Inactivated Pseudomonas aeruginosa inhibits hypoxia-induced pulmonary hypertension by preventing TGF-ß1/Smad signaling.

Pseudomonas aeruginosa is one of the common colonizing bacteria of the human body and is an opportunistic pathogen frequently associated with respiratory infections. Inactivated P. aeruginosa (IPA) have a variety of biological effects against inflammation and allergy. Transforming growth factor-ß (TGF-ß) signaling plays a critical role in the regulation of cell growth, differentiation, and development in a wide range of biological systems. The present study was designed to investigate the effects of IPA on TGF-ß/Smad signaling in vivo, using a hypoxia-induced pulmonary hypertension (PH) rat model. Sprague Dawley rats (n=40) were exposed to 10% oxygen for 21 days to induce PH. At the same time, IPA was administered intravenously from day 1 to day 14. Mean pulmonary artery pressure (mPAP) and the right ventricle (RV) to left ventricle plus the interventricular septum (LV+S) mass ratio were used to evaluate the development of PH. Vessel thickness and density were measured using immunohistochemistry. Primary arterial smooth muscle cells (PASMCs) were isolated and the proliferation of PASMCs was assayed by flow cytometry. The production of TGF-ß1 in cultured supernatant of PASMCs was assayed by ELISA. The expression levels of α-smooth muscle actin (α-SMA), TGF-ß1 and phospho-Smad 2/3 in PASMCs were assayed by western blot. Our data indicated that IPA attenuated PH, RV hypertrophy and pulmonary vascular remodeling in rats, which was probably mediated by restraining the hypoxia-induced overactive TGF-ß1/Smad signaling. In conclusion, IPA is a promising protective treatment in PH due to the inhibiting effects on TGF-ß1/Smad 2/3 signaling.

Antisense expression of Gossypium hirsutum UDP-glucuronate decarboxylase in Arabidopsis leads to changes in cell wall components.

UDP-glucuronate decarboxylase (UDP-xylose synthase; UXS, EC 4.1.1.35) is an essential enzyme of the non-cellulosic polysaccharide biosynthetic pathway. In the present study, using transient expression of fluorescently labeled Gossypium hirsutum UXS (GhUXS3) protein in onion epidermal cells, we observed that this protein was distributed in the cytoplasm. The GhUXS3 cDNA of cotton was expressed in an antisense orientation in Arabidopsis thaliana by Agrobacterium tumefaciens-mediated transformation. Homozygous plants showing down-regulation of UXS were analyzed with northern blots. Compared to the untransformed control, transgenic plant showed shorter roots, earlier blossom formation, and delayed senescence. Biochemical analysis indicated that levels of rhamnose, mannose, galactose, glucose, xylose, and cellulose were reduced in some of the down-regulated antisense plants. These results suggest that GhUXS3 regulates the conversion of non-cellulosic polysaccharides and modulates their composition in plant cell walls. We also discuss a possible cellular function for GhUXS in determining the quality of cotton fibers.

Ion beam transformation with corn DNA alters proteinase expression in rice seedling roots.

Corn DNA was introduced into dry seeds of rice (cv. 'YuJing-6') by ion beam irradiation. Proteinase activities in rice seedling roots were subsequently analyzed by renaturation electrophoresis at pH 4.5, 7.0, and 8.5. Proteinase activity was more pronounced on gels at higher pH. Irradiation of rice seedling roots caused the loss of some proteinase bands at all pH conditions although a novel 50-kDa band was found at both pH 7.0 and 8.5. No new proteinase activity was detected at pH 4.5. However, novel bands and bands showing stronger activity were observed at pH 7.0 and 8.5. The data indicate that the expression of proteinases in rice seedling roots was altered following low energy ion beam mediated transformation with corn DNA.

Selected representative microsatellite loci for genetic monitoring and population structure analysis of miniature swine.

To optimize the combination of microsatellite loci for genetic control of outbred swine stocks, 32 of 100 loci distributed among almost all chromosomes (except 12) were screened out by 1.5% agarose, 8% polyacrylamide gel and capillary electrophoresis scanning among 3 miniature swine outbred stocks, namely Bama (BM), Guizhou (GZ) and Tibeta (TB). The mean total and effective allele numbers among these stocks were 12.1 and 5.9, respectively. The mean heterozygosity for these breeds was 0.5428, 0.6978 and 0.7646, and polymorphism information content was 0.5469, 0.7296 and 0.7663, respectively. Accordingly, hereditary variation from low to high was BM < GZ < TB. This showed that the genetic relationship between BM and GZ pigs was closer, and both were distant from TB. Additionally, the effectiveness of the 32-locus combination for evaluation of genetic quality was demonstrated in Changchun-junmo-1 (CJ-1), a standard outbred Chinese pig stock, in which the mean total and effective allele numbers and mean heterozygosity were 6.1613, 3.8483 and 0.6903, respectively. Since our results were consistent with the breeding pedigrees, the 32 loci could be used for both genetic monitoring within the individual outbred miniature swine stocks and population structure analysis between them.

Analysis of specific sequences in mutant rice generated by introduction of exogenous corn DNA.

Rice variation induced by the introduction of exogenous DNA has become an important method of improving rice varieties and creating new germplasms. In this study, we transferred maize genomic DNA fragments to the receptor of Nipponbare rice using a modified "pollen-tube pathway" method. Material from mutant rice B1 and B2 were acquired and 14 specific bands were obtained from the material using amplified fragment length polymorphism analysis. From the 14 specific sequences obtained, there were 3791 bp, including 144 base mutations with a base mutation rate of 3.80%. Specific bands resulted from base mutation of selective bases or restriction endonuclease recognition sequences, or insertion or deletion of DNA fragments. The frequency of single-base mutations was significantly higher than that of double-base mutations, three-sequential base mutations, and multiple-sequential base mutations. The site frequency of base substitution (87.04%) was significantly higher than that of base insertion (3.70%) or deletion (9.26%). In all cases of base substitution, the frequency of transition (76.47%) was significantly higher than transversion (23.53%). The above results indicate that transferring foreign-species DNA into rice cells can induce base mutations in the receptor, with base substitutions occurring at the highest frequency, and the dominant type of base substitutions being transition. Preliminary analysis reveals that the molecular mechanism of transferring exogenous DNA into rice causes mutations, which provides theoretical data on biological mutagenesis for further research.

Mapping quantitative trait loci with additive effects and additive x additive epistatic interactions for biomass yield, grain yield, and straw yield using a doubled haploid population of wheat (Triticum aestivum L.).

Biomass yield is one of the most important traits for wheat (Triticum aestivum L.)-breeding programs. Increasing the yield of the aerial parts of wheat varieties will be an integral component of future wheat improvement; however, little is known regarding the genetic control of aerial part yield. A doubled haploid population, comprising 168 lines derived from a cross between two winter wheat cultivars, 'Huapei 3' (HP3) and 'Yumai 57' (YM57), was investigated. Quantitative trait loci (QTL) for total biomass yield, grain yield, and straw yield were determined for additive effects and additive x additive epistatic interactions using the QTLNetwork 2.0 software based on the mixed-linear model. Thirteen QTL were determined to have significant additive effects for the three yield traits, of which six also exhibited epistatic effects. Eleven significant additive x additive interactions were detected, of which seven occurred between QTL showing epistatic effects only, two occurred between QTL showing epistatic effects and additive effects, and two occurred between QTL with additive effects. These QTL explained 1.20 to 10.87% of the total phenotypic variation. The QTL with an allele originating from YM57 on chromosome 4B and another QTL contributed by HP3 alleles on chromosome 4D were simultaneously detected on the same or adjacent chromosome intervals for the three traits in two environments. Most of the repeatedly detected QTL across environments were not significant (P > 0.05). These results have implications for selection strategies in wheat biomass yield and for increasing the yield of the aerial part of wheat.

Assessing genetic diversity of cotton cultivars using genomic and newly developed expressed sequence tag-derived microsatellite markers.

Estimations of genetic diversity and of relationships between varieties are crucial for cotton breeding. The genetic diversity of 59 core cotton cultivars, most of which were collected from China's main cotton-growing areas, was analyzed based on genomic and newly developed expressed sequence tag-derived microsatellite markers, using total DNA extracted from fresh leaf tissue. Three hundred and two fragments were detected, of which 255 were polymorphic. The number of amplification products generated by each primer varied from 2 to 14, with a mean of 5.08 bands/primer. The polymorphism information content was 0.50 to 0.90, with a mean of 0.80. The genetic similarity coefficients were calculated and dendrograms were constructed by the unweighted pair group with arithmetic mean method; the resulting distance matrix gave a dendrogram with four main clusters. Some cultivars with similar pedigrees could be clustered. For example, Zhong206 and Shanmian4, both derived from Deltapine15, were clustered. The genetic similarity coefficient of the 59 core cultivars ranged from 0.53 to 0.99, with a mean of 0.72, indicating that there was a relatively high level of genetic variation.