Translationally controlled tumor protein (TCTP), also known as histamine releasing factor (HRF), is found abundantly in different eukaryotic cell types. The sequence homology of TCTP between different species is very high, belonging to the MSS4/DSS4 superfamily of proteins. TCTP is involved in both cell growth and human late allergy reaction, as well as having a calcium binding property; however, its primary biological functions remain to be clearly elucidated. In regard to many possible functions, the TCTP of Plasmodium falciparum (Pf) is known to bind with an antimalarial agent, artemisinin, which is activated by heme. It is assumed that the endoperoxide-bridge of artemisinin is opened up by heme to form a free radical, which then eventually alkylates, probably to the Cys14 of PfTCTP. Study of the docking of artemisinin with heme, and subsequently with PfTCTP, was carried out to verify the above hypothesis on the basis of structural interactions. The three dimensional (3D) structure of PfTCTP was built by homology modeling, using the NMR structure of the TCTP of Schizosaccharomyces pombe as a template. The quality of the model was examined based on its secondary structure and biological function, as well as with the use of structure evaluating programs. The interactions between artemisinin, heme and PfTCTP were then studied using the docking program, FlexiDock. The center of the peroxide bond of artemisinin and the Fe of heme were docked within a short distance of $2.6{\AA}$, implying the strong possibility of an interaction between the two molecules, as proposed. When the activated form of artemisinin was docked on the PfTCTP, the C4-radical of the drug faced towards the sulfur of Cys14 within a distance of $2.48{\AA}$, again suggesting the possibility of alkylation having occurred. These results confirm the proposed mechanism of the antimalarial effect of artemisinin, which will provide a reliable method for establishing the mechanism of its biological activity using a molecular modeling study.
Porous matrices of bioactive polymers such as polyglycolic acid (PGA) or polylactic acid (PLA) can be used as scaffolds in bone tissue growth during bone repair process. These polymers are highly porous and serve as a template for the growth and organization of new bone tissues. We evaluated the effect of PGA and PLA polymers on osteoblastic MC3T3-E1 cell extracellular mineralization. MC3T3-E1 cells were cultured in a time-dependent manner -1, 15, 25d as appropriate - for the period of bone formation stages in one of the five culture circumstances, such as normal osteogenic differentiation medium, PGA-plated, fetal bovine serum (FBS)-plated, PGA/FBS-coplated, and PLA-plated For the evaluation of bone formation, minerals (Ca, Mg, Mn) and alkaline phosphatase activity, a marker for osteoblast differentiation, were measured Alizarin Red staining was used for the measurement of extracellular matrix Ca deposit During the culture period, PGA-plated one was reabsorbed into the medium more easily and faster than the PLA-plated one. At day 15, at the middle stage of bone formation, cellular Ca and Mg levels showed higher tendency in PGA- or PLA-plated treatments compared to non-plated control and at day 25, at the early late stage of bone formation, all three cellular Ca, Mg or Mn levels showed higher tendency as in order of PGA-related treatments and PLA-plated treatments, compared to control even without significance. Medium Ca, Mg or Mn levels didn't show any consistent tendency. Cellular ALP activity was higher in the PGA- or PLA-plated treatments compare to normal osteogenic medium treatment PGA-plated and PGA/FBS-plated treatments showed better Ca deposits than other treatments by measurement of Alizarin Red staining, although PLA-plated treatment also showed reasonable Ca deposit. The results of the present study suggest that biodegradable material, PGA and also with less extent for PLA, can be used as a biomaterial for better extracellular matrix mineralization in osteoblastic MC3T3-E1 cells.
Purpose Principal component analysis (PCA) is a method often used in the neuroimagre analysis as a multivariate analysis technique for describing the structure of high dimensional correlation as the structure of lower dimensional space. PCA is a statistical procedure that uses an orthogonal transformation to convert a set of observations of correlated variables into a set of values of linearly independent variables called principal components. In this study, in order to investigate the usefulness of PCA in the brain PET image analysis, we tried to analyze C[11]-PIB PET image as a representative case. Materials and Methods Nineteen subjects were included in this study (normal = 9, AD/MCI = 10). For C[11]-PIB, PET scan were acquired for 20 min starting 40 min after intravenous injection of 9.6 MBq/kg C[11]-PIB. All emission recordings were acquired with the Biograph 6 Hi-Rez (Siemens-CTI, Knoxville, TN) in three-dimensional acquisition mode. Transmission map for attenuation-correction was acquired using the CT emission scans (130 kVp, 240 mA). Standardized uptake values (SUVs) of C[11]-PIB calculated from PET/CT. In normal subjects, 3T MRI T1-weighted images were obtained to create a C[11]-PIB template. Spatial normalization and smoothing were conducted as a pre-processing for PCA using SPM8 and PCA was conducted using Matlab2012b. Results Through the PCA, we obtained linearly uncorrelated independent principal component images. Principal component images obtained through the PCA can simplify the variation of whole C[11]-PIB images into several principal components including the variation of neocortex and white matter and the variation of deep brain structure such as pons. Conclusion PCA is useful to analyze and extract the main pattern of C[11]-PIB image. PCA, as a method of multivariate analysis, might be useful for pattern recognition of neuroimages such as FDG-PET or fMRI as well as C[11]-PIB image.
Yun, W.I.;Jo, D.W.;Ok, J.E.;Jeon, H.S.;Lee, G.S.;Jung, S.K.;Bae, S.M.;Ahn, H.S.;Yang, M.
Journal of the Korean Crystal Growth and Crystal Technology
/
v.21
no.3
/
pp.110-113
/
2011
In this paper, we propose a new method for the fabrication of GaN microstructures formed only on the vertex of GaN pyramid by using of metal catalysts. GaN pyramidal structures were selectively grown on 3 ${\mu}m$$SiO_2$ dot patterns followed by thin film deposition of Au and Cr only on the vertex area of the GaN pyramids with precisely controlled photolithography. After the metal deposition, the samples were loaded in the MOVPE reactor for the growth of GaN microstructures for 10 minutes. Temperature for the growth of the GaN microstructures was changed from $650^{\circ}C$ to $750^{\circ}C$. Rod type GaN microstructures were grown in the direction of vertical to the six {1-101} facets and the shape of the GaN microstructures was changed depend on the type of metal.
Kang, Hyo Rin;Seong, Mi So;Ku, Bok Kyung;Cheong, JaeHun
Journal of Life Science
/
v.30
no.11
/
pp.947-955
/
2020
The foot-and-mouth disease virus (FMDV), a member of the Aphthovirus genus in the Picornaviridae family, affects wild and domesticated ruminants and pigs. During replication of the FMDV RNA (ribonucleic acid) genome, FMDV-encoding RNA polymerase 3D acts in a highly location-specific manner. This suggests that specific RNA structures recognized by 3D polymerase within non-coding regions of the FMDV genome assist with binding during replication. One such region is the cis-acting replication element (CRE), which functions as a template for RNA replication. The FMDV CRE adopts a stem-loop conformation with an extended duplex stem, supporting a novel 15-17 nucleotide loop that derives stability from base-stacking interactions, with the exact RNA nucleotide sequence of the CRE producing different RNA secondary structures. Here, we show that CRE sequences of FMDVs isolated in Korea from 2010 to 2017 exhibit A and O genotypes. Interestingly, variations in the RNA secondary structure of the Korean FMDVs are consistent with the phylogenetic relationships between these viruses and reveal the specificity of FMDV infections for particular host species. Therefore, we conclude that each genetic clade of Korean FMDV is characterized by a unique functional CRE and that the evolutionary success of new genetic lineages may be associated with the invention of a novel CRE motif. Therefore, we propose that the specific RNA structure of a CRE is an additional criterion for FMDV classification dependent on the host species. These findings will help correctly analyze CRE sequences and indicate the specificity of host species for future FMDV epidemics.
Alkaline protease-overproducing strains of Vibrio metschnikovii were developed by using the molecular evolution from the classical mutants V. metschnikovii L12-23, N4-8, and KS1. Each vapK (Vibrio alkaline protease K) was obtained from the genomic DNAs of mutants by PCR to carry out the DNA shuffling. The modified vapK-1 obtained by DNA shuffling was used again as a template for the error-prone PCR to make the vapK-2. Both genes were cloned in the plasmid pKF3 to construct the recombinant plasmids which have one or two copies of the modified genes. The recombinant plasmids were back-transformed to V. metschnikovii KS1 to construct recombinant V. metschnikovii that expresses the alkaline protease. About 3.9-fold more protease activity was measured in the strain which has the plasmid containing two copies of vapK-2 when compared to strain KS1. When compared to wild type V. metschnikovii RH530, 43-fold more activity was achieved. Comparison of amino acids among vapK, vapK-1, and vapK-2 revealed that the active sites was highly conserved and not changed. However, many amino acids except the active sites were changed. These results suggested that the changes in amino acids might play an important role in the increase of protease activity by allowing the easy access of substrate to active sites of the protease. The fermentation of alkaline protease from the V. metschnikovii KS1 harboring the plasmid that contains two copies of vapK-1 showed the possibility of this strain to be used as industrial producer.
Park, Sang-Hyun;Lee, Soo-Chan;Yun, Il-Dong;Lee, Sang-Uk
Journal of Broadcast Engineering
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v.17
no.1
/
pp.73-80
/
2012
Recently, medical equipments are developed and used for diagnosis or studies. In addition, demand of techniques which automatically deal with three dimensional medical images obtained from the medical equipments is growing. One of the techniques is automatic bone segmentation which is expected to enhance the diagnosis efficiency of osteoporosis, fracture, and other bone diseases. Although various researches have been proposed to solve it, they are unable to be used in practice since a size of the medical data is large and there are many low contrast boundaries with other tissues. In this paper, we present a fast and accurate automatic framework for bone segmentation based on multi-resolutions. On a low resolution step, a position of the bone is roughly detected using constrained branch and mincut which find the optimal template from the training set. Then, the segmentation and the registration are iteratively conducted on the multiple resolutions. To evaluate the performance of the proposed method, we make an experiment with femur and tibia from 50 test knee magnetic resonance images using 100 training set. The proposed method outperformed the constrained branch and mincut in aspect of segmentation accuracy and implementation time.
Antimalarial drugs are an urgently need and crucial tool in the campaign against malaria, which can threaten public health. In this study, we examined the cytotoxicity of the 9 antimalarial compounds chemically synthesized using SKM13-2HCl. Except for SKM13-2HCl, the 5 newly synthesized compounds had a 50% cytotoxic concentration (CC50) >100 μM, indicating that they would be less cytotoxic than SKM13-2HCl. Among the 5 compounds, only SAM13-2HCl outperformed SKM13-2HCl for antimalarial activity, showing a 3- and 1.3-fold greater selective index (SI) (CC50/IC50) than SKM13-2HCl in vitro against both chloroquine-sensitive (3D7) and chloroquine -resistant (K1) Plasmodium falciparum strains, respectively. Thus, the presence of morpholine amide may help to effectively suppress human-infectious P. falciparum parasites. However, the antimalarial activity of SAM13-2HCl was inferior to that of the SKM13-2HCl template compound in the P. berghei NK65-infected mouse model, possibly because SAM13-2HCl had a lower polarity and less efficient pharmacokinetics than SKM13-2HCl. SAM13-2HCl was more toxic in the rodent model. Consequently, SAM13-2HCl containing morpholine was selected from screening a combination of pharmacologically significant structures as being the most effective in vitro against human-infectious P. falciparum but was less efficient in vivo in a P. berghei-infected animal model when compared with SKM13-2HCl. Therefore, SAM13-2HCl containing morpholine could be considered a promising compound to treat chloroquine-resistant P. falciparum infections, although further optimization is crucial to maintain antimalarial activity while reducing toxicity in animals.
The poliovirus is a small, and non-enveloped virus. The RNA genome of poliovirus is continuous, linear, and has a single open reading frame. This polyprotein precursor is cleaved proteolytically to yield mature products. Most of the cleavages occur by viral protease. The mature proteins derived from the P1 polyprotein precursor are the structural components of the viral capsid. The initial cleavage by 2A protease is indirectly involved in the cleavage of a cellular protein p220, a subunit of the eukaryotic translation initiation factor 4F. This cleavage leads to the shut-off of cap-dependent host cell translation, and allows poliovirus to utilize the host cell machinery exclusively for translation its own RNA, which is initiated by internal ribosome entry via a cap-independent mechanism. The functional role of the 2B, 2C and 2BC proteins are not much known. 2B, 2C, 2BC and 3CD proteins are involved in the replication complex of virus induced vesicles. All newly synthesized viral RNAs are linked with VPg. VPg is a 22 amino acid polypeptide which is derived from 3AB. The 3C and 3CD are protease and process most of the cleavage sites of the polyprotein precursor. The 3C protein is also involved in inhibition of RNA polymerase II and III mediated transcription by converting host transcription factor to an inactive form. The 3D is the RNA dependent RNA polymerase. It is known that poliovirus replication follows the general pattern of positive strand RNA virus. Plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA is transcribed into complementary minus strand RNA that, in turn, is transcribed for the synthesis of plus strand RNA strands. Poliovirus RNA synthesis occurs in a membranous environment but how the template RNA and proteins required for RNA replication assemble in the membrane is not much known. The RNA requirements for the encapsidation of the poliovirus genome (packaging signal) are totally unknown. The poliovirus infection cycle lasts approximately 6 hours.
The probabilistic anatomical maps are used to localize the functional neuro-images and morphological variability. The quantitative indicator is very important to inquire the anatomical position of an activated legion because functional image data has the low-resolution nature and no inherent anatomical information. Although previously developed MNI probabilistic anatomical map was enough to localize the data, it was not suitable for the Korean brains because of the morphological difference between Occidental and Oriental. In this study, we develop a probabilistic anatomical map for Korean normal brain. Normal 75 blains of T1-weighted spoiled gradient echo magnetic resonance images were acquired on a 1.5-T GESIGNA scanner. Then, a standard brain is selected in the group through a clinician searches a brain of the average property in the Talairach coordinate system. With the standard brain, an anatomist delineates 89 regions of interest (ROI) parcellating cortical and subcortical areas. The parcellated ROIs of the standard are warped and overlapped into each brain by maximizing intensity similarity. And every brain is automatically labeledwith the registered ROIs. Each of the same-labeled region is linearly normalize to the standard brain, and the occurrence of each legion is counted. Finally, 89 probabilistic ROI volumes are generated. This paper presents a probabilistic anatomical map for localizing the functional and structural analysis of Korean normal brain. In the future, we'll develop the group specific probabilistic anatomical maps of OCD and schizophrenia disease.
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