Yun, Bong Han;Kim, Yong Hwi;Sung, Mu Sung;Han, Ho-Seop;Han, Jeong-Ho;Bang, In-Chul
Korean Journal of Ichthyology
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v.34
no.3
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pp.208-217
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2022
We wanted to develop a real-time PCR assay capable of detecting Liobagrus obesus in environmental DNA (eDNA) extracted from freshwater samples using a pair of species-specific primers and probe for the endangered fish, L. obesus. The species-specific primers and probe were designed in consideration of single nucleotide polymorphisms between 65 species of freshwater fish living in the Republic of Korea within the cytochrome b (cytb) gene of mitochondrial DNA. The species-specific primers and probe, in the real-time PCR assay, showed high specificity as only the L. obesus genomic DNA (gDNA) was found to be positive in the specificity verification using 65 species gDNA of freshwater fish in the Republic of Korea. In addition, in the detection limit analysis using the serial dilution concentrations of L. obesus gDNA, it was found that it was possible to detect up to 0.2 pg, showing high sensitivity. Afterwards, using the species-specific primers and probe, real-time PCR assay was performed on freshwater samples obtained from 8 stations in the mid-upper basin of Geum River. As a result, the cytb gene of L. obesus was detected in total 5 stations including all 3 stations where this species was collected at the time of field survey. Therefore, the species-specific primers and probe developed in present study, and the real-time PCR assay using them, can accurately detect the cytb gene of L. obesus from eDNA samples, which can be utilized to monitor the existing habitats of this species and to discover potential new habitats.
Purpose : To evaluate and compare the accuracy of magnetic resonance imaging (MRI) and ultrasound (US) for detection and estimation of invasion depth of colorectal carcinoma (CRC) by correlation with histopathologic findings in vitro, and to find out the best MR pulse sequence for accurate delineation of tumor from surrounding normal tissue. Materials and Methods: Resected specimens of CRC from 45 patients were examined about tumor detectability and invasion depth of US using high frequency (5-17 MHz) linear transducer in a tube filled with normal saline and MRI in a 8-channel quadrate head coil. The institutional review board approved this study and informed consent was waived. MRI with seven pulse sequences of in- and out-of-phases gradient echo T1 weighted images, fast spin echo T2 weighted image and its fat suppression image, fast imaging employing steady-state acquisition (FIESTA) and its fat suppression image, and diffusion weighted image (DWI) were performed. In each case, both imaging findings of MRI and US were evaluated independently for detection and estimation of invasion depth of tumor by consensus of two radiologists and were compared about diagnostic accuracy according to the histopathologic findings as reference standard. Seven MR pulse sequences were evaluated on the point of accurate delineation of tumor from surrounding normal tissue in each specimen. Results: In specimens of CRC, both imaging modalities of MRI (91.1%) and US (86.7%) showed relatively high diagnostic accuracy to detect tumor and evaluate invasion depth of tumor. In early CRC, diagnostic accuracy of US was 87.5% and that of MRI was 75.0%. There was no statistically significant difference between two imaging modalities (p > 0.05). The best pulse sequence among seven MR sequences for accurate delineation of tumor from surrounding normal tissue in each specimen of CRC was fast spin echo T2 weighted image. Conclusion: MRI and US show relatively high diagnostic accuracy to detect tumor and evaluate invasion depth of resected specimen of CRC. The most excellent pulse sequence of MRI for accurate delineation of tumor from surrounding normal tissue in CRC is fast spin echo T2 weighted image.
Kim, Jin Sung;Ju, Sang Gyu;Hong, Chae Seon;Jeong, Jaewon;Son, Kihong;Shin, Jung Suk;Shin, Eunheak;Ahn, Sung Hwan;Han, Youngyih;Choi, Doo Ho
Progress in Medical Physics
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v.24
no.2
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pp.85-91
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2013
At present, megavoltage computed tomography (MVCT) is the only method used to correct the position of tomotherapy patients. MVCT produces extra radiation, in addition to the radiation used for treatment, and repositioning also takes up much of the total treatment time. To address these issues, we suggest the use of a video image-guided setup (VIGS) system for correcting the position of tomotherapy patients. We developed an in-house program to correct the exact position of patients using two orthogonal images obtained from two video cameras installed at $90^{\circ}$ and fastened inside the tomotherapy gantry. The system is programmed to make automatic registration possible with the use of edge detection of the user-defined region of interest (ROI). A head-and-neck patient is then simulated using a humanoid phantom. After taking the computed tomography (CT) image, tomotherapy planning is performed. To mimic a clinical treatment course, we used an immobilization device to position the phantom on the tomotherapy couch and, using MVCT, corrected its position to match the one captured when the treatment was planned. Video images of the corrected position were used as reference images for the VIGS system. First, the position was repeatedly corrected 10 times using MVCT, and based on the saved reference video image, the patient position was then corrected 10 times using the VIGS method. Thereafter, the results of the two correction methods were compared. The results demonstrated that patient positioning using a video-imaging method ($41.7{\pm}11.2$ seconds) significantly reduces the overall time of the MVCT method ($420{\pm}6$ seconds) (p<0.05). However, there was no meaningful difference in accuracy between the two methods (x=0.11 mm, y=0.27 mm, z=0.58 mm, p>0.05). Because VIGS provides a more accurate result and reduces the required time, compared with the MVCT method, it is expected to manage the overall tomotherapy treatment process more efficiently.
Although early diagnosis of urinary tract infection is important, the radiologic evaluation is still controversial because of the low sensitivity and the lack of cost-effectiveness. This study was carried out to evaluate the clinical utility of high resolution triple head $^{99m}Tc-DMSA$ SPECT imaging in urinary tract infection. We prospectively performed $^{99m}Tc-DMSA$ planar and SPECT imaging, ultrasound of kidney (US), intravenous pyelography (IVP) and voiding cystourethrography (VCU) in all 60 adult patients with UTI [26 with first episode of acute pyelonephritis (APN), 22 with recurrent APN, and 12 persistent asymptomatic pyuria] and 25 normal persons. To assess reversibility of the renal cortical defect (RCD), $^{99m}Tc-DMSA$ SPECT was repeated 1 to 8 months later in those patients with abnormal initial findings. Overall detection rate of $^{99m}Tc-DMSA$ SPECT imaging was 83% (50/60), but planar, US, IVP and VCU showed abnormal findings in 68%, 28%, 32% and 13%, respectively. 25 out of 27 patients with normal or single RCD were all normal in other radioligic studies. Only two patients showed vesicoureteral reflux (VUR) on VCU (grade I) and mild hydronephrosis on IVP. But, high proportion of those with multiple RCD showed abnormal findings on US (17/33), IVP (18/33), and VCU (7/33): 67% in any of these 3 studies. Especially, 3 out 7 patients with VUR showed multiple RCD on $^{99m}Tc-DMSA$ SPECT without any abnormality on IVP or US. 25 normal persons showed normal findings in all studies except one false positive finding on $^{99m}Tc-DMSA$ SPECT imaging. Follow-up $^{99m}Tc-DMSA$ SPECT was done in 28 patients (13 with single RCD, 15 with multiple RCD). All 13 patients with single RCD showed improvement. Those with multiple RCD presented improvement in 4, no change in 10, and aggravation in 1 on follow-up studies. With these results, we conclude: 1) $^{99m}Tc-DMSA$ SPECT imaging is superior to planar imaging, US, IVP or VCU in detection of renal lesion in urinary tract infection. $^{99m}Tc-DMSA$ SPECT is useful as a initial diagnostic tool in adult patients with urinary tract infection. 2) The multiple RCD on $^{99m}Tc-DMSA$ SPECT represent the high probability of irreversible tissue change and need of extensive urological work-up.
Purpose: Although radionuclide cisternography (RNC) is an useful study to detect cerebrospinal fluid (CSF) leakage in the patient with spontaneous intracranial hypotension (SIH), it sometimes fails to demonstrate the site of CSF leakage. The aim of the study is to improve the detection of leakage site of CSF and to reduce time for the study in RNC using modified protocol (m-RNC). Materials & methods : The study consists of 8 studies of 7 patients ($38{\pm}8$ years, M:F=2:5) with SIH, who underwent m-RNC following administration of 185-222 MBq of $^{99m}Tc$-DTPA into the lumbar subarachnoid space. Sequential images were obtained the whole spine with the head including urinary bladder at 10 minute, 30 minute, 1 hour, 2 hour, 4 hour and 6 hour. Radioactivity of extradural space and urinary bladder was evaluated. Results: Leakage site of CSF was identified in all 8 cases by m-RNC. Leakage site was cervicothoracic junction (CTJ, n=3), CTJ with C1-2 (n=2), CTJ with thoracic spine, thoracolumbar spine and lumbar spine (each n=1). All cases presented leakage sites within 1 hour and multiple sites, where CTJ was included in 6 cases. Only one case presented additional site in 6 hour image. Early radioactivity within the urinary bladder was noted in 6 cases, but that was fellowing after identification of the leakage site. Conclusion: Radionuclide cisternography is sensitive to detect the leakage site of CSF and is expected to improve the detection of CSF leakage site and reduce time for the study using modified protocol.
Purpose: RBC blood pool SPECT has been used to diagnose focal liver lesion such as hemangioma owing to its high specificity. However, low spatial resolution is a major limitation of this modality. Recently, ordered subset expectation maximization (OSEM) has been introduced to obtain tomographic images for clinical application. We compared this new modified iterative reconstruction method, OSEM with conventional filtered back projection (FBP) in imaging of liver hemangioma. Materials and Methods: Sixty four projection data were acquired using dual head gamma camera in 28 lesions of 24 patients with cavernous hemangioma of liver and these raw data were transferred to LINUX based personal computer. After the replacement of header file as interfile, OSEM was performed under various conditions of subsets (1,2,4,8,16, and 32) and iteration numbers (1,2,4,8, and 16) to obtain the best setting for liver imaging. The best condition for imaging in our investigation was considered to be 4 iterations and 16 subsets. After then, all the images were processed by both FBP and OSEM. Three experts reviewed these images without any information. Results: According to blind review of 28 lesions, OSEM images revealed at least same or better image quality than those of FBP in nearly all cases. Although there showed no significant difference in detection of large lesions more than 3 cm, 5 lesions with 1.5 to 3 cm in diameter were detected by OSEM only. However, both techniques failed to depict 4 cases of small lesions less than 1.5 cm. Conclusion: OSEM revealed better contrast and define in depiction of liver hemangioma as well as higher sensitivity in detection of small lesions. Furthermore this reconstruction method dose not require high performance computer system or long reconstruction time, therefore OSEM is supposed to be good method that can be applied to RBC blood pool SPECT for the diagnosis of liver hemangioma.
A tactile sensor employs a piezoelectric element to detect contact frequency shifts and thereby measure the stiffness or softness of material such as tissue, which allows the sensor to be used in many fields of research for urology, cardiology, gynecology, sports medicine and caner detection and especially for cosmetics and skin care. In this study, reliability of the tactile sensor system was investigated with its manual application to the muscles susceptible to temporomandibular disorders. Stiffness and elasticity of anterior temporalis, masseter and trapezius muscles were calibrated bilaterally from 5 healthy men with an average of 24.5$\pm$0.94 years. The tactile sensor used in this study had a computer-controlled and motor-driven sensor unit which automatically pressed down on the skin surface over the muscles being measured and retracted, thereby providing the hysteresis curve. The slope of the tangent of the hysteresis curve (${\Delta}f/{\Delta}x$) is defined as stiffness of the muscle being measured and the distance between the two parts of the curve as its elasticity. To determine inter-examiner reliability, all the measurements were performed by the two examiners A and B, respectively and the same examination were repeated with an interval of 2 days for intra-examiner reliability. The results from this study demonstrated high reliability in measuring stiffness and elasticity of anterior temporalis, masseter and upper trapezius muscles using a tactile sensor system. It is suggested that the tactile sensor system can be a highly reproducible and effective instrument for quantitative evaluation of the muscle in head and neck region.
Mulberry leaf sprayed on at the various concentrations of cadmium (Cd) was fed on the just molted fifth instar through the 6th day of the fifth instar and content of cadmium and zinc in the feces, organs and tissues involved were analyzed by spectrophotometer. Along with the effect of those heavy metals on development of the relevant organs and tissues sexwise. 1. The residual content of cadmium on mulberry leaf was about 50% of the total cadmium and the weight of feces disposed from the cadmium fed larvae was less than the control. There was distinctive differences in female-wise disposal of feces between cadmium treatment and control ; cadmium treatment female larvae disposed less feces than female of control. 2. Fourteen percent of the total dry weight of the anterior plus the middle silkgland and of the posterior silkgland in cadmium 100ppm treatment was decreased in female but only 9% of the anterior plus middle silkgland in male was made, showing no decrease the weight of the posterior silkgland in male. 3. Decrease of the dry weight of the silkgland, fat body and hindgut was made in cadmium 100ppm treatment and contrarily the weight of midgut and malphihian tube were increased. The total larval weight of the female and male were decreasd in cadmium 100ppm treatment and the decreasing ratio was higher in famale. 4. There wasa no detection of cadmium in all of the organs and tissues of the control and in those of the cadmium treatment the high residual content was in order of the midgut, hindgut, malphighian tube and head. A slight accumulation of cadmium was shown in fat body and silkgland. 5. Cadmium accumulation in the midgut was higher than in female but in the fat body, malphighian tube, trachea and integument was contrarily high in the female than male. 6. The total zinc content of the larvae was below 60ppm in the control and 80ppm in cadmium 100ppm treantment. The increase of zinc content in cadmium treatment was made on silkgland, malphigian tube and midgut but contrarily the decrease of zinc content in fat body was resulted. 7. In cadmium treatment, much more cadmium accumulation took place in the female and zinc accumulation was made much more in the male.
The release of hazardous waste materials into the environment poses serious risks in humans and ecosystems. The risk assessment of environmental pollutants including hazardous chemicals requires a comprehensive measurement of hazard and exposure of the chemicals that can be achieved by toxicity evaluation using a biological system such as biomarkers. In this report we have tried to develop a biomarker used to elucidate a molecular basis of, and to monitor abnormal behaviors caused by diazinon in Japanese medaka (Oryzias latipes) as a model organism. First, an attempt was made to clone tyrosine hydroxylase gene from Japanese medaka that would be a candidate for a biomarker for neuronal modulations and behaviors. For monitoring experiments at behavioral and molecular biological levels, the fish were treated under different sublethal conditions of diazinon and their behavioral responses were observed . In this study we have successfully cloned a partial TH gene from the medaka fish through PCR screening of an ovary cDNA library. DNA sequencing analysis revealed that the amplified fragment was 327 bp encoding 109 amino acids. Comparing the DNA sequence of medaka TH with other species, TH gene revealed the DNA sequence was completely identical to that of rat TH. In the RT-PCR, 330 Up of mRNA was consistently amplified in all the treated samples including control There were no significant differences in the TH expression level regardless of treating concentrations (1∼5,000 ppb) and time (0∼48 hr) The reason appeared to be that RT-PCR was not performed using through a quantitative analysis normalized against an actin gene expression. Organ or tissue - specific detection of TH activity and mRNA as biomarkers will be a useful monitoring tool for neurobehavioral changes in fish influenced by toxic chemicals. Furthermore, quantitative analysis of locomotive patterns and its correlation with the neurochemical and molecular data would be highly useful in measuring toxicity and hazard ofvarious environmental pollutants.
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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