Insect cell culture has greatly increased in part due to the widespread use of insect virus-based vectors for efficient expression of foreign proteins. Insect cells such as Sf9 cells are susceptible ...to arboviruses which may pose a safety concern by adventitious introduction during the production process. The objective of this study was to establish techniques for viral clearance validation of insect cell-derived biotechnological products using Japanese encephalitis virus (JEV) as a model, since JEV is a member of arthropod-borne flaviviruses that are known to be infectious in insect cells. Here we report the development of a quantitative assay for JEV RNA using real-time reverse transcription-polymerase chain reaction (RT–PCR). The assay was performed using LightCycler and RNA amplification kit SYBR Green I. The JEV specific primer was selected from the 3′ untranslated region, and the expected band size was 323 base pairs (bp). The sensitivity of the assay was calculated to be approximately 15 TCID
50per reaction. Highly reproducible standard curves were obtained from experiments performed on three different days. JEV clearance was determined during the purification process of rHPV-16 L1 VLPs by CsCl equilibrium density centrifugation. The comparative results obtained by real-time RT–PCR assay for JEV and infectivity titrations suggested that the real-time RT–PCR assay could have an additive effect on the interpretation and evaluation of virus clearance, especially during the virus removal process.
With two ultrasonographic and two CT films of three cases of thyroid tuberculosis, we evaluate the ultrasonographic and CT findings and correlate them with the pathologic findings. They are ...demonstrated as heterogeneous hypoechoic mass on ultrasonogram and peripheral-enhancing low-density abscess on CT scan with regional lymphadenopathy. Ultrasonography (US) and CT can help the diagnosis of thyroid tuberculosis.
Behavioral sequences of the medaka (Oryzias latipes) were continuously investigated through an automatic image recognition system in response to medaka treated with the insecticide and medaka not ...treated with the insecticide, diazinon (0.1 mg/l) during a 1 hour period. The observation of behavior through the movement tracking program showed many patterns of the medaka. After much observation, behavioral patterns were divided into four basic patterns: active-smooth, active-shaking, inactive-smooth, and inactive-shaking. The “smooth” and “shaking” patterns were shown as normal movement behavior. However, the “shaking” pattern was more frequently observed than the “smooth” pattern in medaka specimens that were treated with insecticide. Each pattern was classified using a devised decision tree after the feature choice. It provides a natural way to incorporate prior knowledge from human experts in fish behavior and contains the information in a logical expression tree. The main focus of this study was to determine whether the decision tree could be useful for interpreting and classifying behavior patterns of the medaka.
Application of Next Generation Sequencing to Investigate Microbiome in the Livestock Sector Kim, M., Animal Nutrition and Physiology Team, National Institute of Animal Science, Wanju, Republic of Korea; Baek, Y., Animal Nutrition and Physiology Team, National Institute of Animal Science, Wanju, Republic of Korea; Oh, Y.K., Animal Nutrition and Physiology Team, National Institute of Animal Science, Wanju, Republic of Korea
Journal of Livestock Housing and Environment,
20/Sep , Letnik:
21, Številka:
3
Journal Article
The objective of this study was to review application of next-generation sequencing (NGS) to investigate microbiome in the livestock sector. Since the 16S rRNA gene is used as a phylogenetic marker, ...unculturable members of microbiome in nature or managed environments have been investigated using the NGS technique based on 16S rRNA genes. However, few NGS studies have been conducted to investigate microbiome in the livestock sector. The 16S rRNA gene sequences obtained from NGS are classified to microbial taxa against the 16S rRNA gene reference database such as RDP, Greengenes and Silva databases. The sequences also are clustered into species-level OTUs at 97% sequence similarity. Microbiome similarity among treatment groups is visualized using principal coordinates analysis, while microbiome shared among treatment groups is visualized using a venn diagram. The use of the NGS technique will contribute to elucidating roles of microbiome in the livestock sector.
In 58 patients with mast cell disease (MCD) and three with basophilic leukemia, bone marrow (54 cases) or skin tissue (four cases) was studied immunohistochemically for expression of Kit (c-
kit ...protein), the different isomers of transforming growth factor-β (TGF-β), basic fibroblast growth factor (bFGF), and their respective receptors. Kit was expressed in all cases of MCD but in none of basophilic leukemia. Expression pattern of cytokines and their receptors was variable in systemic MCD with (SMCD-HD) or without (SMCD) associated hematologic disorder. However, type I TGF-β receptor (TGFβ1R) was not expressed in 30% of SMCD-HD patients or in patients with mast cell leukemia, but the remaining cases of MCD showed near uniform expression. The associated hematologic disorders in TGFβ1R-negative cases of SMCD-HD were prognostically less favorable than those associated with TGFβ1R-positive cases of SMCD-HD. The results confirm the diagnostic value of Kit immunohistochemistry in MCD and suggest a biologically relevant heterogeneity in TGFβ1R expression among patients with SMCD-HD.
This paper presents a novel Maglev moving-magnet-type synchronous PM planar motor (SPMPM) that has a simple and thin stator composed of several PCBs. In order to study the feasibility of the proposed ...SPMPM, the magnetic flux density and thrust force equations are estimated analytically and verified using the finite-element method (FEM) and experimental works.
