Antioxidative Effects of Astaxanthin against Nitric Oxide‐Induced Oxidative Stress on Cell Viability and Gene Expression in Bovine Oviduct Epithelial Cell and the Developmental Competence of Bovine IVM/IVF Embryos

The aim of the present study was to elucidate the fundamental mechanism of bovine oviduct epithelial cell (BOEC) co‐culture on developmental capacity of bovine in vitro oocyte maturation/in vitro fertilization (IVM/IVF) embryos. We examined the effects of astaxanthin against nitric oxide‐induced oxidative stress on cell viability by MTT assay, lipid peroxidation (LPO) by using thiobarbituric acid (TBA) reaction for malondialdehyde (MDA) and the expression of antioxidant genes (CuZnSOD, MnSOD and Catalase) or apoptosis genes (Bcl‐2, Caspase‐3 and Bax) by RT‐PCR in BOEC. We also evaluated the developmental rates of bovine IVM/IVF embryos co‐cultured with BOEC pre‐treated with astaxanthin (500 μm ) in the presence or absence of sodium nitroprusside (SNP, 1000 μm ) for 24 h. Cell viability in BOEC treated with SNP (50–2000 μm ) lowered, while astaxanthin addition (50–500 μm ) increased it in a dose‐dependent manner. Cell viability in astaxanthin plus SNP (1000 μm ) gradually recovered according to the increase in astaxanthin additions (100–500 mm ). The LPO in astaxanthin group (50–500 μM) gradually decreased in a dose dependent manner and among SNP or astaxanthin plus SNP group, SNP alone and astaxanthin (50 μM) plus SNP shown a significant increase than other groups (p < 0.05). Expression of apoptosis or antioxidant genes was detected by RT‐PCR. Bcl‐2 and antioxidant genes were detected in astaxanthin or astaxanthin plus SNP group, and Caspase‐3 and Bax genes were only found in SNP group. When bovine IVM/IVF embryos were cultured for 6–7 days under co‐culture system such as BOEC treated with astaxanthin in the presence or absence of SNP, the developmental ability to blastocysts in 500 μm astaxanthin group was the highest of all groups. These results suggest that astaxanthin has a antioxidative effect on cell viability and LPO of BOEC, and development of bovine IVM/IVF embryos due to the induction of antioxidant genes and suppression of apoptosis genes.     

Electrospun nanofibers of poly(NPEMA-co.-CMPMA): Used as Heavy metal ion remover and water sanitizer

Homo and copolymers of monomers 2-(N-phthalimido) ethylmethacrylate (NPEMA) and 4-Chloro-3-methyl phenyl methacrylate (CMPMA) were prepared in N,N-dimethyl formamide (DMF) solution at 70 °C using 2,2-azobisisobutyronitrile (AIBN) as initiator. The solution of poly(NPEMA-co.-CMPMA) in 20 % DMF was used to fabrication electrospun nanofiber by electrospinning technique. IR data were primarily employed to characterize polymers. The formation of nanofibers was identified by SEM study. The metal ion uptake capacity of copolymers and nanofibers were obtain by batch equilibrium method using different metal ion solution. The antimicrobial activity of the copolymers, Polymer nanocomposites and their nanofibers were tested against different microbial organisms by using quantitative method. The main objective of this investigation was to find whether nanofiber are better remover of metal ions compared to copolymers. It was also aimed to study the efficacy of nanofibers of copolymers and copolymer composite with nano Ag as water sanitizer.     

Cryopreservation of Canine Platelets

Background: Platelet cryopreservation allows long-term storage and immediate availability of transfusion products.
Hypothesis: The addition of a preparation inhibiting platelet activation (Thrombosol, in 2% dimethyl sulfoxide [DMSO]) will enhance in vitro function and prolong in vivo survival of cryopreserved platelets compared with those preserved in 6% DMSO.
Animals: Thirty-three research dogs.
Methods: Prospective study. Eleven fresh canine apheresis platelet concentrates (PCs) were each split into 3 units: fresh and cryopreserved in 6% DMSO or Thrombosol. Platelet analysis, performed 1–10 weeks postfreezing, included in vitro functional testing and in vivo survival assessed by administration of biotinylated platelets.
Results: Platelet aggregation was diminished in cryopreserved PC. Cryopreserved platelets could be activated, as based on mean thrombin-stimulated P-selectin expression (6% DMSO, 23.0%; Thrombosol, 18.4%), although to a lesser extent than fresh PC (49.1%) ( P < .0001). The mean maximum in vivo platelet recovery for fresh PC was 80.3%, significantly greater than recovery for 6% DMSO (49.2%) and Thrombosol PC (43.7%) ( P ≤ .001). The half-life (days) of fresh PC (3.8 ± 0.4) was significantly ( P < .002) greater than that of 6% DMSO (1.9 ± 1.0) and Thrombosol (2.4 ± 1.1) PC, with no difference ( P = .3) between cryopreserved PC.
Conclusions and Clinical Importance: Cryopreservation of canine platelets using Thrombosol did not provide any advantage over preservation using 6% DMSO. Cryopreserved platelets can be activated in vitro and provide therapeutic benefit when fresh platelets are unavailable. Further studies are needed to assess their in vivo hemostatic function.     

Synthesis of some new thermally stable reactive dyes having 4(3<Emphasis Type="Italic">H</Emphasis>)-quinazolinone molecule for the dyeing of silk,wool, and cotton fibers

Diazotized 3-{4-[2-chloro-4-amino-5-carboxybenzyl]-5-chloro-2-carboxyphenyl}-6-iodo-2-phenylquinazolin-4(3H)-one (4) was coupled with various p-nitro anilino cyanurated coupling components (6a-j) to give the corresponding quinazolinone based reactive dyes (7a-j) in reasonable yields. All the reactive dyes (7a-j) were characterized by spectroscopic technique and elemental analysis. These dyes were applied to silk, wool, and cotton fibers as reactive dyes and their spectroscopic data, colorimetric data, antimicrobial activity, thermal stability, and fastness properties were evaluated.     

Characterization and application of recombinant β-glucosidase (BglH) from <Emphasis Type="Italic">Bacillus licheniformis</Emphasis> KCTC 1918

β-Glucosidase (β-1,4-D-glucoside glucohydrolase: EC.3.2.1.21) catalyzes the hydrolysis of β-glucosidic bonds between saccharides and aryl or alkyl groups. A gene encoding β-glucosidase from Bacillus licheniformis KCTC 1918, an anaerobic spore-forming soil bacterium, was cloned and characterized. The structural gene for the β-glucosidase consists of 1410 bp encoding 469 amino acid residues, and has a molecular weight of 53.4 kDa as estimated by sodium dodecyl sulfate polyacrylamide gel electrophoresis with 12% separating gel. The enzyme activity was determined against pNPG as a substrate. The enzyme was optimally active at pH 6.0 (citrate-phosphate buffer) and 47°C. β-Glucosidase retained 100% of its original activity for 24 h. The activity of the enzyme was stimulated by glycerol and urea and was decreased by Ca²⁺, Cu²⁺, Hg²⁺, Mg²⁺, and Mn²⁺. In particular, Cu²⁺ had the strongest negative effect on β-glucosidase activity. The purified β-glucosidase was active against pNPG and cellobiose. When the β-glucosidase was tested for cellulose hydrolysis, the supplement of β-glucosidase with cellulose increased the glucose yield from pine wood powder by 139.8%.     

10年生府竹丛生竹的离体无性繁殖

用10年生丛生竹的节片通过离体腋笋增殖和生根产生出府竹小苗。不同季节对无菌培养物的建立、腋芽发萌和初代培养的影响很大。3片丛生林中,无菌培养物的建立有差别,但腋芽发萌没有差别。用加有31.06uM BA和2.85um IAA的MS液体介质,最大的增值率为3.18;用加有20~25um IBA MS液体介质,离体生根率为66.7~77.8%。用不同的植物生长素,生根所用的天数也不同,而IBA液能在短期促进生根,仅仅为2-3周。成功炼苗后,在装有土壤、沙子和农家肥塑料袋里,离体繁殖的小苗成活率为90%,大约有2000株小苗用于野外栽植。     

Exogenous DNA Uptake of Boar Spermatozoa by a Magnetic Nanoparticle Vector System

The sperm‐mediated gene transfer method is applicable to transgenesis in many species that use spermatozoa for reproduction recently, which has been shown various results. In the current study, we show that transgenic porcine embryos can be efficiently produced by employing a simple transfection method that uses magnetic nanoparticles (MNPs). The complexes formed between plasmid DNA and MNPs were bounded on ejaculated boar spermatozoa at a higher efficiency compared to methods using DNA alone or lipofection. Using confocal microscopy, rhodamine fluorophore‐labelled MNPs were detected on external surfaces of the spermatozoa membrane, which were bounded on zona pellucida of in vitro maturated oocyte during in vitro fertilization. Electron microscopy revealed that clusters of MNPs were detected in inside of plasma membrane and nucleus of the spermatozoa head. Additionally, we found that magnetofected boar spermatozoa could be fertilized with oocytes in vitro and that the resulting gene of green fluorescent protein was detected in fertilized eggs by genomic PCR analysis. Taken together, these results suggest that MNPs can be used to efficiently introduce a transgene into embryo via spermatozoa.     

Culture-dependent and culture-independent microbial investigation of pine litters and soil in subtropical Australia

Background, aim, and scope  Forest plantations, widely grown for wood production, involve the selective promotion of single-tree species or replacement of natural species by exotic tree species. Slash pine (Pinus elliottii) has been chosen for reforestation of infertile sandy soils in southeast Queensland, Australia. These exotic pine plantations minimize soil and water losses and are important scientific study sites. The soil environment of these plantations, though devoid of sufficient nutrients, organic carbon and other factors, harbors innumerable bacteria that may play a crucial role in maintaining soil quality and ecosystem functions. These soil microorganisms also have the potential for use as sensitive biological indicators to reflect environmental changes. It is therefore essential to understand the interrelationships among bacterial communities and their environment by assessing their structural and functional diversity and their responses to disturbances. The main aim of our investigation was to determine the diversity of bacterial communities in forest litters and soil during the forest leaf litter decomposition using culture-dependent and culture-independent techniques. Materials and methods  A 25-cm (diameter) × 40-cm core sample was collected and fractionated into three subsamples designated E1 (L leaf litter layer), E2 (F leaf litter layer), and E5 (0–10 cm soil layer). Both culture-dependent and culture-independent methods were applied in this study. In the culture-independent study, a strategy of whole-community DNA extraction, polymerase chain reaction (PCR) amplification followed by cloning and 16S rDNA sequence analysis was used; for culture-dependent study, the strategy included sample plating and bacteria isolating, DNA extraction, PCR amplification, and 16S rDNA sequence analysis. The diversity similarities between two bacterial communities and two methods are quantified using Jensen–Shannon divergence. Results  From culture-dependent study, 336 colonies in total were isolated and grouped from the three subsamples, and the 16S rRNA sequence analysis from a representative isolate from each morphogroup (21 isolates) indicated that they belonged to the phyla Actinobacteria, Firmicutes, and Proteobacteria. Culture-independent assessment based on 16S rRNA gene library comprising 194 clones revealed that members of the phylum Actinobacteria were absent in the culture-independent studies. Clones in libraries from E1 consisted exclusively of members of the Firmicutes. The majority of clones from E2 were related to Firmicutes (79%) and Proteobacteria (21%). Clones derived from E5 were mostly affiliated with Acidobacterium (42%), followed by unclassified bacteria (27%), Verrucomicrobiales (12%), Proteobacteria (11%), and Planctomycetes (8%). Discussion  This study showed that bacterial culturabilities in different fractions of leaf litters were similar, and both of them were higher than the bacterial culturability in the soil. Unculturable bacterial diversity in the soil, however, was much higher than the leaf litter bacterial diversity. The bacterial diversity on the top layer of leaf litters was slightly less than that on the bottom layer of leaf litters. This might indicate that forest soils are a more complex environment than leaf litters are and also that they might inhabit more unculturable microorganisms in the forest soils, which would need to be further investigated. The leaf litter layer samples also demonstrate the significant difference between the bacterial community diversity discovered by these two methods in this study. The information provided by assessing the different fractions of leaf litters and forest soil has improved our understanding of the bacterial community distributions within the forest soil and the above-leaf litters in an exotic pine plantation of subtropical Australia. Conclusions  This study represents the first attempt to examine the bacterial community in the different fractions of forest leaf litters and soil in subtropical Australia. The data from this study show that the 16S rDNA clone libraries provided more comprehensive phylogenetic diversity in the soil and leaf litter samples than the culture collections provided, and both the culture-dependent and culture-independent studies revealed that the bacterial diversity present in the leaf litters was very different to that present in the soil. The comparative analysis of bacterial communities in different fractions of leaf litters and soil samples has also provided important baseline information about the bacterial diversity and composition in the exotic pine forest plantations. Recommendations and perspectives  The experimental data provided important information on the bacterial diversity in forest leaf litter and soil samples, though additional surveys and comparisons at different locations would be needed to further characterize. In addition, combined methods that can provide different parts of information on bacterial diversity are encouraged to be used in bacterial community study. The established libraries of diverse 16S rRNA gene fragments from slash pine leaf litters and forest soil can be used to construct specific DNA primers and probes to target bacterial groups of interest. It may then be possible to study the ecology of these bacterial communities and the role of specific bacterial groups that contribute to the many interesting properties of these environments.     

Analysis of relationship between crop growth parameters,yield and physical environment within the crop canopy of sesame (Sesamum Indicum) at different sowing dates: Untersuchung der beziehungen zwischen wachstumsparametern,ertrag und physischer umwelt im pflanzenbestand von sesam (Sesamum Indicum) bei verschiedenen aussaatterminen

Relation between crop growth parameters of sesame (Sesamum indicum) and the physical environment within the crop canopy at different sowing dates was studied during the summer seasons of 1999 and 2000. The maximum leaf growth rate (LGR) and leaf area index (LAI) was obtained from February 19 sown crop. About 34.4% variation in LGR could be explained through the variation in the physical environmental elements within the crop canopy. The LAI was depressed in the later months of sowing. The February 19 sown crop produced significantly, the highest dry matter production (DMP) in all the stages of crop growth. The regression model indicated that the crop growth rate (CGR) was adversely affected by the ambient temperature and photosynthetic active radiation (PAR) within the crop canopy. Crops sown on 19 February and 1 March produced statistically similar yields. The cultivar Rama produced higher yields than B-67 and Kanke-1. Regression models suggested that the temperature profile and PAR within the crop canopy produced 69 and 39% variation in yield, respectively.