The Principal component analysis (PCA) is a well-known data analysis method that is useful in linear feature extraction and data compression. The PCA is a linear transformation that applies an orthogonal rotation to the original data, so as to maximize the retained variance. PCA is a classical technique for obtaining an optimal overall mapping of linearly dependent patterns of correlation between variables (e.g. neurons). PCA provides, in the mean-squared error sense, an optimal linear mapping of the signals which are spread across a group of variables. These signals are concentrated into the first few components, while the noise, i.e. variance which is uncorrelated across variables, is sequestered in the remaining components. PCA has been used extensively to resolve temporal patterns in neurophysiological recordings. Because the retinal signal is stochastic process, PCA can be used to identify the retinal spikes. With excised rabbit eye, retina was isolated. A piece of retina was attached with the ganglion cell side to the surface of the microelectrode array (MEA). The MEA consisted of glass plate with 60 substrate integrated and insulated golden connection lanes terminating in an 8${\times}$8 array (spacing 200 $\mu$m, electrode diameter 30 $\mu$m) in the center of the plate. The MEA 60 system was used for the recording of retinal ganglion cell activity. The action potentials of each channel were sorted by offline analysis tool. Spikes were detected with a threshold criterion and sorted according to their principal component composition. The first (PC1) and second principal component values (PC2) were calculated using all the waveforms of the each channel and all n time points in the waveform, where several clusters could be separated clearly in two dimension. We verified that PCA-based waveform detection was effective as an initial approach for spike sorting method.
An isocratic high performance liquid chromatography (HPLC) method for routine analysis of deoxynivalenol in noodles was validated and estimated the measurement uncertainty. Noodles (dried noodle and ramyeon) were analyzed by HPLC-ultraviolet detection using immunoaffinity column for clean-up. The limits of detection (LOD) and quantification (LOQ) were 7.5 ${\mu}g$/kg and 18.8 ${\mu}g$/kg, respectively. The calibration curve showed a good linearity, with correlation coefficients $r^2$ of 0.9999 in the concentration range from 20 to 500 ${\mu}g$/kg. Recoveries and Repeatabilities expressed as coefficients of variation (CV) spiked with 200 and 500 ${\mu}g$/kg were $82{\pm}2.7%$ and $87{\pm}1.3%$% in dried noodle, and $97{\pm}1.6%$ and $91{\pm}12.0%$ in ramyeon, respectively. The uncertainty sources in measurement process were identified as sample weight, final volume, and sample concentration in extraction volume as well as components such as standard stock solution, working standard solution, 5 standard solutions, calibration curve, matrix, and instrument. Deoxynivalenol concentration and expanded uncertainty in two matrixes spiked with 200 ${\mu}g$/kg and 500 ${\mu}g$/kg were estimated to be $163.8{\pm}52.1$ and $435.2{\pm}91.6\;{\mu}g$/kg for dried noodle, and $194.3{\pm}33.0$ and $453.2{\pm}91.1\;{\mu}g$/kg for ramyeon using a coverage factor of two which gives a level of statistical confidence with approximately 95%. The most influential component among uncertainty sources was the recovery of matrix, followed by calibration curve.
Park, Bong-Wook;Byun, June-Ho;Ryu, Young-Mo;Hah, Young-Sool;Kim, Deok-Ryong;Cho, Yeong-Cheol;Sung, Iel-Yong;Kim, Jong-Ryoul
Maxillofacial Plastic and Reconstructive Surgery
/
v.29
no.3
/
pp.197-205
/
2007
Angiogenesis is a essential part for bone formation and bone fracture healing. Vascular endothelial growth factor (VEGF), one of the most important molecules among many angiogenic factors, is a specific mitogen for vascular endothelial cells. VEGF-mediated angiogenesis is required for bone formation and repair. However, the effect of VEGF on osteoblastic cells during osteogenesis is still controversial. In recent days, substantial progress have been made toward developing tissue-engineered alternatives to autologous bone grafting for maxillofacial bony defects. Periosteum has received considerable interest as a better source of adult stem cells. Periosteum has the advantage of easy harvest and contains various cell types and progenitor cells that are able to differentiate into a several mesenchymal lineages, including bone. Several studies have reported the bone formation potential of periosteal cells, however, the correlation between VEGF signaling and cultured human periosteal cell-derived osteogenesis has not been fully investigated yet. The purpose of this study was to examine the correlation between VEGF signaling and cultured human periosteal-derived cells osteogenesis. Periosteal tissues of $5\;{\times}\;20\;mm$ were obtained from mandible during surgical extraction of lower impacted third molar from 3 patients. Periosteal-derived cells were introduced into the cell culture and were subcultured once they reached confluence. After passage 3, the periosteal-derived cells were further cultured for 42 days in an osteogenic inductive culture medium containing dexamethasone, ascorbic acid, and ${\beta}-glycerophosphate$. We evaluated the alkaline phosphatase (ALP) activity, the expression of Runx2 and VEGF, alizarin red S staining, and the quantification of osteocalcin and VEGF secretion in the periosteal-derived cells. The ALP activity increased rapidly up to day 14, followed by decrease in activity to day 35. Runx2 was expressed strongly at day 7, followed by decreased expression at day 14, and its expression was not observed thereafter. Both VEGF 165 and VEGF 121 were expressed strongly at day 35 and 42 of culture, particularly during the later stages of differentiation. Alizarin red S-positive nodules were first observed on day 14 and then increased in number during the entire culture period. Osteocalcin and VEGF were first detected in the culture medium on day 14, and their levels increased thereafter in a time-dependent manner. These results suggest that VEGF secretion from cultured human periosteal-derived cells increases along with mineralization process of the extracellular matrix. The level of VEGF secretion from periosteal-derived cells might depend on the extent of osteoblastic differentiation.
The fermented soybean product, cheonggukjang, is favored by many people, partly due to its bio-functional ingredients. Since the fermentation process of cheonggukjang is mediated by enzymes, including proteases, produced by microbes, analysis of the proteome profile changes in cheonggukjang during fermentation would provide us with valuable information for fermentation optimization, as well as a better understanding of the formation mechanisms of the bio-functional substances. The soluble proteins from cheonggukjang were prepared by a phenol/chloroform extraction method, in order to remove interfering molecules for high resolution 2-D gel analysis. Proteomic analysis of the cheonggukjang different fermentation periods suggested that most of the soluble soy proteins were degraded into smaller forms within 20hr, and many microbial proteins, such as mucilage proteins, dominated the soluble protein fraction. The proteomic profile of cheonggukjang was very different from natto, in terms of the 2-D gel protein profile. Among the separated protein spots on the 2-D gels, 50 proteins from each gel were analyzed by MALDI-TOF MS and PMF for protein identification. Due to database limitations with regard to soy proteins and microbial proteins, identification of the changed proteins during fermentation was restricted to 9 proteins for cheonggukjang and 15 for natto. From de novo sequencing of the proteins by a tandem MS/MS, as well as by database searches using BLASTP, a limited number of proteins were identified with low reliability. However, the 2-D gel analysis of proteins, including protein preparation methods, remains a valuable tool to analyze complex mixtures of proteins entirely. Also, for intensive mass spectrometric analysis, it is also advisable to focus on a few of the interestingly changed proteins in cheonggukjang.
The anticancer and immunomodulatory activities of low molecular weight (Mw 11 kDa) fucoidan isolated from Hizikia fusiforme (H. fusiforme) via the ultrasonification extraction process were assessed in this study. Low molecular weight fucoidan improved the growth of human B and T cells, up to approximately 40% as compared to the controls (untreated) and 30% for commercially available fucoidan (Mw 150 kDa). IL-6 and TNF-$\alpha$ were secreted from human B cells at levels of $7.8\times10^{-4}$ pg/mL and $7.2\times10^{-4}$ pg/mL, respectively, and these levels were higher than the levels measured in the controls and with other high molecular weight fucoidan. It was also determined that the cytokine from human B and T cells cultivated with added fucoidan enhanced the growth of human NK cells. The fucoidan isolated from H. fusiforme showed low cytotoxicity, approximately 19%, after the addition of 1.0 mg/mL, the highest tested concentration. The growth of human lung cancer cells (A549) and human breast cancer cells (MCF-7) were inhibited by 69.8% and 83.3%, respectively. These results demonstrated that the low molecular weight fucoidan isolated from H. fusiforme has potential as a new functional food component that evidences immunomodulatory activities and anticancer activity. One of the primary positive features of this fucoidan is that low molecular weight polysaccharides can be readily handled during processing.
The 100 l culture system was made on the basis of LED light, and Nannochloropsis oculata was cultured in f/2 medium at light intensity ($100{\mu}mol/m^2/s$), culture temperature ($20^{\circ}C{\pm}1^{\circ}C$) and LD cycle (12hr). As a result, the maximum biomass of 1.07 g/l was cultured as a result of 100 l mass culture at $100{\mu}mol/m^2/s$ and 24 mg/l nitrate concentration in LED blue (475 nm). The extraction was carried out using sonicator, homogenizer and chemical method 0.5M HCl shredding method. The contents of chlorophyll a, chlorophyll b and carotenoid were 1.6, 0.5 and 0.3 mg/g cell. When using homogenizer, it was measured at 1.0, 0.6 and 0.2 mg/g cell. The chemical breakdown method of 0.5M HCl, chlorophyll a, b, and carotenoid contents were measured as 0.9, 0.8, 0 mg/g cell. The highest amount of biomass during the distruption time was measured at 3.6 mg/g cell at 15 min disintegration and acetone, 3.6 mg/g cell of acetone, methanol, and ethanol were measured as effective solvents. Concentration was measured by using microfilter, disk type continuous centrifuge and tubular type continuous centrifuge were 16.0, 1.1 and 0.5 g/l, respectively. Four kinds of equipment such as hot air dryer, vacuum dryer, spray dryer and freeze dryer were tested to optimize the drying process. As a result, the recovery rates of spray dryer and freeze dryer were 80% and 60%.
Park, Yong-Sun;Kim, Kyung-Wook;Lee, Jae-Hoon;Kim, Chang-Jin
Journal of the Korean Association of Oral and Maxillofacial Surgeons
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v.27
no.5
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pp.373-384
/
2001
Cellular proliferation is an intricately regulated process mediated by the coordinated interactions of critical growth control genes. Two of these factors in mammalian cells are the p53 and mdm-2 genes. A protein product of the mem-2 oncogene has been recently shown to associate with the protein encoded by the tumor suppressor gene p53. The p53 tumor suppressor protein is stabilized in response to DNA damage and other stress signals and causes the cell to undergo growth arrest or apoptosis, thus preventing the establishment of mutations in future cellular generations. Mutation or loss of p53 is a very common event in tumor progression. It occurs in about 50% of all tumors analysed including of colon, lung, breast and liver. The cellular mdm-2 gene, which has potential transforming activity that can be activated by overexpression, is amplified in a significant percentage of human sarcoma and in other mammalian tumors. Proteins encoded by the mdm-2 gene are able to bind to the p53 protein and, when overexpressed, can inhibit p53's transcriptional activation function, thus mdm-2 can act as a negative regulator of p53 function. Experimental study was performed to observe the relationship between p53 gene mutation and mdm-2 protein expression and apply the results to the clinical activity. 36 golden syrian hamster each weighing $60{\sim}80g$ were used and painted with 0.5% DMBA by 3 times weekly on the right buccal cheek(experimental side) for 6, 8, 10, 12, 14 and 16 weeks. Left buccal cheek(control side) was treated with mineral oil as the same manner to the right side. The hamsters were sacrificed on the 6, 8, 10, 12, 14 & 16 weeks. Normal and tumor tissues from paraffin block were examined for histology and immunohistochemistry observation, and were completely dissected by microdissection and DNA from both tissue were isolated by proteins K/phenol/chloroform extraction. Segments of the hamster p53 exons 5, 6, 7 and 8 were amplified by PCR using the oligonucleotide primers, and then confirmational change was observed by SSCP respectively. The results were as follows : 1. Dysplasia at 6 weeks, carcinoma in situ at 8 weeks and invasive carcinoma from 10 weeks could be observed in experimental groups. 2. p53 mutations were detected in 10 of the 36(28%) and the exons 6(6 of the 10 : 60%) was the most hot spot area among the highy conserved region(exons 5, 6, 7 & 8). 3. Immunohistochemical study confirmed 22 of the 36(61%) of p53 expression involving 10 of p53 mutations. 4. mdm-2 expression of was showed in 3 of the 36(8%) involving 1 of the 22 of p53 expression and 2 of the 14 of p53 non-expression. From the above results, mutation of p53 gene or expression of p53 protein may have the influence of the DMBA induced carcinoma of hamster buccal pouch but the expression of mdm-2 protein may not have relationship with tumorigenesis.
Recently, use of natural medicine is getting more attention, and some of them are believed to be effective in the treatment of periodontitis. Among them, the seeds of safflower(Carthamus tinctrorius L.) have been proven to be effective through its use in bone diseases such as fracture and osteoporosis. During the last few years, studies using the seeds of safflower gown in Korea have been active, and it has been reported that safflower seed extract increase the proliferation and the alkaline phosphatase(ALP) activity of human periodontal ligament fibroblast(hPDLF), osteoblast, and that they promote the mineralization process. In animal studies, when safflower seed extract were administered orally new bone formation was promoted. Recently, in an effort to find out the most effective osteogenic components, among many components of the safflower seed, various safflower seed fraction extracts were obtained by multistep extraction of the safflower components using various solvents. Among these, saf-M-W fraction extracted by methanol and water was most effective in increasing osteogenic potential of osteoblasts. In this study, the effect of safflower seed fraction extract, saf-M-W, on the growth and differentiation of hPDLF and MC3T3-E1 cell was investigated. The toxicity of saf-M-W on both cells was measured using M'IT(3-(4,5dimethylthiazol-2-y1)-2,5-diphenyl tetrazolium bromide) test, and ALP activity was measured using the colorimetric assay of hPDLF. In addition, in MC3T3-El cells, the expression of ALP, bone sialoprotein(BSP) mRNA was observed using Northern blot, and the mineralized nodule formation Was observed using von Kossa stain and phase-contrast microscope. 1. In concentrations below $10{\mu}g/ml$, saf-M-W didn't show any toxicity on hPDLF and MC3T3-El cell. 2. The change in saf-M-W concentration had no effect on the ALP activity of hPDLF. 3. In MC3T-E1 cells, mRNA expressions of ALP and BSP were greater in the experimental group treated with $10{\mu}g/ml$ concentration of saf-M-W compared with the control group. 4. In MC3T3-El cells, abundance of mineralized nodules were formed in the experimental group treated with $10{\mu}g/ml$ Concentration of saf-M-W, while no mineralized nodule was formed in the control group. These results suggest that safflower seed fraction extract, saf-M-W. didn't show any toxicity on hPDLF and MC3T3-E1 cell at concentrations below $10{\mu}g/ml$ and effectively enhanced the differentiation and osteogenic potential of MC3T3-El cell.
There have been many studies on the application of the reciprocal advantages of multimodality image to define accurate target volume in the Process of radiation treatment planning. For the proper use of the multimodality images, the registration works between different modality images should be performed in advance. In this study, we selected chamfer matching method and mutual information method as most popular methods in recent image registration studies considering the registration accuracy and clinical practicality. And the two registration methods were analyzed to deduce the optimal registration method according to the characteristics of images. Lung phantom of which multimodality images could be acquired was fabricated and CT, MRI and SPECT images of the phantom were used in this study. We developed the registration program which can perform the two registration methods properly and analyzed the registration results which were produced by the developed program in many different images' conditions. Although the overall accuracy of the registration in both chamfer matching method and mutual information method was acceptable, the registration errors in SPECT images which had lower resolution and in degraded images of which data were removed in some part were increased when chamfer matching method was applied. Especially in the case of degraded reference image, chamfer matching methods produce relatively large errors compared with mutual information method. Mutual information method can be estimated as more robust registration method than chamfer matching method in this study because it did not need the prerequisite works, the extraction of accurate contour points, and it produced more accurate registration results consistently regardless of the images' characteristics. The analysis of the registration methods in this study can be expected to provide useful information to the utilization of multimodality images in delineating target volume for radiation treatment planning and in many other clinical applications.
In recent years, packed column has been widely used in separation processes, such as absorption, desorption, distillation, and extraction, in the petrochemical, fine chemistry, and environmental industries. Packed column is used as a contacting facility for gas-liquid and liquid-liquid systems filled with random packed materials in the column. Packed column has various advantages such as low pressure drop, economical efficiency, thermally sensitive liquids, easy repairing restoration, and noxious gas treatment. The performance of a packed column is highly dependent on the maintenance of good gas and liquid distribution throughout a packed bed; thus, this is an important consideration in a design of packed column. In this study, hydraulic pressure drop, hold-up as a function of liquid load, and mass transfer in the air, air/water, and air-NH3/water systems were studied to find the geometrical characteristic for raschig super-ring experiment dry pressure drop. Based on the results, design factors and operating conditions to handle noxious gases were obtained. The dry pressure drop of the random packing raschig super-ring was linearly increased as a function of gas capacity factor with various liquid loads in the Air/Water system. This result is lower than that of 35 mm Pall-ring, which is most commonly used in the industrial field. Also, it can be found that the hydraulic pressure drop of raschig super-ring is consistently increased by gas capacity factor with various liquid loads. When gas capacity factor with various liquid loads is increased from 1.855 to 2.323 kg-1/2 m-1/2 S-1, hydraulic pressure drop increases around 17%. Finally, the liquid hold-up related to packing volume, which is a parameter of specific liquid load depending on gas capacity factor, shows consistent increase by around 3.84 kg-1/2 m-1/2 S-1 of the gas capacity factor. However, liquid hold-up significantly increases above it.
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