Background: Though infections of Helicobacter pylori (H. pylori) are closely associated with activation of host angiogenesis, the underlying mechanisms, as well as the strategy for its prevention, have not been identified. Here, we investigated a causal role of H. pylori infection in angiogenesis of gastric mucosa and a potent inhibitory effect of a gastric proton pump inhibitor (PPI) on the gastropathy. Materials and Methods: A comparative analysis of CD 34 expression in tissues obtained from 20 H. pylori-associated gastritis and 18 H. pylori-negative gastritis patients was performed. Expression of $HIF-1{\alpha}$ and VEGF were tested by using RT-PCR. To evaluate the direct effect of H. pylori infection on differentiation of endothelial HUVEC cells, we carried out an in vitro angiogenesis assay. Results: H. pyfori-associated gastritis tissues showed significantly higher density of $CD34^+$ blood vessels than did H. pylori-negative gastritis tissues, and the levels were well correlated with expressions of $HIF-1{\alpha}$. Conditioned media from H. pylori-infected gastric mucosal cells stimulated a tubular formation of HUVEC cells. We also found a significant inhibitory effect of PPI, an agent frequently used for H. pylori eradication, on H. pylori-induced angiogenesis. This drug effectively inhibited the phosphorylation of MAP kinase ERK1/2, which is a principal signal for H. pylori-induced angiogenesis. Conclusion: The fact that PPls can down-regulate H. pylori-induced angiogenesis suggest that anti-angiogenic treatment using PPI may be a preventive approach for H. pylori-associated carcinogenesis.
Background : The emergence of multidrug-resistant strains of Mycobacterium tuberculosis presents a significant challange to the treatment and control of tuberculosis, and there is an urgent need to understand the mechanisms by which strains acquire multidrug resistance. Recent advances in molecular methods for the detection of M. tuberculosis genetic targets have approached the sensitivity of culture. Furthermore the prospect of determining resistance in mycobacteria at the nucleic acid level particulary to first-line drugs like rifampin, isoniazid has provided a glimps of the next generation of sensitivity test for M. tuberculosis. Previous studies in RMP resistant M. tuberculosis have shown that mutation in $\beta$subunit of RNA polymerase is main mechanism of resistance. Method : In this study, rpoB gene for the $\beta$subunit of RNA polymerase from M. tuberculosis of 42 cultured samples (32 were RMP resistant and 10 were sensitive cases) were isolated and characterised the mutations. Direct sequencing data were compared with the results of INNO-LiPA Line Probe Assay (LiPA, Innogenetics, Belgium), commercial RMP resistance detecting kit using reverse hybridization method. Results : All of the RMP resistant samples were revealed the presence of mutation by LiPA. In 22 samples (68.8%) out of 32 RMP resistant cases, the mutation types were confirmed by the positive signal at one of 4 mutation bands in the strip. The most frequent type was R5 (S531L) which were 17 cases (77.3%). Results of direct sequencing were identified the exact characteristics of 8 mutations which were not confirmed by LiPA. S522W type point mutation and 9 base pair deletion at codon 513~515 were new identified mutations for the first time. Conclusion : Mutations in rpoB gene is the main mechanism of RMP resistance in M. tuberculosis and LiPA is a very useful diagnostic tool for the early diagnosis of RMP resistance in M. tuberculosis.
Background: Phospholipase C(PLC) plays a central role in cellular signal transduction and is important in cellular growth, differentiation and transformation. There are currently ten known mammalian isozymes of PLC reported to this date. Hydrolysis of phosphatidylinositol 4,5-bisphosphate($PIP_2$) by PLC produces two important second messengers, inositol 1,4,5-trisphosphate($IP_3$) and diacylglycerol. PLC-${\gamma}1$, previously, was known to be activated mainly through growth factor receptor tyrosine kinase. Other mechanisms of activating PLC-yl have been reported such as activation through tau protein in the presence of arachidonic acid in bovine brain and activation by $IP_3$, phosphatidic acid, etc. Very recently, another PLC-${\gamma}1$ activator protein such as tau has been found in bovine lung tissue, which now is considered to be AHNAK protein. But there has been no report concerning AHNAK and its associated disease to this date. In this study, we examined the expression of the PLC-${\gamma}1$ activator, AHNAK, in lung cancer specimens and their paired normal. Methods: From surgically resected human lung cancer tissues taken from twenty-eight patients and their paired normal counterparts, we evaluated expression level of AHNAK protein using immunoblot analysis of total tissue extract Immunohistochemical stain was performed with primary antibody against AHNAK protein. Results: Twenty-two among twenty-eight lung cancer tissues showed overexpression of AHNAK protein (eight of fourteen squamous cell lung cancers, all of fourteen adenocarcinomas). The resulting bands were multiple ranging from 70 to 200 kDa in molecular weight and each band was indistinct and formed a smear, reflecting mobility shift mainly due to proteolysis during extraction process. On immunohistochemistry, lung cancer tissues showed a very heavy, dense staining with anti-AHNAK protein antibody as compared to the surrounding normal lung tissue, coresponding well with the results of the western blot Conclusion: The overexpression of PLC-${\gamma}1$ activator protein, AHNAK in lung cancer may provide evidence that the AHNAK protein and PLC-${\gamma}1$ act in concerted manner in carcinogenesis.
Kim, Tae-You;Park, Jong-Kook;Ryoo, Baek-Ryeol;Im, Yung-Hyuck;Kang, Yoon-Koo
Tuberculosis and Respiratory Diseases
/
v.47
no.6
/
pp.797-806
/
1999
Background: About 20% of small cell lung cancer(SCLC) patients have bone marrow(EM) metastasis at the time of diagnosis and the remaining patients are also considered with micrometastasis. In an attempt to detect EM micrometastasis, we used cytokeratin(CK)-20 as a molecular marker, which is specific for epithelial cells. Method: A sensitive RT-PCR assay was used to compare CK-20 expression both in SCLC cell line H209 and normal leukocyte and to evaluate EM aspirates of 28 SCLC patients. Result: H209 cell line showed CK-20 expression but normal leukocyte did not, suggesting CK-20 expression is lung tissue-specific. Of 28 patients(11 limited disease, 17 extensive disease), only 2(1/11, 1/17) samples tested revealed positive signal for CK-20. Two patients with CK-20 expression had EM metastasis or multiple bone involvement during follow-up. Conclusion: Although circulating tumor cells were detected in EM of small portion of patients with bone metastasis, CK-20 doesn't seem to be a reliable marker for the detection of micrometastasis in SCLC. This study emphasizes that identification of more specific marker for micrometastsis is mandatory prior to clinical application.
H2AX, a crucial component of chromatin, is implicated in DNA repair, cell cycle check point and tumor suppression. The aim of this study was to identify direct binding partners of H2AX to regulate cellular responses to above mechanisms. Literature reviews and bioinformatical tools were attempted intensively to find binding partners of H2AX, which resulted in identifying two potential proteins, breast cancer-1 (BRCA1) and BRCA1-associated RING domain 1 (BARD1). Although it has been reported in vivo that BRCA1 co-localizes with H2AX at the site of DNA damage, their biochemical mechanism for H2AX were however only known that the complex monoubiquitinates histone monomers, including unphosphorylated H2AX in vitro. Therefore, it is important to know whether the complex directly interacts with H2AX, and also which regions of these are specifically mediated for the interaction. Using in vitro GST pull-down assay, we present here that BRCA1 and BARD1 directly bind to H2AX. Moreover, through combinational approaches of domain analysis, fragment clonings and in vitro binding assay, we revealed molecular details of the BRCA1-H2AX and BARD1-H2AX complex. These data provide the potential evidence that each of the BRCA1 nuclear localization signal (NLS) and BARD1 BRCA1 C-terminal (BRCT) repeat domain is the novel mediator of H2AX recognition.
Objetives : Identification of target genes for ethanol in neurons is important for understanding its molecular and cellular mechanism of action and the neuropathological changes seen in alcoholics. The purpose of this study is to identify of altered gene expression after acute treatmet of ethanol in rat gliom cells. Methods : We used high density cDNA microarray chip to measure the expression patterns of multiple genes in cultured rat glioma cells. DNA microarrays allow for the simultaneous measurement of the expression of several hundreds of genes. Results : After comparing hybridized signals between control and ethanol treated groups, we found that treatment with ethanol increased the expression of 15 genes and decreased the expression of 12 genes. Upregulated genes included Orthodenticle(Drosophila) homolog 1, procollagen type II, adenosine A2a receptor, GATA bindning protein 2. Downregulated genes included diacylglycerol kinase beta, PRKC, Protein phosphatase 1, clathrin-associated protein 17, nucleoporin p58, proteasome. Conclusion : The gene changes noted were those related to the regulation of transcription, signal transduction, second messenger systems. modulation of ischemic brain injury, and neurodengeneration. Although some of the genes were previously known to be ethanol responsive, we have for the most part identified novel genes involved in the brain response to ethanol.
Kim Do-Il;Kim Sung-Hyun;Ho Dong-Su;Choe Bo-young;Suh Tae-Suk;Lee Jae-Mun;Lee Hyoung-Koo
Progress in Medical Physics
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v.15
no.4
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pp.202-209
/
2004
Flat panel based digital radiography (DR) systems have recently become useful and important in the field of diagnostic radiology. For DRs with amorphous silicon photosensors, CsI(TI) is normally used as the scintillator, which produces visible light corresponding to the absorbed radiation energy. The visible light photons are converted into electric signal in the amorphous silicon photodiodes which constitute a two dimensional array. In order to produce good quality images, detailed behaviors of DR detectors to radiation must be studied. The relationship between air exposure and the DR outputs has been investigated in many studies. But this relationship was investigated under the condition of the fixed tube voltage. In this study, we investigated the relationship between the DR outputs and X-ray in terms of the absorbed energy in the detector rather than the air exposure using SPEC-l8, an X-ray energy spectrum model. Measured exposure was compared with calculated exposure for obtaining the inherent filtration that is a important input variable of SPEC-l8. The absorbed energy in the detector was calculated using algorithm of calculating the absorbed energy in the material and pixel values of real images under various conditions was obtained. The characteristic curve was obtained using the relationship of two parameter and the results were verified using phantoms made of water and aluminum. The pixel values of the phantom image were estimated and compared with the characteristic curve under various conditions. It was found that the relationship between the DR outputs and the absorbed energy in the detector was almost linear. In a experiment using the phantoms, the estimated pixel values agreed with the characteristic curve, although the effect of scattered photons introduced some errors. However, effect of a scattered X-ray must be studied because it was not included in the calculation algorithm. The result of this study can provide useful information about a pre-processing of digital radiography.
Purpose : Because adenocarcinomas of the uterine cervix have lower 5-year survival rate than squamous cell carcinomas due to early lymph node metastasis and local extension, scrutiny of lymph node metastasis and local extension by radiologic examination is necessary in case of clinically diagnosed or suspected adenocarcinomas. The purpose of this study is to evaluate whether there are specific findings of these tumors, compared with squamous cell carcinomas, through the analysis of magnetic resonance (MR) imaging findings. Materials and Methods : Of 21 pathologically proven cervical adenocarcinomas, MR imaging findings of 18 tumors (histologic staging : two Ib, four IIa, two IIb, one IIIa, and one IIIb) were retrospectively analyzed and compared with those of 40 wquamous cell carcinoma in consecutive patients as a control group. T1-wetighted and fast spin echo T2-weighted images were obtained on the axial and sagittal planes, using a 1.5-T MR scanner. The largest diameter, location, signal intensity and degree of contrast enhancement contour, shape and longitudinal extent of the tumor and associated findings on MR image were analyzed. Results : The largest diameters of cervical adenocarcinomas ranged from 0.8 to 4.1 cm(mean, 2.2 cm). Of 18 adenocarcinomas, nine were of endocervical type. All adenocarcinomas were isointense to surrounding cervical stroma on T1-weighted images and hyperintense(homogeneous in ten, inhomogeneous in eight) on fast spin echo T2-weighted images. Adenocarcinomas enhanced on contrast study in all patients (homogeneous in six, inhomogeneous in 12 with hyperintese enhnacing rim in two). Eight adenocarcinomas had smooth contours and ten had irregular ones. The shape of adenocarcinoma was irregular in eight patients, barrel shape in six, papillary/polypod in three, and nodular in one. All adenocarcinomas involved lower half of the uterine cervix and six tumors extended up to the upper half. Pelvic lymph nodes of more than 1.5cm in diameter in two adenocarcinomas pateints and no detectable small pelvic lymph nodes on MR imaging in one patient were pathologically positive. Hydrometra was associated in two adenocarcinomas patients, and hematometra in one patient. Compared with squamous cell carcinomas, more frequent MR findings of endocervical type and barrel shape in cervical adenocarcinomas were statistically significant. Conclusion : Cervical adenocarcinomas had more frequent MR findings of endocervical type and barrel shape, compared with wquamous cell carcinomas. Adenocarcinoma of the uterine cervix may be suspected on MR imaging, when a cervical carcinoma is of barrel shape along the endocervical canal and tends to involve lymth nodes in earlier stages.
Lee, Sun Joo;Lee, Sang Yong;Kang, Kyung Pyo;Kim, Won;Park, Sung Kwang
Investigative Magnetic Resonance Imaging
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v.17
no.3
/
pp.181-191
/
2013
Purpose : To evaluate the usefulness of in vivo magnetic resonance (MR) imaging for tracking intravenously injected superparamagnetic iron oxide (SPIO)-labeled human umbilical vein endothelial cells (HUVECs) in an acute renal failure (ARF) rat model. Materials and Methods: HUVECs were labeled with SPIO and poly-L-lysine (PLL) complex. Relaxation rates at 1.5-T MR, cell viability, and labeling stability were assessed. HUVECs were injected into the tail vein of ARF rats (labeled cells in 10 rats, unlabeled cells in 2 rats). Follow-up serial $T2^*$-weighted gradient-echo MR imaging was performed at 1, 3, 5 and 7 days after injection, and the MR findings were compared with histologic findings. Results: There was an average of $98.4{\pm}2.4%$ Prussian blue stain-positive cells after labeling with SPIOPLL complex. Relaxation rates ($R2^*$) of all cultured HUVECs at day 3 and 5 were not markedly decreased compared with that at day 1. The stability of SPIO in HUVECs was maintained during the proliferation of HUVECs in culture media. In the presence of left unilateral renal artery ischemia, $T2^*$-weighted MR imaging performed 1 day after the intravenous injection of labeled HUVECs revealed a significant signal intensity (SI) loss exclusively in the left renal outer medulla regions, but not in the right kidney. The MR imaging findings at days 3, 5 and 7 after intravenous injection of HUVECs showed a SI loss in the outer medulla regions of the ischemically injured kidney, but the SI progressively recovered with time and the right kidney did not have a significant change in SI in the same period. Upon histologic analysis, the SI loss on MR images was correspondent to the presence of Prussian blue stained cells, primarily in the renal outer medulla. Conclusion: MR imaging appears to be useful for in vivo monitoring of intravenously injected SPIO-labeled HUVECs in an ischemically injured rat kidney.
High-resolution shallow marine seismic surveys have been carried out for the resources exploration, engineering applications and Quaternary mapping. To improve the resolution of subsurface structure image, multichannel digital technique has been applied. The quality of the image depends on the vertical and horizontal resolution and signal to noise (S/N) ratio which are associated with the data acquisition parameters such as sample interval, common midpoint (CMP) interval and CMP fold. To understand the effect of the acquisition parameters, a test survey was carried out off Yeosu and the acquired data were analyzed. A 30 $in^3$ small air gun was used as a seismic source and 8 channel streamer cable with a 5 m group interval was used as a receiver. The data were digitally recorded with a shot interval of 2 s and sample interval of 0.1 ms. The acquired data were resampled with various sample intervals, CMP intervals and CMP folds. The resampled data were processed, plotted as seismic sections and compared each other. The analysis results show that thin bed structure with ${\~}1m$ thickness and ${\~}6^{\circ}$ slope can be imaged with good resolution and continuity and low noise using the acquisition parameters with a sample interval shorter than 0.2 ms, CMP interval shorter than 2.5 m and CMP fold more than 4. Because seismic resolution is associated with the acquisition parameters, the quality of the subsurface structure can be imaged successfully using suitable and optimum acquisition parameters.
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