Decomposition of coffee residue in soil

In this study the changes in the chemical properties of coffee residue during decomposition in soil were investigated. 1) Coffee residue contained about 20 g kg^-1 N. The main N compound was a structural protein-No However, the content of easily decomposable N (soluble protein-N and nonprotein-N) extracted by 800 mL L^-1 ethanol and 0.1 mol L^-1 phosphate buffer (pH 7.5) was very low. 2) The rates of nonprotein-N and soluble protein-N which remained reached values of 59 and 66% at 314 d after application of the coffee residue (DAA) to soil, respectively. However, the amount of structural protein-N reached a value of 96% at 314 DAA. 3) The lipid fraction of the coffee residue decreased from 186 to 47 g kg^-1 by 314 d after application. Coffee residue has a high content of lipid fraction and a large fraction corresponding to structural protein-No Therefore, the decomposition of the coffee residue is slower than that of other organic materials.     

Oolong tea theasinensins attenuate cyclooxygenase-2 expression in lipopolysaccharide (LPS)-activated mouse macrophages: structure-activity relationship and molecular mechanisms

Oolong tea theasinensins are a group of tea polyphenols different from green tea catechins and black tea theaflavins. The present study reports the inhibitory effects of oolong tea theasinensins on the expression of cyclooxygenase-2 (COX-2) and underlying molecular mechanisms in lipopolysaccharide (LPS)-activated murine macrophage RAW264 cells. The structure-activity data revealed that the galloyl moiety of theasinensins played an important role in the inhibitory actions. Theasinensin A, a more potent inhibitor, caused a dose-dependent inhibition of mRNA, protein, and promoter activity of COX-2. An electrophoretic mobility shift assay (EMSA) revealed that theasinensin A reduced the complex of NF-κB- and AP-1-DNA in the promoter of COX-2. Signaling analysis demonstrated that theasinensin A attenuated IκB-α degradation, nuclear p65 accumulation, and c-Jun phosphorylation. Furthermore, theasinensin A suppressed the phosphorylation of MAPKs, IκB kinase α/β (IKKα/β), and TGF-β activated kinase (TAK1). These data demonstrated that the down-regulation of TAK1-mediated MAPKs and NF-κB signaling pathways might be involved in the inhibition of COX-2 expression by theasinensin A. These findings provide the first molecular basis for the anti-inflammatory properties of oolong tea theasinensins.     

Detection of telomere length and oxidative stress in Chondrichthyes

Telomeres, repeating TTAGGG sequences at the ends of eukaryotic chromosomes, increase genomic stability. Telomere shortening occurs not only during DNA replication associated with cell division but also under oxidative stress, where reactive oxygen species damage DNA. Therefore, changes in telomere length can be used to evaluate chronic cost or stress responses incurred by individuals. The phylogenetically unique Chondrichthyes are among the least-studied groups of marine vertebrates. Telomere data are limited and have only been reported in a few Chondrichthyes species. In this study, we measured telomere length and quantified oxidative stress and antioxidant power in 17 Chondrichthyes species whose telomere length has not been measured before. The presence of telomere sequences?>?30 bp and lower values of oxidative stress were confirmed in most species. Average telomere length was not correlated with oxidative stress and antioxidant power in 15 species for which both measurements were available. It would be desirable in the future to elucidate the nature of telomeres in Chondrichthyes, and their direct relationship with reactive oxygen species.

     

Simple method for isolation of glyceraldehyde 3‐phosphate dehydrogenase and the improvement of myofibril gel properties

Porcine glycoliytic enzyme, glyceraldehyde 3‐phosphate dehydrogenase (G3PD) was prepared effectively by a combination of ethylene diamine tetra‐acetate (EDTA) pretreatment and affinity purification. After salting out of porcine sarcoplasmic proteins (SP) with ammonium sulfate at 75% saturation, the obtained supernatant (SP‐f3) was treated with EDTA, leaving G3PD in the supernatant (G3PD‐E) and most other SPs in the precipitate. At that time, the separation of G3PD‐E required more than 20 mmol/L EDTA. G3PD‐E was then subjected to affinity purification by batchwise method using blue‐sepharose CL‐6B, and purified G3PD (G3PD‐AP) was obtained using 2 mol/L potassium chloride (KCl) as an eluent. Texture analysis showed that the hardness, adhesiveness and gumminess of the myofibril gel at 0.2‐mol/L NaCl increased with the addition of G3PD‐AP. Scanning electron microscopy revealed that the G3PD‐AP reinforced the gel network of the myofibril. However, scanning electron micrograph analysis showed that the network‐structure of the gel by the addition of G3PD‐AP developed in a different manner from that by adding 0.6 mol/L NaCl. These results showed that glycolytic enzyme, G3PD, contributes to the improvement of the rheological properties of meat products.     

An expanded palette of genetically encoded Ca²⁺ indicators

Engineered fluorescent protein (FP) chimeras that modulate their fluorescence in response to changes in calcium ion (Ca(2+)) concentration are powerful tools for visualizing intracellular signaling activity. However, despite a decade of availability, the palette of single FP-based Ca(2+) indicators has remained limited to a single green hue. We have expanded this palette by developing blue, improved green, and red intensiometric indicators, as well as an emission ratiometric indicator with an 11,000% ratio change. This series enables improved single-color Ca(2+) imaging in neurons and transgenic Caenorhabditis elegans. In HeLa cells, Ca(2+) was imaged in three subcellular compartments, and, in conjunction with a cyan FP-yellow FP-based indicator, Ca(2+) and adenosine 5'-triphosphate were simultaneously imaged. This palette of indicators paints the way to a colorful new era of Ca(2+) imaging.     

Leaf margin phenotype-specific restriction-site-associated DNA-derived markers for pineapple (Ananas comosus L.)

To explore genome-wide DNA polymorphisms and identify DNA markers for leaf margin phenotypes, a restriction-site-associated DNA sequencing analysis was employed to analyze three bulked DNAs of F1 progeny from a cross between a ‘piping-leaf-type’ cultivar, ‘Yugafu’, and a ‘spiny-tip-leaf-type’ variety, ‘Yonekura’. The parents were both Ananas comosus var. comosus. From the analysis, piping-leaf and spiny-tip-leaf gene-specific restriction-site-associated DNA sequencing tags were obtained and designated as PLSTs and STLSTs, respectively. The five PLSTs and two STSLTs were successfully converted to cleaved amplified polymorphic sequence (CAPS) or simple sequence repeat (SSR) markers using the sequence differences between alleles. Based on the genotyping of the F1 with two SSR and three CAPS markers, the five PLST markers were mapped in the vicinity of the P locus, with the closest marker, PLST1_SSR, being located 1.5 cM from the P locus. The two CAPS markers from STLST1 and STLST3 perfectly assessed the ‘spiny-leaf type’ as homozygotes of the recessive s allele of the S gene. The recombination value between the S locus and STLST loci was 2.4, and STLSTs were located 2.2 cM from the S locus. SSR and CAPS markers are applicable to marker-assisted selection of leaf margin phenotypes in pineapple breeding.     

Repeated applications of farmyard manure enhance resistance and resilience of soil biological functions against soil disinfection

The effect of repeated application of farmyard manure (FYM) on resistance and resilience of some of soil biological functions was compared between two soils [chemically fertilized soil (CF soil) and chemically and organically amended soil (CF+FYM-soil)]. The impact of chloropicrin and metam sodium disinfection on organic matter (glucose and chitin) decomposition activity and number of ammonia oxidizing bacteria in the soils was measured periodically from just after disinfection to evaluate stability of soil biological functions to soil disinfection. Glucose and chitin decomposition activity and number of ammonia oxidizing bacteria was suppressed by soil disinfection more seriously in the CF soil than in the CF+FYM soil. The decomposition rates recovered in the disinfected CF+FYM soil 2 to 12 weeks after disinfection, but not in the CF soil during 12 weeks incubation. These results demonstrated that soil resistance and resilience of the selected biological functions may be higher in the CF+ FYM soil than in the CF soil. The ratios of fungal and bacterial contribution to glucose decomposition activity were evaluated by the substrate-induced respiration method. Chloropicrin and metam sodium disinfection decreased fungal contribution throughout the incubation period (12 weeks) in the CF soil. A similar tendency was observed in the CF+FYM soil, but the fungal contribution tended to recover more rapidly in the CF+FYM soil than in the CF soil. Repeated applications of FYM may be an effective way to enhance soil functional stability.     

Use of enoyl coenzyme A hydratase of Mycobacterium avium subsp. paratuberculosis for the serological diagnosis of Johne's disease

Johne's disease (JD), caused by Mycobacterium avium subspecies paratuberculosis (MAP), remains difficult to control because of the lack of specific and sensitive diagnostic tests. In order to improve the specificity of sero-diagnosis for JD, the phage display library derived from genomic DNA of MAP was immunoscreened to identify novel antigenic targets. We selected a clone using antibodies from MAP experimentally infected cattle, and annotated its coding sequence as MAP1197 in the MAP genome, which encoded “echA12_2” in the MAP protein (Map-echA) belonging to Enoyl-CoA hydratase, known as a crotonase enzyme. The Map-echA was expressed in Esherichia coli and purified as a histidine-tag recombinant protein (rMap-echA), and the diagnostic potential of the protein was further evaluated by enzyme-linked immunosorbent assays (ELISA). Antibody responses to rMap-echA were higher in MAP-infected cattle than in uninfected cattle. The specificity of the Map-echA ELISA was also confirmed by evaluation with hyper-immune sera against various kinds of Mycobacterium species. Furthermore, in all experimentally infected cattle the antibody against rMap-echA was detected 2–7 months earlier than by a commercially available ELISA kit. These results suggested that Map-echA can be used as a specific and sensitive serological diagnostic antigen for the detection of MAP infection.     

Effect of medium additives during liquid storage on developmental competence of in vitro matured bovine oocytes

Our aim was to improve the developmental competence of bovine oocytes during their liquid storage by using additives. In vitro matured oocytes were stored for 20 h at 25°C in HEPES buffered TCM 199 medium (base medium). After storage, in vitro embryo development after in vitro fertilization was compared to those of non‐stored (control) ones. Addition of 10% (v/v) newborn calf serum or 10.27 mmol/L pyruvate alone to the base medium did not improve blastocyst formation rates in stored oocytes; however, their simultaneous addition significantly improved the rate compared with those stored in base medium (P < 0.05). Supplementation of the holding medium with dithiothreitol (DTT) at any concentrations did not improve embryo development from stored oocytes. Although supplementation with cyclosporine A (CsA) significantly reduced apoptosis and membrane damage rates during storage, it did not improve the developmental competence of oocytes. 1,2‐bis(2‐aminophenoxy) ethane N,N,N’,N’‐tetraacetic acid tetrakis‐acetoxymethyl ester and ruthenium red had no effect on oocyte apoptotic rates. Blastocyst formation rates in all stored groups remained significantly lower than that of the control. In conclusion, pyruvate and serum had a synergic effect to moderate the reduction of oocyte quality during storage, whereas mitochondrial membrane pore inhibitor CsA and the antioxidant DTT did not affect their developmental competence.