Detection of porcine parvovirus using a taqman-based real-time pcr with primers and probe designed for the NS1 gene

A TaqMan-based real-time polymerase chain reaction (PCR) assay was devised for the detection of porcine parvovirus (PPV). Two primers and a TaqMan probe for the non-structural protein NS1 gene were designed. The detection limit was 1 × 102 DNA copies/μL, and the assay was linear in the range of 1 × 102 to 1 × 109 copies/μL. There was no cross-reaction with porcine circovirus 2 (PCV2), porcine reproductive and respiratory syndrome virus (PRRSV), pseudorabies virus (PRV), classical swine fever virus (CSFV), or Japanese encephalitis virus (JEV). The assay was specific and reproducible. In 41 clinical samples, PPV was detected in 32 samples with the real-time PCR assay and in only 11 samples with a conventional PCR assay. The real-time assay using the TaqMan-system can therefore be practically used for studying the epidemiology and management of PPV

Antroquinonol Exerts Immunosuppressive Effect on CD8 +

Antroquinonol was investigated as antioxidant and inhibition of inflammatory responses. Our study was to evaluate its immunosuppressive effect on CD8+ T cells and protective effect on depigmentation. CD8+ T cells were treated with antroquinonol in vitro, and C57BL/6 mice were treated with antroquinonol with or without H2O2 in vivo for 50 consecutive days. We found antroquinonol could inhibit proliferation of CD8+ T cells and suppress the production of cytokines IL-2 and IFN-γ and T cell activation markers CD69 and CD137 in vitro. H2O2 treatment induced depigmentation and reduced hair follicle length, skin thickness, and tyrosinase expression in vivo. Whereas, antroquinonol obviously ameliorated depigmentation of mice skin and resisted the reduction of hair follicle length, skin thickness, and tyrosinase expression induced by H2O2. Antroquinonol decreased CD8+ T cell infiltration in mice skin, inhibited the production of IL-2 and IFN-γ, and decreased the expression of CXCL10 and CXCR3. Summarily, our data shows antroquinonol inhibits CD8+ T cell proliferation in vitro. It also reduces CD8+ T cell infiltration and proinflammatory cytokine secretion and suppresses the thinning of epidermal layer in vivo. Our findings suggest that antroquinonol exerts immunosuppressive effects on CD8+ T cell proliferation and activation to resist depigmentation induced by H2O2

Imobilizacija tirozinaze na pustu od ugljičnih vlakana s (3-aminopropil)trietoksisilanom za protočnu elektrokemijsku detekciju fenolnih spojeva

Tyrosinase (TYR) was covalently immobilized onto amino-functionalized carbon felt (CF) surface via glutaraldehyde (GA). Prior to the TYR-immobilization, primary amino group was introduced to the CF surface by treatment with 3-aminopropyltriethoxysilane (APTES). The resulting TYR-immobilized CF was used as a working electrode unit of an electrochemical flow-through detector for mono- and di-phenolic compounds (i.e., catechol, p-cresol, phenol and p-chlorophenol). Additionally, flow injection peaks based on electroreduction of the enzymatically produced o-quinone species were detected at −0.05 V vs. Ag/AgCl. The resulting TYR/GA/APTES/CF biosensor responded well to all compounds tested with limits of detection range from 7.5 to 35 nmol l–1 (based on three times S/N ratio). Moreover, such modified electrode exhibits good stability and reproducibility for catechol. No serious degradation of the peak current was found over 30 consecutive injections. This work is licensed under a Creative Commons Attribution 4.0 International License.Tirozinaza (TYR) je kovalentno vezana na aminiranu površinu pusta izrađenog od ugljičnih vlakana (CF) s pomoću glutaraldehida (GA). Prije imobilizacije tirozinaze primarna amino-skupina uvedena je na ugljična vlakna (3-aminopropil)trietoksisilanom (APTES). CF s imobiliziranom tirozinazom upotrijebljen je kao elektroda u protočnom elektrokemijskom detektoru jednostruko i dvostruko hidroksiliranih fenola (katehol, p-krezol, fenol, p-klorfenol). Pri − 0,05 V (u odnosu na Ag/AgCl) uočeni su protočni injekcijski signali elektroredukcije o-kinona nastalog enzimskom reakcijom. Biosenzor TYR/GA/APTES/CF dobro se odaziva za sve ispitane spojeve uz detekcijski limit od 7,5 do 35 nmol l–1 (tri puta veći signal od šuma). Modificirana elektroda stabilna je i pokazuje dobru reproducibilnost za katehol. Jakost struje signala nije se značajno smanjila ni nakon 30 uzastopnih injektiranja. Ovo djelo je dano na korištenje pod licencom Creative Commons Imenovanje 4.0 međunarodna

Tracing children's vocabulary development from preschool through the school‐age years: an 8‐year longitudinal study

In this 8‐year longitudinal study, we traced the vocabulary growth of Chinese children, explored potential precursors of vocabulary knowledge, and investigated how vocabulary growth predicted future reading skills. Two hundred and sixty‐four (264) native Chinese children from Beijing were measured on a variety of reading and language tasks over 8 years. Between the ages of 4 to 10 years, they were administered tasks of vocabulary and related cognitive skills. At age 11, comprehensive reading skills, including character recognition, reading fluency, and reading comprehension were examined. Individual differences in vocabulary developmental profiles were estimated using the intercept‐slope cluster method. Vocabulary development was then examined in relation to later reading outcomes. Three subgroups of lexical growth were classified, namely high‐high (with a large initial vocabulary size and a fast growth rate), low‐high (with a small initial vocabulary size and a fast growth rate) and low‐low (with a small initial vocabulary size and a slow growth rate) groups. Low‐high and low‐low groups were distinguishable mostly through phonological skills, morphological skills and other reading‐related cognitive skills. Childhood vocabulary development (using intercept and slope) explained subsequent reading skills. Findings suggest that language‐related and reading‐related cognitive skills differ among groups with different developmental trajectories of vocabulary, and the initial size and growth rate of vocabulary may be two predictors for later reading development. “In this 8‐year longitudinal study, we traced the vocabulary growth of Chinese children, explored potential precursors of vocabulary knowledge, and investigated how vocabulary growth predicted future reading skills. Three subgroups of lexical growth were classified, namely high‐high (with a large initial vocabulary size and a fast growth rate), low‐high (with a small initial vocabulary size and a fast growth rate) and low‐low (with a small initial vocabulary size and a slow growth rate) groups. Low‐high and low‐low groups were distinguishable mostly through phonological skills, morphological skills and other reading‐related cognitive skills. Childhood vocabulary development (using intercept and slope) explained subsequent reading skills. Findings suggest that language‐related and reading‐related cognitive skills differ among groups with different developmental trajectories of vocabulary, and the initial size and growth rate of vocabulary may be two predictors for later reading development.”Peer Reviewedhttp://deepblue.lib.umich.edu/bitstream/2027.42/109871/1/desc12190.pd

Classification, replication, and transcription of Nidovirales

Nidovirales is one order of RNA virus, with the largest single-stranded positive sense RNA genome enwrapped with membrane envelope. It comprises four families (Arterividae, Mesoniviridae, Roniviridae, and Coronaviridae) and has been circulating in humans and animals for almost one century, posing great threat to livestock and poultry，as well as to public health. Nidovirales shares similar life cycle: attachment to cell surface, entry, primary translation of replicases, viral RNA replication in cytoplasm, translation of viral proteins, virion assembly, budding, and release. The viral RNA synthesis is the critical step during infection, including genomic RNA (gRNA) replication and subgenomic mRNAs (sg mRNAs) transcription. gRNA replication requires the synthesis of a negative sense full-length RNA intermediate, while the sg mRNAs transcription involves the synthesis of a nested set of negative sense subgenomic intermediates by a discontinuous strategy. This RNA synthesis process is mediated by the viral replication/transcription complex (RTC), which consists of several enzymatic replicases derived from the polyprotein 1a and polyprotein 1ab and several cellular proteins. These replicases and host factors represent the optimal potential therapeutic targets. Hereby, we summarize the Nidovirales classification, associated diseases, “replication organelle,” replication and transcription mechanisms, as well as related regulatory factors

Potential of genotype VII Newcastle disease viruses to cause differential infections in chickens and ducks

Newcastle disease (ND), caused by ND virus (NDV), is one of the most infectious and economically important diseases of the poultry industry worldwide. While infections are reported in a wide range of avian species, the pathogenicity of chicken-origin virulent NDV isolates in ducks remains elusive. In this study, two NDV strains were isolated and biologically and genetically characterized from an outbreak in chickens and apparently healthy ducks. Pathogenicity assessment indices, including the mean death time (MDT), intracerebral pathogenicity index (ICPI) and cleavage motifs in the fusion (F) protein, indicated that both isolates were velogenic in nature. While these isolates carried pathogenic characteristics, interestingly they showed differential pathogenicity in ducks. The chicken-origin isolate caused high (70%) mortality, whereas the duck-origin virus resulted in low (20%) mortality in 4-week-old ducks. Intriguingly, both isolates showed comparable disease pathologies in chickens. Full-genome sequence analysis showed that the virus genome contains 15 192 nucleotides and carried features that are characteristic of velogenic strains of NDV. A phylogenetic analysis revealed that both isolates clustered in class II and genotype VII. However, there were several mutations in the functionally important regions of the fusion (F) and haemagglutinin-neuraminidase (HN) proteins, which may be responsible for the differential pathogenicity of these viruses in ducks. In summary, these results suggest that NDV strains with the same genotype show differential pathogenicity in chickens and ducks. Furthermore, chicken-origin virulent NDVs are more pathogenic for ducks than duck-origin viruses. These findings propose a role for chickens in the evolution of viral pathogenicity and the potential genetic resistance of ducks to poultry viruses

Supplementation of Vitamin E Protects Chickens from Newcastle Disease Virus-Mediated Exacerbation of Intestinal Oxidative Stress and Tissue Damage

BACKGROUND/AIMS: Newcastle disease virus (NDV) causes a highly devastating and contagious disease in poultry, which is mainly attributed to extensive tissue damages in the digestive, respiratory and nervous systems. However, nature and dynamics of NDV-induced oxidative stresses in the intestine of chickens remain elusive. METHODS: In this study, we examined the magnitude of intestinal oxidative stress and histopathological changes caused by the virulent NDV infection, and explored the protective roles of vitamin E (vit. E) in ameliorating these pathological changes. For these purposes, chickens were divided into four groups namely i) non supplemented and non-challenged (negative control, CON); ii) no supplementation of vit. E but challenged with ZJ1 (positive control, NS+CHA); iii) vit. E supplementation at the dose of 50 IU/day/Kg body weight and ZJ1 challenge (VE50+CHA); and 4) vit. E supplementation at the dose of 100 IU/day/Kg body weight and ZJ1 challenge (VE100+CHA). In all groups, we analyzed concentrations of glutathione (GSH), malondialdehyde (MDA), nitric oxide (NO), total antioxidant capacity (T-AOC), and activity of glutathione S-transferase (GST), superoxide dismutase (SOD), catalase (CAT) using biochemical methods. The virus loads were determined by quantitative RT-PCR and antibody titers by hemagglutination inhibition assays. We also examined the histopathological changes in the duodenal and jejunal mucosa at 3 and 5-day post infection (dpi) with NDV. RESULTS: A significant elevation in the NO level was observed in NDV challenged chickens compared to the CON chickens at 2 dpi. The MDA contents were significantly increased whereas GSH was significantly decreased in NDV-challenged chickens compared to control. Furthermore, activities of GST, CAT, SOD, as well as the TOAC were markedly decreased in challenged chickens in comparison with control. Virus copy numbers were higher in NDV infected NS+CHA group compared to other groups. Severe histopathological changes including inflammation, degeneration and broken villi were observed in the intestine of NDV challenged chickens. However, all these malfunctions of antioxidant system and pathological changes in the intestine were partially or completely reversed by the vit. E supplementation. CONCLUSIONS: Our results suggest that NDV infection causes oxidative stress and histopathological changes in the duodenum and jejunum of chickens, which can be partially or fully ameliorated by supplementation of vit. E. Additionally, these findings suggest that oxidative stress contributes to the intestinal damages in NDV infected chickens. These findings will help to understand the pathogenesis of NDV and further investigation of therapeutic agents for control of Newcastle disease

Production, characterization, and epitope mapping of a monoclonal antibody against genotype VII Newcastle disease virus V protein

Newcastle disease virus (NDV) V protein is crucial for viral interferon (IFN) antagonism and virulence, determining its host range restriction. However, little information is available on the B cell epitopes of V protein and the subcellular movement of V protein in the process of NDV infection. In this study, the monoclonal antibody (mAb) clone 3D7 against genotype VII NDV V protein was generated by immunizing mice with a purified recombinant His-tagged carboxyl-terminal domain (CTD) region of V protein. Fine epitope mapping analysis and B-cell epitope prediction indicated that mAb 3D7 recognized a linear epitope 152RGPAELWK159, which is located in the V protein CTD region. Sequence alignment showed that the mAb clone 3D7-recognized epitope is highly conserved among Class II genotype VII NDV strains, but not among other genotypes, suggesting it could serve as a genetic marker to differentiate NDV genotypes. Furthermore, the movement of V protein during NDV replication in infected cells were determined by using this mAb. It was found that V protein localized around the nucleus during virus replication. The establishment of V protein-specific mAb and identification of its epitope extend our understanding of the antigenic characteristics of V protein and provide a basis for the development of epitope-based diagnostic assays

Prosedur penyelesaian pembiayaan bermasalah pada akad mudharabah dalam rangka meminimalisir resiko di BMT Amanah Usaha Mulia Magelang

Permasalah kehidupan perekonomian yang sulit, membuat masyarakat berinisiatif untuk membuka usaha sendiri. Mereka membutuhkan suatu bantuan berupa dana untuk memperlancar usahanya, maka BMT Amanah Usaha Mulia Magelang ikut untuk mengembangkan produknya yaitu pembiayaan mudharabah sesuai perkembangan dunia perbankan dalam target peningkatan keuntungan dan menyejahterakan masyarakat. Dengan diberikanya pembiayaan tersebut, terkadang muncul adanya pembiayaan bermasalah dikarenakan ada beberapa faktor diantaranya ketidakmampuan anggota untuk membayar tepat waktu atau jatuh tempo pembayaran diakibatkan karena usaha anggota yang kurang lancar dan lain sebagaianya. Tugas Akhir ini berjudul “ Prosedur Penyelesaian Pembiayaan Bermasalah pada Akad Mudharabah Dalam Rangka Meminimalisir Risiko” Berdasarkan judul tersebut dapat diambil rumusan masalah yaitu apa penyebab terjadinya pembiayaan bermasalah pada BMT Amanah Usaha Mulia Magelang dan bagaimana prosedur penyelesaian pembiayaaan bermasalah pada akad mudharabah di BMT Amanah Usaha Mulia Magelang. Penelitian ini merupakan penelitian lapangan dimana sumber data yang digunakan berasal dari data primer dan sekunder yang diperoleh melalui metode wawancara dengan manajer, bagian pembiayaan dan dokumentasi. Metode yang digunakan dalam penelitian ini adalah deskriptif kualitatif yang bertujuan untuk menggambarkan secara sistematis dan akurat mengenai objek penelitian. Berdasarkan hasil penelitian dapat disimpulkan bahwa penyebab terjadinya pembiayaan bermasalah yaitu faktor internal meliputi kurang telitinya petugas BMT dalam menganalisi data calon anggota, kurang disiplinya dalam penagihan dan eksternal meliputi karakter anggota yang kurang baik, usahanya bangkrut dan terjadinya bencana alam yang tidak terduga. Adapun prosesdur yang digunakan BMT Amanah Usaha Mulia dalam menyelesaian pembiayaan bermasalah pada akad mudharabah dengan cara kekeluargaan atau musyawarah dengan anggota, penjadwalan kembali (rescheduling), persyaratan kembali (reconditioning), pengambilan jaminan (eksekusi), dan write off final. Di BMT Amanah Usaha Mulia dalam penyelesaian pembiayaan bermasalah jarang menngunakan jalur hukum, tetapi sering menggunakan cara kekeluargaan yang dianggap lebih efektif dan eksekusi jaminan apabila anggota tersebut sudah mengalami macet atau bermasalah