), when it comes to absolute temperatures. There is significant variation in body temperature readings depending on the site at which the temperature was taken. NCIT may still may have, in that it does seem to be able to reliably detect when temperatures are more than that which is clinically normal, and does not seem to give falselyelevated readings in clinically normal horses. In horses that resent having their temperatures taken rectally, the NCIT seems to offer the equine clinician a quick and easy way to at least determine whether the horse's temperature is at or more than normal, even if it does not agree with DRT results.     

Antimicrobial resistance, virulence-associated genes, and pulsed-field gel electrophoresis profiles of Salmonella enterica subsp. enterica serovar Typhimurium isolated from piglets with diarrhea in Korea

Salmonella enterica subsp. enterica serovar Typhimurium was isolated from diarrheic piglets in 2 periods, 2000-2001 (n = 25) and 2005-2006 (n = 17). To compare the characteristics of the isolates collected during the 2 periods, all isolates were tested for antimicrobial resistance, the presence of virulence genes, and pulsed-field gel electrophoresis (PFGE) patterns. All 42 isolates were resistant to at least 1 of the 20 antimicrobials tested, and 39 (93%) were resistant to 2 or more antimicrobials. One isolate was resistant to 12 antimicrobials. Profiles of antimicrobial resistance revealed 20 resistance types. Several isolates were also resistant to quinolones and expanded-spectrum cephalosporins. Ten isolates (24%) were resistant to ampicillin, chloramphenicol, streptomycin, sulfonamides, and tetracycline (ACSSuT); only one isolate had been isolated in 2000-2001, indicating that this type of resistance has rapidly disseminated. Polymerase chain reaction (PCR) assays revealed that all the isolates carried invA. Among the 25 strains isolated in 2000-2001, all carried the sipA, sopA, sopD, sopE2, and ssaR genes, and 96% carried sopB and sifA. Among the 17 strains isolated in 2005-2006, all carried sifA, and approximately 90% carried sipA, sopA, sopB, sopD, sopE2, and ssaR. However, only 6 (14%) of the 42 isolates carried spvC. By PFGE analysis, all 42 strains were classified into 4 major clusters, basically by collection period. The genetic similarity according to PFGE suggests that the strains isolated from diarrheic piglets of this region within the same period may be closely related.     

Molecular epidemiology of rabies virus circulating in South Korea, 1998-2010

We analyzed the nucleotide sequences of the G-L (glycoprotein-large protein) intergenic non-coding region of 33 strains of the rabies virus (RABV) isolated in South Korea in 1998-2010 and compared the sequences with those of previously reported non-Korean strains. The similarities of the nucleotide sequences of the G-L region among all Korean RABV isolates ranged from 97.1 to 100%. Based on the phylogenetic analysis of the G-L region, the Korean RABV isolates were classified into three distinct subgroups with high similarity and were most closely related to the non-Korean NeiMeng1025C isolate, which was isolated from a rabid raccoon dog in eastern China, suggesting that the Korean RABV isolates originate from a rabid raccoon dog in northeastern Asia. Our results indicated that G-L region, as a useful phylogenetic indicator, is equivalent to the nucleoprotein (N) or glycoprotein (G) gene for study of RABV molecular epidemiology and that the Korean RABV isolates showing a few substitutions in the G-L region are continuously circulating in South Korea.     

Mitochondrial dysfunction influences apoptosis and autophagy in porcine parthenotes developing in vitro

Mitochondria are important regulators of both apoptosis and autophagy. One of the triggers for mitochondrial-mediated apoptosis is the production of reactive oxygen species (ROS), which include hydrogen peroxide, superoxide, hydroxyl radical, nitric oxide and peroxynitrite. Recently, several studies have indicated that ROS may also be involved in the induction of autophagy. In the present study, we used H(2)O(2) to induce mitochondrial stress, examined apoptotic- and autophagic-related gene expression and observed LC3 protein (autophagosome presence marker) expression in porcine parthenotes developing in vitro. In porcine four-cell parthenotes cultured for 5 days in NCSU37 medium containing 0.4% BSA, the developmental rate and mitochondrial distribution did not differ from that of the group supplemented with 100 μM H(2)O(2) but was significantly decreased in the group supplemented with 500 μM H(2)O(2) (P<0.05). Transmission electron microscopy (TEM) indicated that whereas normal shaped mitochondria were observed in blastocysts from the control group, abnormal mitochondria (mitophagy) and autophagic vacuoles were observed in blastocysts from the group that received 500 μM H(2)O(2). Furthermore, addition of H(2)O(2) (100 μM and 500 μM) decreased cell numbers (P<0.05) and increased both apoptosis (P<0.05) and LC3 protein expression in the blastocysts. Real-time RT-PCR showed that H(2)O(2) significantly decreased mRNA expression of anti-apoptotic gene Bcl-xL but increased pro-apoptotic genes, Caspase 3 (Casp3) and Bak, and autophagy-related genes, microtubule-associated protein 1 light chain 3 (Map1lc3b) and lysosomal-associated membrane protein 2 (Lamp2). However, the addition of H(2)O(2) had no effect on mRNA expression levels in nuclear DNA-encoded mitochondrial-related genes, cytochrome oxidase (Cox) 5a, Cox5b and Cox6b1, in blastocysts. These results suggest that H(2)O(2) leads to mitochondrial dysfunction that results in apoptosis and autophagy, which is possibly related to porcine early embryo development.     

Immune responses to new vaccine candidates constructed by a live attenuated Salmonella Typhimurium delivery system expressing Escherichia coli F4, F5, F6, F41 and intimin adhesin antigens in a murine model

To construct a novel live oral vaccine candidate for the prevention of pathogenic Escherichia coli infections in neonatal piglets, an expression and secretion plasmid and an attenuated Salmonella delivery system were used. The individual E. coli genes K88ac, K99, FasA, F41 and intimin adhesins were inserted into pBP244 containing asd, lepB, secA and secB, and these plasmids were transformed into a Salmonella Typhimurium ΔcpxR Δlon Δasd. Forty female BALB/c mice were divided into four groups, A to D (ten mice per group). Groups A and B were administered with the mixture containing all constructs and the S. Typhimurium containing pBP244 only as a control, respectively. Groups C and D were primed and boosted with the mixture and the S. Typhimurium harboring pBP244 only, respectively. Each recombinant adhesin secreted from the individual candidates was confirmed by Western blot analysis. The serum IgG and secretory IgA (sIgA) titers to individual adhesins in all immunized groups were higher than those in control. Furthermore, IgG and sIgA levels in group C were higher than those in group A, and the IgG1 titers were increased in Group C but IgG2a titers were similar or decreased in Group C compared to Group A. In addition, the vaccine strains were not detected in fecal samples of any immunized mice. The novel vaccine candidates are not only highly immunogenic, but also safe for vaccinated mice and environment. In addition, the immune responses can be more efficiently induced through the booster-administration.