Sapelovirus A (SV-A) is a positive-sense single-stranded RNA virus which is associated with acute diarrhea, pneumonia and reproductive disorders. The virus capsid is composed of four proteins, and the functions of the structural proteins are unclear. In this study, we expressed SV-A structural protein VP1 and studied its antigenicity and immunogenicity. SDS-PAGE analysis revealed that the target gene was expressed at high levels at 0.6 mM concentration of IPTG for 24 h. The mouse polyclonal antibody against SV-A VP1 protein was produced and reached a high antiserum titer (1: 2,048,000). Immunized mice sera with the recombinant SV-A VP1 protein showed specific recognition of purified VP1 protein by western blot assay and could recognize native SV-A VP1 protein in PK-15 cells infected with SV-A by indirect immunofluorescence assay. The successfully purified recombinant protein was able to preserve its antigenic determinants and the generated mouse anti-SV-A VP1 antibodies could recognize native SV-A, which may have the potential to be used to detect SV-A infection in pigs.
Osteocalcin is a major non-collagenous component of the bone extracellular matrix and is considered to be an indicative factor of osteoblast differentiation. In the present study, we detected osteocalcin expression in different antler areas and growth phases by immunohisto- chemistry. Osteocalcin was highly expressed in all areas during the mineralization period and in mesenchymal cell and chondrocyte areas during the rapid growth period. The nucleotide sequence of the osteocalcin gene in sika deer antler was determined. The open reading frame was 303 bp encoding a protein of 100 amino acids. The estimated molecular mass of osteocalcin was 10.38 kDa and the theoretical isoelectric point was 5.37. The osteocalcin gene with a 6× His-tag at the C-terminus was cloned into the pGEX-4T1 vector and expressed in Escherichia coli under optimal conditions. The recombinant soluble protein fused with GST was purified with Ni-NTA resin. The purified osteocalcin protein exhibited a significant increase in HA adhesion and promoted antler chondrocyte proliferation. Osteocalcin is an important factor in regulating the rapid growth and differentiation of deer antlers.
Every part of the human body goes beyond the anatomy-physiology limits to reach deep contents and symbolic meanings. We can identify a range of verbs (which constitute a part of idiomatic expressions) that indicate different alterations of the body’s integrity. As for their figurative use, they serve to describe a mental state. The parts of the body linked to the sensory, motor and intellectual spheres tend to be accompanied by adjectives that are part of the terminology of the psychiatric past. We come to the conclusion that some medical terms (in this case the parts of the body) have entered into everyday speech and have assumed symbolic meanings. From the interlingual point of view, it is whereas possible to see considerable differences between Italian and Polish. It follows that the linguistic picture of the world helps to understand the generally accepted statements in a certain community.
Polychlorinated biphenyls (PCBs), dichlorodiphenyltrichloroethane (DDT) and its metabolite, dichlorodiphenyldichloroethylene (DDE) can disturb the secretory function of the ovary and both contractions and secretory function of the uterus during the estrus cycle and pregnancy. Additionally, PCBs can pass through the placental barrier into allantoic and amniotic fluid. The presence of PCBs in these fluids is associated with higher frequency of spontaneous abortions and premature births in humans and animals. Therefore, the effect of PCBs, DDT and DDE on the connexins (Cx26, Cx32 and Cx43) and keratin 8 (KRT8) expression in bovine placentomes was investigated. The placentome slices from the second trimester of pregnancy were incubated with PCB153, 126, 77, DDT and DDE (each at doses of 1, 10 or 100 ng/ml) for 48 h. Then, the slices were stained using immunohistochemistry. The density of Cxs staining was measured with Axio- Vision Rel. 4.8 software in fetal-maternal connections and binuclear cells (BNC). None of the tested xenobiotics (XBs) affected the localization of Cxs and KRT8 in the fetal-maternal connection area, but the XBs affected the density of Cxs in fetal-maternal connections and binuclear cells (BNCs). Depend on the doses, in fetal-maternal connections all used PCBs changed the protein expression of different Cxs, while in BNCs, all tested XBs except DDT increased the expression of Cxs. None of investigated XBs affected on KRT8 expression. In summary, used XBs affect the expression of Cxs and change the quantitative relationships between them. Therefore, XBs can unfavorably influence function of the utero-placental barrier in cows.
Protein digestion in insects relies on several groups of proteases, among which trypsin plays a prominent role. In the current study, larvae of Pieris brassicae L. were fed radish leaves treated with 1 mM concentrations of three specific inhibitors of trypsin: AEBSF.HCl [4-(2- -aminoethyl)-benzenesulfonyl fluoride, monohydrochloride], TLCK (N-a-tosyl-l-lysine chloromethyl ketone) and SBTI (Soybean Trypsin Inhibitor) to find their potential effects on gene expression of trypsin. Initially, RT-PCR analysis revealed a gene of 748 bp responsible for synthesizing the digestive trypsin in P. brassicae larvae. Also, qRT-PCR data indicated a statistically greater expression of trypsin gene in the larvae fed 1 mM concentrations of AEBSF.HCl, TLCK and SBTI than the control. Results of the current study indicated that synthetic inhibitors can not only negatively affect the gene expression of P. brassicae trypsin, but also the insect can activate a compensatory mechanism against interruption of protein digestion by inducing more expression of the gene and producing more trypsin into the midgut lumen.
In vitro embryogenic callus is a critical factor for genetic transformation of rice, especially for indica varieties. In this study, we investigated the relationship between polyamines, including putrescine (Put), spermidine (Spd) and spermine (Spm), and callus browning, and we studied the effect of exogenous Put on callus regeneration and on the content of endogenous polyamines. In addition, the expression levels of arginine decarboxylase gene (Adcl) and S-adenosylmethionine decarboxylase gene (Samdc) in embryogenic callus were studied by quantitative Real-time PCR analysis. The results showed that the contents of endogenous Put and Spd in the browning callus were significantly lower than those in normal callus. Exogenous Put could effectively improve the growing state of callus of indica rice and enhance the development of embryogenic callus. The content of endogenous polyamines in embryogenic callus, especially Spd and Spm, was increased after addition of exogenous Put. Additionally, exogenous Put also had an obvious impact on the expression levels of Adcl but partial effect on the expression levels of Samdc gene. This study could increase the knowledge of both embryogenic callus induction and polyamine catabolism in callus in indica rice.
Arbuscular mycorrhizal fungi are the most widespread root fungal symbionts, forming associations with the vast majority of plant species. Ectomycorrhizal development alters gene expression in plant symbionts. In this work we examined the effect of arbuscular mycorrhizal fungi spores on the growth and development of Brassica and on the expression of BnMT2 in winter rape. In a pot experiment, rape seedlings growing on different types of sterile and nonsterile soils were inoculated simultaneously with mycorrhizal fungi spores of Acaulospora longula,Glomus geosporum, Glomus mosseae and Scutellospora calospora. As compared with control plants growing in the absence of spores, ten-week-old seedlings of Brassica napus L. in sterile soil inoculated with arbuscular spores had longer shoots and higher fresh biomass of above-ground plant parts. In other types of substrates enriched with mycorrhizal fungi spores, the plants were smaller than non-inoculated plants. The presence of AMF spores stimulated the elongation growth of hypocotyls in both analyzed substrates. BnMT2 expression was highest in plants growing on the sterile substrate. Generally, the presence of mycorrhizal fungi spores appeared to have an adverse effect on the growth of rape plants.
The study was aimed to develop an indirect enzyme-linked immunosorbent assay (ELISA), which can detect specifically Feline herpesvirus type 1 (FHV-1). The primers were designed based on the conserved sequence of FHV-1 glycoprotein B gene. The recombinant protein with reactogenicity was purified as coating antigen of the assay. The indirect ELISA, characterized by high sensitivity showed no cross-reaction with two types of feline virus, had detection limit at 1:2000 dilution. The positive rate of the assay, according to the determined cutoff value (0.25), was basically consistent with Feline Herpes Virus Antibody ELISA kit. In conclusion, the indirect ELISA with high repeatability and reproducibility can be used for detecting FHV-1, and can provide necessary support to related research.
We are exploring the relationship between accents and expression in piano performance. Accents are local events that attract a listener's attention and are either evident from the score (immanent) or added by the performer (performed). Immanent accents are associated with grouping (phrasing), metre, melody and harmony. In piano music, performed accents involve changes in timing, dynamics, articulation, and pedalling; they vary in amplitude, form, and duration. We analyzed the first eight bars of Chopin Prelude op. 28 n. 6. In a separate study, music theorists had marked grouping, melodic and harmonic accents on the score and estimated the importance (salience) of each. Here, we mathematically modeled timing and dynamics in the prelude in two ways using Director Musices (DM) - a software package for automatic rendering of expressive performance. The first rendering focused on phrasing following existing and tested procedures in DM. The second focused on accents - timing and dynamics in the vicinity of the accents identified by the theorists. In an informal listening test, 10 out of 12 participants (5 of 6 musicians and 5 of 6 non-musicians) preferred the accent-based formulation, and several stated that it had more variation of timing and dynamics from one phrase to the next.
We explored the use of the medicinally important plant Centella asiatica for expression of hemagglutinin-neuraminidase (HN) protein of Newcastle disease virus (NDV) strain AF2240. HN protein is the principal target for subunit vaccine development against NDV. The full-length HN gene was cloned into a plant expression construct driven by the CaMV 35S promoter and C-terminal fusion of green fluorescence protein (GFP) as reporter system. The recombinant expression construct was transformed via particle bombardment into C. asiatica callus. Transformants were screened using GFP and selected on MS medium supplemented with 15 mg/l hygromycin. The ~1.8 kb HN mRNA transcript was detected on the putative transformants using RT-PCR. The presence of HN protein expression was further confirmed through dot blot analysis using anti-NDV chicken serum. Here we report, for the first time, the use of a novel medicinal plant as a new platform for HN protein expression.
This paper presents methods for optimal test frequencies search with the use of heuristic approaches. It includes a short summary of the analogue circuits fault diagnosis and brief introductions to the soft computing techniques like evolutionary computation and the fuzzy set theory. The reduction of both, test time and signal complexity are the main goals of developed methods. At the before test stage, a heuristic engine is applied for the principal frequency search. The methods produce a frequency set which can be used in the SBT diagnosis procedure. At the after test stage, only a few frequencies can be assembled instead of full amplitude response characteristic. There are ambiguity sets provided to avoid a fault tolerance masking effect.
The article discusses the circumstances surrounding the founding of the Association of Polish Writers Abroad (SPPzG) by Dr. Alina Siomkajło in London in 2010, oppositional to the Union of Polish Writers Abroad (ZPPnO), also in Great Britain, which had been active since 1945. The method of qualitative analysis was used to review the anti-communist content, the right-wing magazine.
This article analyses the amendments of January 2018 to the Act on the Institute of National Remembrance (INR) of 1998, which has raised doubts in light of in ternational law and provoked diplomatic tensions between Poland on one side and Germany, Ukraine, United States of America and Israel on the other. The INR is a national in stitution whose role is, among others, to prosecute perpetrators of in ternational crimes committed between 1917-1990. The article proves that the wording of the amendments is in consistent with in ternational law, as it ignores the principles of in ternational responsibility, definitions of in ternational crimes, and disproportionately limits freedom of expression. In consequence, it cannot be expected that third states will cooperate with Poland in the execution of responsibility for violation of the newly adopted norms.