• Title/Summary/Keyword: Detection Technique

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($P16^{ink4}$ Methylation in Squamous Cell Carcinoma of the Oral Cavity. (구강 편평세포암종에서 $P16^{ink4}$ 유전자의 Methylation에 대한 연구)

  • Kang, Gin-Won;Kim, Kyung-Wook;Lyu, Jin-Woo;Kim, Chang-Jin
    • Maxillofacial Plastic and Reconstructive Surgery
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    • v.22 no.2
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    • pp.164-173
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    • 2000
  • The p16 protein is a cyclin dependent kinase inhibitor that inhibits cell cycle progression from $G_1$ phase to S phase in cell cycle. Many p16 gene mutations have been noted in many cancer-cell lines and in some primary cancers, and alterations of p16 gene function by DNA methylation have been noticed in various kinds of cancer tissues and cell-lines. There have been a large body of literature has accumulated indicating that abnormal patterns of DNA methylation (both hypomethylation and hypermethylation) occur in a wide variety of human neoplasma and that these aberrations of DNA methylation may play an important epigenetic role in the development and progression of neoplasia. DNA methylation is a part of the inheritable epigenetic system that influences expression or silencing of genes necessary for normal differentiation and proliferation. Gene activity may be silenced by methylation of up steream regulatory regions. Reactivation is associated with demethylation. Although evidence or a high incidence of p16 alterations in a variety of cell lines and primary tumors has been reported, that has been contested by other investigators. The precise mechanisms by which abnormal methylation might contribute to carcinogenesis are still not fully elucidated, but conceivably could involve the modulation of oncogene and other important regulatory gene expression, in addition to creating areas of genetic instability, thus predisposing to mutational events causing neoplasia. There have been many variable results of studies of head and neck squamous cell carcinoma(HNSCC). This investigation was studied on 13 primary HNSCC for p16 gene status by protein expression in immunohistochemistry, and DNA genetic/epigenetic analyzed to determine the incidence, the mechanisms, and the potential biological significance of its Inactivation. As methylation detection method of p16 gene, the methylation specific PCR(MSP) is sensitive and specific for methylation of any block of CpG sites in a CpG islands using bisulfite-modified DNA. The genomic DNA is modified by treatment with sodium bisulfate, which converts all unmethylated cytosines to uracil(thymidine). The primers designed for MSP were chosen for regions containing frequent cytosines (to distinguish unmodified from modified DNA), and CpG pairs near the 5' end of the primers (to provide maximal discrimination in the PCR between methylated and unmethylated DNA). The two strands of DNA are no longer complementary after bisulfite treatment, primers can be designed for either modified strand. In this study, 13 paraffin embedded block tissues were used, so the fragment of DNA to be amplified was intentionally small, to allow the assessment of methylation pattern in a limited region and to facilitate the application of this technique to samlples. In this 13 primary HNSCC tissues, there was no methylation of p16 promoter gene (detected by MSP and automatic sequencing). The p16 protein-specific immunohistochemical staining was performed on 13 paraffin embedded primary HNSCC tissue samples. Twelve cases among the 13 showed altered expression of p16 proteins (negative expression). In this study, The author suggested that low expression of p16 protein may play an important role in human HNSCC, and this study suggested that many kinds of genetic mechanisms including DNA methylation may play the role in carcinogenesis.

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PCA­based Waveform Classification of Rabbit Retinal Ganglion Cell Activity (주성분분석을 이용한 토끼 망막 신경절세포의 활동전위 파형 분류)

  • 진계환;조현숙;이태수;구용숙
    • Progress in Medical Physics
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    • v.14 no.4
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    • pp.211-217
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    • 2003
  • 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.

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Immunization Effect on Naegleria fowleyi Infection in Splenectomized Mice (비장절제 마우스에서 Naegleria fowleri 감염에 대한 면역효과)

  • Han, Gwang-Hyeop;An, Myeong-Hui;Min, Deuk-Yeong
    • Parasites, Hosts and Diseases
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    • v.26 no.1
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    • pp.39-44
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    • 1988
  • A pathogenic free-living amoeba, Naegleria fowleri, is a causative protozoan parasite of primary amebic meningoencephalitis in human and experimental animals. It is known that humoral and cellular immunity contribute as the defence mechanism of host against this organism. Recently splenectomy has been argued on its effect on host defence mechanisms. The present study was aimed to observe the enact of immunization in splenectomized mice. For immunization, $5~10{\times}10^5$ trophozoites of Naegleria fewleri o 359 were intraperitoneally inoculated once a week for two weeks to BALB/c mice, and $5~10{\times}10^4$ of ameba trophozoites were intranasally inoculated for infection after splenectomy and/or immunization. ELISA technique was applied for the detection of seum IgG antibody levels. Experimental animals were divided into 4 groups; I. splenectomized and immuniEed; ll. splenectomized only; III. immunized only; IV. not splenectomized nor immunized. The results obtained were as follows: 1. Mortality rates of splenectomized and immunized mice in group I (38.1%) and immunized only in group III (25.0%) were lower than those of not immunized mice in group II (50%) and control group, IV (46.4%). 2. Survival times of mice in group I, II, III and IV were $20.1{\pm}3.6$, $17.3{\pm}4.5$, $20.4{\pm}7.0$ and $19.6{\pm}7.6$ days respectively, and there were no significant differences between them. 3. ELISA values (absorbance at 492nm) of group I (1, $10{\pm}0.29$) and group III ($1.31{\pm}0.28$) were significantly higher than that of group IV($0.24{\pm}0.37$) at day 31 of infection (p<0.05). Conclusively, it is presumed that humoral immunity against N. fowleri may operate as ever, after immunization, even though the mouse was splenectomized.

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Developmental Genetic Analysis of Avian Primordial Germ Cells and the Application to Poultry Biotechnology

  • Kagami, H.
    • Korean Journal of Poultry Science
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    • v.28 no.2
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    • pp.135-142
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    • 2001
  • A novel sterategy has been established to determine the origin of the Primordial Germ Cells (PGCs) in avian embryos directly and the developmental fate of the PGCs for the application to Poultry biotechnology. Cells were removed from 1) the centre of area pellucida, 2) the outer of area pellucida and 3) the area opaca of the stage X blastoderm (Eyal-Giladi & Kochav, 1976). When the cells were removed from the centre of area pellucida, the mean number of circulating PGCs in blood was significantly decreased in the embryo at stage 15 (Hamburger & Hamilton, 1951) as compared to intact embryos. When the cells were replenished with donor cells, no reduction in the PGCs number was observed. The removal of cells at the outer of area pellucida or at the area opaca had no effect on the number of PGCs. In case, another set of the manipulated embryos were cultured ex vivo to the hatching and reared to the sexual maturity, the absence of germ cells and degeneration of seminiferous tubules was observed in resulting chickens derived from the blastoderm in which the cells were removed from the centre of the area pellucida. It was concluded that the avian Primordial Germ cells are originated at the center of area pellucida. Developmental ability of the cells to differentiate into somatic cells and germ cells in chimeras were analyzed. Somatic chimerism was detected as black feather attributed from donor cells. Molecular identification by use of female - specific DNA was performed. It was confirmed that the donor cells could be differentiated into chimeric body and erythrocytes. Donor cells retained the ability to differentiate into germline in chimeric gonads. More than 70% of the generated chimeras transmitted donor derived gametes to their offspring indicating that the cells at the center of area pellucida had the high ability to differentiate into germ cells. A molecular technique to identify germline chimerism has been developed by use of gene scan analysis. Strain specific DNA fragments were amplified by the method. It would be greatly contributed for the detection of germline chimerism. Mixed- sex chimeras which contained both male and female cells were produced to investigate the developmental fate of male and female cells in ovary and testes. The sex combinations of donor and recipient of the resulting chimeras were following 4 pairs; (1) chimeras (ZZ/ZZ) produced by a male donor (ZZ) and a male recipient (ZZ), (2) chimeras (ZW/ZW) produced by a female donor (ZW) and a female recipient (ZW), (3) chimeras (ZZ/ZW) Produce by a male donor (ZZ) and a female recipient (ZW), (4) chimeras (ZW/ZZ) produced by a female donor (ZW) and a male recipient (ZZ). It was found that genetically male avian germ cells could differentiate into functional ova and that genetically female germ cells can differentiate into functional spermatozoa in the gonad of the mixed- sex chimeras. An ability for introduction of exogenous DNA into the PGCs from stage X blastoderms were analyzed. Two reporter genes, SV-$\beta$gal and RSV-GFP, were introduced into the PGCs. Expression of bacterial/gal was improved by complexing DNA with liposome detectedcc in 75% of embryos at 3 days embryos. At the embryos incubated for 1 day, expression of the GFP was observed all the embryos. At day 3 of incubation, GFP was detected in about 70% of the manipulated embryos. In case of GFP, expression of the transgene was detected in 30 %e of the manipulated embryos. These results suggested that the cells is one of the most promising vectors for transgenesis. The established strategy should be very powerfull for application to poultry biotechnology.

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Camparison between the 1 Day and the 2 Day Protocols of Lymphoscintigraphy and Sentinel Node Biopsy using Subareolar Injection in Breast Cancer Patients: A Retrospective Study (유륜하 주사에 의한 유방암 환자의 전초림프절 스캔과 전초림프절 생검에 있어서 당일검사와 전날검사의 비교: 후향적 연구)

  • Seok, Ju-Won;Jun, Sung-Min;Nam, Hyun-Yeol;Kim, In-Ju
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.1
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    • pp.55-59
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    • 2009
  • Purpose: Lymphoscintigraphy and sentinel node biopsy are used in detection of axillary lymph node metastasis in breast cancer patients, but standardized technique is not established. We compared the results of the injection the morning of surgery (1 day protocol) with the subareolar injection the day before surgery (2 day protocol) with the subareolar injection in patients with breast cancer having lymphoscintigraphy and sentinel node biopsy. Materials and Methods: This study included 349 patients who underwent the breast cancer operation during 2001-2004. One hundred seventy one patients (1 day protocol, 1 hour) was injected 0.8ml of Tc-99m Tin-Colloid (37 MBq) by subareolar injection on the morning of surgery. One hundred seventy eight patients (2 day protocol, 16 hour) was injected 0.8 ml of T c-99m Tin-Colloid (185 MBq) on the afternoon before surgery. Lymphoscintigraphy was performed in sitting position and sentinel node localization was performed by hand-held gamma probe during operation. Result: In the 1 day protocol, 153 cases (89.5%) of the sentinel node were localized by lymphoscintigraphy and 150 cases (87.7%) were localized by gamma probe. In the 2 day protocol, 159 cases (89.3%) were localized by lymphoscintigraphy and 154 cases (86.5%) were localized by gamma probe. There was no significant difference in localization of sentinel node between the 1 day and the 2 day protocol by lymphoscintigraphy and gamma probe (p>0.05, p>0.05). Conclusion: There was no difference the result of localization of sentinel node with subareolar injection between the 1 day and the 2 day protocol in breast cancer patients. Because the 2 day protocol allows the enough time of performing lymphoscintigraphy, it is more useful in localization of sentinel node in breast cancer patients.

Epidemiological Studies on Giardia Infection Associated with environmental Pollution (Giardia에 의한 환경오염(環境汚染)과 감염(感染)에 관한 역학적(疫學的) 연구(硏究))

  • Lee, Keun-Tae;Kim, Seok-Chan;Song, Jong-Sool;Chung, Pyung-Rim
    • Journal of agricultural medicine and community health
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    • v.9 no.1
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    • pp.56-66
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    • 1984
  • Giardia lamblia is a pathogenic flagellate causing intestinal disorders such as diarrhea, abdominal pain and malabsorption of nutrients. Giardia is mainly infected by the ingestion of contaminated foods per os. Craun (1979) has recently reported that mass infection of this flagellate through the contaminated water supply systems is one of public health hazards. Also, so-called traveller's diarrhea is sometimes caused by Giardia infection (CDC, U.S.A., 1971). However, a few epidemiological studies figuring out the mode of infection or control measures of Giardia infection has been done so far in Korea. The present study was aimed to know the prevalence of Giardia infection in several Korean populations, detectability of this flagellate in water systems and the viability of the cysts against sewages and disinfectants applying to drinking water. In the present study, 388 stool specimens from orphanage children in Chun-joo, Chung-joo, On-yang and Chun-an areas and 538 stool specimens from inhabitants in Woo-do, In-chon, and Chun-joo were examined by formalin-ether concentration technique to detect out Giardia cysts. On the other hand, water samples from 14 sites of Han River and its tributaries were collected in May through July, 1984. Fifty liter of water sample in each sampling site was then filtered through water filtering system deviced by U.S. Environmental Proutection Agency and the sediments rinsed out from the thread rolls, a part of water filtering system, were examined to detect out the Giardia cysts. In order to observe the viability of Giardia cysts in the sewage samples, the cysts were treated in it at $4^{\circ}C$ or $25^{\circ}C$ for 7 through 28 days. For this purpose, the cysts were also exposed to various concentrations of disinfectants such as chlorine, iodine and ozone gas for proper time intervals. After treatment, the viability test of the Giardia cysts were carried out by method of Rice and Schaefer (1981) with minor modification. The results obtained in this study were as follows : 1) The detection rates of G lamblia cysts in the stool specimens were 18.3% in orphans and 4.3% in general examinees. 2) The prevalences of Giardia Infection were higher in the young age groups than in-adults. The highest positive rate was 18.4% in the age group less than 10. 3) Of 14 water specimens sampled from Han River system and its tributaries around the Seoul area, the Giardia cysts were detected from 4 samples, and no cyst was found in the water supply systems. 4) The cysts treated in the sewage survived for 28 days at $4^{\circ}C$ and for 13 days at $25^{\circ}C$. 5) The cysts were completely destroyed within 60 minutes by exposure to 8 mg/l of residual chlorine at 4g and within 30 minutes by exposure to the same concentration of chlorine at $25^{\circ}C$. 6) The cysts were all dead when exposed to 1 mg/1 of iodine for 60 minutes at $4^{\circ}C$ or $25^{\circ}C$. 7) The cysts were destroyed after 10 minute exposure in 0.15 mg to 0.25mg of residual ozone gas per liter. Summarizing the above results, it is considered that Giardia infection is regarded as water-borne disease and the cysts are able to be controlled by the application with the disinfectants in the water supply systems.

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Assessment of DNA damage and Chromosome aberration in human lymphocyte exposed to low dose radiation detected by FISH(fluorescence in situ hybridization) and SCGE(single cell gel electrophoresis) (FISH기법 및 단세포전기영동기법을 이용한 저선량 방사선에 의한 DNA 상해 및 염색체이상 평가)

  • Chung, Hai-Won;Kim, Su-Young;Kim, Byung-Mo;Kim, Sun-Jin;Kim, Tae-Hwan;Cho, Chul-Koo;Ha, Sung-Whan
    • Journal of Radiation Protection and Research
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    • v.25 no.4
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    • pp.223-232
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    • 2000
  • Comparative study was performed for the assessment of DNA damage and Chromosomal aberration in human lymphocyte exposed to low dose radiation using fluorescence in situ hybridization(FISH) and single cell gel electrophoresis(SCGE). Chromosomal aberrations in human lymphocytes exposed to radiation at doses of 5, 10, 30 and 50cGy were analysed with whole chromosome-specific probes by human chromosome 1, 2 and 4 according to PAINT system. FISH with chromosome-specific probe has been used to be a valid and rapid method fer detection of chromosome rearrangements induced by low dose radiation. The frequencies of stable translocation per cell equivalents were 0.0116, 0.0375, 0.040f, 0.0727 and 0.0814 for 0, 5, 10, 30 and 50cGy, respectively, and those of dicentric were 0.00, 0.0125, 0.174, 0.0291 and 0.0407 respectively. Radiation induced DNA damage in human lymphocyte in a dose-dependent manner at low doses from 5cGy to 50cGy, which were analysed by single tell gel electrophoresis(SCGE). From above results, FISH seemed to be useful for radiation biodosimetry by which the frequencies of stable aberrations in human lymphocyte can be observed more easily than by conventional method and SCGE also seemed to be sensitive method f9r detecting DNA damage by low dose radiation exposure, so that those methods will improve our technique to perform meaningful biodosimetry for radiation at low doses.

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Improved Device Performance Due to AlxGa1-xAs Barrier in Sub-monolayer Quantum Dot Infrared Photodetector

  • Han, Im Sik;Byun, Young-Jin;Lee, Yong Seok;Noh, Sam Kyu;Kang, Sangwoo;Kim, Jong Su;Kim, Jun Oh;Krishna, Sanjay;Ku, Zahyun;Urbas, Augustine;Lee, Sang Jun
    • Proceedings of the Korean Vacuum Society Conference
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    • 2014.02a
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    • pp.298-298
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    • 2014
  • Quantum dot infrared photodetectors (QDIPs) based on Stranski-Krastanov (SK) quantum dots (QDs) have been widely explored for improved device performance using various designs of heterostructures. However, one of the biggest limitations of this approach is the "pancake" shape of the dot, with a base of 20-30 nm and a height of 4-6 nm. This limits the 3D confinement in the quantum dot and reduces the ratio of normal incidence absorption to the off-axis absorption. One of the alternative growth modes to the formation of SK QDs is a sub-monolayer (SML) deposition technique, which can achieve a much higher density, smaller size, better uniformity, and has no wetting layer as compared to the SK growth mode. Due to the advantages of SML-QDs, the SML-QDIP design has attractive features such as increased normal incidence absorption, strong in-plane quantum confinement, and narrow spectral wavelength detection as compared with SK-DWELL. In this study, we report on the improved device performance of InAs/InGaAs SML-QDIP with different composition of $Al_xGa1-_xAs$ barrier. Two SML-QDIPs (x=0.07 for sample A and x=0.20 for sample B) are grown with the 4 stacks 0.3 ML InAs. It is investigated that sample A with a confinement-enhanced (CE) $Al_{0.22}Ga_{0.78}As$ barrier had a single peak at $7.8{\mu}m$ at 77 K. However, sample B with an $Al_{0.20}Ga_{0.80}As$ barrier had three peaks at (${\sim}3.5{\mu}m$, ${\sim}5{\mu}m$, ${\sim}7{\mu}m$) due to various quantum confined transitions. The measured peak responsivities (see Fig) are ~0.45 A/W (sample A, at $7.8{\mu}m$, $V_b=-0.4V$ bias) and ~1.3 A/W (sample B, at $7{\mu}m$, $V_b=-1.5V$ bias). At 77 K, sample A and B had a detectivity of $1.2{\times}10^{11}cm.Hz^{1/2}/W$ ($V_b=-0.4V$ bias) and $5.4{\times}10^{11}cm.Hz^{1/2}/W$ ($V_b=-1.5V$ bias), respectively. It is obvious that the higher $D^*$ of sample B (than sample A) is mainly due to the low dark current and high responsivity.

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A Study on GPU-based Iterative ML-EM Reconstruction Algorithm for Emission Computed Tomographic Imaging Systems (방출단층촬영 시스템을 위한 GPU 기반 반복적 기댓값 최대화 재구성 알고리즘 연구)

  • Ha, Woo-Seok;Kim, Soo-Mee;Park, Min-Jae;Lee, Dong-Soo;Lee, Jae-Sung
    • Nuclear Medicine and Molecular Imaging
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    • v.43 no.5
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    • pp.459-467
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    • 2009
  • Purpose: The maximum likelihood-expectation maximization (ML-EM) is the statistical reconstruction algorithm derived from probabilistic model of the emission and detection processes. Although the ML-EM has many advantages in accuracy and utility, the use of the ML-EM is limited due to the computational burden of iterating processing on a CPU (central processing unit). In this study, we developed a parallel computing technique on GPU (graphic processing unit) for ML-EM algorithm. Materials and Methods: Using Geforce 9800 GTX+ graphic card and CUDA (compute unified device architecture) the projection and backprojection in ML-EM algorithm were parallelized by NVIDIA's technology. The time delay on computations for projection, errors between measured and estimated data and backprojection in an iteration were measured. Total time included the latency in data transmission between RAM and GPU memory. Results: The total computation time of the CPU- and GPU-based ML-EM with 32 iterations were 3.83 and 0.26 see, respectively. In this case, the computing speed was improved about 15 times on GPU. When the number of iterations increased into 1024, the CPU- and GPU-based computing took totally 18 min and 8 see, respectively. The improvement was about 135 times and was caused by delay on CPU-based computing after certain iterations. On the other hand, the GPU-based computation provided very small variation on time delay per iteration due to use of shared memory. Conclusion: The GPU-based parallel computation for ML-EM improved significantly the computing speed and stability. The developed GPU-based ML-EM algorithm could be easily modified for some other imaging geometries.

Perspective on Rapid and Selective Method for Detecting Microbiology in Dairy Industry: A Review (낙농산업에 필요한 미생물 검사방법과 전망: 총설)

  • Chon, Jung-Whan;Kim, Hyun-Sook;Kim, Hong-Seok;Kim, Dong-Hyeon;Song, Kwang-Young;Yim, Jin-Hyuk;Choi, Dasom;Lim, Jong-Soo;Jeong, Dong-Gwan;Kim, Soo-Ki;Seo, Kun-Ho
    • Journal of Dairy Science and Biotechnology
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    • v.33 no.2
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    • pp.119-127
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    • 2015
  • To date, detection of microbial populations in dairy products has been performed using culture media, which is a time-consuming and laborious method. The recently developed chromogenic media could be more rapid and specific than classical culture media. However, the newly developed molecular-based technology can detect microbial populations with greater rapidity and sensitivity than the classical method involving culture media and chromogenic media. This molecular-based technology could provide various options for monitoring the characterization of different states of bacteria and cells. Thus, it could help upgrade the processing system of the dairy industry so as to maintain the safety and quality of dairy foods. Among the various newly developed molecular-based technologies, flow cytometry can potentially be used for monitoring microbiological populations in the dairy industry if official international standards are available for this purpose. When omics technology would have biomarker identification, it could be regarded as the rapid and sensitive analytical methods. Methods based on PCR, which has become a basic technique in microbiological research, can be developed and validated as alternative methods for quantification of dairy microorganisms. This review discusses methods for monitoring microbiological populations in dairy foods and the limitations of these studies, as well as the need for further research on such methods in the dairy industry.

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